Document O3Xb9Q86Mo3w2My7bQmw245Bj

& C \P > \7 O - r Reprinted by the U.S. DEPARTMENT OF HEALTH. EDUCATION. AND WELFARE PUBLIC HEALTH SERVICE f r o m CRC CRITICAL REVIEWS IN TOXICOLOGY, J a n u a r y 1974 THE TOXICITY OF POLYCHLORINATED POLYCYCLIC COMPOUNDS AND RELATED CHEMICALS i Author: Renile D. Kimbrough* Center for Disease Control Atlanta, Ga t Referees: M. M. Barnes J. B. Greif Medicei Research Council MRC Toeicwlofy Unit Medical Research Council Laboratories Cirstialton. Surrey England INTRODUCTION Only the small group of the polychJorinated polycyclic compounds given in Figures 1 and 2 will he reviewed in this article. Several years ago when tHis laboratory was the Atlanta Toxicology Branch of the Food and Drug Administration, concern was voiced over the possible long-term effects of polychlorinated biphenyls. 2nd we began to study the toxicity of Aroclor 125-4 and Aroclor 1260. About this lime a follow-up study was performed on a group of workers in New Jersey who w-ere engjged in the production of 2.4.5-T.1 These workers had very severe chloraene and had previously been shown tu have porpnvna.' During that same lime, a petition was filed with the Food and Drug Administration for a residue level of hexachlorophene on certain food crops HcxachJorophene. as well as 2.4.5-T. is made from trichlorophenol. although the chemical reactions are quite different. When we cheeked the available literature on the toxicity of hexachlorophene we fell that adequate toxicity data by present standards w'err not available, particularly not if the compound was meant to be ingested. Further more. Larson3 had pointed out that hexachiorophene was more toxic than commonly acknowl edged. Hexachlorophene and similar chemicals behave differently from such polycyclic polychlorinated compounds as the chlorinated biphenyls (PCBs.f Hcxacluorophene in particular is acutely much more toxic than the PCBs and is fairly rapid*) eliminated from the body, wlule the PCBs have * very low acute toxicity, but are very poorU metabolized, particularly those with mure thar four chlonne atoms on the phenyl rings The longterm effects are therefore more critical fc*> compounds such as l*CBs. The chlorinate a dibcnzodioxins and furans. some of which at: extremely toxic on an acute basis, may 2ko she* persistence and their long-term effects ntas > equally as important. These observations stv thai it would be erroneous to make predictions cr lung-term effects from results obtained in a ,. toxicity studies. l-ormerly wnh ihc Environmental Protection Arctic)'. Crwmblte lo n co lc* ) Laboratory, -177(1 Bulord Highway. Clumblev. Cr ?l'}4 |. January 197 NPC00016172 207858 -i n i i i w m i C h lo rln attd D1bniofurns Chlor1nit*d D1benzod1ox)n Polychlorinated Naphthalenes FIGURE I The materials shown in this figure are mixtures of compounds in which several but usually not all of the positions (-X) are chlorine and the rest are hvdrogen. The properties of the materials depend on the degree of chlorination of the mixture All are insoluble in water but soluble in varying decrees in organic solvents and in fall. The trade name for the U.S. product (Monsanto) of the chlorinated biohenvls and terphenvls It Aroclor. The trade name is followed by the number. For the biphenyls, the first 2 digits of this number are 12. and for the terphenvls, 54. In each instance the last 2 digits give the percentage of chlorine t OH OH r.4,4' tncMorc- 2`*yflroy-<1ie,'e"yl etnar ITrlelosan * (nnssn 0eJO3) UfTJCLia l .i ' TMo-bll(4-ehleropH''0l) FIGURE 2 The compounds in his figure are while crystalline solids with clcarlv defined meliiot points ab '^ e lb*C They jre practically insoluble in " ter and soluble in varying amounts in organic solvents and in fats Those containing jn -OH group, afl the above excepting j.+rd'-tnehlor.Qurfranilidc. are soluble in aqueous alkali. i i 446 CRC Criticcl Reviews itt Toxitvltigy NPC0001617 3 207859 NPC0001617 4 * V The present review is an attempt to correlate has been provided by Broadhursl 1n With a peak various toxic effects produced by the chemicals production of 42.527 short tons of PCBs in 1970. :} <(.$1! listed in Figures I and 2 in order to better understand and appreciate their behavior Monsanto is the soie producer of Aroclors (U S tradename for PCBs) in the U S Yearly PCB The subjects covered in this article have lately production figures for the period 1960-1971 were Jenjoyed great interest and a rapidly growing number of scientific reports are available. Since a reported by Monsanto.11 The production increas ed steadily until 1970 and dropped to 20.236 '.n number of the chemicals discussed produce similar or related effects, their toxicology is discussed short tons in 1971 due to a voluntary restriction of the use of PCBs by the company. Trade names together rather than listing each compound sepa in other countries include Kanneclor and rately. which would have led to a great deal of Sant ha term in Japan, Clophen. Flcndur, repetition Phenoclor, and Sovol in Eurupc. A detailed discussion of the various chemical The polychlorinated capacitor liquids, com reactions, analytical methods for the determina monly called askarels. are mixtures of chlorinated tion of the compounds, is not given The review biphenyls and chlorinated benzenes. A variety of t has been written with the toxic effects of the other industrial applications10 have been found various compounds as the focal point and the for PCBs. They have served as fluids fur hydraulic, literature cited serves to illustrate these points. No gas turbine, and vacuum pumps, as heat transfer attempt has been made to present a complete fluids, as plasticizers, adhesives, textile coatings, review of the literature in this area. surface coatings in paints and varnishes, as seal ants. and as part of formulations to prepare DEVELOPMENT. USAGE. pressure sensitive records and color copying AND OCCURRENCE papers. The extent of the current use of PCBs in printing processes is unknown. Other suggested A description of the synthesis of chlorinated uses over the years have included catalyst carrier biphenyls (PCBs) can be found in Liebig's Annalen for polymerization of olefins, conversion of water as early as 1881 4 Successful production of permeable suil to a nonpermeable state, and diphenyl in commercial quantities was not report combined insecticide and bactericide formulations. ed until 1930 5 At that time the chlorination of Mixtures of chlorinated biphenyls and chiunnatcd technical biphenyl was also described * Biphenyl naphthalenes have been employed to insulate itself is fungistatic and used as a preservative for electric wires and cables, particularly when they" citrus fruits1 Chlorinated biphenyls arc very were used in the mining industry and on stable, and have been used for protective coatings, as plastic resins or gums for varnishes, and for waterproofing and flameproofing of wood. The excellent dielectric properties of these compounds were noted as early as 1930 Until the advent of the PCBs. mineral oil was used as a cooling and dielectric liquid impregnant for high voltage electric apparatus such as transformers and capacitors. Since minera! oil is flammable and llammablc gases could also be produced, restric tions for the use and installations of these irans. tourers, hadlo be set. paiticulaily for indoor warships.11 Residues of polychlorinated biphenyls have been reported in the environment by a number of authors in different parts of the world. These repom were usually from arcus where industrial pollution was responsible lor the environmental contamination Holden,13 after finding PCBs in marine fish and moJIusks fiom Scottish waters, noiiced that ihey were generally highest in samples Irom ihe Firth ot Clyde and he discovered that the PCB contamination probably originaied from industrial sewage. The sewage sludge was trans installation When mine'rjl oifs wire npl. -b^ ported Irom two Glasgow purification works by the Aroclors. (he fire and explosion hazards were boats and cIuinpccTin deep water six miles south ot eliminated This was recognized by the National Garroch Head. The disposal ol sewage sludge un Board of Foe Underwriters and the National land could lead to Hie introduction of PCB Electric Code was modified J Use ot Aroclors also residues into the Terieslnj|'trco>ys(em by transfer-; greatly reduced the physical size ol the through food chains. So far PCB residues have not capacitors.* been delected in liesh water fish in Scotland. A recent summary ol the varied uses of PClts PwlychJurmuled biphenyl* have also been lound 207860 January 1974 CM-- L jj. -it -- -- . _________ J L in wildlife samples collected along the Outch coast nd the Rhine Rjver14 and in fish and seal in different parts of the Baltic Sea along the Swedish coast.15 Rjsebrough et al.16 compared the presence of PCBs with that of DDT and its metabolites in various birds These authors found very low concentrations of PCBs in wildlife species in Baja, California, Mexico, and the Guif of California, which are very remote areas with a high ratio of DDT to PCB of approximately 9 or 10 In sea birds from the Pacific, the ratio was between 5 and 10 and at that lime. PCBs were not found in eggs of the Adeiie penguin from Cape Crozier, Antarctica Relatively high PCB concentrations were found in San Francisco Bay, Puget Sound, and San Diego Bay and the ratio of DDT to PCBs in wildlife samples from this area was between 1 and 2. This is again emphasized by the findings of Dustman et al.17 Twelve AJaskan bald eagle eggs had median PCB residues of 1.65 ppm while II eggs collected in Maine. Michigan. Minnesota, and Honda showed i median concentration of 9 7 ppm. Zitko1* found low concentrations of PCBs in different types of frsh taken from several lakes and the St John River System. New Brunswick. Canada, and the Nova Scotia banks. Fish from the Milwaukee River and Lake Michigan in the mid west part of the U.S.. where the river enters into the lake, contained varying amounts of PCBs19 while fish taken from various small lakes in Wisconsin without industrial discharge failed to show PCBs. Large predators from Lake Michigan contained as much as 10 to 25 ppm PCBs on a fresh weight basis expressed as Aroclor 1254 and salmon eggs contained as much as 12 to 17 pom PCBs on a f-esh weight basis. Further studies by Veith:0 of the concentration of PCBs in different rivers emptying into Green Bay indicated a trend towards a decrease in PCB concentrations since the voluntary partial ban on PCB sales by Monsanto was imple mented in September of 1970. If PCB discharges are very high a fishkill may result such as the one in Escambia Bay. Florida in 1969 31 In that particular episode the I'CBs had entered a plant's effluent through accidental leak age of heat exchange fluid. The PCB found in this case resembled Aroclor 1254 most closely. The peak amount found in water of the Escambia River it the outfall amounted to 275 ppb and th** peak concentration found in oysters from Escambia Bay was 3.0 ppm. Aroclor residues in sediment samples taken near (fie outfall reached 486 ppm. When the leakage from the plant was corrected. Aroclor levels gradually decreased in this area. The wildlife portion of the U.S. National Monitoring Program analyzed starlings, mallard and black ducks, bald eagles, and fish of several species for PCBs According to Stickci.*1 monitor ing for terphenyls is also done but not for any of the other chlorinated compounds in this report Research surveys that include fishcating birds, insectivorous birds, ospreys, woodcocks, and cer tain ducks also monitor only for PCBs and for pesticides. Oysters (Crassvstrea vtrginica) are a good indicator of PCB and other chlorinated hydrocarbon concentrations in water and a nation al monitoring progiam m these mollusks in 15 coastal areas has been developed.1s So far. mcllusks have only very occasionally contained low levels of PCBs. In summary, the PCB concentrations in water are low or nonexistent in remote lakes and fresh water streams and can be as high as 50 yg/1 (fcppb) 'w or higher in polluted nvers such as the Hudson River.14 The PCB concentration is usually higher in sediment from these polluted waters because PCBs are only very slightly soluble in water.*5 Since PCBs are stored in living matter, they are concentrated in the food chain and the usual " biomagnification" is encountered as wc know it from other poorly metabolized and excreted sub stances. The concentration of PCBs in fish, par ticularly those in the upper end of the food chain from polluted rivers and lakes, is usually estimated in the ppm range and fisheating birds will have somewhat higher levels of PCBs than ihc-tlsh from the same geographic location.J` Young animals may contain less PCBs than their older counter parts of the same species in a particular location PCBs arc also infrequently encountered in dairy products, meat. fish, and poultry in ready-to-eat human food 37,1 * Occasionally, human food and domestic animal feed have been more extensively contaminated with PCBs. In these instances, the contamination could usually be traced to locai misuse of transformer fluid, leakage of PCBs from heat exchange fluid, or contamination of recycled paper with PCBs where the paper had been usee fr-r fond packapng such as cereal boxes 39 Th U.S. Food and Drug Administration has rccentl> set temporary tolerances for PCBs in certain too. products (see Table I ). 448 CRC Crttual Heviewi in Tuxicofttxy NPC00016175 207861 TABLE 1 Temporary Tolerance} Expressed as Par per Million Ippm) Established by L'.S. Food and Drug Administration (1) Milk (fat basis* (2) Dairy products (fat bans) (3) Poultry (fat basis) () Lpx* (5) Complete and finished animal feeds (6) Animal feed components (?) Fish and shellfish (edible portion) (8) Infunt and junior food (9) Taper food-packaging material 2.3 2.5 5.0 05 02 2.0 5.0 0.2 10.0 Note: The tolerance for paper food-packaging material shall not apply to such material separated from the packaged food by a functional barrier impermeable to PCB migration The Food and Drug Administration will provide upon request the analytical methods it will use for enforcing the temporary tolerances Ail of these aspects of PCBs in the environment have been the subtect of a number of reports, reviews,30 and conferences 3I`35 Terphenyls and chlorinated terphenyls are used as reactor coolants. The terphenyls consist of mixtures of ortho-, meta-, and paraterphenyls. Chlorinated terphenyls are produced by Monsanto and termed Arodor, followed by numbers. The first 2 dipts of these numbers are 54 and the lait 2 indicate the percent of chlorine such as Arodor 5460. Aroclor senes 25 and 44 consist of mixtures of polychlorinited biphenyls and polychlorinated terphenyls.3* According to Zitko et al.,31 poly chlorinated terphenyls have been found in environ mental samples. Polychlorinated terphenyls do not elute from GLC column* under conditions used for PCBs,38 which may explain why they have only been detected occasionally, but the way they are used probably also predudes their extensive spread in the environment. Chlorinated dtbenzodioxins and chlorinated dibenzofurans may occur as contaminants of cyclic chlorinated hydrocarbons In 195V Tomita et al.39 reported on the synthesis of polyhalodibcnzo-p-dioxins. According to these authors, chloiophenols. when heated to about 200C or more under alkaline conditions and pressure, will form chlorodibenzodioxins. as shown below Cl. - C C Cl N: *0 -- * q-- f S C @ - a' 2 NiCl Under similar conditions a chiorodibenzofuran may also be formed. OH H Cl N;0 Cl * H,0 * NiCl Chlorinated dibenzodioxins and chlorinated dibenzofurans are formed, or can be formed, as contam inants in the manufacture of a variety of products wluch use chlorophenojs and chlorobenzenes as starting materials, such as 2,4,5-T, pcnlachJorophcnol, nilrofen. hexa- and penlachlorobenzcne. and tri- and tctrachlorophenol. Polychlorinated biphenyls may also be contaminated with chlor inated dibcnzofuians.40'* 1 and chlorinated dibenzolu ran s may result fiom polychlorinated biphenyls under LTV irradiation 43 The toxiciiy of these products vanes depending on the position and number of chlorines attached to die phenyl ringj. Products from dillerenl manufacturers may show a great deal of difference in the type and amount of contamination and different lots of the same product may vary in composition, making toxicological evaluations of these products very difficult. Rigorous production control and cleanup procedures need to be Implemented so that the composition of different lots manufactured by the same company becomes more uniform. Villanueva et al.43 and Jensen and Renberg44 reported (he presence of chlorinated dibenzo-pdioxin m p en tac h lu ro p h e n o l. HydroxynonaehJorodiplienyl ether was also found and referred to as "predioxin" under the assumption that il represented a precursor of dioxin. January 1974 449 mT- **" NPC00016176 207862 u. range of 4 to 10 g. These two compounds proved (o be more toxic in weanling rats. The single o 'ai LD*e for weanlings was 1295 mg/kg, and 13 i 5 mg/kg for Aioclor 1254 and 1260, respectively, and the lowest lethal doses were 1000 mg/kg for Aroclor 1254, and 1200 mg/kg for Aroclor I 2 i \ The intravenous l.DJ0 for Aroclor 1254 in adult female rats was 358 mg/kg.94 A number of short-term studies.have been reported where tire PCBs were given to anixais over a period of one or two weeks. When the-PCBs were given in this fashion they were somewhat more toxic than when they were given in a siiwie dose. These studies are reviewed tn great detail i > Nelson et al.31 and will not be reported here since thr.v do nni give a great deal of additie" -1 ' y.'>p changes were also observed in the livers and the treated skin. Vos and Nntenboom-Rant91 compared ihc dermal toxicity of the isomer 2,4,S,2',4',S'^hexachlorobfphcnyl with that of a PCB mixture (Aroclor 1260) in rabbits. Their dermal application resulted in early microscopic skin lesions in the Arodot group. The lesions in the 2.4.5.2'.4'.5'. hexachlorobiphenyl group appeared later and were less severe. Both experimental groups showed liver changes. The liverdamage was more pronounced in the animals that had been exposed to hcxachlorobiphenyl than in those that had been exposed to the Aroclui. In inhalation studies with Aroclor 1243 and 1254. rats, mice, rabbits, guinea pigs, and one cat in. ;!! il ii ____ i MB* Jj ,:Tv. ai i *>ji i J I * COOO^ According to Johnson et th: predom presence . of dioxins in fish taken from four inant dioxjn in a commercial pentachlorophenol locations in Vietnam in Military Region III, whuh was octacKlorudibeiizudioxin while trace amounts includes Saigon and several provinces lu the of hexachlorodibenzodioxin and no let;* .-hlorodi- north.' 9 The concentrations ranged frum !fi :o benzodioxln were found. 5ince the technical 814 ppt. penttchlorophenol was in some respects more Zilko" wai not* able to demonstrate chi'" , toxic in animal studies, than the pure product, the inoted dibenzodiuxins and dibenzofuram ;n production process of pentachloropScnol was, aquatic animats from various locations in Canaca changed to minimize the dioxin concert.'ation and ` Chlorinated mpftr/ia/ertcs may so tat hrt A a new product has now been registered ender the remained "unidentified since they intcrfeie n 7Tv Federal Insecticide Fungicide and Rode. ;icide Act other chJorinated compounds in any niulmes.c, with a lower dioxin content. . analysis and are detected only when n u st ipr . Technical 2,4,5-T has also been f. .d to be trometry is also performed.56 contaminated with a dioxin. In' ihi: instance, The various germicides given in Figure hi-? unfortunately, the very toxic 2,3,7,5 'utrachloro- been used as antiseptics, deodorants, and p^ - . , dibenzodioxin.48"4 * was present. The contsmi- lady, as far as hexachlorophene is cuncernes j .* nation of the2,4.5-T with 2,3.7,S-u; :hlorodi- as preservatives for various cosmetics, soap* < ' benzodloxtn ranged frorn,'<~l ppm To m u clr --Motions. A listing, jif different produ.M 32 ppm over the years. In,newly manufactured contained hexachlorophcne prior to the ne * **. 2.4.5- T in the L'.S, the content of tetrachlorodi- lations was given by tite American Pharma *j i benzo-p-dioxin has been substantially reduced and Association.17 Regjstratioh. -cLJiiihionv Is usually kept below 0.1 ppm. Whether other cancelled in the U.S." because it p -o u ^ rj dioxins or furans have ever been detected -in photosensitivity reactions in humans usmg *r* 2.4.5- T in the U.S. has not been ref -::d in the that contained this product. None of the . literature. chlorinated polycyclic compounds used as r - Chlorinated dioxins have also be - found in __ cities have been shown to accumulate - '-* so-called "toxic fat," which pruducc:. .:tick edema _ environment. Hexachlorophene hus addtti*.--* '" *** wh#n fed tu chickens. After several years as a fungicide on ornamental plants, on ..t. . According to Johnson et nl.,4! the predom presence .o f dioxins in fish taken from four inant dioxjn In a commercial pentaehiufuphenol locutions in Vietnam in Military Region HI, which was octichiurudibeir/odioxin while trace amounts includes Saigon and several provinces to the of hexachlorodibenzodioxin and no tetnichlorodi- north.*4 The concentrations ranged from 18 to bcnzodfoxln were found. Since the technical 814 ppt. pentachlorophcno! was in some respects more Zitko** was not able to demonstrate chlor toxic In animal studies, than the pure product, the inated dibenzodioxins and dibenzofurans in production process of pentachlorophenol was. aquatic animals from various locations in Canada. changed to minimize the dioxin concentration and ' CwmhluonriHinHaftaeWd nraaupyhintihiamlecnNes imhqaty 3sUo fuatr hMaflvyeg . ^ a new product has now been registered under the _remained unidentified since they interfere with "7 T Federal Insecticide Fungicide and Rodenticide Act other chlorinated compounds in any niulliresidue with a lower dioxin content. analysis and are detected only when mass spec Technical 2.4,5-T has also been found to be trom etry is also performed/* ~ contaminated with a dioxin. In' this instance, The various germicides given in Figure 2 have unfortunately, the very toxic 2.3,7,8 tetnchloro- been used as antiseptics, deodorants, and particu dibeniodioxln,**'4a was present. The contami larly, as far as hexachlorophenc is concerned, also nation of the 2,4,5-T with 2,3,7 ,S-letrach]orodi- as preservatives for various cosmetics, soaps, and benzodtoxln ranged frum.