fl \ V I y — — 7, '_ __. _,—__._ Pl _ — —— — — — MM MCIDENCE OF SALMONELLA CARREERS AMONG MAMMALS IN THE LANSiNG AREA ‘E‘hocés {as the Degree of M. S. MICHEGAN STATE COLLEGE .i'azsafa Campeéanéco WQB :45 Jan- 273?; thesis entitled presented by J osei‘a Campodonico of the requirements for This is to certify that the daster'a degree in Bacteriology Among Mammals in the Lansing Area Incidence of Salmonella Carriers has been accepted towards fulfillment WBt 3 g 1948 I a Jo {Cg . 9., I. ‘ Told 4.... . a“ .1 a???“ .. .. (to-.. .- at Mt. I-Jyti. .- , I‘ V. c m \ a. , u .. .. cud . . 7.? av 13 I J ’5, Sp- .1 k). ’N I cf..\ [.9 u . 1 «a . it. a .. , .. \ , Jx, _. rt INCIDENCE OF SALMONELLA CARRIERS AMONG MAMMELS IN'THE LANSING AREA .A.THESIS By Iosefa ggnpodonico Submitted to the School of Graduate Studies of Michigan State College of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE Department of Bacteriology 1948 THERS ACKNOWLEDGMENT The writer wishes to acknowledge her sincere appreciation to Doctor H; I. Starseth under whose supervision these studies were planned and executed. She also wishes to thank.unse Lisa men who helped in one way or another to make this study possible. ******** *****# stat ** 4: 207%")? TABLE OF CONTENTS . . Page INTRODUCTION..............o...o..............o.....oo REVIEW OF LITERATURE Human Carriers.................................... 1 Animal Carriers..u....u....................u... 4 EXPERIMENTHL‘WORK Matorlals..uu....no...“....................uo 11 Procedure..uo.........u....................u.n 12 Hemltao.........u..o..........................o. 13 Humanauuuuuu.no......................n. 14 S'ine.........ou...uou....................u 14 cattheeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeoeee 14 Horseseeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeoo l4 Sh.°peoeeeeeeeeeeeeeeeeeoeeeeeeeeeeeeeeeeeeeee. 14 mtseeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 14 DOSSeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeee 14 Cats............o..........................o... l4 Rabbitsoeoeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeeoo l4 Minkaooeoeeeeeeeeeeeeeoeeeeeeeeeeeeeeeeeeeeeee. l4 Chinchillas.................................... 14 Mice........................................... l4 RatBOCCOOOODOOOOOOOOOO0.0.0....00.00.000.000... 14 DISCUSSIONOOCOOOCOO00......0....OOOOOOCOOOOOOOOOOOOOO 15 Sm‘mRYOOOOOOOOOCOOOOOOOOOOOOIOOOOOOOOOOOOOOOOOOOOOOO 16 BIB-IIMRAPIIYOOOOOOOIOI.OOOOOOOOOOOOOOOOIOOOOOOO0.0... 17 INI'RODUCTI ON A carrier is an individual who harbors specific organisms of a disease in his body without manifesting any clinical symptoms. Human and animal Salmonella carriers are a potential danger to susceptible individuals in the neighborhood. Human carriers are a serious public health menace, because of their many contacts with other people, especially if they are employed as food handlers. Animal carriers endanger the health of other animals, and pets and food animals which harbor organisms patho- genic for human beings are likewise serious threats to public health. Salmonella carriers may have subclinical infections or they may be convalescent carriers or chronic carriers. In some cases persons and animals are considered as carriers because they harbor some extra-intestinal localized infection. Such infections are not accompanied by intestinal disorder. Because of the importance of Salmonella carriers, the following experi- ment was undertaken for the purpose of obtaining some informa- tion concerning the incidence of Salmonella carriers among mammals in the Lansing area. REVIEW OF LITERATURE Human Carriers Wolff (1947) stated that only a few Salmonella types occur exclusively in man. The majority of the known types occur pri- marily as animal and bird pathogens but attain importance in human medicine because of their obvious adaptation as human pathogens. Welch, Dehler, and Havens (1925) examined 1,076 persons in the dairy industry in Alabama and found 55 carriers (5.1 per cent). The incidence of infection was as follows: Salmonella typhosa 39, Salmonella paretyphi is and Salmonella schottmalleri, 3. Of the typhoid and paratyphoid carriers, 18 and 5, respec- tively, were discharging the organism.in the urine. Salmonella schottmfilleri was isolated only from feces of the 3 carriers. Schiff and Saphra identified (1941) Salmonella hazgna_from an outbreak of meningitis in a maternity hospital in Havana. Twenty-one infants born within the hospital and an additional number born outside develOped symptoms of severe purulent menin- gitis. The source of the infection could not be determined. Sera of three servants of the hospital agglutinated the organism in a dilution 1:150, but the organisms were not isolated from the stools. Bernstein, Saphra, and Strauss (1941) at the New Yerk Sal- monella Center isolated the following organisnm from the stools of healthy carriers, three of whom were food handlers: _S_. schottmiilleri, _S_. typhosa, Salmonella oranienburg, Salmonella _de_rb_z, Salmonella typhimurium, Salmonella cheater, Salmonella montendeo. Stone (1943) reported that from two thousand food handlers in the Panama Canal Zone, 14 species of Salmonella were isolated from 44 of the individuals. Galton and Quan (1943) in Florida isolated the following Salmonellae from normal persons: _S_. 513121 from a nun, age 5'7; Salmonella am from a foodhandler and from the feces of a normal female applying for a job as foodhandler. Edwards and Bruner (1943) reported a study of 3,000 _S_a_l_- monella cultures isolated from man and animls in the United States and its possessions between 1934 and 1941. The following species were isolated from normal humn carriers: Nb. of Salmonellg_0arriers paratyphi B paralyphi B var java typhi-murimm derby california bredeney oranienburg bareilly montevideo tenessee newport oregon manhattan litchfield bonariensis typhi panama Javiana give anatum nemington senftenberg simsbury northington mississippi p POI p to P‘F‘F‘alh'firh'h‘uiaiklklr4kl(DFJCDtfltutobdto-a Seligmann, Saphra, and Wessermann (1943) reported that bacteriological examination of 1,000 persons by the New Yerk Salmonella Center, showed 89 to be healthy carriers. or these, 44 were