< 1 ppm to as much as lotions. A listing of different products that 32 ppm over the years. In newly manufactured contained hexachJorophcne prior to the new regu 2.4.5- T in the U.S. the content of tetrachlorodi- lations was given by the American Pharmaceutical benzo-p-dioxin has been substantially reduced and Association.31 Registration of bithionol was Is usually kept below 0.1 ppm. Whether other cancelled in the U.S.1* because it produced dioxins or furans have ever been detected in photosensitivity reactions in humans using soaps 2.4.5- T in the U.S. has not been reported in the that contained this product. None of the poly literature. chlorinated polycyclic compounds used as germi Chlorinated dioxins have also been found in cides have been shown to accumulate in the so-called "toxic fat," which produces chick edema environment. Hexachlorophene has additional uses disease when fed to chickens.. After several years as a fungicide on ornamental plants, on conveyor of study by industry and the Food and Drug chains as a mildcwstat, In laundry rinses as a Administration, Flick et al.49 were able to mildewstat, in shoes and air filters and assorted separate a purified crystalline product which industrial uses as a fungistat, and on leather, paper. produced chick edema disease. Two .years later, . and textiles,39 U is also registered as a pesticide Cantrell et al.1 .announced that 1.2,3,7.8.9- for use on cucumbers, peppers; and tomatoes,60 chlorodibenzo-p-dioxin was- one of the' toxic and for cotton. compounds that produced chick edema. The origin Dichhrophcnc is used as a mildew preventive of the chlorinated diben/.odioxin in "toxic fat" on cotton and is recommended as a fungicide ami was not clear. It was suspected that it arose from bactericide for the protection of textiles and chlorophenols which were used lo preserve.hides materials from molds.6 1 ' for the manufacture of leather. Tallowy and greases were obtained from these hides51 and a CHEMISTRY: METHODS FOR more recent outbreak of chick edema disease was CHEMICAL DETERMINATION traced to the cohlummaiion wiih'dUorophenois ui AND PHOTOLYSIS soapstock used as feed fats. Higginbrillarti e ta l.5i showed that chlorophenols and their salts, when Much' o f the chemistry of polychlorinated healed, undergo condensation reactions anti form biphenyls has recently peen reviewed.33`14 chlorinated derivatives of dibenzo-p-dioxin; Commercial chlorinated biphenyls arc mixtures of furthermore, chlorinated dioxins and ptedioxins biphenyl groups with different numbers of were originally present in some of the chlorinated chlorine atoms (Figure 1:. mien hydrogen atoms phenols. Generally no efforts have hecn made to are replaced by chlorine on the biphenyl ring, a del ermine whether various chlorinated dioxins and large number of substitution combinations can ~furans atc-prcscnj in.lhc environment However. arise since the number of chlorine atoms on the Baughman and MeseIson*1 recently rcpofilt! W "* rntriecuies can vary and the chlorines can also be 450 CHC Critical Rrt icws in IrtxAvtltisy NPC00016178 207864 '----- . -1r iltachcd to the molecule in different positions, relationship the theoretical response of the resulting in a. variety' of isomers. Even if the detector to eath cldonna ted biphenyl was calcu | chlorination may be' the same, different commer lated. When this approach was used, the levels of cial batches may vary in the relative concentration PCBs were lower than when the PCB levels were i of isomers. The PCBs produced in the U.S. by Monsanto fall under the trade name Aiodor. The compared to the Aroclor 1254 standard. Because of these difficulties in quantitation, the PCD levels various Aroclor mixtures have a great number of reported from different laboratories may vary constituents, and when the biphenyl is chlori greatly. The various problems encountered in the nated, 210 different chemicals are theoretically an aly tical methodology of polychlorinated possible. Aroclor 1254, for instance, yielded 69 biphenyls and their quantitation are also described constituents while the fractionation of Aroclor in a U.S. FDA report which can be obtained upon 1242 and .1260 yielded 45 and 78 components, request from the U.S. Food and Drug Adminis respectively.84 tration in Rockville, Md. The tide of the report is The chlorinated biphenyls (PCBs) are chemi Analytical Methodology fo r Polychlorinated cally very inert, resistant to corrosive chemicals, Biphenyls (Feb. 1973). 'it ipiolttble in water, and have a low vapor pressure. Aroclor 1221, for instance, has a boiling point of Polychlorinated terphenyls (Figure 1) are not eluted from gas chromatography columns under 278C. The boiling point increases with increasing conditions used for polychlorinated biphenyls.38 chlorination. The dilorinatcd biphenyls are deter By altering their gas chromatograph column, Zitko .-a mined in biological and environmental samples by et al.37 were able to detect polychlorinated a combination of electron capture gas liquid terphenyls. chromatugraphy and mass spectrometry. Nuclear rChhrinated naphthalenes (Figure 1) elute at magnetic resonance spectroscopy is used in the same time and cause interference with the addition to the above two procedures, in some determination of chlorinated biphenyls and chlori^ H laboratories. PCBs were first identified-in environ nated hydrocarbon pesticides in general, unless mental samples by Jensen in 1966,41 They were mass spectrometry is pertormed to establish thtT detected as interfering peaks on GLC analysis of identity of the various chemicals involved.34 _ environmental samples that were analyzed for The methods presently available for the deter chlorinated pesticide residues,*3 mination of trichlorocarbanilidc and other germi The PCBs interfered with routine pesticide cides (Figure 2) arc not very well developed. analyses for chlorinated hydrocarbons. Various Graber et al.69 outlined a thin layer chroma authors have therefore published separation tography method fur the determination o f these methods44 and Reynolds34 discussed this exten chemicals in soap and very briefly mentioned sively in a review. othet methods that have been employed in the According to Safe and Hutzinger,65 the use of past. mass spectrometry for structural studies of PCBs is Several methods for the determination o f chlor limi ed because the chlorine atoms may be ran inated dibenzodtoxins and chlorinated dibenzo- domized between the phenyl groups when the furans have been described. The goal o f these PCBs are fragmented by electrons. The only methods is .two-fold. First o f all. quite a number of exception to this is 2,2'-dichlorobiphenyl. technical polychlorinated organic compounds may The quantitation of polychlorinated biphenyl contain one or several of these compounds70-' *1 as in environmental samples has met with difficulty 1contaminants and methods have been developed to since the PCBs represent mixtures. Several workers determine them in various technical ptoducts. have compared PCB components in field samples Other methods dealt primarily with toxic fats to Aroclor 1254 because the sample chroma where many attempts were made to isolate and tograms were most similar to these mixtures.44-4 8 identify the compounds responsible for chick Rote and Murphy48 'quantitated individual peaks edema disease.73 Some of these methods were of different Aroclurs by means of a response described in the papers that will be cited in curve-. The scmilogaiithmic relationship of cnnncction with the description of chick edema detector response (total peak area/16 ng) to disease in this article. average chlorine .content was obtained for each When preparing the various chlorinated diben- Aroclor by the method of least squares. From this zodioxins and dibenzofurans for toxicity studies in January 1974 .45 & in NPC00016179 207865 itif i. order to obtain pure products, methods also need to be available for.their separation and identifi cation.14 One of the problems with tetrachlnrodibenzo- syn the fixed a few PCB isomers, namely, 2,4,6,2*.4*.6*-licxachlorobiphenyi, 2.4,5.2',4r,6'hexachlorobiphenyl. 2.4.l',4'-tctrachlorubiphcnyl, 2,5,2* .5'-letrachlornhiphenyl. and irradiated the dioxin as weti as leiracmowtiibtnzoluran is their various isomers with UV light. A mercury vapor ^ r ,,severe toxicity in th<jnicrogram J n y which wtll_ lamp with quartz tillers was used as a light source. probably not result in an accumulation within the The UV spectrum of the synthetized chlorinated ppm in biological tissues, for instance, for residue biphenyls only shows significant absorption below analysis. Baughman and Mcsclson*3 have recently 300 nm. When these compounds were irradiated in described a method for the determination of these a solvent such as hexane, acetone, methanol, or a materialsTwilhin the ppt range. Hexachlorophene \ is made from trichJuto- mixture of methanol and water, dechlorination and polymerization of the compounds was phenol. A process for its pi eduction was patented observed. When the PCB isomers were irradiated in by Cump.7i Additional methods for the produc perfluorinated dimelhylcydohexunc, .in inert tion and purification for hexachlorophene have medium, isomerization, and cldurinatkm decldor- been published and are cited in Chemical ination were observed. These experiments indicate Abstracts. that, under environmental conditions, photolysis Recently several methods76-7 9 have been products may result with a lower chlorine content developed fur the determination of small amounts but small amounts of higher chlorinated biphenyls of hexachlorophene in blood and biological may also occur in low concentrations. Sab and tissues. .Some methods were based on extraction of Hutzingcr"* reported that the Eradiation of hexachlorophene with ether or ethanol or a ^ . t . ^ 'A'^-hexachlorobiphenyl at 310 nm mixture of ether and ethanol. An acetyl or methyl resulted in compounds of lower chlorine content, derivative was formed and the hexachlorophene such as di-. tri-. tetri*, and pentachlornhiphenyls. was determined with electron capture gas chroma In these invest) calkins hexane and methanol were tography.79 In an earlier method described by used as solvents. It is possible that new compuunds Bachman n and Shetlar.80 simple benzene are formed by loss of chlorine. Rearrangement and extraction was used and no derivative of hexa- . condensation may lead to chlorohiphenyls in chloiophene was made, which led to very variable environmental samples that arc not present in results. The various advantages and diradvantages commercial PCB mixtures. The 24-hr madia lion of of these methods were discussed by Ulsamer.79 In 3 ,3 *,4 ,4 * -teirach lo ro b i p h e n y l, 2 .2 , ,6,6*- earlier work my co-workers used ether for tetrachlorobiphenyl, 2,2\5,5'-tetrachJorobtphenyl extraction and prepared a dimethyl ether deriv dissolved in hexane resulted in about 7 0" loss of ative of hexachlorophene.*1 Recovery with this the starting material, while with 2.2\4,4',5.5'- method was only about 759 and we have recently hexachlorobtphenyl, only 3.k'5- remained and less altered our method of extraction to obtain better than I% of 2,2*.3.3\4,4\5.5*-orlachlorobiphcnyl recovery.83 A method developed by Kabacoff ct remained. The irradiation of A ro d o rl2 S 4 in al.7a for the determination of hexachlorophene is. hexane resulted in a change uf the composition of ] according to the authors, also suitable for deter the Amdur mixtuie. The gbs chrumalogram mination of diehlorophene and Guicnmann and obtained from the inadiated Arcelor showed Lisk" J have described a method for the determin shorter retention times than lh standard Arod or ation of hexachlorophene in several agricultural 1254. The irradiation of Aiuclur 1254 in a products. dioxanc water mixture' also cimtuming sodium The production of hexachlorophene from bicarbonate did not lead (o the formation of trichlorophenol proceeds at low temperature and dilurodiben/ufuruns or hydroxychlorobi phenyls. under acidic conditions which makes the produc The compounds that were ohscrvctl corresponded tion of a chlorinated* dibenzodioxin as an impurity to PCtl molecules to which water was added ami a highly unlikely. more polar "carboxylic" acid fraction. Irradiation Since chemicals that are found in the environ of Arodor as a thin Hint with a frequent addition ment are exposed to sunlight, the effect of W of water led to newly formed hydroxy kited light on the compounds under discussion has been compounds and a more polar "carboxylic" given considerable attention. Hustcrl and K o n c'4 fraction. The results obtained from irradiating 451 CRC trinca/ Rt^^rv^ in Toxit nlngy ! NPC00016180 207866 if .*#, i. .m .......... i i i I |j PCBs therefore depended greatly on the solvents a fungistatic agent which is used as a preservative r. used and the pH of the solution in addition to the for citrus fruits, and is usually impregnated into ! > $ light source used and may result in dechlorination, the wrapping paper. The protected citrus fruit formation of polymers, hydroxyiation, and contains a certain amount of biphenyl residue. In y j carboxylic products.** different countries the amount allowed as a Exposure of various chlorinated dibenzodioxins residue on citrus fruit varies between 70 to 110 r to summer sunlight or fluorescent UV light with ppm of biphenyl. Booth et al.90 found a reversible < an intensity of roughly 100 fiw/cms in the nephrotoxic effect of biphenyl on the rat kidney r pretence of organic solvents such as methanol led when rats were fed 0.57 or 1% (10,000 ppm tn rapid photolysis of 2,3,7,8-tclr3chloro- and biphenyl in the diet). None of these effects was r 2 , 7 - d i c h l o r o d i b e n z o d i o x i n . w h ile observed on diets containing 0.1% (1000 ppm 1,2,3,4,6,7,8,9-oclachlorodibenzo-p-dioN in biphenyl). Deichmann et al.91 showed that pro produced a series of chlorinated dioxins with longed exposure to air containing diphenyl in high 'Sfi uniformly decreasing chlorine content. Some evi concentrations of 5,40, and 300 mg/m3 produced dence was also found that reactions other than liver and kidney injury well as broncho reductive dechlorination occurred."1 When pulmonary lesions in mice and rats. In general, . 'it 2.3.7.8-tetrachlorodibenzodioxin was spread on a diphenyl has always been considered to be 1 glass plate in a methanol solution and the meth relatively nonloxic. In' 1969, in Finland, a man ; i anol was allowed to dry', quantitative recovery of who had been Intensively exposed to diphenyl for r/J the dioxin was possible even after 14 days of liV 11 years became ill and died.91 Diphenyl impreg ?` irradiation. -Similar results were obtained when nated wrapping paper had been manufactured in 2.3.7.8-tclracliiorodibenzodioxin was dissolved in Finland for about 15 years in the plant where the methanol, pul on a glass plate, covered with soil, man was employed. The equipment used in the and irradiated.81 These results demonstrated that impregnating process was cleaned 5 to 10 times a under favorable environmental conditions, when year with tri- or tetrachloroelhylene. The cause of organic hydrogen donors are present, photolysis death of the worker in this particular report was will take place. Bare surfaces of soil, concrelc, or not satisfactorily explained and neither were the 3* water arc not effective inducers of photolysis. complaints of 31 other workers engaged in the i Chlorinated dibenzodioxins could at' limes prove same factory. Since chlorinated ethylene com * I: to be quite persistent. Partial dechlorination may pounds were used for cleaning, it should be also occur with chlorinated diphenyls or the higher established whether organic solvents containing chlorinated dibenzodioxins and may result in less halogens could lead to the production of toxic tr chlorinated compounds. substances when (hey are brought in contact with The possible long lasting presence of the diphenyls. If this proves to be the case, then care i -I dioxins is also emphasized by Kearney et al.,88 should be taken that this situation is avoided. who recovered 56 and 6YJc o f originally applied The / `CBs are also not very toxic when given as lelrachlorodibetizodioxin after one year in a single or a few repeated doses to - birds and Hagerstown and Lakeland, Md. soils. mammals. However, the juvenile pink shrimp is Tc trachtorosalicvlan itid v and sim ilar very sensitive to FCBs in water, while trout and i compounds split off chlorine atoms when exposed catfish can tolerate higher doses; insects are also tk fV tu sunlight. This is briefly mentioned in connec not very sensitive to PCBs. Pin fish are not quite as tion with photosensitivity reactions of the skin sensitive tu a single dose, but will die when * Shaffer et al.89 irradiated hcxachhrophene in exposed for 14 to 45 days lo 5 pph of Aroclor absolute ethanol with UV light and obtained a 1254 VJ PCBs are of a low order of toxicity to rats * number of dcchlotinuicd bisphcnols resulting from and rabbits when given as a single dose. The acute loss of chlorine in the ortho and para positions oral toxicity decreases in rats with increasing relative to the hydroxy group. cldonre content, while this is nut is ohvious when the dermal toxicity is tested in rabbits.33 Tills GENERAL TOXICITY AND j EFFECT ON REPRODUCTION *l"l 1 may be due to poorer absorption o f the higher chlorinated compounds. In studies with adult Sherman strain rats, wc found that the oral LD(0 T4 Diphcnvt itself, without any chlorine atoms, is for Aroclor 1254 and 1260 was somewhere in the * if Januiry1974 . *53 V >- i NPC00016181 I 207867 u ;.* range of 4 lo 10 g. These two compounds proved changes were also observed in the livers an. to be more toxic in weanling rats. The single oral treated skin. LDjq for weanlings was 1295 mg/kg. and 1315 Vos and Notenboom-Ram91 compare.: ;;-,p mg/kg for Aroclor 1254 and 1260, respectively, dermal toxicity of the isomer 2,4,5,2'/' : and the lowest lethal doses were 1000 mg/kg for **hcxachlorobphenyl with that of a PC3 Aroclor 1254, and 1200 mg/kg for Aroclor 1260. (Aroclor 1260) in rabbits. Their dermal appiu.i,. The intravenous LDj0 for Aroclor 1254 in adult resulted in early microscopic skin lesions female rats was 358 mg/kg,*4 Aroclor group. The lesions in the. 2,4,5.2'.-- . A number of short-term studies have been reported where the PCBs were given to animals over a period of one or two weeks. When the-PCBs were given in this fashion they were somewhat more toxic than when they were given in a single dose. These studies aie reviewed in great detail by hexachlorobiphenyl group appeared later an-j . less severe. Both experimental groups showe-:. i . changes. The liver damage was more pronounce-, the animals that had been exposed to hexs-rh,.biphenyl than in those that had been exp-'v-.-d : the Aroclor. Nelson et al.12 and will not be reported here since In inhalation studies with Aroclor 1242_ and (hey do not give a great deal of additional 1254' rats, mice, rabbits, guinea pigs, and one cat information. were exposed to PCB vapors five days a week for Vos and Koeman9 studied the toxicity of three PCB preparations in chickens, Phenodor DBG. a French product, Clophen A6G. a German product, and Aroclor 1260. a product from J Monsanto in the U.S. The birds were fed 400 nom of PCBs in their diets for 60 d a w the Phehuclnr < and Clophen were much more toxic than Aroclor. The birds fed the two European samples, died between 12 and GO days after onset of exposure. The findings at autopsy consisted of hydropericardium, ascites, subcutaneous edema, and liver necrosis. Only 3 of 20 birds fed Aroclor 1260 died during this time with hydropericardium. None of the Aroclor fed birds showed liver necrosis. Several weeks. The concentration of Aroclor 1242 1 vapor was 1.90 to 8,63 mk/I and no ill effects werT^ observed, The exposure of. animals to the vapor of Aroclor 1254 in the concentration of 5.4 or l .5 fig/l produced enlarged livers in rats. An inteicurrenl respiratory infection in some control as well as experimental rats made it difficult to interpret the results.98 Since it is now known that the chronic toxicity of PCBs is a more important parameter m establishing their effects than short term exposuie.s. further studies with lower levels over longer periods of time w/ould give mut meaningful information. Fortunately, most of ihr compound; under discussion have a low vapor pressure so Hint exposure by inhalation prohabl^ docs not present a great piublem. _With the help of mass spectiometric and micro A number of reproditciit.111 studies in various cuuiomtUlt JUJlyiet. 'HKTHmUtTr'S11f ueiermmeT" species have been conducted with PCBs'; usual;-, that the two European samples were contaminaTed embryo toxicity was observed but there were iv- with tetra- and penlJchiurodiheruoftirgm. As an malformations. IL'ath et al 99 were unable jrdditional contaminant, chlorinated naphthalenes dcmo*\t:yie an effect on rrprtuiuciivn with me were found. ary levels of 25 and 50 ppm of Aroclor Dermal toxicity studies with the three com milliards and bnlnvhiies. Art increase in egesht! pounds, Clophen, Phenodor. and Aroclor 1260. in " cracking or reduenon in >liel! thick revs was jl. rabbits96 liter two days led to reddening of the no< observed while DDIi, which was used j skin where the materials were applied. -The skin posiijvc control, induced significant shell ihun.n-t reaction was more pronounced after one week, and cracking and-a reduction in hatching sm.cc".' desquamation of the external epidermal layers of in nvj.lli*Mls hut not in hobwhilc. the skin was observed, the hair grew at a reduced In Held s'lidics with brown pelican shell' rate, the skin became thickened, and prominent found Mini llunmng could be besl eorrei u transverse wrinkles developed. The lest animals DDF residues rather than dtddrin or K B> 1- showed a gradual loss of weight. and one of the finding indicates; according lo Blue el al..10 ` rabbits given Phenodor and three of those given PCH- arc nol responsible loi eggshell llnnn -Chvphnr died. AJLruit_oitc rabbit showed fluores mo-.t buds followingcnvnonmvnial e.\|umu. cence of the liver under UV light, nod microscopic"'' 1* 'Fcakall10 '*led ringdoves 10 ppm Aroclo' : 1 454 CRC C'itucl Aicv.-cn-v in Tnxivnhtgy 7 ; . f . - NPC00016182 207868 & for 6 months. He injected olhers with 160 mg/kg intraperituncully for 4 days, and evaluated the ashed eggshell'weights of these birds. He fuund nu difference in eggshell thickness between the experimental and the cumred groups. In a pheasant reproduction study with Aroclur )254,IOJ,M,J a significantly greater difference in the number of eggs that were pipped but not hatched was fuund in the group of hens that received 50 mg of PCBs weekly. Egg production and hatchabilily was lower in this gruup, the survival of chickens that did hatch at six weeks of age was reduced, and the ones that did survive were significantly lighter than the controls. Arodor 1242 at 10 ppm or 100 ppm and Aroclor 1254 at 100 ppm in the diets of chickens did reduce egg production and hatchabilily and caused thin eggshells, while Aroclor 1242 at 1 ppm, Aruclor 1254 at 10 ppm. and ArocJor 1260 at 100 ppm did not affect reproduction in chickens.' 04 Ip a rat reproduction study, decreas ed survival of pups at a dietary level of 100 ppm uf either Aroclor 1242 o r 1254 was observed. The results obtained from the reproduction studies in birds particularly seem to indicate that the lower chlorinated biphenyls affect reproduction more than the higher chlorinated biphenyls, and it is very important to determine whether the lower chlorinated biphenyls shut have been suggested as replacements for presently employed chlorinated biphenyls have an effect on reproduction. In anoihcT rat reproduction study94 the lowest dose of Aroclor J254 that affected reproduction was 20 ppm fl.5 mg/kg/day) which'manifested itself in a decreased number of litters and less pups per litter. Neither 1 ppm nor 5 ppm of Aroclor 1254 had an effect on reproduction in the Sherman strain tat; however, an exposure uf the dams to the Aroclurs at 5 ppm or higher increased the livcr-to-body weight ratios in weanling rats of both sexes. At l ppm the increase in liver weight was observed only in Fla and Flh weanling marc rats. The dietary levels of 100, 20, and 5 ppm of Arodor 1260 did not affect reproduction-, how ever, the liver-io-fcody weight ratio or 21*<Jay-oId pups was increased at all exposure levels. Mink are highly susceptible to the toxic effects uf PCBs. A daily dietary intake of 30 ppm resulted in death in about 6 months. F.xposure to 5 ppm Aroclor 1254 in the daily diet severely affected, reproduction,' 115 . .Unfonjinujcly.. it J often very difficult to compare the various dietary levels that affect different species, since the food consumption may vary decisively I'rutn one animal species to anuthcr ami the daily fnntJ intake in g/kg body \vi. is often not included in the scientific report. Some mice strains, for instance, will be exposed to as much o i a given substance that is present in the diet at the concentration of 300 ppm as rats on a diet containing 600 ppm or more. With some species it is of course difficult or almost impossible to measure the food consumption, but a greater effort in establishing these very basic facts wouldbe helpful for the interpretation o f toxic effects in different species. Not very much is knuwn at present about the toxicity of teryhenyis or chlorinated terphenyh. These materials have not been studied to any great extent. The few studies that arc available only describe very specific effects such as microsomal enzyme induction in the liver and give no overall evaluation of their toxicity. The most toxic of th e . various chlorinated dihenzod'tcjxins and dibenzafurans is 2,3,7,8-tetrachlorodibenzodioxin which, in the German litera ture. is sometimes referred to as 2.3,6,7-tetruchhwudjbenzodioxin. Single oral ~LD< ^ values are female rats: 44,7 mt/kg: male rats: 22.5 uxdkg-_ male guinea pigs: 0,72 pg/kg: rabbits: < 3 0 jtg/kgt and dtnfs: betwecn_.30 and 300 ug/kg.106 It L possible that young animals are more susceptible tu the toxic effects of this compound than older . animals and LDS0 values should therefore also be determined in weanling animals. One of the characteristics of this compound was that it caused delayed death, sometimes as delayed as 40 days after a single dose. During the period of intoxica tion, the animals lost a great deal of weight,'06 , On the other hand. 2.0 g/kg uf dlchluiodibenzo-p-dioxin and 4.0 g/kg body weight uctachlorodibenzo-p-dioxin were not lethal to rats. Hexa-, hepta-, and trichlurudibenzo-p-dioxin were more toxic.106 Schulz'07 in earlier experiments found that oral doses of 20 to 50 pg/kg/body weight or 2,3,7,8-tetraehlorudibenzudioxin resulted in fatal liver necrosis in rabbits. More recently Milnes10" reported that lOpg/kg was lethal to rabbits. The 2,3.7,8-tclrachlorodibcnzodinxin has a variety uf specific toxic effects. Whether these effects vary in different species is presently not well defined. With the interest in (lus'`and the other related compounds, more information on January IS 74 " *53 NPC00016183 207869 I i 1ii f 1 |( i1 i i ii pq P: r- nit -.ii (heir toxicity probably will soon become available. Ug/kg/week died wilhi:. 32 days ami a The problems inherent in studying ihese com weekly dose of 0.2 ".id their weight, pounds are the unavailability uf some of f-ern, the Of the various animal spf the guinea pig fact that they may not be completely pure, seemed to be most susc*. * the toxic effects particularly the hex- and hepta- preparations, and of TCDD. of course with respect to the letrachluroc-oenzu- Reduced thymus a.-, weights were dioxin, its extreme toxicity, which n.-f.es it observed In rats that re . rngle dose of 25 difficult to handle. fig/kg TCDD. The rats : j at the various In contrast to PCBs, teratogenic as well as dosage levels showed ir.av' r.orrhages into (he felotoxfe effects have been reported foi `..3,7,8- heart, liver, brain, adren..1 . . and gastrointesti- tetrachlorodibenzodiuxin. Sparschu et a! * 7 gave nal tract. Necrosis a n d _;- of the elimduiar 0, 0.03. 0.125, 0.5. 2.0, and 8.0 Jsry of part of the stomach w; . `ed. Occasionally 2,3,7,8-tetrachlorodibenzo-p-dioxin to pegnant organized thrombi were jv. and the liver reveal- rats on days 6 to 15 of pregnancy. With the 0.03 ed necrosis of.parenchyr; :s. Sublethal doses pg/kg dosage level, an effect on the fetus -.<as not of TCDD produced mi;; changes in liver. observed, but doses from 0.125 tig/kg on up kidneys, and thyroid, in ir . joeys the collecting resulted.in fetal mortality, early and late resorp tubules were primarih . ... ::?d. Doses of 0.1 tions, and fetal intestinal hemorrhage. Kr.ers and jrgfkg/day for 31 days, i ,? dose of 5 jug/kg. Roddick110 gave doses ranging from ! i : i 0.125 and multiple weekly d-. .f 1.0 and 0.2 pgfkg /igfkg to Wistar rats on days 6 to 15 of y- cnancy. did not produce chang-., >> . as discernible with These authors also observed pronounced iciotoxi- the light microscope. city. at all dosage levels except 0.12; u. kg. The Toxicity studies we:. <. : .`.,,`ted in our labors- I affected fetuses showed cerebral and intestinal tory with tlichloroph. - ` lire oral LDJ0 fur hemorrhage as well as subcutaneous edema at sdult female rats wh>. nerial was given in autopsy. The offspring of mothers given 0.5 and l peanut oil by stomach cs 1,660 me/kg and pg/kg of the teirachlurodibenzo-p-dio - gained in males it was 1.5C ; The only signs of less weight during their suckling pcricJ ir./ their toxicity ubserved in ; e were diarrhea and survival to weaning was reduced. This v'd Ivave some depression. W'r mg/kg was applied been caused by the intrauterine ef:;*; of the daily in propylene g.!>. . i* skin of a clipped diuxin or by excretion of the dioxin in 1 :ni)k uf area of the back of lull rats as a I2.5& the dams given the material during j vgnancy, solution, local irriio:. ,'.e skin with some Courtney and Muuie111 observed ci?f: . .hates in ulceration resulted :. was no systemic i 3 strains of mice given 3 yg/kg TCD1 .utane- toxicity. The acute . ously from days 6 to IS of pregr- ; and an male rats was 16-8 ;>us LDjo in adult .iL-n it was given in a increased incidence of kidney anom: liydro- saline lecithin s.usper. j iwo generation tat nephrotic kidneys were also proriuz-. mouse reproduction study, . eveI as high as 1,000 pups that were nursed by dams treated u TCDD ppm (50 mg/kg/dav . r effect. Cross and during pregnancy or at time of parturr ` 11 microscopic examir." ' (he various organs The 2.3.7,8-teirachlorodib.enz-. , :-;oxm is including rhe brains rrents as well as the extremely toxic in the chick embry 'sav.*1 In offspring did not rr. -rpiiological changes the rat. hcxachlorodibenzodioxir. caused fatotoxic that could be related u . 'u'uruphene exposure. effects at I and lOyg/kgund teratogenic effects at These findings indicated ; dichlorophcnc was doses of 100 pg/kg,10* Harris rt gave not very toxic to s. . 'h e great difference female rats daily doses of 0.1. I. 10 itgfkg between the oral .v. intravenous LDSo TCDD for 31 days. Rats given lOug.'-v tost weight suggested cither pin:: oiion or rapid break- *, iI and most of them were moribund w. *m 3 weeks. Weight gain was also less at the I tg/kg dietary level. The rats receiving 0.) pg/kg were not down o f the materi.i. ;.. gastrointestinal (racl. Marlins*1 lists the scuu -l>so lor guinea pigs as t,250 mg/kg and for dog; . 2.000 mg/kfi. R.ils fed i affected. In experiments with guinea pics. of 10 fur hO days on a d.c Zvataming 2,000 ppm animals died after a single dose of '3 /Jg/kg TCDD. showed no evidence of ! Mty, H3 while guinea pies receiving a single dose of 1 pg/kg The acute toxic 1 . ' icitlnrophctu' is much gained less weight. All guinea pigs receiving 1 greater ami varies r: v" M animal species and is Jj- i56 * CRUt'iiirvI in Tit.xivuI.KY, NPC00016184 207870 m it .*. - .............. -- J l also dependent tin the route of administration and the solvent in which it is given. Gump*15 lists the acute oral LDi0 in mice as 168 mg/kg and cites 1-DJ0 values reported by other investigators rang-' ing from 80 mg/kg tn 2)5 mg/kg. Dogs and sheep were more susceptible to the toxic effects of hexacltlorophenc. The merest in the effect of hexacldoropliene on sheep in particular stemmed from the fact that it is effective against liver flukes116 and advocated for its anthelmintic properties. This even led to the treatment of children with orally administered hexachlorophene in China.113 Intravenously administered hexachlorophene in rats resulted in an LDjo value of 9.1 mg/kg and in rabbits of 8.5 mg/kg. More recently we found that the ural LDi0 (single dose) for adult male rats was 66 mg/kg and for adult female rats 56 mg/kg; for weanlings it was 120 mg/kg when the material was given in peanut oil by stomach tube to Sherman strain rats. The acute intravenous LDJ0 was 7.5 mg/kg in adult male rats.111 Nakaue el a).1** reported similar oral LD50 values in Wistar rats. The single oral I.Djo fui sheep and cattle lies between 30 and 60 mg/kg.130 Single dermal LP(0 values have not been reported in the literature. We found in rats1' that single applications of 600 mg/kg hexachlorophene in 95" ethyl alcohol solution caused death in 1 of 10 rats. Dermal application of 24 and 4S mg/kg/day for 30 days in propylene glycol or in a detergent as a 37c solution caused ulceration of the skin and affected tite white mailer of the central nervous system m at least half of the rats. A dose of 12 mg/kg/day as a 1.5& solution caused only mild erythema and desquamation of the skin or rats. No effect on the central nervous system was observed. Whether a single dose given in either propylene glycol or a detergent would be more* toxic than one given in ethyl alcohol v.-as not established. The signs of acute hexachlorophene poisoning following a single otal dose varied with the age of the rats.111 Adult rats suffered from severe depression and diarrhea, but showed few signs oT leg weakness Weanlings, on the other hand, developed posterior leg weakness as well as de pression. The leg weakness became more apparent in the adults when they were given repeated doses of hexachlorophene. Death was usually preceded by convulsions in rats dosed intravenously. In rats killed with liexachloinphene. rigor mortis developed very rapidly and wus very prunouncrd. Body temperature elevation following hexachloro- phene poisuning has also'been recorded in rats.1 Both changes, the pronounced rigor mortis and the temperature elevation, may be related to the fact dial hexachlorophene uncouples oxidative phos phorylation. Additional symptoms of hexaehloru- phcnc poisoning particularly in other species me discussed under neurotoxicity. Nakaue et ul.l ,? fed 400 ppm hexachloro- pheite (2S.9 mg/kg/day) to Wistar rats. They developed leg weakness within 5 days, diarrhea, emaciation, and they died in 7 to 10 days. No deaths occurred in the rats that were fed 200 ppm (about 13 mg/kg/day.) for 16 weeks. ' In a two generation rat reproduction study which we conducted in our laboratory. \vc found a reduced survival in the offspring or the F, generation fed 100 ppm (11.8 to 5.5 mg/kg/day) in their diet for 54 and 166 days. This was statistically significant in tiie second breeding of the F'i generation. In the second generation the Jitters were slightly smaller, but the survival to weaning was not significantly reduced.11* A dietary level of 20 ppnt (2.3 to 1.1 mg/kg) did not influence reproduction. Thorpe121 reported that the oral administration of hexachlorophene to male rats as well as single doses of 25 and 50 mg/kg hexachlorophene to sheep produced de generation of spermaiogenic cells: multinudeated cells were observed within the tubules of the testes. The doses given by Thorpe wcie higher than the ones we used in the reproduction study, and microscopic examination of the testes of the rats used in'our reproduction study did not reveal any abnormalities. Toxic doses of hexachlorophene given to the dam during pregnancy may result in malformations of the offspring. Thus far, two studies have been reported in the literature; both studies were conducted in rats.113,1 73 It should be determined whether malformations can also be produced in other species and with lower doses before any conclusion can be drawn from these findings. ** A number of human poisoning cases have been reported in the literature and some or these were fatal. Most of (he poisoning cases in humans have either been caused by accidental ingestion, because a hexachlorophene detergent solution was mistaken for milk of magnesia, or by the applica tion of hexachlorophene to large areas uf btirneJ or otherwise damaged skin.11**'14 Larson3 was ennary 197* <37 NPCOOOI6185 207871 't i* V' '1 K '** Bk&. '& l the first to recognize that burned patients observed in these patients were mainly confined to developed convulsions and other toxic symptoms the central nervous system and to the gastro after repealed exposure to hexachlorophene which intestinal tract. The group that received 3 doses of was applied to burned areas of the skin. The 20 mg/kg/day had the most severe reaction. One incidence of convulsions in burned patients patient in this group became comatose on the decreased when the use of hexachlorophene was fourth day with loss of light reflex and positive discontinued/3* pyramidal signs. Fundus examination of the eyes Eleven cases have been reported*36,131 in revealed mild papillary edema. A low grade fever which a tingle dose of 3% hexachlorophene deter* was also observed. Complete recovery occurred gent solution had been taken orally. Castro-. within three days. One half to two thirds of (he intestinal symptoms including nausea and vomiting patients had diarrhea within 1 to 2 hr after were observed in ten of these cases; the eleventh medication and in some cases a marked depressive case, o child who drank about 230 mg/kg hexa effect on the central nervous system was also chlorophene, became comatose and died. One observed. An additional case of poisoning after additional death occurred in this group, however, oral ingestion was reported by Korlof. and this was most likely due to ocher causes. A Wjnsten139 in Scandinavia and two fairly recent 17-day-old infant was accidentally given daily oral cases with fatal outcome c-urred in the U.S.*10 doses of 37 mg/kg hexachlorophene for 7 days. On Hcrter131 was probably the first to report a the third day the child started to have loose stools. human poisoning case caused by the dermal On the fourth day of hexachlorophene administra absorption of hexachlonplicne. Because of a tion the child developed spasms of the extremities language barrier, the mo:hc: of a newborn infant with twitching of the face, sucking movements of applied hexachlorophn.e as a lotion after the the lips, and lateral shaking movements of the infant's bath without rinslrg the baby. After four head. It was also noted in the case report that the days, excoriations appear s or, the child's face and child refused to take the medicine, there was buttocks, the infant d;* .;iped convulsions, his drooling and frequent vomiting, and it is not quite face twitched and his e>.: ties'jerked. A roving clear whether the child received 37 mg/kg/day. nystagmus was present. *:..s cltiM recovered with The child survived. A week after the hexachloro in 16 days after expos*-" : c hexachlorophene had phene suspension was discontinued, persistent been discontinued. spastic' flexion of the fingers, hands, and arms was In addition to the tv* * already mentioned still observed. After a month the hands were in which poisoning r .red'' because of oral slightly flexed at the wrists but were not spastic. ingestion, Mullick*30 * sported four cases - Horizontal nystagmus was observed but cleared children who .died T o n g the treatment for later. The child was discharged when it was two bums in two instan- :,d the treatment for months and three weeks old. At this point it had severe ichthiosis with . hexachlorophene bath. shown great improvement and when checked at six Larson1 reported , burned patients who months the child was healthy looking/38 Pilopil developed central ;s system symptoms mentioned in his discussion a number of other following the derm. ,:-.>sure to hexachloro unpublished cases, some of which were reported to phene. All six pat* recovered. Korlof and the National Clearing House for Poison Control Winsten119 report: cases of poisoning Centers. Among these was the case of an infant following the dermaration of hexachloro who by mistake got a spoonful of PhisoHex in phene. Both paOf.- :re fairly extensively each feeding bottle for approximately two weeks, burned, and devslc. .entral and pcriphcial when it suddenly died. nerve damage as we!; ne kidney impairment. - Chung el al.117 treated a total of 105 panents Ixckhart*33 me;;:. ; six deaths that were 5 to 15 years old with hexachlorophene for reported to the r.l -jused by the topical chlonorchiasis sinensis infestation. Sixty of these application of he-., ..wophenc on burned patients received t dose of 20 mg/kg, 32 received patients.' In all of .e cases the skin was 2 doses of 20 mg/kg/day, 8 patients received 3 damaged, probably er* ring absorption of hexa daily doses of 20 mg/kg, and 5 patients received chlorophene, and a!- skin docs represent a 20- Trig/kg body weight -on,the. first day_ and 10 barrier, it is not impr tble --a fact wc lend to mg/kg on the second day. The toxicity reactions "Tbtget at times. Darri_r.. or abnormal skin may be SB CRCCntit el H n irw u n Taximftfty NPC00016186 207872 w <1 more permeable, and the skin of prematures, SPECIAL TOXIC EFFECTS newborns, ar.c children in general mav liave- a t sorption characteristics other than those of Skin Reactions . adult human beings', another factor that is often * Chioracne. This occupational skin disease can be produced hexacJtlurophene poisoning have been reported in by a number of chemical compounds.1'11 Poly Trance. The deaths occurred in infants who were chlorinated biphenyls as wil ac ih .i treated with a powder to which 67a hexadiloro- naphthalenes, a few isomers of the chlorinated phene1had been added by mistake. This powder dibenrodioxins. and chlorinated diben/ofuram ar was mainly applied in the diaper area where the all able to produce chloracnc in a certain pro- skin became inflamed, this as well as the diaper _ pt)Ttlun~~oI the human Bunnlminn--Oilv skin nnrf which served as an oedusjve dressing, probably en large pores seem to predispose to the disease while hanced absorption. These cases will not be made the opposite is (rue for smooth, tender skin and it public until litigation problems have been settled. is possible that young children might be less susceptible because of the nature of their skin. Chloracnc has also occurred in workers engaged in the production of 2,4,5-T1. During the indus Little published information is available on the trial production of trichlorophenol a sudden rise in toxicity of the germicides in Figure 2 aside from temperature within the reactor has occasionally hexachlorophcnc and dichlorophene. The acute resulted in accidents. In one instance this exo oral LDSo for triulosan (TCC) in mice and rats is thermic reaction caused an explosion.*08 When about 4,000 mg/kg.,JS The acute subcutaneous the me in temperature occurred, 2.3.7.8-letra- toxicity for rats was 14.700 mg/kg am! the acute clilorodibenzodioxin was formed and severe out dermal toxicity was about 9.3 g in rabbits. breaks nf chioracne were observed among the. Concentrations of I to 5% solutions caused eye workers.1314 irritation in rabbits. Daily oral doses ofTCC up to The, term chioracne was first used by Herxheimer 1,000 mg/kg/day for 4 weeks did not produce an in 1899,l3e who thought it was caused by effect in rats. When 2.5 and SSrsuspenskms in gum free chlorine generated in coitmn factories. Wauer arabic were applied dermally to rats for 4 weeks, in I91813' and Tricky in 1927140 suggested the the animals did not gain as much weight. Un term ``Pwnakrankheil.'' Aanther term sometimes published material cited in the minutes of the OTC used is halogemvaxacnc. (Over the Counter Drugs! Panel listed a "no As the word chioracne implies, part of the effect" level as 170 mg/kg/day in rats and for a lesion of this occupational skin disease resembles 90-day feeding study in rabbiis,438 mg/kg/day. In adolescent acne; however, it is generally mere baboons a daily oral dose of 100 mg/kg/day did severe and the distribution of the lesions is not not produce a toxic reaction. The minutes also consistent with adolescent acne, although it may mentioned some work where lower doses showed be superimposed on adolescent acne. Chioracne is an effect on rats and dogs. Since some of these described in the literature as consisting of the results are conflicting, further studies are indicated formation of comedones with m without cysts anr! and should be performed with products used as pustules. The follicular orifices are filled wnh ingredients in soaps; a chemical analysis of'these sebaceous and keratinous material. Melanosis and a products is necessary to determine whether im secondary inflammatory reaction may also exis< purities might be responsible for varying results. In addition to lliis description, skin changes owu* Chioroaniiine as a result of healing during manu that have been discussed less frequently in the facture could be present as such an impurity. Of more recent literature. These skin changes arc the anilines, according to Hamblin,'3b only p- and usually referred to as cable rash ur cable itLh m-chIonian line may cause methemoglobinemia if Cable rash has been observed in workers who lu^r absorbed through the skin. The o- and m-chloro- insulated cables with various mixtures of chlonr anilinc may cause liver and kidney damage. Tn- ated naphthalenes and chlorinated biphenyls . closun itseir is .also absorbed through the skin and in workers handling these ariiles.14 1 It has beer absorption is greater when applied to damaged observed also in laciorics that make i.*" skin. denser*.141l43 According in llratm .'41 a lew ' January.1974 -is jr. NPC00016187 207873 the eases with m.'ftst chloracne started with changes n the of the face resembling photo sensitivity and - rearers of these lesions suffered severe puritus jj Y; areas of the skin lesions. This lesion termed c. :c rash occurred predominantly on the checks-;.ot also on the skin of the arms. The outcome of cable rash varied; it could either disappear or could develop into full-blown chioracnc. In addin;: the pustules and comedones of typical chiorj-- .*. brownish kcratinizalion of the skin was obi; -td in some patients. Braun141 pointed out . ; the workers, particularly those with poor h> . transmitted the disease to their spousesand cl; .d-e;i through direct contact. Many of t ic occupational cases of chloracne that have bcr. -.-ported in the literature have been reviewed pic-.* jjiv.137,144 Jones and Alden145 reported one . *. ;c first outbreaks of chloracne in a company tr.j- v.as engaged in the manufacture of chlorinated biphenyls. They described one typical case and mentioned 24 men working on the manufacture of chlorinated biphenyls; 23 had had an aeneform eruption un the face and body.' Particularly during the 1930s and 1940s large outbreaks of chloracne occurred in a number of factories. Much discussion has been devoted in the literature to the problem of whether the chloracne was produced" by external contact alone or whether it was caused by systemic absorption, particularly the inhalation of vapors. One question asked was whether internal administration could cause chloracne. and" if so what part u played in any particular case. As we know now from outbreaks of poisoning due to the consumption of rice oil tn.o was contaminated with polychlor inated bipj.i --Is in Japan (see under " Yusho"), chloracne ct .-i be produced by the systemic absoip- tion of chlorinated biphenyls. Jhe experiments bv Shelley ami Kheman1*4 and Plewi^147 showed that the skm lesions could also be pruduced_b^ applying -be chloracncgcmc compound to the akin.-- After occupational chloracne manifested itself, the patten's also complained of systemic effects such as loss if appetite, nausea, edema of the face and hand-. .-dominal pain, vomiting.and burning and soreness of the eyes. Particularly with the chlorine.. iphlhalcncs. hepalotoxii: effects have, also be;' r~-T*gfvcd. Application nf chemicals to the sktr "van hfad" ru their absorption anil c a u ^ systemic toxicity. Whether a svstcniic toxic effect results will depend on the skin surface area covered, the amount of the toxic substance applied, and the ability of the skin to absorb it. If dermal absorption is poor, a single application will usually *not be very toxic, fiowever. if the com-, pound is not easily metabolized and excreted.' repeated dermal application mav_ result in suffi cient accumulation in the body t came systemic toxicity. Crowe'44 has pointed out that chloracne due to chlorinated naphthalenes has decreased because these compounds have been used less since the Second World War, However, Weber in 1969'48 reported un outbreak of chloracne in u company engaged in the production of electric coils that were coaled with a mixture of chlorinated naph thalenes with the tradename "Nibrenwax." Detailed examination by several German invest igators indicated that the capacity of 2,4.5trichlorophcnol as well as 2A5-T (2.4.5.trichlorophenoxyaectic acid)149'50 to produce chloracne was due to contamination with 2,3,7.K-tetruchlCTodibenzofuran or 2.3.7.8-lctrachlorodibenzodioxin. Schwartz et al.1* 1 indicated that the most potent chloracne producing agents were the chlor inated naphthalenes, chlorodi phenyls, and cltlorodiphenyloxides. Persons who worked with chlorinated naphthalenes usually developed acne after a month or more of exposure. Light microscopic observations made of the chloracne lesions oT humans varied somewhat, depending primarily on the length of tunc the lesions had existed at the lime of exan ation. The curliest change encountered consisted of ucantholic widening of the external root sheath of the hair follicle. Above n layer of basal cells o to 10 layers of large round cells were observed, followed by a layer of 2 to 3 cell rows of granular cells. In later skin biopstcs, folding of the basal layer occurred ami comedo formation was observed, resulting in dilatation of the follicle and atrophy of the epithelium surrounding the com edo. The contents of the follicles consisted predominantly of keratinous material rathei than sebum. The sebaceous glands also underwent changes. Within the alveoli of the jdands the basophil cells increased in ritunher and basal cell by|>crphisia was noted, the ducts I (In: sebaceous glands were filled with :i solid mass ol basoplul epithelial cells. The sebaceous glands gradually 460 c - Ki/I R c v m m n Tnxicnhtxy NPC00016188 !'*! 207874 r* \ \- A: ....... jJ i disappeared and in later secretions were only be investigated In chloracne patients who manifest g recognised as solid protrusions of the comedo hyperpigmentation. * i % wall. Aside from atrophy of the sebaceous glands, In none of the human case reports on chloracne *, hyperplasia lias also been described and many large have the mucuus membranes been studied to any ii sebaceous glands forming huge cysts filled pri great extent. Vaginal smears in women and sputa, 4 marily with keratinous material have been corneal, and nasal scrapings may give helpful observed. Hyperkeratosis and acanthosis o f the information in this respect. Since in anjraali_ surrounding epidermis has usually accompanied exposed to polychlorinated.tiolvcvclic eomyounds these lesions. Foreign body graiiulomata which are ~Iow vitamin A lcve|s_Jayg_JagiLJhsmcdJfl-llig- thought to result from rupture of the follicular J h ^ jtjs ^ jo s s ib lc ih a iih e jm m ^ ^ cysts have been found in the skin.'*7,131 In the jyjtantin A absorption and storage. and_itdgamifi. Japanese poisoning incident (Yusho). the predom utilisation in the liver and elsewhere in the bod^ inant skin lesion was marked hyperkeratosis of the "if thejesion is PioSucedTv inadequate utilization epidermis, cystic dilatation of the hair follicles. of vitamin A .thcn administration of_vjUmiiLAjI 3nd ait inercasc'in the melanin pigment in the basal se may nut necessarrlj^aUer the course of .the cells of the epidermis. Foreign body granulomala jtisM Sj^Prnreindcfie^ frvtm. A were also observed. The contents of the cysts vicamtnosis A in th aiitaftcH s^mnsgmt of vitamin \ TM consisted partially of keratin. In a stillborn child the hyperkeratosis was even more pronounced, accompanied by atrophy of the epidermis: cystic *A^ThjLjvgt^CHC190% of the available vitamin A ^ar^Mhcse^sigre^jjja^ IffllMitiJI ttyrr rii The microscopic appearance of the skin lesions dilation of hair follicles was observed, especially in resembles m some respects observations made in the skin of the head. According to earlier skin of patients suffering from vitamin A reports131 some differences exist in the micro deficiency, In vitamin A deficiency, hyperkeratosis scopic changes observed in the skin following of the hair follicles is observed. Multiple firm exposure to the various chlorinated cyclic com papules may develop as a result of the keratin pounds that can produce the lesion. It is, of plugs in the sebaceous glands. Dryness and course, possible that these histopathologic vari scaliness as well as furunculosis of the skin may ations arc in reality only differences that exist also be present. In contrast, night blindness has between an early, a well-developed, and a not been reported as a symptom in patients subsiding lesion. suffering from chloracne. Unfortunately, chlor . Many theories have been advanced to explain acne outbreaks are often not well investigated and, the development of chloracne. None is satis because of problems with litigation, accounts are factory. The concept of mechanical plugging of often only published many years after the out the hair follicles on skin exposure to polychlor break has occurred if they arc published at all. inated polycyclic compounds is not supported by Chlotacne is a very persistent skin disease and the fact that chloracne develops following in the Japanese outbreak of poisoning with chloio- ingestion of these compounds. It is possible that diphenyls, the skin changes were still present in a ilic skin lesions are caused by a toxic effect on the .number or patients three years after exposure lu epithelium of the skin and its appendages which is the chlorodiphcnyls had been discontinued. Wlwi manifested in proliferation and loss of normal needs to be investigated is whether this persistence function of the cell. Since the chemicals are lipid of the skin disease is caused only hy the fact lhat soluble they could occur in *j more concentrated once the lesion has developed it regresses very form in sebum and therefore affect sebaceous slowly, or whether it may be sustained by the glands mure .than sweat glands or salivary glands. ehlor.iene-producing chemicals which have been The'reports on the Yuslio outbreak suggest that absorbed and stored in the body, particularly the proliferation of the epithelium of the mucous adipose tissue, and arc only very gradually membranes may also occur. The increased pigmen excreted In patients with chloracne. adipose tissue tation of the skin and mucous membranes may biopsies should be analyzed fur the presence of the indicate a functional disturbance resulting in offending polycyclic pulychlorinated compounds. increased melanin production. Ifypcrpigmcnlation If adipose tissue is unobtainable, the sebum of the is also observed when the adrenals arc durtlagcih as '.ar-etKial or the cement ,,of the skin lesions may in Addison's disease, and adrenal function should represent a good substitute. g i a rr TM <61 NPC00016189 Apparently there is no known effective treatmerit far chlorucnc and the best control measure is prevention. The' manufacture of chlorinated hydrocarbons and (he coating of wires and con densers with insulating materials containing them should be done in totally enclosed areas so that the fumes of the insulating substances and of the solvents if any are used do nut come in contact with the workers. The workers should be provided' with clean overalls and underclothes daily. These should be laundered at the plant and cleaned in such a way that no chlorinated hydrocarbons remain on them. Shower baths aftci work should be compulsory and supervised and there should be_ sufficient shower facilities provided so that the workers are not unduly delayed after work. Special synthetic wetting agents'51 seem to remove the materia] from the skin more effectively Ilian ordinary soaps The experimental induction of chloraene in men has also been accomplished with 2 appli cations of 10 tre J.-M.^-leirachlorotlibenzo-pT .dioxin* ^ and wuh Halt}wav lfl14 nentachloro-nmhlhalcne, and hexachloinnanfithalene 1 1 Experimental work in animals has shown that most species do noi develop the same type of dermatitis. The rabbit's skin.t s l particularly the inner aspect of the ear lobe, provides a good test medium for the development of acncfonn dermathis. The rabbits, in addition to the development of hyperkeratosis of the ear. will also show a hepatoloxic effect.150 Microscopic examination of the changes in the rabbit ear were described by Jones and Kri/ek.11'"1 Inagami and Koca*54 were able to produce hyperkeratosis in hairless mice that were fed rice oil contaminated with Kaneclcu 400. These mice also showed hepatomegaly. Microscopic examination of the skin revealed cystic dilation and hyperkeratosis of hair MhcJes and sweat glands. Hairless mice may be another suitable experimental animal for testing the effect of these chemicals on the skin. X-discase in A ninials Ch!oracne*typc lesions do not usually occur in- other species except for the rabbit ami the hairless mouse, already mentioned. However, in N47 Olafson155 described a disease in cattle which was designated as X-Uiseaxt- ar hyperkeratosis. Symptomatology included excessive lacrimalion. diarrhea, polyuna. marked salivation, and dis* cltargc from the nostrils. The animals developed a chronic cough, poor appetite, numerous red maculae in the buccal mucosa: in addition, hyperkeratosis of the skin developed. The skin was haul, and fissured particularly across the withers and sides of the neck. Many wart-likc pruiileratiuns on the lips, longue; and hard palate were also observ ed. The abomasum was greatly inflamed, swollen, and edematous with many superficial ulcers.'Sf' In the cows that had X-discasc, a low vitamin A plasma level was also observed, which again indi cates that vitamin A may play a role in the etiology of these various disease entities. The disease was first thought to represent a virus infection and other factors were also suspect ed. After numerous investigations it was finally established that X-dlsease could be produced in cattle by the ingestion of highly chlorinated naphthalenes, and also by petroleum products such as crankcase oil. Dell151 tested the ability of the various compounds of the chlorinated naphthalene group to produce X-diseasc. He demonstrated in his studies that dichlurinaud ami IrichJnrinalPd naphthalenes did not result in X-diseasc, while tctfuchloronaphlhalene li3tl an effect and the higher chlorinated naphthalenes sucli as pcntachJoronaphthalcne. hexachloron a p h th a le n e , hepiachloronaphthalcnc. and ocncfdnrunaphihalcnc caused severe disease. Octachloronaphthalerie was less toxic than hexaand heplachlomiiaplithalcnc.AVufi^ncr1:iB report ed the occurrence of hyperkeratosis m callfc ill Germany'between l_9_4b and WAS. JThe cause of this outbreak was traced to a wood preservative which was used in postwar Germany. The .unhm.^ ^proved experimentally that animals housed in _ cahins-pariiicd with the wood preservative develop- 1 cd ivoica! X-Jisease- The poisonous ingredient In the wood preservative appeared only in certain lots of this product ant! later lots did not contain it. The poisonous ingredient was never identified by chemical analysts Animal experiments weie conducted with the toxic ingredients of rice, htan oil that produced Yusho in Japan. Those toxic substances consisted . of a mixture of chlorinated binltenvl'j f-pninminp chlorine, with a trace of O.OlTLor nanluluIcnes.^50 Whrn mice were exposed to them fot 3 to 4 months, they showed eczematous changes of the skin around the eyelids, erosion, ulceration and perforation of the car laps, loss of hair, and , erosion and ulceration of the skirt around lire neck, forelegs, and sides of the chest. The authors i 462 CFC t r i m at Revn-*'i i/i T/t.xivfUtyy ..................................! " ( NpC000l 6 l9 o 207876 Jk. '> staled that, at the end of [he experiment, some changes in the skin similar to those in the experimental mice were seen in some of the coni mi mice also, which makes it difficult in interpret their findings. In ffi57 a disease (hat will be discussed in greater detail later in this article occurred in a large number or chickens. Tdfn' disease and was .found to be caused hv ccrlair. toxic fat, Allen and Carstens16.p-fcd this suhsiar.cc, _ ^tojm jnke^ an d fa u n d jfa t. amunfl yther chance-. ^ tiic h ^ w i llti e ^ jis c u ^ ^ laler the m onk;-'. _ deveJoC^^MncralizedaloCji^u^lJitiiiaflfiUUi__ dema one to two months before death. Schoetlh et al. `"piciduced hyperkeratosis of (he skin in rats by feeding them hexachloronaphlhalene. Young sw'ine'81 and sheep'69 did not develop skin lesions as a result of exposure to chlorinated naphthalenes. roducts to which the skin had been - iuus brominated and chlorinated s. -.re irradiated in vitro in an approp;:. ic resulting breakdown products cair. .ic reaction on contact, without exp.. These studies were performer ,f. .forma (ion provided in earlier studk if.,' *' who had irradiated bron->: :Jurinated salicyianilides and found to, impounds gave off halogens. With 3,5 sub-, jlicylanilides. the halogen substituted ... -vsilion was lost. Tctrachlorosalicylanilidr nsitized subjects reacted to 3', 4', 5-irici-. ;ylanliide and to 3', 4'-dichJorosalkyL;. reaction to nonhalugcnaled saiicyliimhdc .a- .. observed. Halogens on the anilide ring were ti.:- jtfeefed by UV radiation, while the ha.c. .-r salicyl ring were replaced rtepwhc - * gen upon irradiation. Photocontact Dcrnvtitis and Irritation o f the 5kin As was pointed out earlier, the hyperpigmenta- tion observed in patients with chloracne as well as the reaction described as cable rash could also be (he result of a photosensitive reaction. This photo sensitive reaction is the predominant lesion in the skin produced by a number of germicides, namely, bithiono) (figure 2), telrachlorusalicylanilicte (TCSA), and a number of other halogens tec phenolic compounds including fenliclor. Patients who develop photodermatitis complain initially of a burning .sensation, and this is followed by itching. A rash develops which in the beginning is erythematous and associated with varying degrees of edema. As it subsides, pigmentation is observ ed.1*4 Wilkinson165 was the first ore to describe cases where patients developed a photo allergy wlucli was due to the incorporation of TCSA ini' popular soaps. He observed that the outbreak oi dermatitis was confined to the areas that usually were exposed lu light. Most of the cases reported from England were transient and as suor. as exposure to the compound was discontinued the reaction subsided. Jillson and Baughman166 and Jillson.'*1 on the oilier hand, reported that their cases were more persistent. Wiljis and Khgnun16* demonstrated that the phutocunnrci allergy ofthe skin with these various compounds was simply a contact sensitisation The reason that allergic reactions of the skin developed only after exposure to the sun was due to the pholodccompositiun of the various Among the patients who developed phot--i.*.. tivity to tetrachlomsalicylanilide' repyr.:.: : Wilkinson,1*5 a gioup of patients m ^ factory was also included. This same Uuibr:.u- *. reported by Jones.'10 who found that 3*i.'-ns. -'* employees who used the same soap very f ; ; . -.. iy, 3 \% were affected by a dermal' i i *. contact with tctrachlomsalicylanilide p re v r. the soap. Additional cases of photodermatins c-. exposure to lelrachlorosalicylanilide v. by Calnan ct al.,111 who pointed c - : pronounced edema of the eyelids was also ><"- ed when involvement was severe. Some c z m persistent or developed acute relapse: >* exposure to sunlight without further exp...-re the offending chemical. Frenk' 73 reported at; additional nine eases of acute contact dermatitis ; the hands due to iclrachlorosalicykm Epstein11,3 described a photoalJergic reaction r: two patients to tiibromos:rlicyianinde (TBS). With natch tests these patients showed a cross reaction iu TCSA. A similar crux* reaction pattern in a number of halopenaled salicylandides w ji also observed by Osmundsen114 in patients who had developed photodcrmalitis following exposure iijbromusahcylanilidc. Because of the phuloscnsilivity reactions and ihi cross reactions that were also found wr.h bithicTi-'l, which is similar in structure to icua- chh'Tos.ilicylanilids (Figure 21. the U.S. Food and Drug Administration, on October 24. Ilj(i7, with- dtew all new drug applications of all drugs that January 197* *6J w* NPC00016191 207& 77 i> 1 contained bilhionol. In the notice of withdrawal of approval in the Federal Register, the Commissioner of the Food and Drug Administra tion pointed out that bilhionol may in some instances -cause a very persistent photosensitization and severe dermatitis may occur with exposure to sunlight without further contact with tit; sensitiz ing articles, and that bilhionol may produce cross photosensitization with other commonly used chemicals such as certain halogcnatcd saticyianitides and hexachlorophcne. Hexachlorophenc preparations may also cause irritation of the skin if these preparations arc used frequently. A true allergic reaction to hexachlorophene is apparently rare.115 The .more severe reactions were usually confined to the skin of the scrotum and consisted of a primary irritant contact dermatitis.118*'77 The cases reported hy Baker c( al.1' * also included a 33-monlh-old Caucasian female who developed a rash around the buttocks, vulva, and upper thighs fallowing (he use of a hexachlorophcne preparation in her belli water. We observed severe skin irritation and ulcera tion in rats that were exposed- dermally to a 3% hexachlorophcne solution in propylene glycol or a detergent. The skin reaction was not as severe when the hexachlorophcne concentration was reduced to 1.5^.*1* Boutwell et al.17" reported considerable skin irritation in mice that had 5 ing/mouse applied twice weekly for 21 weeks. Harber el al.179 were able to induce contact photosensitivity with TCSA or TBS. These authors also found that cross contact sensitivity and photosensitivity to hexachlorophenc existed in guinea pigs with primary photosensitivity to TCSA and TBS. Photosensitivity to trichlorocurbanilidc (TCC) was not produced in guinea pigs. The guinea pig may represent u suitable animal species to study contact and photosensitivity or these and related compounds. After exposme to the photosensitizing agents has been discontinued, humans may still be sensitive to sunlight, a fact that needs further study. To a certain extent this could be due to a persistence of the chemical compound on or in the skin. D tc/iinm pfivm ' nsctl as an antimicrobial preserv ative may also cause contact dermatitis, ('msssensitivity in hcxachlorophune was not demonsirated.1 Chick Edema Simpson et al.181 and Sanger el a l.'83 describ ed a new highly fatal disease in chickens. The (iisi outbreak occurred in 1957 in Gcoigni. Large numbers of birds from approximately tluec weeks of age to adult laying hens were affected. Clinical signs consisted o f dyspnea, reduced body weight gain, stunting, subcutaneous edema, paleness, and sudden death. In young chickens, gasping was the first noticeable sign. This was followed by a waddling, unsteady gar. Gross inspection of the birds revealed a pale hca:i. fluid was noticed in the pericardial sac, and the livers were p.ilo, mottled, and had an irregular granular surface. In advanced stages of the disease the cliickens had large distended abdomens ilm were filled with fluid. Ecchymotic hemorrhages were present in the skin beneath the wings, on the legs, and aver the Q bone in a few cliickens. The kidneys were pale and swollen. Following the first outbreak, no addition al cases were reported until late summer and early fall of 1957, when extensive losses of chickens and turkeys occurred throughout the southeastern part of the U.S. Suspicion soon developed that losses were caused by the consumption of rations con taining certain lots of ,,nimal fat.18' Hie animal fats contained tallows and greases obtained a: by-products from hides preserved with chloro phenols prior to leather manufacture.11 This material was excluded from coinin'1rim5 puultiy lations and no additional cases occuh- ; where animal fats were responsible. Strips'"! * al.181 came to the conclusion (hat the edom the chickens was the result of vascular lln r:-* that seemed to be.the primary lesion. The vast-air lesion,* or endothclinsis. consisted of prr.jitr'-" and hypertrophy of the endothelial u lu - arterioles and small arteries in most t i w n p- nounced proliferative: changes in endi'lh.'lnr the glomerular capillaries were observed by - significant changes were noticed in th* ;* epithelium of the kidney. Necrosis of ihe pj-rr chymnl cells of the liver was also noted In some reports it was nuimed-- exposure to toxic h i exulted in a d c c i. ^ total scrum protein and_ti shift_it globulin ratio with a decrease in die . *T7?TTion. this could, of course, lead t ___ ,, ^functyigiiyyjjjm cT i^ lu\ - ^ * suggested that increuscd permeability diavascular bed contributed to the cU v . ' ' ~ * " ' '*"*. %mm 464 CtiCCnriral KiTton* iti ToMi'il'W NPC0001619 2 207878 - -a ..A.k '* ^ Uun.u c , , " i In other studies (he serum protein concentration did not differ from that of (he controls1*4 ' 1.1'* ml increased permeability of (Jic cardiovascular bed was advanced as the sole cause of the fluid accumulation. In addition, pulmonary edema was observed in diseased chickens and perivascular lymphocytic infiltration as well as edema of the cardiac muscle with interstitial lymphucytic infiltration has been noted.1"* Electron microscopic examination of the cardiac muscle revealed degeneration and loss of mito chondria.1*4 It is possible tliat the heart may play a primary role in the development of chick edema, but 'hepatorenal syndrome could be the undolying cause as well. Studies that investigate the development of the lesion more closely are needed to elucidate these problems. The type of diet also influenced llie development of the disease. It could be brought on more rapidly with a semipurified diet than with a natural Brain ration; sodium chloride in the diet was a prerequisite for the development of the syndrome.'8 7 If low rations of the toxic fat were fed to chickens they did not develop chick edema but reproduction and hatchxbility were drastically reduced. Toxic fat was not the only product (hat caused the chick edema syndrome. Chlorinated biphenyls caused the disease in chickens',B and in Bengalese finches.3` A mixture of penlachloronaphthalene and hexachloronaphlitalene fed to chickens also resulted in chick edema disease.'8* Cantrell cl I*1' 0 announced in 1967 that one of the toxic compounds capable or producing chick edema found in toxic fat was 1,2,3.7,8,9-hexachIurodibenzo-p-dioxin. Then Higgenbotham e f ul.** showed that the chick edema factor was pruduced by two principal compounds isolated from toxic Tuts: 2,3,7-iM chlorodibenzo-p-dioxin. and 2.3,7,8-tetrachiorodibenzo-p-dioxin. They also suggested fats and fatty acids that contain commercial chlotophenals as possible sources of contamination. When crude Tats and tallows arc heated to produce fatty acids, clilurophcnol residues may be converted to a chick .edema factor. Whether these Iasi three compounds mentioned are the only ones that produce chick edema disease is questionable unless the chlori nated naphthalenes us well as chlorinated biphenyls that produced chick edema were also contaminated with them. Anuthcr outbreak of chick edema disease occurred in 1967." (it this incident millions of briu- .n North Carolina were involved. The nutbrcj'. 1;jced to a vegetable oil refinery where le;:,' . ecume contaminated with "chloruphcnob," j company formula ted the ctriuruphcmils at . . . rubia! water b a t men products An u:;. .< .d pipeline from the "pesticide" plant IcJ j;n used to collect by-product fatly acids: reed fats. Flick el at-1** iiav- ,iv .iy shown that pyro lytic pruducts of 2.3 : jdrJoruphcuol were toxic to chicks and ;. i chick edema if SO ppb were fed for ; The product fed consisted of 53% he;:. hep la-, and 25% ociachloro-p*dibe...'i<' ,ti. The pyrolytic product of pentach - a! consisted of a mixture of 5% h>. . t -. hepia-. and 33% ociachlorodibcnzo-p*. hat pruduced mild edema and increase.. ,.y when fed a diet containing 1 ppm ('J ><ai ingestion) of the material fur 21 days i mixture of tri- and teirachlorudibcnzn-n-ci. js fed to the animals at a dietary cuneen i-j.-.-r 0.01 ppm (a calcula ted intake level of ! 7 \ the birds developed edema and 83% of ih; : :ri. Further studies with pure compounds ar-' vjrv to cunfirm these findings. The effect o PH re chicken was again demonstrated when ' :,r ihat were used in the U.S. as heat lu m i a ; ,2*; in food processing plants contaminated Lr. juantines-oT fisti meal that was processed for _ ...a! feed in a plant in Wilmington. North Ca:v n-.19' From about April 1971 until-July 1 9 V t Ohs leaked-into the fuh meal and 16 ihousurr. cm of contaminated fish meal were distributed ,L more than 60 companies in 10 stales. One of ' c purchasers of the fish meal, The 'Holly F- <. me nation's largest poultry producer, ha-' daughter 77,000 fowls after discovering con e* . arimi of the chickens with PCBs, Some of the ; a ls found in the adipose tissue o f the animus ^ : i : as high as 40 ppm. The Holly Farms first cis-. -^r.'d that the chicken meal was contaminated bc:jv (he hatchability of their eggs was drastically redo- ed lo F82. of nornul In February and Ma.=,ii uf 1968 an outbreak of chick edema also uc. c--.5 in Japan, where mure than 400.000 clu.- - : reportedly died. The disease was traced to iru- contamination of animal feed with Knticclrin' 400. a Japanese PCD. The chick edema discuse <.a.arred simultaneously with an outbreak of puiycn .mated biphenyl poisoning in people that wj; din1 to (lie wwlannnaiiuii of ury 1914 46J ft NPC00016193 207879 k r.m rice bran oil, also contaminated with Kancchlor 400 and produced by the same com pany.'W1 Metcalfe191 suggested that chick edema factor ,,Cflul^J^lpiiurni^edjLiieiiimRs grease from hid preserved with technical pcntachlorophcnols con-_ taming chlorinated.dibenzodioxins were no longer used fur too^T ruquels. 11 was also pointed out jthat the use of glue emulsions containing chloro^hcrKnrTwen5crlng upcrat7ons*Ts^nothcr possible source o f tlicToxK feeim. Whether elimination T these sources from animat food products will jjrcveiujr^c^^^^T ^^^rm cs^rcm am sJoJjcjicen^ jsince polycnTontcTbiDhiiiylLiiiai-iaa^_h-j^nlaminated with chlorinated dibenzofurans have _ajgg_BIpduced chick edema disease. Yusho During the summer-of 1968. 13 cases of chJoracne were observed in the western part of Japan, centering around Fukuoka prefecture. The incidence of the disease seemed to be familial and it became apparent that all patients had used the same brand of cooking oil (canned Kancmi rice oil). These findings were picked up by, the news papers and a strong demand was voiced to elucidate the cause. Therefore, a Yusho study group was established by Kyushu University.1' ' 1 The term Yusho stands- for rice oil disease. Aftci extensive investigations it was found that several illnesses occurred in a number of households that had used common cans of rice bran oil. When a total number of 89 males and 100 females tlut suffered from Yusho were examined, a variety of symptoms were fount! in these patients. The incidence of the different symptoms that varied a great deal is given in Table 2. The epidemic outbreak was finally, al ter very extensive investigation, pinpointed (u Kancmi oil. manufac.tured on February 5. 1968. Chemical analysis of this oil established that it was contaminated with 2,000 ppm or Kanechlor. 400. a chloiobiplienyl that contains 48% chlorine. It was also demonstra ted that most of the components of the cltioru paiignts for^uiu^a Iona time and nassed tlumndT happcnctlJu be nregnant.1^ The amount of oil necessary to produce disease was found to he at least 2.5 1. The younger the patient, thr higher the morbidity rate with u smaller total amuunt of ingested oil. The average amount of ehlmobiphenyi consumed 6y individual .patients was calculated in be a total of 2 g.|fl' (Various reports written on lids subject--give, Jind tlic-amuinii can at best only be cslim^ctU When the rice bran oil was healed under reduced pressure, Kancchlor leaked from Ihc old pipes in which holes were subsequently discovered. The Yusho outbreak spread not only over Fukuoka. Japan, but also over 20 oilier prefectures in the western pxiTt of Japan. A total of 1.057 poisoning cases resulted according to the latest tabulation; (August 1971, Ministry nr Welfare) 1,6 Tin: most common initial symptom experienced by 1.16 patterns with Yusho was increased eye discharge and swelling of the upper eyelids (38.3%) followed by aeneform eruption, follicular accentuation (35.1%), and pigmentation of the skin (9.6%). Itching of the skin and "stiffening" of the soles of the feet and the palms of the hands were alio observed. Fjgnnmtcihmncus_ membranes we_rg_noticed in many of the patients; a small percentage of1 them devcioned tlmmlicL. ""many complained of a fccline of weakness an headaches; occasionally vomiting. diarrhea. fever also occurred. Various neurological symptoms manitested themselves such as transient visual disturbances, numbness and spasms in th e , limbs, and hearing did tail tic*. Most patients showed acnefike skin eruptions that resembled the chioracue observed after occupational exposure to chlorinated naphthalenes or chlorinated biphenyls. When [lie age and sex specific incidence rates woo analyzed it was found thal a significant sex difference was not present, but a lower risk fm both males and females in the age grutips over 60 yeais was noted, lire proportion of severe cases among those age 15 to 29 was significantly larger than tliat of other age groups. During the Yusho epidemic. 11 women wiih Yusho and 1 wives whose husbands had Yusho. but who themselves did nut show- the disease, delivered 10 livcbom and 2 stillborn babies.-Nine of these babies had unusually greyish. d:ok brown slumcd skm. and dark pigmentation o tin* gingiva and nails was noted m Five of then Must intunis also hud heavy eye discharge. INsmioeioI exami nation of a stillborn ten is showed a tnaiked lis perkci.iuisis and atrophy of the epidermis and cystic dilution ofliuir follicles, especially of those of ilie head. A marked increase of melanin pigment in the basal cells of the epidermis-was also -imtvib. Mosl loloses ilnu vera born were smaller 466 CMC Crturel Ri *tewi in Tttxmifaxy *r "t HPC00016194 207880 ft f ; H' . u. TAIU.K 2 Symptoms (complaints) of Patients (89 nule*. 100 P em il , at of October 31, 1968)* Symptoms Mates 7 Uluckcninc at* n. O Ulack poli in .i. t:\ccwivc sweu(. 1 tettai* Aenelike skm c l^ US Rett ipoO tin ,: 1telling Clunp: in s^m -, Stvellitiy ni e., ; >j icci llardcniiv. >' ! . .J hands Pt/mcniaDi.i `cons me nib ranos Svbutn (cui;; . .'in in eyes) Hypciciiiu vi in ' '.'" is tr.timbrane* in eyes Teitipiif.iiv .in s'* --ycstylU Jaurnii.. Swelling e' . i eiids Scnc ! Numbnes i-: ! ' and feel t :e*r Hearinf d-"!-. i, Spasms n: ie na.- n- lect Headache Vmnii id" Dialitica H3.I 64.0 50.6 87.6 20.2 42.7 75-3 ' 2(U 24.7 56.2 88.8 70.8 56.2 11.2 71.9 58.4 32.6 16.9 18.0 7.9 30.3 23.6 19.1 Reprinted uiit: ri-rnniiun I'khh K urjtsiine et al.'*' 1-emales % 75 0 56.0 55.0 82.0 16.0 52.072.0 41.0 29.0 47,0 K3.0 71.0 <5.0 fj b -ij/, ; 2 1 1 . . than the national standar.1. .. <! : j or of them were Siiull for gestalionhI age tv:: res- As (hey grew ulder tilth skin giadually betaine ; . .\'.-r over a pet loti of three months. Okuniura and Katsuki11' <Jied a group of 24 patients whose main cumpla... d Seen ucucform skirt eruplions and who he-j been diagnosed as dennitely having Yushn. Eighteen of these were adults and six were children. Irrespective of the seriousness of the disease in a)] patients, le initial symptoms were eye discharge associated with edema of the eyelids, defects in vision, generalized fatigue, anorexia followed by comedo and acneforni eruptions of the face, and pigmentation and flattening of the nails. Ttic principal subjective symptoms were fatigue, numbness of the lower limbs, and intolerance for sakv, which is a Japanese alcohola beverage made from fermented rice. Three of twelve females had an irregular menstrual cycle Seven of nine, patients with serious disease showed a iow giatlc lever, Movt laboratory tesis conducted on these patients were normal, inciutielevation in aik. over half uf the the a , globulin : noted with a albumin fracu--. were within i . patients had : glycerides ram-i.24 cases, v/!-.. remained utten. somewhat low c men is in rabb> showed that is within several .previously no:r sure had been The scrum . usually elevate disease (Vein plasma triglywi." in fssaifial /.,*. of I( ir ' 77 - ------ N PC 00Q 16195 207881  1\ ! t``j h 'd t; u 'i 4it i r. i i i *4 1 i i) i I1 i i > - genital .; been o l i but the . course ,,>. . consist e uf the IV . of trig!;, arteries ' .: Anoi.. cpider.ii. Yusho. {.'.* ncurop^; ... hypoes'i * areflexia ; would .< experimen;: further. Samples from p!iti?'. ' chlorinated ... Tlie concert'. higher the:. ( . reported or suffered fStill suffermade ih those oLm the patir' '. ` ; remaining more thu; Inuminosi;. Several possibly ; and lu ran animals, A` white cou v; _\Si * i-'.. ............ II `.v.g'yceridejnia has mber of families. not knuwn. The* . Aiy benign uiui is : ' 99 Observation ic mere elevation -.reuse (lie risk of this poisoning ,.-.es with obvious sgns of a sensury ness, pain and case also showed rtc sensory nerves i-jus in animal .1 be investigated : tissue collected :\>ho contained .si analysis.1*i-iue was much jisune et at.196 ';c patients who he patients were *-V*n the findings .. compared with and tliat liatf of icullv, while the improvement and f:d become woisc. .j that PCDs and ti dibenzodioxins : ine response in A rtfureA m l.1 i s followine the well as extensive hemorrhages intu various organs including the adrenals. Reduced thymus and spleen weights were also observed in ruts that received a single dose of 25 yg/kg TCDIJ and m nice the cell mediated immunity was found to be suppressed.503 Tlie various findings listed heredo not conclusively prove that the immune response is specifically altered by these compounds as we encounter it following radiation exposure or therapy with alkylating agents. In most studies relatively high doses of the chemical compounds were given, resulting in general toxicity accompanied by. weight loss. Weight loss in itself may lead to atrophy of the spleen. In Unmans maintained on a low caloric intake with negative nitrogen balance, a general reduction oflymplioid tissue results which is not specifically related to the immune response and vitamin A deficiency is also accompanied by a reduction in lymphalic tissue. In this respect it is also of interest that when monkeys were fed "toxic fat" containing polycyclic polychlorinated hydrocarbons. Tlie germinal centers in the lymph nodesand spleen as well as islands of hematopoietic cells-in hone marrow were exirsi-iciy sparse.1i>,) This is more extensively discussed in connect inn with chick edema. The fae; hut a combination of duck hepatitis vi-i- jnd 1'Ctis is more toxic to 10* day-nld d u c k i h a ; ! me virus alone suggests a reduced nn::`.i;ie reip* use but could also merely represent an addk.v.* effect on the livei, made mort* susceptive to infection by the toxic effects o! Pt.L: Su convincing evidence of "wasting d'.fi esuli of exposure tn chlorinated impounds has been reported in n. -, / other species in the literature. and Denta.mm _ 'f ^ ., to Arot loi tn u c iiti'. .* However. organisms h . mallard cu: . (Aruclor !. Interperit ti r.t significar.'." :. received v v subletlial * ; r betuodu..;*. thymus lymphoid ... : '"Tslt, exposure j days led to .kin lesions91 ce to isolate scii lO-day-old otitaining I'CHs p.sully injected : cpatitis virus. ;har. wlicn they . guinea pigs. ' * etrachlorotli* upiiy nf tlie iymphucytcs. lymph nodes, as reduces ih .. .. . serum antib.-j..-. referred to : . i . Such anime'"- . foreign skin . the mus: v - disease. At . influence : and alter P. . .order to pro*.. A... dibenzodiuxr immuns resc. on newborn tested v. i .ur. . e animals to produce . i wasting disease. nko homologous disease. ncapable o f rejecting :> probably rcpicsents for liojti(jlogons :r:c response could -id growth of cancer in infectious. In eu as chlonnated kcc influence the ` `- f these substances species should be tors such as skin 468 ' 'T tfT * - NPC00016196 207882 grafting and the measuring of the development of antibodies. Whether the oilier chlorinated polycylic compounds in this article also affect the immune response has not been suggested or reported. Biochemical Effects and Chemical Toxicology Porphyria Porphyria cutanea tarda 'in humans is an acquired defect in hepatic porphyrin metabolism, characterized by uroporphorinmia, pUotcsensitivity as manifested by blisters, and mechanical fragility of the skin. The hepatic porphyrin (Porphyria cutania tarda) which is responsible fot the increase in porphyrins and the skin photosen sitivity can be produced experimentally by a number of drugs; all of these have the ability to stimulate the activity of the initial enzyme in the heme synthesis -aminolevulinic acid synthetase (ALA). In addition to this acquired porphyria, an acute intermittent porphyria crihropoielica and an acute intermittent porphyria hepaticn, which are both hereditary, exist together with a number of subgroups which will not be considered Imre. In 1964, Bleiberg el al.1 reported studies of 2 workers in a 2,4-D (2.4-dichlorophcnoxyacetic acid) and a 2,4.5-T (trichlorophrno.xyacetic acidl factory. Many of the workers had chloracne and 11 had abnorm al excretion of urinary' uroporphyrins. Many of the workers with uropor* phyrinuria had hirsutism, hyperpigmentation, and increased skin fragility. Liver dysfunction was noted in two hospitalized patients. Poland ct al.! reinvestigated the workers of the same plant studied by Bleiberg et al.3 They studied a total of 73 male employees; chloracnc was found in 60?* of the workers. In this study six years after the first report, clinical porphyria was not observed and only one worker had persistent moporphyrinuria. In the Japanese poisoning epidemic (Vusho), increased uroporphyrin excretion was not reported. Apparently polychlorinated biphenyls, as well as tetrachlorodibenzod toxins, produce hepatic porphyria in animals. Experimental hepatic porphyria was observed in Sherman strain rats when the rats were exposed to Arudor 1254 in the diet. The porphyria resembled hexachlorobenzene poisoning and human porphyria cutanea tarda.' 01 When female fats were fed 100 ppm Amclor 1254 in the diet (approximately 7.9 mg/kg/day) they became porphyric after a delay of about 2 to 4 months. At this time the urinary porphyrin ex cretio n increased rapidly. Uroporphyrin excietiun was elevated to a maximum of 550-fold, prnphobilinogcn 27-fold, and -aminolevulinic acid 18-fold. The major porphyrins found in urine from Aroclor treated rats were S-carboxyporphyrin (73% ) and 7-carboxyporphyrin (16%), Substantial increases in amounts of 5 and 6 carboxcylic porphyrins were also observed. A 10-fold increase in the excretion of coproporphy rins was observed, but the increase in uroporphyrin excretion was 1,400-fold. In control rat urines, coproporphyrin (81%) represented the major porphyrin fraction. A good correlation between urinary porphyrins and liver porphyrins was also observed. Only a trace of porphyrin was found in liver samples in control rats. When feeding rats 100 ppm Aroclor 1254, -aminolevulinic acid synthetase was not increased in the first month of exposure. After this time aminolevulinic 'acid synthetase was elevated 3tu 6-fold in porphyric rats but was normal in nonpor pilyric Aroclor treated rats. Cytochrome P-450 and microsomal heme were also increased throughout the study but the increase was maximal in one week despite the absence of an increase of the rate limiting enzyme in heme synthesis -aminolevulinic acid synthetase. When a large single dose of Aroclor 1254 (500 mg/kg) was given, a '4-fold induction of -aminolevulinic at id synthetase could be demonstrated in the liver 5 hr after dosing the animals. These results suggested .that -aminolevulinic acid synthetase induction occurred when porphyria had developed iT low r levels of Aroclor 1254 were fed.over a period bf time, lull when high doses of Atoclor were given it was induced immediately. Vos and Koeman,J reported that chemn-,*! porphyria was produced by rite two European products. Pitenoclor DP6 and Clophen A60 n welt as the American product, Aroclor 1260 Ih r porphyrugenic action was observed in chickem quail, and rats. It lias not been establisJifj^OJAL. nated bi phenvls or contamination will cMlorinnlei production n ftig tia lic ntnwhurl.-i finim-a f r ""^ToUmd and Glover106 used fertile chicken se at a developmental stage of 15 to 20 da1. \ injected them with 25 pi of the hatou?*(henrn-p-dioxins. They found that 2,3.7> i f . chlorodibcnzo-p-dioxin produced a dose rela te January 1914 *< NPC00016197 207883 1 i i f I. I a aaa t i ii i / i i h y p ertro p h y after (ceding polychlorinated cantly in male rats for a period o f up to 28 days. f f biphenyls fAroclor 1248) or a polychlorinated Cytochrome b-5 content was also increased. biphenyl fAroclor 5460) for three 'months to Hydroxylation of aniline was induced and amino- primates. The increase in liver size was attributed pyrine demelhylation rales were dct-icascd. Micro ' to the increase in smooth endoplasmic reticulum somal proline was increased by approximately 1`*IH.1 and induction oT microsomal enzymes was 15%. The induction of UDl' glucuronyltransferase observed. Proof of the induction of microsomal was most striking. Oxidative phosphorylation rates enzymes in birds is the finding made by Lincer and in rat liver mitochondria using succinate as a Peaka]l1,ft that livers obtained from American substrate were unchanged. The enzyme induction kestrels exposed to either Arcelor I254 or Aroclor was observed as early as one day - the peak 1262 showed an increase in the in vitro breakdown induction was retched between three and ten days of oestradiol to a more polar metabolite. This after treatment. The enzyme induction was accom effect on sex hormones is also emphasized by panied by tn artrease in smooth endoplasmic I'latonow et al.31' Aroclor 1254 given. to reticulum and i -mild increase in rough endo :Jr.!" Yorkshire boars reduced the urinary excretion of plasmic reticulum.214 These changes weic first *r gonadal steroids: observed three days after the injection of the -I JJoth the polychlorinated biphenyls as well as letneblnrndibciizodioxin. Thus, enlaigement oi f; the chlorinated naphthalenes affect the vitamin A the liver during exposure to various chiurinated content of the liver. This was particularly studied compounds is accompanied by ah increase in with the chlorinated naphthalenes since hyperker smooth endoplasmic reticulum. atosis and other abnormal keratin formation of the Furthermore, if "inclusions*' or "hyaline bodies" skin is often accompanied by vitamin A arc seen within the' cytoplasm with the light deficiency. Low vitamin A plasma levels were microscope, concentrically arranged membranes observed in cattle with X-disease and young that often surround lipid vacuoles can be seen with swine.*11,161 A decrease iri vitamin A storage-in- the electron microscope (Figure 6). Hexachlorophene, on the other hand, does not A rociorT^^Tw asTabo demonstrated.7 1w_SiaCfi_ increase the size of the liver.118 The livers show no u ltra s tru c tu ra l changes and .p'obably ispecjjhould be further investigated to determin microsomal enzymes are not induced. Whether other compounds listed in Figure 2 affeci liver ^sw qndarvtojdtam inA jtgiijjjflj^3 microsomal enzymes is not known. Hcxachloi- Ecobichon and Johnstone110 studied the effect ophene has an effect on a variety of other enzyme of various PCD isomers on the hepatic enzyme systems than those mentioned thus far. Cammei system. Biphenyl itself did nor cause induction of and Moore225 and Caldwell el a!.136 have shown hepatic drug metabolizing enzymes and neither did th a t hexachlorophcne uncoupled oxidative monochlorobiphcnyl (4-0). Substitution of chloro phorphuiylation, inn cased adenosine' triphos groups at the three and fuur positions of the di- phatase activity, but did not affect the activities nf and telrachloiobiphenyls caused induction of .electron transport enzymes when compared to microsomal nwnouxygenases. Induction of micro controls. Gould et al.J 11 demonstrated that liexa- somal monooxygenases occurred also when hexa- chlorophenc inhibited succinoxidase activity of rai > and octachlorobiphcnyls were given to the rats. heart, liver, and kidney preparations. According to Nitroreductase and carboxylcsterase activities were unpublished material -cited by Gluck,118 licxa- not affected while all compounds produced a chlorophene is conjugated to the glucmonidr marked induction of the sulfohromophthalcin conjugate which is excreted in bile. The conjugating system. Tcrphcnyh as well as chlori proportion of conjugated hexachlorophene to nated terphenyls.increascd the pentobarbital meta- unccmjugated hexachlorophene in blood may be bulism in the u i with corresponding increase in moic significant in determining the toxicity of smooth endoplasmic reticulum which suggested hexachlorophene than the hexachlorophene blood i microsomal en/yinc induction.131 *333 levels as they are.presently determined. Lucicr ct al.: 13 reported that a single oral dose of 5 or 25 "uc-'kg body weight of tetrechbrodi- Methem agiobincmia A - 'hetiwduucut iudueciL. cytochrome R:45U signifi- In the early l?(*Os outbreaks of methcmnplnb **iuary 1974 <11 NPC00016198 207884 inemias were reported from hospital nur and the least in plasma. Brain, liver, and kidney series.11*, i 30 One outbreak involved a total or 18 had levels of the same order of magnitude hut infants with a level of methemoglobiu from 4.9 to were lowci. If, for instance, !.t>00 mfi/ky body 30% of the total hemoglobin. An epidemiological wei||ji<^if_Aror^ JIM were given bv stomach investigation traced the outbreak to 3.4,4'- lube, 24 hr later rhe mean concent ration in _ tridiiorocarbaniiidc fTCCj that was used as a 2% jjfffi&KTTiijfcjfe was TTl46 onm. The concentrations solution in a rinse in the laundering process, after observed in individual animals varied greatly. which the laundry was neutralised with an acid Similar levels were found in adipose tissue when and then dried in an autoclave. Twelve of the 3,200 mg/kg or Aroclor 1200 were given by eighteen cases in this report were prematures who stomach tube to adult female rats. The levels of were probably longer exposed to the offending Aroclor 1254 found in the brain showed a mean of chemical than term infants since their hospital stay 138 ppm pnd the values observed for Aroclor 1200 is usually longer. It is also possible that chemicals' .were 145 ppin. All o f the values were calculated ^ penetrate more easily ihruunKj h c skin of prema? on a wet weight basis. The same concentrations in tu res^ ppm were found with both compounds even In another outbreak, methemoglobinemia though only half of the amount of Aroclor 1254 occurred among a group of custodial patients in an was given. This is probably due to the fact that institution. These patients were given soap suds only a certain amount of Aroclor is absorbed at a enemas with boiled soap that contained 2% given time. TCC.111 When 100 ppm of. Aroclor 1254 was fed to It is possible that by heating TCC a breakdown male weanling rats for 58 days the mean concen product occurred which produced the methemo tration of Aroclor derived materials found in the globinemia. P-chloroanilinc and 3,4-dicbIoroanilinc feces varied roughly between 11 and 35 ppm. have been suggested as breakdown products. None After the animals had been removed from the of the other compounds discussed in this article - experimental diet for a week, the excretion in the have thus far been shown to produce methemo feces was ft ppm and after 16 days' recovery an globinemia. average of 2.5 ppm was found in the feces. FCDs at the concentration of about 2 ppm were still Storage, Distribution, Metabolism, and Kxcretion observed in the feces 24 days after the animal; As wc progress further in our. study on the ' were removed from the experimental diets. During d is trib u tio n , me la but ism, excretion, and the entire study the amount of PCB drrived absorption of compounds such as chlorinated . material recovered from the urine remained bclmv biphenyls, chlorinated terpher.yls. the chlorinated 1 ppm While PCBs were fed to the rats, the gas dihenzodioxins, and chlorinated naphthalenes, we chromatogram obtained from the feces was super- will probably find that since they arc lipid soluble imposable nn the one obtained from the standard they are stored in quite the same way as the more Atocloi 1254 that was fed lu the rats. Once the studied persistent pesticides such as DDT and animals were no longer fed Aroclor 1254 the dieldrin. From the evidence presently available on number of peaks observed in the gas chroma the chlorinated biphenyls it can be assumed that tograms that resulted from the analysis of the fecal the turnover of these rum pounds in the body, material decreased. Fewei peaks were also particularly of those that have a high chlorine observed in. the gas chromatograms obtained for content, may be slower than- some of the the tissuev and from urine. During the 58-dav persistent pesticides. The turnover rale will vary feeding period, a gradual increase in the concen with the degree nT chlorination. Hcxaclilorophene, tration of the PCI) derivatives was observed in on the other hand, is fairly rapidly excreted and tissues. The highest concentration was found in presently no information is available on bilhionol adipose tissue: after 5S days it reached mean and related compounds that have a germicidal valurs of ilmui 500 ppm. The concentration in the effect. All of these compound* arc not very well liver only amounted to u mean ut 12 ppm at that absorbed by the gastrointestinal tract or the skin. time. Following a recovery period of 7) Jays when Analysis of various tissues of rats given j single l lie anim als wnie fed plain chow, the dose of Die PCI) Aroclor showed the conccnnation of the IVI) derived material in highest^ concentration of PClls in adipose tissue adipose tissue had not decreased appreciably and " *'r%,t 472 CMC Cnnia! KfviVu'v m ` V NPC00016199 207885 dill showed values in Ihe neighborhood of about j a s chromatufirams si. jtfu'nn in 400 ppm and 4.7 ppm was found in-the liver. The ihcjiunihjfr of pea?.'* r.:ivs on a concentiaiion in flic brain never readied very high control flic! only 4 .1 n o tio n s proport ioas; if ranged from about 2 to 4 ppm. ' remain, wliich Mimcstn .. Following a 7 1-day recovery period Ihe amount in metabolism of some ti iililiili. .j of the (he brain was still about 2 ppm. When Aroclor 6 In c s -dy. Heath 1254 was fed for 240 days at ihe dietary level of ct al.i,g analyzed 2 mallards 100 ppm, the concentration of the HCB derived fed 25 ppm Aroclor 1. - 2 years material observed in adipose tissue had reached and found 56 and 33 p; tit. The values of about 1,000 ppm. if, on (he other hand, gas chromatogram sho ! diniin- J00 ppm of DDT is fed to male rats for 2 years, the concentration in adipose tissue is about 95 ution of the eariy pea/ (he standard, suggesting . r-arod to . . Auction ppm.333 in a separate study where 500 ppm of of the components vm concern. Aroclor 1254 was fed to male rats for 6 months tratinns. T h l s i s i b p ; jommmeenitHalall and the exposure to FCB containing diets was . am piesJ^w erorB O B r*1 rjinPiisn V s subsequently discontinued for an additional 10 months, the concentration of the PCBs ranged J o w e ^ S lo n n a tio ^ S jj^ , from 924 to l ,688 ppm in adipose tissue with an In mammals, excreta . milk and arithmetic mean of 1,192 ppm. The concen transplacental passage ir- v: : Feeding trations in the liver ranged from 17.3 ppm to cows 200 mg/day of A1 i <k: day's1* 26.24 ppm with an arithmetic mean of 22.65 resulted in higher PCB _pp_m_ . 13* body fat. At 30 days i* .> ilianin :'pm was Further studies are necessary to determine found tn milk fat and 2 i . :'.;at 60 whether PCBs arc stored in tissues to a certain .days' exposure, 66.7 p ; , ::ulk fat point when a steady state is reached and the and 44 ppm in body fv ssposure concentration does not increase further. The given to PCBs was disconliii .snt; at ion in concentration at which this equilibrium is readied mjlk fat sank below tha*. in adipose tissue would vary with the amount Ted Transplacental pas.. 1 k also to the animals and the type of compound. The occurred.337,33 * Shcnr. effect of starvation or illness when the amount of Aroclor. 1254 in peat.; r-- ^ii-iiiL ^ ..h rube adipose tissue in the mammalian body is suddenly during davs 7 to_L5 of cc^ . - mu-.-*M'20.60 reduced also needs to be investigated further. ppm PCB derived material ` J i:. 'cir milk Would this lead to increased excretion of the when 10 mg/kg/day was ci.*, ujrni during material or to redistribution or both? Dahlgren et ' pregnancy, while those' . rv-cjkg/day at.i , s who determined PCB tissue levels in excreted a mean of 66.3-i . *t : r nik on a pheasants, found that a concentration of 300 to wet weight basis. Fetus * jesarean 40,0 ppm of the PCB derived material was usually section bn day 20 of pre^r / mean found in the brains of birds that had died, while ol'0.G3 ppm and 1.38 p;u> naively. live birds that were fed the same amount of PCBs With a Fivefold difference in d-. ; . ii: concen had much lower concentrations in their brain tration in Teluses showed a tw oiok ; .c ;m-e and tissue. in milk a threefold dlffcrenc;.7' * .Studies in boh white quail showed that relative. The hyperpigmcnicd tun.;:. .= --d in the extracUwer^rinulaM othcArodorsla^ partitioning with acetonitrile-hexane when the. Vuslio incidents in Japan repres. : ...iuitional circumstantial evidence of transp!:.. passage their carcasses were analyzed soon after exposure Polychlorinated biphenyls lu vc > tcciecl in human adipose tissue. K This suggests absolution of all components of analyzed (vm htuuan adinose risM.c : *j winch ' fltoefor. The pslroiniestinal tract, liver, skin. "contained 200 and' 600 umn PC B- 'T7 ffCIlli-- whirs, and feet had been removed prior to pentachlorobiplictnri to deracliiori'1-::- analysis^ When hohulikc ouail were fed Arorlnr _ Vice and Welch* g found 100 mini I'CUs in - 1254 for 14 Lavs pr.d ihen allowed to recover. Qie_ "anoTKrrTnjinanadipusc tissue samp]'' .According Jamurv 1974 473 N PC 00016200 207886 to Price and Welch;140 41 to 45% i f r ^ v - j ! population in ihc U.S. contain I n . r "m e PCBs in their adipose tissue. Amor . ' ' - :acs they discussed the food chain ar.v .. . Just which, from residences in southwestern contained up to 180 ppm PCBs. P c . : - i:d biphenyls were found in 31% of ' sum / ,,if human adipose tissue that were colic ' <ie penerai population of the U.S.. . per! ;he Human Monitoring Survey. Of trie r ..-if Jyzed, 5% contained more than 1 p p m .1, K'iJi have also been found in human adipose tisi'.-, of the general population of Scar.;- avij7'1* ;r'.d Japan,7*4-3*1 and in human m: man adipose tissue of (lie general . of Germany.** '1 In general, low coiu\ .u b, of PCBs were stored in adipose tissue in a c.:;u;n proportion of the general population. This is, jirohably^ more prevalent i^ th c highly indnsfi'1)-. rountries unless most of the % w - -if imtwirH from nnlluled Jffjas- H i? i-.v !f . / PCBi *rg Jiajin lcss^ as faf as is* - >m; . JiMLieni. however. the occasional : 1jp. th c lG 5 "t^ T !^T g pin ranget-lvc : i cm j: sv^uTsPS^ofbitcrest to establish ... _-c adipose tissue levels increase with a?.* ,:r those in certain professions such - printers, textile workers, workers n. c- lories, and those engaged in the manu:.- jf transformers and capacitors had higher PC i* than the geneial population. Reports un the absorption, storage, and i m . : olism of chlorinated naphthalenes, tcrpliemv.. dichlorophcne, bilhionols, fenticlor, tridosan. anti tetrachlorosalicylanilide arc almost nonexistent m the literature. Some unpublished results on some of the compounds used us xermiutics arc nieiv tioned in the summary minutes of the Over the Counter Drug Panel on antimicrobials. Cnp'cs .m be obtained by writing to Dr. Mary Bruch, ft iv-u. of Drugs, Rockville, Md. Approximately .9% of triclnian, also - ; j irgasan DP300 and CH 3S65 (2.4.4'-[rich: hydioxydiphcnyl ether), is ubsorhed whe applied dcrmally in a soap solution. The ha!* the chemical was 20 hr when applied dermal . . .J 10 hr when given intravenously Humans 65% in mine and 20% in feces after intro " * injection. Il rs excreted either as a free cor-* c or as a gtucuronidc. In dogs nearly 100% of the injected n:-1 is recovered as the gluciiroi ;,Ic or solfjti in tuine and feces over a 5-day period, tl has been shown (lint ICC is absorbed llirouei: the skin. Tissue levels of the chlorinated terphenyl Arocli't 5460 were deter::.tried in codfish.1** In this study II was found that Aroclor 5460 was apparently poorly absorbed from the gastrointesti nal Had, il was stored in all tissues that weic analyzed. The highest an: central ion was found in the liver and following a single dose of 1 g sonic of it was still present in the codfish after 70 days, an indication of slow excretion. The authors pointed out that with the method used to determine the chlorinated tcrphctiyls, it would be difficult to distinguish.chlorinated biphenyls from chlorinated terphenyk. A comparison of the chromatograms of the standard with those obtained from chlorin ated terphenyk extracted from tissues revealed more of the earlier.peaks in material obtained from tissues and fewer. oT the later peaks. A variation in the intensity of the peaks was also observed. There seems to be a change in the chlorinated terphcnyls within the living organism; however, part of this could also have been intro duced as an artefact by the analytical methods or by differential absorption from the gastrointestinal tract. Calves bom to cows with X-discasc developed hvpcrkcralQfris aiiri. it__was--ju u m l__(|ral_|)icl' chlorinata.} naphthalenes were excreted in lenes may well follow similar storage and excretion . patterns as_HCI!s^_ Qxtorinatvd dtbenzodinxim ami furani ait probably also stored in tissues of animals and men who arc exposed to them, but again nut much infmrnation is * presently available. Firestone el al.1* 3 Ted the unsjpunifiahlc fraction isolated from "toxic fat" or 3% "toxic fat" itself to young cockerels. The "toxic ret" contained chhm.dibenzodioxins with 2 to 8 chlorine atoms. Wien the excreta were analyzed it was found that a certain percentage of the Uexachlorodibcn/.odioxins had been absorbed from the gasirointestitial tract, while all of the oclachlorodiheii/odioxins remained in the gastrointestinal tract, and only very little o f the licpladitorociiben/.odioxin was absorbed. B e vau se o fth e j)^ ^ fcri)i|' co n ip in icn tsTn irT in cklissu c^ i i j f r ^ ^ ^ coultl not be exHtnmcJ r`u' touf'r ^hiarihau the organs, the highest concentration! was found in the Mycijand an appreciable am o u n tyr , . _47-i, C* C' t'rjlkoIJt rw'rWf W 7; ;L.;hjyy NPC00016201 207887 was also found in the bon-, im-j skin, while the other organs contained ve; few dioxins. Some evidence for i possible metabolism of Iicxj- and Iteplaclilorudibcniodiuxin by the chick was also observed. Zilko and Wildish14 h fed di*. iri-, tcira-, aiid octachlorodibcnznfwun to fish and delected only oelachlorodibenzoforan in muscle and gut of dead fish. One problem with metabolism, storage, and distribution studies of chlorinated diberundiuxins and furuns is if-.3i not all uf the com pounds are readily available and. if they arc available, any long-term feeding studies in large animals arc expensive. The very toxic dioxin. p v n n '^ n lv al su c h tow levels TTiatuieyescapc deicctjorL^i^iiiiJiwiULj^ tor>Mveoeiccteo Williams el il.i4> fed rats a total of 22.7 and 120.7 pg of octacliloiodibenzodioxm. They re covered a great deal of the compound in Lite feces. Very small amounts were recovered in the liver at both douge levels and in adipose tissue al (he higher dosage level. Further studies arc needed to establish the absorption, excretion, and storage of these compounds, and their insistence. Studies with labeled material may eventually give us some information on their metabolism. Piper el al.1**1 gave a single oral dose .of C14*4 labeled 2.3,7,8tetiachlorodibcnzo-p-dioxin to rats and found that 1 about half of the radioactive material was re-, covered within 21 days in the feces and only l'A' in urine and 3% in expired air. Most of the remaining radioactive material was found in the liver. Jn another study1" 100 rig of Cl'*oetacliinindibcnzodioxin was given daily In rats by stomach tube for 21 days. Less than 0.3% m* the material was absorbed. Subsequently the rats received a control diet foi 7 weeks and still showed 20% of the absorbed material found at 21 ` days in their tissues. The in vivo metabolites of the various com pounds under discussion have hi must instances not been identified. Block and CWnisIr **' studied (he urinary excretion of biphenyl and 4chloiobiphcnyl in rabbits. Iliphenyipluensiiltirunic add and 4-hydmxybiphenyl were isolated from _urine of rabbits fed biphenyls. 1he rabbits led the 4 - c h l u r o h i p h c n y 1 e x c r e t e d 4 -1p chloiuphenylpphcnol and 4-dilotubiphenyl ghi- cosiduroniJe. Twice as m.ich 4-cldoiobi|*iIicnyl as biphenyl was excreted as ihe glucusidmunic acid derivative.. It is possible that other low ehlorinaled biphenyls and naphthalenes me excreted in tniuc in a similar manner; wi.nikei they would be detected by piesenl gas chromatographic nit'I hods would have to be checked by combining the identification of'radio labeled material with gas chroniarography. Hut/ingcr ct al.IW found evidence dial some chlorinated biphenyls are uydroxylaicd bi some species. ApgarerHjvJ^oc^ ^ not able to livihuxyi late ^IflooMirimoiivl. 4.- Jichlorolupheii^l. mid 2.2 .5,5'-tetradilorobiplir. .*, wIdle the rat and the Pigeon both founeiLnntn- y-fT YVhif" , ;,i,%T^ " JVom4^hIorobi|jjj^L I:, addition. the rat also produced dihydrochluiobi phenyl. Large quantities of unchanged material were excreted by the rat when 4,4'-diclilorobipiienyl and 2,2 .?,?'letrachloiubiphcnyl were given to them arid only a monohydroxyderivative was identified in their unne. Pigeons followed a similar paltern hot no dihydrochlotuhiphcnyi was observed in the bird excreta fed 4-ehlorobi phenyl. No hydroxy metab olites could be identified' in the excreta of the three species when they were given 2.2\4,4'5,v' hexachlorobiplieuyl. No evidence _of rcductiic. dechlorination was observed in any species Only veiy few studies have been conducted with radiolabeled hcxtu-hiorophcnc to snnlv i\ absmpiion. disinhmion. am! metabolism. Wit mi. Van Gemlercn7" gave mts and rabbits Itrv chloroplicne-CM (2.2'-meihylone-C14 bi>f3 - < trichloruphcnol). Complete ' recovery o!' radio activity in excreta after a yuglc dose of 5' or 1' nig/kg of radioactive hexaililmuphcm.' w.i* to. served in rabbits within 5 days. Roughly on.' <k v was excreted in urine and leccs.- respeem -tv .. unchanged hcxachioiuphene and .the inn..... . was excreted in the feces us unidcniiTied ru-i olites. Tiro metabolites could nut be cm-j ... 1`roiii the feces with ethanol. The hiotogical ! . calculated from flic experimental resU * found to be approximately 17.K l.r. In ui< , two thirds of ihc rudioa; ij'-ilv was rct."i-i Icces within seven days and only ve:> i>. urine. A eoinpleie ri'covcv of radioaci:*-i: > ' mil obtained in these su.d i- wii*.:a . Five cows given a suiplt . ; -weight o f licxuchluiopiten.' s -:m-. ic.' two ihiuls in llreir feces w .'.-a. . little was excreted m um - NPC00016202 207888 Mctaholitcs were not identified and tissue levels weie not determined in this study. Tlic dutlmrs cited a study by K ok,TM wlio did determine hexaclilompiienr in various organs of rats. Carroll et al.1J* studied llie absorption of C-14 labeled hexachlorophcnc on the tail skin of Lite rat. HcxachJoinpherte in a detergent solution was absorbed by the intact skin of the tail at the average rate of 1.7 pg/cnr of exposed surface arca/lir. Immediately after the infliction of second degree bums, (he absorption rate increased 2.5 times that of the normal skin surface. After 24 In ` lhr absorption rale dropped to normal or sub normal levels.* Immediately after traumatizing the tail, the absorption rate of hcxadduroplitmc was even greater that) following the infliction of a burn wound. Maiiowil/ and Johnston2iT found that hexacbJoropbrnr was deposited on the skin when used in.various formulations and could be extracted from the skin by soaking ihc'exposcd areas (hands arid forearms) in alcohol. The amount recovered increased in proportion to the amount applied. These authors were unable to confirm earlier investigations which suggested (hat the quantity of hexachlorophcnc retained by the skin reached a plateau level after a number of washings and remained relatively constant thereafter. Arms were washed and rinsed every 20 min. When extractions with alcohol were made on different arms after different numbers of washings, it was found that the imount of hexachlorophcnc recovered in creased with the number of washings and no plateau was reached. A greater amount of hexaclilorophciic was also deposited on the skin with increasing length of time of a single washing. Since we now know that hexachlorophcnc is also ab sorbed from the intact skin and some of if is probably also lost from the surface of the skin, the time interval between Ihc washings plays it role in the amount of Itexachlmophcnc found to accumu late on the skin. Also of interest is (bat bathing wilier Containing 4 mg/l of hexachlorophcnc de posited approximately the same amount of the chemical on the skin as a single washing with a 2% hexuchlnrophcnc soap. Since hexachlorophcnc wax used very exten sively and often unknowingly by the general population, we investigated whether it was present in (he blood of a lew random samples of the general population. We found Unit the arithmetic mean concentration of hexachlorophenc in udufir. was 0.028 ppm (tp/g) of ,, -mod of a total oi 14 samples. In infanls v U .cere washed with hexachlorophcnc in the : y the amount of hexachlorophenc found t> ;? bfood after the infanls had anywhere \ to 14 washes averaged 0.1 O'1 ppm (pf-v. . -hAc blood). Blood obtained from the corJ '.he lime of birth contained an arithmetic -i nf 0.022 ppm hexachlorophcnc.2,B*2' " In the rats that were m a reproduction study and were fed 100 p; hexachlorophcnc in the diet fur 258 days, the i;.",it:tilorophene blood levels averaged* 1.21 ppm, v the blood levels of rats fed 500 ppm of hcxitc phene fur 55 days who had severe symptoms r - risoning amounted to 8.5 ppm.15*-1** At the ; we reported these blood levels, our recover-, ? was only about 75%. All blood levels .. at that lime were determined in whole blood. is assumed at the moment that blood Icvcix . equivalent to half the amount of that found .. *cn plasma is ana lyzed.) A study undertake-t ::r the request of Sterling Laboratories1*1 hi : ted that newborn rhesus monkeys following to..; Sidy bathing with a 5% detergent solution f. ` week hud lilnod levels of about I,5m/ih> In subsequent months a plateau effect .'served and at <30 days the blood level wa; . .-cut to 1.1 pg/tnl. TIicsc monkeys showe" .-nongiosiis itf ihc white mutter of thc-b.\..'.. indicates that at least in the monkey, b:t" n .in-ls of about 1.5 pg/ml whole blood v, ere r.isier.t wfth fluid .accumulation in the - m u t h of the central nervous system. Lockhart112 cited u shed studies by Cluck, who found t!:.- iotophene blood levels in term infants n -:t;ng with a 3% hexachlnrophene dete^ fort were sonic- whal higher than thosr , -is with a mean concentration of 0.5-11 . . whole bloodl. This dilTercitcc could ! ;c to a differ ence in washing procu. -tier recovery rate in the analytical pc, study, c; both. Prematuir infants > *'*.J h.'s chlorophene mote read . :ni that were washed w,:' . ' ph-. detergent solution, ` tration amounted to 0 Abt el al.261 using a difft- . - a mean hcxachlorophr t In ] K hr after the ini: . f 476 CRC ( ntH t/l R i \ icw\ in Tuxirnhw .r ,'. NPC00016203 207889 A. ** ** [ L whole blood) and on the 7 to 9th day of lift*, 24 to weigh l/day of hexachiorophene for 38 days y- 48 hr aflci the second or thiid application of showed a mean hcxaddorophcnir amccntiutiun in v to hexachiorophene, the mean blond level annum led Ihe brain of 0.38 ppm and in adipose tissue the to 0.287 ppm frig/g). The amount found in cord licxacllluroplicnc concentration was 0.94 ppm.3*1* bluod of J6 infants was 0 .0 13 ppm (pg/g). When The recovery rate in- this study was only 75^, '1 hexachiorophene was applied only on the day of These squirrel monkeys had no overt clinical signs delivery, the hexachiorophene blood levels gradu of toxicity, hut showed mild status spongiosus of f al; ally decreased from a mean of 0,145 ppm on the the while matter of the brain, lilccimcnccphalu- 'first day to a mean of 0,057 ppm on the ninth graplnc changes had also been observed in these day, following an increase to U.J94 ppm on the pnmates. Ulsampr et al.1"*' found hcxaeliloro- fourth day. 'Die authors suggested that picmature phene brain levels of ah mu I ppm in newborn ink .infants may absorb more hexarhlorophcne with sigio of central nervous system toxicity. In 1 through their skin since in one infant 29 weeks of suspected case of human liexachloiophvnc gestation crnl weigliing 1.42 kg the blood level was poisoning in a child, the hexachiorophene brain 1.17 ppm 24 hr after the initial wash and 0.655 level was about 2.2 ppm.4 6 1 Adult rats dying after J ppm at 4S haute. Alder er al.:6J reported mean the administration of a single dose of hc.xnehloro- ;*j hexachiorophene blood levels of 0.18 ppm in plicnc showed concentrations of 6 to 9 ppm of '! infants who were dusted with a powder containing hexachiorophene in the brain.3*6 Sume variation 0.33% hexachiorophene. of the conceit Iration of hexachiorophene in the Adults using hexachiorophene preparations also brain which can be related to neuiotoxic *<*' absorbed h via the skin.'33 The blood levels symptoms must be expected in dirfcieui species obtained m adults that exposed their total body and perhaps also with age an.d sex. Whether the > surface to i 3% hexachiorophene preparation fluid accumulation which may occur in the myelin twice daily for 60 days, 5 min each time, showed sheaths after. repeated exposure to hexaddoro- blood levels of approximately 0.68 ppm (fig/ml phene would alter hexachiorophene brain levels whole blood) with a range of 0.25 to 1.08 ppm has thus far not been reported. | , ftig/ml whole blood). If only hand washing was With present methods. v.*c arc probably only I employed for a period o f 28 days, 4 limes daily, a determining free hexachiorophene while the conju : mean blood level of 0.07 ppm was reached and in gated portion remains undetected. The toxicity of j a face hand washing study the mean blood level hexachiorophene will vary with the ability of the :i after 28 days was 0.196 pg/ml. organism to conjugate, metabolize, and excrete I Blood levels obtained in 3 patient with burns this material. ,{ who died between 24 and 48 hr after the last We found in rats that hexachiorophene is hcxachhm.'phene application were 2.2 ppm (qg/g excreted in milk of dams fed hexachiorophene at whole bluod). The blood levels that have been the rate o f 20 and 100 ppm in the diet (2.3-1.1 ' reported ail indicate that following dermal hexa- nig/kft/day md 11.8-5,5 mg'kg/day. respectively', chloropheiic exposure the chemical can be found for 241 days). Electron capture, pas-liquid chroma in blood, and from information through animal tographic determinations revealed levels of 0.07 studies .i' well as some human exp elienee. levels of and 0.33 ppm.in the milk. Since hexachiorophene approximately 1.5 to 2 ppm fpg/g) in whole blood in human' are presently' considered to represent toxic levcz- if the analysis for blood was performed did not seem to accumulate to a great extent similar values would probably have been obtained, had the levels in milk in these animals been I with the methods mentioned elsewhere in this measured eailier. Although wc now have some j article. information on the absorption and distribution as Probably a better way of assessing u toxic dose well as excretion of hexachiorophene. the possible i '*. would be to measure the amount of hexachior- storage of the material as well as its metabolism opliene in tne brain, but for obvious reasons that is not possible. Female rats fed hexachhnn phene at needs further investigation. Since hexachiorophene was found in cord blood of infants, jt can be the rate of 500 ppm (25 mp/kg body weight/day) assumed that transplacental passage also occurred. I for 55 days with very.dc(tiuic lesions had about 4 ppm liexachi-.-rophcnc in their brain and 3.91 ppm Morphologic EfTect on the Liver , in the liver. Squirrel monkeys given S mg/kgbody Some of the chlorinated naphthalenes, chlori- < d "l ry 1974 477 . *. i -v\ NPC00016204 207890 tfic chlorinigjL^iplitnyJi^s_v!^lJ_jii_ihg_dlliiliZ [ H l H trrn lim iu lo 1 1^ ... -I*....| ,, ,, 11,,- liv e r lO- various species. cnumartd tnc toxieitv of differently ellIrnk nalcd naphthalenes auiLa biphenyl that contained (5% chlorine. Tri-cliloromiplillialcric apparently was leu hcpatmoxic than iJic higher chlorinated compounds and the chlorinated (Iiphenyl was (ftc most toxic of jll. The similes were done in rats and the exposure was by reeding as well as inhalation. The findings described in the liver consisted of liver necrosis, fat accumulation, and the presence of hyaline bodies in the cytoplasm of the liver-cells. A marked deposit of finely divided granular yellow or yellow-brown pigment was observed in KuplTci cells, particularly in chose animals that had been exposed to lire chlorinated biphenyls. Tliij pigment apparently did not slain characteristically for either hemosiderin or hcinofudisin O f^am ae- all livers were enlarged macroscopieallv m addition, these authors described nmlifeuiivc changes in occasional bile dorl^ particularly gllfr fcsftinjj mixjpf^ uf dilori--- naled naphthalenes and chlurinalrH binhe.nvU The number of bile duels was increased in certain areas and the epithelial cells lining the bile ducts showed mitotic figures. In inhalation studies wiih low concent rations uf PCUs which consisted of either a IG-hr daily exposure lu average concentrations of 0.57 mg/m3, or to 8 hr daily with an average concen tration of 0.9.t tng/ntJ . neither group of rats appeared ill. The periods oT exposure varied from 37 to 143 days. When the animals were sacrificed the livers were pale or slightly yellow and some what mottled. Microscopic examination of the livers revealed simitar changes as had been observed after feeding the material, and an increased vadrrolalion of the liver cells. Micro scopic evidence uf recovery was not observed in the livers of rats that had been exposed to the chlorinated diphenyl for a total of 105 days and were removed from exposure for 2 months. Similar findings were made hy Miller1** who studied the effect of a PC'B which contained approximately 42',>- chlorine. This author observed varying degrres u! liver damage alter subcueaneons injection, as well as oral ingestion or the toxic substance in guinea pigs. rats, and rabbits. The liver damage consisted ul fatty degeneration and atrophy of the central lobular cells. In ih, rat. hyaline bodies within the cytoplasm of the liver cells were also noted. Mosi of the liver damage was found in the guinea pig. less in die rabbit, and least in the rat. Nislii/.umi.Ifl J who studied the effects of a Japanese biphenyl Kancclor 400 with 407c chlorine in monkeys and mice, described enlarge ment of liver, fatty changes, granular cytoplasm of the licpalocyics, increases in the size of Kupffer celts and hepatecytes, and a brown pigment in some or these cells. Me aIso studied (he livers under the electron microscope and found an inciease in smooth endoplasmic reticulum, vari ation in the appearance of mitochondria, and an increase in the number of inicrabodics. In addition, "myelin figures'* were observed iu the cytoplasm orhepatocytes. Vos and Koenian't5 observed liver necrosis in chickens that were dosed with European PCUs. while i Monsanto product with 60% chlorine did not produce this effect, [netcased amounts of iron were demonstrated with Pcil's iron slain in the livers of all chickens exposed to the European as well 'a* the American PCD sample. Vos and Reerns"' studied the PCU-induced lesions in the liver of rabbits, following dermal exposure to Aroclor 1260. Fhciiuclor. and Clophcn. The micro scopic changes observed in Ihc liver included fatty degeneration, focal necrosis, ccnlrolobular liver cell atrophy, eernid pigment in Kupffer eells. periportal fibrosis, and cytoplasmic hyaline degen eration. The feeding of Arnclor' 5460, i chlorinated tcrplicnyl. Aroclor 1254. or a chlorinated dibenzop-dioxm, not fuither specified in the article,*led to liver hypertmphy of .Sprague*Daw!ey rats within eight days to three weeks. Ultmslrucniral alter ations were similar in the three groups and consisted of numerous multilayered, concentric membrane arrays and proliferated smooth endo plasmic reticulum. In many instances these mem brane arrays surrounded lipid droplets.1"'0 Rabbits given 300 mg of Aroclor 1242 and 1254 once a week for 14 weeks developed enlarged livers and the ones given Arodni 1254 showed midzonjl necrosis of the hepatic lobules. The livers o! lahhits fed Aroeloi 1221 in a similar fashion did not show any histologic changes.111 We conducted studies to compare the toxicity of Aroclor 1254 and Aroclor 1260 in Shcrnint strain rats. The .mute eft eel of these two mix tuic on the liver was negligible. If rats were given c 478 (,`K C ( r itn v t / ir n ru t m NPC0 0 1 6 2 o5 PKiUR!-. 3. Set. lion e'* liver of a rat led Aroclur 1261* for PC*i!.i> <ThH fiiriire illustrate* .1 utiioMe figure anti a hypcidiromalk nucleus (arrows) II & I. X 50*> sin(Jc dose of 10.000 mg/kg in peanut oil by Mmmch lube, and were sacrificed the next Jay, llie liver was essentially normal microscopically. Duses of 1.000 ppm (72 mg/kg) of Aroclor 1260 for .3 months produced only an increase in mitotic figures (Figure 3) and enlarged hepalocylcs. When groups of rats were fed dietary levels of 20. 100. 500, and 1,000 ppm of Aroclor 1260 and 20. 100. and 500 ppm Aruclor 1254 for 8 mouths, the liver changes were pronounced.555 The hcpaiocytes were enlarged hr many of the rats of the experi mental groups and lipid accumulation, foamy cytoplasm, and a brown pigment which stained partially positive for hemosiderin were observed in the livers. The pigment was primarily observed in macrophages and Kup ffer cells. Inclusions (Figure 4) that stained slightly more eosinophilic than the surrounding cytoplasm were present in the cells. These inclusions within the cytoplasm have been described for many compounds such as niirex11*1 ( d u d ecn ch lo ro -o ctah y d ro -1 .3 .4-mclheiu> 211* cyclohuta |ctl] pcntalcnc). DDT511. l.l.l-itu* chlom-2,2-bis(p-ch]orophcny!) ethane dieldrir: 7,<. - 1.2 ,2 ,4 ,10, l 0-hex achloro-6,7'C poxy I 5 .6 .7 .7 ,8 a - oe l a h y d ro - 1,4-ciido-cx u* v.8- d i me l h a n on aphih.i|flr . and pipcronyl but n v id e5 to-iZ(-buio\>Tt!ui\yh'|lio.'iy)*4.5* rnelhvlenedioxv-Z propyltoluen;:). They me identi cal to the so-called hyaline bodies that were described in the earlier liieriilmc by Ucnnell el ah5" 7 and Miller.5'1'1 Aside fiutii these general changes. extensive, grayish-white, linn, glistening areas were noticed in the livers of u numbfi nl the experimental annrutv particularly at the high dietary levels. Miciowopk- examination of these grayish-while arcus showed that the hr palie parenchyma lud been replaced by glandular pale staining epilhilial cells that IdMUL'd ducts and were suinmndcd by proliferating fibrous tissue fFigure 5). Larger lesions often had extensive fibrosis and also contained collagen and the ducts th.rt were formed by rpi!luli.d pale' staining cells were markedly dilated and tout.lined hcltoUc debris nr mucus. The lev.... has been classified as adenofibiosrs for tin* nr.v bcini: It has been discriheii in detail by lidwards and White1 who observed it in rats that were fed butler yellow (p i!mioihyt;tmiili-a/.o* bcn/enc) 1 licse lesions enlarge peripherally by the (ornia'ion of new i't->ndular slruclures. Hepatic January I97i 479- NPC00016206 207892 .. .. !I * 'i 'I i f I i. 3 i i ,t i i ii V*! *' WCVRL 4. Section of liver from rat fed Arurior 1254. A duster of inclusions is present in the cvtopWsrn of one hepuloe> tc farrow). Usually indivKtiint cells contain only one oT these form alions al a timr.. Tolutdine blue stain rf marcrcluss embedded m utcrul X 300. tissue may be trapped between the glands of the peripheral area of adcnofmrosis. The older cenlnd pans of the lesions sometimes contain pigmented avascular collagcnizcd stroma wjth atrophic epi thelial cells lining dilated .'ducts that appear atrophic and dead. The glands of actively growing adenofibrosis show atypical cpilhelium with large pleomorphic nuclei. The epithelium can occasion ally become sirati Tied. Parts of the lesion may be (pule extensive. while in a'iiliiion, in many areas of the same liver sinaii foe of adenofibrosis exist simultaneously in vlo;s o-kuiunship lo porial areas. It has therefore t*.*'a assumed that these lesions arise from hue eh..;-. but they can-also be primarily confute:! ! egion of the hepatic veins. The absemv : ' ..- ducts around hepatic veins Jed to the that a precursor, other than bile .ft.vis. existed from which adenofibrosis dcvelopeCi However some investigators of the lesions ha'- jvf'mcd that it represented an atypical bile Uv .; rooiiferjlinn. also referred to as cholangiofibroM; .*7* lr zduitron to this lesion we also observed s s .--Tets of glandular cells within areas of uiii.mifibrosis as well as adjacent lo bliHui vessels that were surrounded by relatively normal appear tug hepatic parenchyma with stain ing characteristics of salivary gland tissue.'These clusters of cells very closely icsemhled pancreatic tissue.2711 The distinction between adenofibrosis and adenocarcinoma is difficult. The glands ate quite atypical in adenofibrosis but it may be helpful to remember that in adcnofibiosis, in contrast to adenocarcinoma.. the it regularity observed is limited and duplicated with consider able exactness from lesion lo lesion.. Mucus if. always present in adcnofibiosis. hut nut always in adcnor.ucmotn:i. The struma of adenofibrosis is concern neatly'arranged, young and cellular at th-: periphery and old and hyalinized towards th* center of the lesion.211 Of course a liver showing adenofibrosis may also contain carcinomaioi; Icsinns. So fin copclusivcevjiifliieris_^ inth liier;itUT^^Tut*^^i]ot'ihtriiLL-iimiS^JJsc to carv u o m ^ iumever. it lias been found lo coexist whi carcinoma oT the liver in rats and some invest gators feel that the lesion is in fact u precursor i cancer.2"" Choline deficient rats apparently. < addition lo other changes in the liver, also develo. *8.0 .. .CltC-L'iiiif o! V'>.. 'i 1i,Tu.*fnei* I NPC00016207 207893 tf i 4 {4 J ' i.: ' v hTTr *'-' r^V ,, T. i ' .-I' , - f v . : . . ' r -.>! 0 -V..USk^'^> I-ICIURI* 5, Section thus tt;i 1cs an aren of mK* j1- -' ni Ilie liver in' u ra t led A io c Uk 1254. Note the cpilliflial le lh I'tirinirY sin.tll ducts. Tl- . .fiuin e d tu b i di-bris and are tum nuukd >1 by Tibiniis which cun beenme ipiiii* p i o m m r u x d .i 1 adenofibrusis,3* 1 However, these mis were fed 'i peanut meal and soymeal and ihctefure the posst* bilily exists that the experimental diet was con,, laminated with a mycoioxin. Bentret cl ;rI.TS 7 also y desetibed the lesion (atienolih insist in tats when a mixluic of chlorinated naphthalenes and chluron diphenyls was fed to them. A cm teal review of the histopaihogcnciis of this lesion was made by Siewerl and Snell.*r1 Ultrastruciuial changes of the livers of animals exposed to the Articlor consisted of an increase in - smooth endoplasmic reticulum and atypical mitochondria Many lipid vacuoles were observed, I: particularly al Ihe higher dietary levels. They were al limes surrounded by conceit Iricjlly arranged membranes. The "inclusions" or "hyaline" bodies observed in the cytoplasm with the light micro scope ctit/espund to those formations when they arc examined under the electron microscope. Their ultiaslructural appearance has led to their being '! refer red lo as "finger prints" (Figure M. The ! epithelial component of the adenofihrosis outlineJ 7, consisted of cubmdaf or columnar cells with the j- - liee-surface lined by niicuivilli, gjaoujaf cytoplasm Many ribosomes, and a few endoplasm membranes. Some of the cells contained a greadeal of mucus and resembled guide! cells. Tonofilaments and terminal bars were also observed These vanuus observations suggested th.v. icpuhehn component of adenufibrusis had all hum pholouicai characteristics of biie dtc pit helium. Chickens with clink edema disease and cat:.' with 'he X-disease. already mentioned. usui-h showed chances in the live* which consist : predominantly of liver cell necrosis and fat .. cumulation. The hcpatotuxic cflecl orchlorine ir; pint',aienes has also been described i:i ra:s.: < ' lt> a number of reports- on the occupat:*.. occurrence of chloracn; in workers that w e.Mivd to, chlorinated naphthalenes. chiorina, d'piisr.yis. `or ^.J.^-i. and tcchnict;) tnehicpl'cti. reference is made to the hspaton- effect of these compounds. This hepatoic cite;! can manifest itsoif only in a decrc; : `i-'ranee for alcohol; it may also develop m* hepatic deficiency with acute yellow aliophy a:., ijiumlice Apparently the effect on the liver w January 1974 ' NPC00016208 207894 i i HClfRH 6. tlcciron mk-ropiaph of a portion of ! maopcd airays of membranes tiich surround m'tiplci >. *s:v .-< :.* wlut;h a'^uionQaTecnianhniaTcnc^^^ Colter2*3 reported seven esses of pciuachie-t: 3red naphthalene [owiming in workers who engaged in manufacturing wire cable during WWai 11 lor the navy. Four of the workers (level p* cd jaundice and two died. Microscopic exummalion of the liver of the two workers that t!ie:! showed complete loss of liver evils in some ii .. the cenlrolobular areas were hemorrhagic, prominent bile duel proliferation was seen periphery. AngtMLgrotip of compounds that h:ive_a- _*ome^rfrcse^ Ifly i-aure a iwpalr . effect ?are j|r . chlorinated dibcnzofuiam.__- .ingle or^ihrscs_jjf_J_lo 10 ugfkg " tctm-hloaidihiirizifdit'xiniw n to rabbit;- r ' jcdjL{lI2aiAllS12iliLiJkiL-ALli^ t 0^r<. the imi'mrtlc .lw>l j|ml lfie lower dose:. had rc^ . 20-50 f/g/kg b. , .! .. .rosis in rabbits. eared with applying a o w tf -p.-i-.t of both cars :.* n was 7.05 fig * obits were sacrifices . . iv'bi application of '.he v-v.c significantly l3rp.cr . . > Microscopic examinatirr. ' wiiarccd hcpatoeyies. Some o-` .' ,,vies were muJimuelcaicd. Tire cyow-i> foamy or vacuolated. A light bo*..T; . . m some liver cells as well as Kupffer `i observed. Inclusions were seen in . ...m in some river cells, slight inters!' :<' > s present, anil two of three rabbi! ; *J showed foci of necrosis th*J! -led by fibrosis.23,1 .'.J.-es with "toxic fat." the cause ot . . ease. showed that it had a neon *82 CHC Crifivat firvt'oi'i i>i ToxKnhtfy NPC00016209 207895 effect on the liver in chickens. As we now know; membranes witht- the toxic agents responsible for chick edema that smooth erw disease are chiurinaled dibenzu-p-dioxins. Toxic . hypoaciivc was fats also altered liver morphology in Mncacca a!.**9 mulatta monkeys.*** Hcpatocytei.were enlarged, Fibrosis oT to multinucicaicd and fucaj necrosis in Lhc centro- adenoftbrosis or lobular zone was observed. Many liver cells were should dcfiniivi vacuolated ami stained positive for neutral Tat. lesion.^UsualWj^^ In our studies of rats given hcxachlorophcnc, - with hepatoma? we did not observe any liver pathology with the livers. A p;l" light microscope and no change with the electron Baba**9" show*.8 - microscope at the dosage levels (hat wc in rats with employed.11* Thorpe111 on the other hand, studies mav shot- observed periportal fatty changes in livers of sheep hepatomas in rod;1 given three or four doses of 50 nig/kg hexachloro- phcnc and histochemicai changes were also observ Neurotoxicity ed in the liepalocytes. Pugh and Crowley2*6 also In some of the rr? observed hcpalotoxicity in sheep after giving them the Yusho inrs-Jc". hexnchlorophene. mentioned as v. c:. Of the other germicides discussed in this article, part of the symi. only Iriclosan (Irgasan DP-300) (2,4,4-tricliloro- most reports no- : 2'-hydroxydiphenyl ether) has been reported to the Yusho epiohave a toxic effect or. the liver at a dose of 125 formed mure ; mg/kg and enlarged the liver of rats at a dose of 25 admilied consn-'i t mg/kg, according to the summary minutes of the Hospital in (he OTC Panel, U.S. Food nd Drug Administration. It the patients co: was not stated whether these were single or the peripheral . repeated doses. Oral administration.of triclosan lu five cases, hypr. beagle dogs produced jaundice and severe liver hypesthesia were * damage was observed. The effect that these com tion of *litc sensi pounds have on the liver will have to be studied the radial and : further. . number of these p.. The accumulation of fal in the liver indicates that either, (attransport "or iit meiqfy^m k impaired; if the insult to the iiver is removed, the liver gradually i m a c n unless the lesion has . nervous system sy! poisoning episode company where ; biphenyl.91 _heamcjrreversible. The increase in smooth cndti^. The only chctr.i* afra. Dulvcvglic _ aisn disamaai after -j ccuain amount uf time if. renurt that bus a :- "expusure to the matcri:ilJLiliiCflniuuuLii|lL-I!hL effect m animals as _s[enificance of the increase in smooth endoplasmic. diene. Adult fen1,,1' _ieticulmo_md_ iligjiyajine bodies with a concumit- hcxaciilorophcne if ' ant rise in liver microsomal enzyme jkuvU^js 30 m 18 mg/kg b- presently nut uniteismmhThcse chanties are con- weakness after 12 to _sidcrrd adaptive a_mLfacnSaLbut in addition to weakness which w mclahyli/.iiie unwanted rfn-micais at a hielter rate, progressed to p a - the stimulated liver also metabolizes some medica autopsy the brat" tion.1: as well as sut)stani-cs_~ enlarged; they wci.- nruduccd hv the bud^^uft :is steroids. Uliclhcr the control brains lhc inclusions within the cytoplasm represetsI the Microscopic exam!' so-called inactive smooth endoplasmic rciiculunt normal gray my'.;-. which is deposited in the form of arrays of . spinal cord in the * ''rrn^m not known, The fact :'jlum may beeume 'ed by Hutterer et .1 as necrosis and the "CU exposure in rats ilcred a very serious .curs concomitantly .iicinomas in rodent ;i bv Ktmura and . uinas can be induced -00. Chronic feedine n chioracne and also in .-hsral neuropathy is ic alterations' *190 as ; of poisoning, but in r.sis was put on it. In -d Kuroiwa390 per.nations in 21 cases Kuyushu University : of Japan. Seven of ..umbness or pain in ;*.r extremities and in . palgesia, and thermowing as well as reduc..-induction velocity in was obsetved in a -..jache and peripheral - also reported in a red. in a Finnish ' :;;>regn.Hcd with. of colvchk'-ic-. ij , -*. ** NPC00016210 207896 f s ft. H t.U R I; 7. Electron niter rrpntph fn>n ^ empty appcarinc vauiolc! lined by myelin O' a. A - M iin $ s i.t t its tp o n E H 'W l . N o t e i h e i j 34 200r a n y l a c e t a t e X , . t cord was examined. The white rii.uie- . ..! lined by myelin. The axons ilu contrary seemed widened and snowed m i: appeared usually to be quite norma- spaces Ilia: appeared empty and were 1; strands of myelin had separated : strands of materia) that stained po:- e vacuoles. The vacuoles were on- myelin. The vacuojes were very tr.i-r..- ..except for some elcclrni) dense grs;,. varied greatly in size giving the tissue a soo... . Tlcure 7>. No other alterations v.i i I t appearance. The brains in these studies were ,:.k -ca white matter. This lesion is I usually fixed in either Bourn's solution ur buffeteJ formalin for light microscopic study. .wucidurophene; oilier chemicals status spongiosus as we have cm;* Brain tissue, particularly that of rodents, m- ' ,de irtcihyUin.7'' isoniaztd.19* t nurmully show small vacuoles that arc cimsidr: - -nzanilide. 2-acelu\y-4-ch]oro-: to represent an artefact.797 These vacuole; : i : . 79S and raloxanide.796 w :\. smaller, usually very uniform, and at limes con:,., ';:i-3,*chloro-4,-{p*chloro*phcnt.: a homogeneous pink staining material. They ' octhyltin seemed to produce usually separated by a wider area of norry ..h lower dietary level and r*. appearing while matter than the vacuoles in sij* . uthcr chemicals mentioneJ. ; spongiosus. Brains that show this artefact do ; a. Vrophcnc. weigh 'sipnificanlly more than those that do ..jl studv. after feeding rats r - show it. ppn phuic in the diet for 10 week: Electron microscopic examination of b:.r;n- Iv. .ng (he exposure to hexavii. rats given hcxachloruphenc showed that the v.i. l : began to return in the h- lation in the white matter of the brain res... . /etl rats after 2 week's and w: 404 CH C Critical R n r i \ i>i T "X irn l'> fr NPC000162U 207897 6 weeks the raU had utmost , /ecu'-' clinically. However, iTu'crusc.; t-i: lire brains of these rats shuwie - . *' spaces were still present f ' . exposure lo hexachiotophei': - is-, r linuctl. In weanling rats, but r. brain lesion and symptoms ir _... produced with a single orai d js .. body weight given by stcrn.:-;i o il.*1a Part of the reasen produce the brain lesion in ; - < < a single dose may be relate_ . . ;i<c weanling was able to survive ,, _osc than the adult rat. Other species that have ,v.u and other neurological deficits alter hexachlorophcne are rabbits,197 | . , cats.159 Sheep become ace.'.;. and Malone.150 Hanig el a' : i y : the neurological deficits produced in .r.orc detail. Early symptoms consist/ weakness and ataxia of lhe h;:-. righling reflex, patellar hypenefu:... . .er developed into hyporeflexia. uprv<- and eventually complete flacoV. cranial nerve functions remained /l preteiminally. In these cats an i n c r e a s i , . uai cercbialspinal fluid pressure was In our first studies we con tin-a \ hexachlurophene for a period n.i . wc had designed lhe experiment ; , i approximate 90-day iVcdin sic.;- ' necessary to feed rats for this l*n; ! wc determined later, microscopic . observed in rats after a feeding i: weeks when the animals were e v v .. ' high enough lo produce lhe Jesicn i- ; . tion can already be observed ? *. - onset of exposure to hcxaehloiop!-.*-< s-. rats. When lower dietary levels - : v. reproduction study,118 the diet:: ppm (2 .4 -0 .9 mg/kg body v. :t/r . . effect on the brain. At the die1- : i s . 00 ppm (1 1.8 -4.6 mg/kg body v.-cy.t * . foca: areas or vacuolation of the white ' ' he brain were observed in some of the rats. were never extensive. The rats in this*: dietary levels did not show my clinical r.etj , cal symptoms. The brain lesion observed in rats v-.p described in monkeys following the export). .. hexachlaiuphene ,, Newborn,monkeys washe^ .* , 1 ,, Jeicicnt soluiiuu developed . oils'" .'.Jtjs of lhe white matter ol i . t : icitt5'"*,3,>1 was ilile to ; . r.'in.- i m of lhe white maticr. the si , ' - s;n'..;:v.i;us. by injecting squirrel .. i `.ih.i-:jncously with 5 mg hexa* c; .\-.y 1 : cy weight foi 37 days. When j.w r. - r-.uchlorophenc they did not it.-, 'c on but showed elect ru- r * . '-..rites which need (uniter i.i u The monkeys that showed the -> :r.jo >truled elcctrocticephalo- . a v have apparently alco shown v s t ic white matter according to . ' ll` it:pcrl et al.:' nJ were able lo i " nice us well as'rats. These . .. the morphological changes : . frozen sections of bruins t-'1.'; .' .'.i , - -.rogen. Trypan blue injected :!). ` not stain the brain, an indrea- .rum bjrrier was intact as far 'verned. -.onsistent with status spnngj- * .ruphenc exposure have also :t h':inam. paiiicularly children. the brains,of six human . Itexachlorophenc poisoning, v, tliis group did not show -!.tto matte) of the brain. They <18 hr alter exposure to v The four children (two with . two'witli burnsi hnd at least :i -. * of exposure to hexaehloro* -li a Vw hexnchlorophene 1 The final concentration of : the built walet wa; not . ; J'cn showed status spunposu; rr The brain weight given in one -jv-old child, was <125 g. Thi. `or a chiid that age is 382 g . a retrospective blind stud; nitrate tha the incidence t :> the brain stem reticul. ' - -s who had died from a vane1. .undated to hcxachlorupher- found ?l cases in a series t .dreu. Of these cases. S w i weighing less than 1.400 g a- K w icks' gestation. All hot 2 .< more total washes with v :v .* the group of cases that wr<- ; .. :`; :iber of siiilborns who did i nuaiy W 74 48! NPC00016212 207898 show the lesion and t>f course h.id not had derma! hexacliluruphcne exposure. Shuman el al.30** mentioned catiicr cases.uf Lclierer Siwe's disease in which they had ohserved status spongiustis of the wliitc matter. Each of the children with Lclierer Siwe's disease had had pronounced expo* sure to hcxuuhlurophenc because of the skin lesions wludi had been vigorously setuhbed with a detergent containing the chemical. The various observations show dial, in patients with abnormal skin as well as the skin of the premature, Jicxj* chiorophene may be absorbed in sufficient amounts to cause status spongiosus. The fact that blood levels in premature infants washed with hcxacblorophene tend to be higher than in mature infants also substantiates these findings. Almost identical observations were made by Powell cl al.30* In premature infants weighing less than 1.400 g, 7 of 13 that had 4 or more exposures to hexachloruplicnc washes showed status spungiu$us of the myelinated tracts oT the brain stem. If this group was further subdivided according to the number of exposures, 6 of 9 in fruits that had 9 or more exposures to hexnchlurophcnc washes showed the lesion, and I of 4 that was washed 4 io 8 limes. Four 51illburns and 13 infants washed 0 to 3 limes with hexachloro phenc and weighing less than 1.400 g did nut show the. spongy change. Electron microscopic examina tion showed Liul the spongy change observed in the premature infants was due to a split in the myelin sheaths. . In August of iy7Z the press reported a number of deaths of infants in France that occuired because a baby powder to wluch hcxachloraphcnc hail accidentally been added at a concentration of 69E was used on the infants. The final death lull amounted to 41 infants and young children3fif (New York News. Dee. 13, 1972J. Detailed accounts have not been published of this incident because of litigation problems. As we pointed uu: earllci, undei acute toxicity, the higher the con centration of hexudiluruphcne is in a preparation applied to the skin, the more likely it is that skin damage will result. Skin damage also occurred in the French incident. In addition to this. (lie powder was probably used in many instances where some diaper rash already existed and v.-js then well covered with a diaper, which may have contributed to increased ahsmpiiun in ihis area. The brains and spinal cord of u few infants from, the French poisoning uuthtcak that were examined micioscopicHlIy showed status sponp'osus of the white matter.31*6 These various accounts show that status spon giosus can also he produced in humans and probably represents a nonspecific reaction which can be elicited bv a number of chemicals. The tesion status spongiosus of the white matter as far as we now know seems to represent a specific or restricted type of brain edema where the fluid accumulation is confined to the inside of (he myelin sheath and myelin is not 3Ctualty damaged or at least breakdown products of myelin have" not been observed. The term status sponpiosus or spongy change has been used as a descriptive term and a spongy change'of the gray matter ns;]>' also occur in certain diseases. Status spongiosus together with other morphological changes can be observed in the brain in a variety of many etiologically different diseases and the reader is referred to Kla(7.o,301 Ardonalo and Lamport."'* and a recent editorial in the British Medical Journal.3'1 From clinically observed poisoning cases as well as animal experiments it is evident that the biain lesion is to a great extent reversible. However, in our own studies as well as those by Utmpert ct al..303 microscopic changes in 1odenis did not completely regress and the fact that sheep do rot recover their eyesight after hexaclilnroplivnc- poisoning110 indicates that repair irtay nnt always be complete. No reasonable explanation exists at the moment to indicate why certain chemicals produce Hits very specific edema con fined to ceilaiu structures of the centra nervous system Gastrointestinal Lesions_____ un 1y Twotvpcs 0f compound s under discu have produced lessens in the castromter.inji 1^. TCT observe!! TiMui^tudywitl) Sherman suain > that high single oral doses of 3.000 mg/kg A.-v I3M an^l 12^ or mote caused ulceration castTie and duodenal mucosa ( Figure b~. IV: : " the Cl tract wer.- not observed in the i-'..;-.: feeding studies with 500 ppm cr lea:., changes in Mir gastrointestinal tract we:: 1 Kiiiecr et ill.,11,5 in their repmduutr.ii:-;. . mink, observed that Aroclor 1254 prbd..-:J .. orrhape in the pashoiiitesiinal tract . ~ Jr.~. spring. Allen and Nenback*110 observed : and dysplasia nf the gastric mucosa 1 : monkeys. Six of these iliesus monke;.; -. 4fib CHC C ndnil ftrwe j 01 T u m o lvty NPC00016213 207899 FtCl - tnc*% - . ~;.ir p o r tio n o r th e s to m a c h o r a rat g iv en a jin g le d o se o r 3 .0 0 0 c illustrate) the focal loss o f m ucosa and the infiltration- by 50. 1 300 ppm At ppm Arocir.; - .';i Si' after exposure to ^2.3^8-tetrachlflmdihcfl2Q:__ jjhoxig. At autopsy a ruptured duodenal ulcer and The glandith . ir rt.i a gastric ulcer were found. Whether any relatfon- i atypical; pe, and invasion epithelium wl mucosa in~ : ' which cor: ' : mucosae .ne mucosal ,.J ..-a"Mens'is on -`h^. gastric inulatra, ts eastric slup existed between the ulcers and the exposure to the poisonous chemical was not mentioned in the paper. fih o u ld b c e s ta b lish c tn n i^ ^ also occurT ^m ^^ftetarv^|vglfr ftyrLudiglliec-iL-. mucosa and ; ] " 'the toxic ' know, con*.? (V 1 It has *-- -- *-*u" "ie mucosa. ,;:r^ *- we now imiter. sms. :< observed . -I vrhaging them. Valuable information might be obtain: from retrospective studies of workers followm. occupational exposure. Whether (he ulcers a* occurred in #'ll roinii1- *.1 i'set of rat ' caused by a direct effect of the chemical on thr fetuses who .is'-c :c. 2. .'cchlorodi* mucosa of the gastrointestinal tract or whelk - . bcn7.o-0-dioxi:: ti-TO. it >-' r*w. tied in the stress plays a primary role in their etiology sin' - * reports whet it;- *.i gast- '"liorrhages to ulcers that may occur in burned patients fo; were caused by . citation 1. I l 0,1 I 1 instance (Curling's ulcer) is noi clear. Ulceration of ? . !: tr.d'iL-r o* <rduJr rat -i stomach was ah' -.re:veil. ' .Patients wis!: of loss of appe- 'gtained COMMENTS The acute toxicity of compound^ such as ' i f% trial pain, an.with a foil.cv'j - ............... ... * .w n d u c monihf Tcncs. and the chlorinated terpbenyis rs very _ . . ,, _r . % ^ January 1974 ~3 NPC00016214 207900 (>n Ilie other hand, they are poorly broken down can apparently occur.1 it is also not known if the in the environment and the mclahnlisni oi ttic combined effect- of different porphyria inducing biphenyls with more than four chlorine atoms agents is cumulative. Whether the photosensitivity seems to be negligible. Recause of these character observed in humans with some of the germicides istics, the use, wherever possible, of chlorinated discussed in this paper is related to an effect on biphenyls that consist of mixtures of compounds (lie porphyrin metabolism was not evident from with one to four chlorines has been proposed. the published literature. Neither the_chlprinatcd naphthalenes no; Hie So far the mechanism that causes chick edema, chloracne.'X-disease, and liver disease has not been . present a pyflinmn mhlpm satisfactorily demonstrated. Cluck edema disease Mliclher the various disease entities reviewed in has been thought to be caused by proliferation of this article arc produced by all the different the Vascular endothelium. This might explain the products mentioned in connection with them is fluid accumulation observed in-diseased animals, not established with certainty. For instance, some but so far the sequence of event* that Jed to this of tire compounds that produce cltiorccne or chick disease is not really clear and factors other than edema have been shown to be contaminated with the embarrassment o f the vascular system may various chlorinated dibenzodioxins and chlor play a role in its etiology such as a toxic inated dibenznfurans The possibility remains that myocarditis, decrease in serum proteins, and renal the compounds illustrated m Figure ! are contami failure. More detailed study of (he pathology of nated with these chemicals and tire products the various disease entities is needed for a belter themselves are r.ot really responsible for the understanding of the development of the various observed symptoms. On the other hard, a number lesions mentioned. of structurally unrelated compound-, may produce The chlorinated polycyclic hydrocarbons seem the same disease entity. tu alter The immune response oi certain animal Some of the chlorinated dibenzodioxins and species and perhaps also of these very the chlorinated dibenzofurans are highly toxic. in ip o rta iU fn d in g w rio be pursued further. The most toxic is 2.3.7,8-ietrachtorodibcnzo- Thus far it has not been established beyond doubt dioxins, while the dibenzodioxins without any wherlvrr this is caused by a general debilitating chlorine and those with right chlorines are much effect bn the animal ftl^llilJIEEliy- f-the less toxic. The difference in toxicity of these lymphoendothelial systejrL_nas^ttJMlfaBMBtftftw. compounds may be partly caused by the fact that they arc very poorly dissolved and absorbed. wc know _it--frnm-ihg--eflgclL^Q-iadialiQl]--iilL- Intravenous toxicity data are thus far not available exagsi--IP--gM_ain alkylating agents. In this on most of these compounds. ""respect not enough attention has been paid to*the We also'do not know at present if the porphyria function of the adrenal, and several of the which develops after prolonged exposure to a observed symptoms in humans such as the hyper number of technical compounds such as hexa- pigmentation suggest hypofunction of the adrenal chlorobenzene, chlorinated biphenyls, chlorinated cortex. c j> e n d jn ja n riie in ^ .obtained dibenzodioxin, and 2,4,5-T is in many instances from these studies .a tolerance level of 5 ppm in , due to tile contamination of these products with some food products ai-S cL i iiltfJljL J-aad and chlorinated dibenzodioxins: of course it may be T ^ u ^ ^ d jn in istia tio n may have t u b e revised possible that one or more chlorinated dibenzo particularly sin c p n o lo rt| term low lew! feeding furans also produce porphyria. It has also not been study lias been reportedin animals.___ established whether all compounds that produce Many questionsTrav^ccently been raised about porphyria iij animals will do so in humans as well. the use of germicides in various toiletries, common In. general, the exposure of the various popu soaps, etc. The objection to their use in the lation groups to chlorinated biphenyls and Other specific case of hexachloiophene is its neurotoxic cyclic chlorinated hydrocarbons ts at too low a effect, that can also occur in humans, particularly range to induce porphyria. Whether this can also if the skin is defective in some way. Of course be said for occupational exposure is not known. limited localized exposure, such as handwashing or When production conditions and general hygiene treatment oT a small wound, will not produce of*thc -workers aw poor, porphyria -cutanea arda_ neurotoxicity. 488 CRC Critical Hcvtcu s m Toxicology NPCQ0016215 207901 In infant nurseries in hospitals, one o'' ihe greatest problems is the constant threat :; ,i staphylococcus infection of epidemic p ro p r:./ ' In order to prevent outbreaks of staphylu... infections, a routine was developed i: r.i-ity hospitals in the U.S. and elsewhere in t>.. t.-.riy 1.960*5 where Infants were washed once da,- v-ilh a .1C hexachlorophcne deterge l solution, and in addition the diapet area was washed several times a day. The detergent was subsequently rinsed of*' more or less vigorously, depending upon the philosophy of the different nurses. In some hos pitals the detergent was directly applied to Hit infant, in others it was first diluted in water. Since the neurotoxie effect of hcxachlorophene has .become known, many hospitals have changed their policies on the use of hexachlorophene. Another drawback to this routine, in addition to a possible neurotoxic effect, is that hexachloroplienc keeps not only the staphylococcus or certain phace types from growing on the skin, but reduces all Gram-positive bacteria,111 which is highly unde sirable and may lead to disease. Recently Evans ci al.313 have pointed out that a marked annuel variation exists in the prevalence of several bac teria in the nares as ell as the umbilical cord of newborns. A decline of staphylococcus aureus was observed in the fall; the prevalence of /tithcrkluj enti and streptococcus also varied. Staphylococcus epidermis and Emcrobacter organisms did not show this fluctuation. Any studies of efficacy undertaken in a clinical setting should take these normal fluctuations into account and i* is possible that good general housekeeping practices arc ultimately more effective in, preventing bacterial disease in newborns than various germicides. Unfortunately, breaks in technic do occur which, in many hospitals makes the use of germicides on infants desirable. .............. - . -J i.. . - ihal the tu... tffc. -, vi w r.'\ muled compounds .re c n n t l T v f T " " ~ 'r vj i / if.' .e ' ~i -M'.'1' . . .. ______ ,i acc.-m panted ay a* great deal oT emot,- distorted facts, arid often g-eat ecor.orr.. > Pmdciice. better judgm-'m,: ;r ire usage of chemt'-ais . ..i- - an evaluation of whether in s p .. *: s ;... - - they 'jis necessary nr not. n v ,, these suuaunns ir. the future. ; ;U' . a* *1.. he rcid U: avu.Jrr.e inv.s-. ev . . ... 1 'o chemicals that we ..now , ir :: r '. *.r it:// react differently frur.i a: dr* an. ; . ' . . nc-iertn exposure is concerned, '.ir-;-- v-. be ; : most vulnerable. ACKNOWLEDGMENT My sincere thanks go to my colleagues former co-workcrs who have made this re\i-;-' possible, particularly to Drs. R. E. Zrhr. .* Suggs, V. \V. Burse. R. E; Jennings. R. E. Lin:' V. . Sedlak, J. A. Goldstein, fi. C. Villancuev*, and T D. Gaines for letting me incorporate som/ of the unpublished lesults into Ibis manuscript. REFERENCES t. Poland. A. I'.. Snti.lli, D., Melier, G..and 1'iwick. P.. A|-.ej:T'. c> (*l workers ina 2.4-D and 2,4.5-T plant. Arch fcVii'irrw, 22.316. 1971. 2. Blcibcrji. J.. "alleo. M - Brodkin. R.. and Applcbaum, T I .. Indu'in-illy acquired porphyria. .-In-h. Dcrmatut. 8?, 793.1964 3. Larson. [X L.. Studies s)>uw lu.-Mjchlc'ruplii'm: eau*t * i u>;i v*> i j - I m , Hmp. /f.tioc . 4 2 .6 3 , 1968 J. Schmid I , It. ami Schulz. C,. t.'ber Diphenv Ib.i'cn, ,1ie ri. 'u *. .*S I ntc | 5, Penning, C\ II.. fliyMi'al chjucKiiMK* and cnninn-i.i. i ' nl chloiinjled tliphirnvl. Jntl. 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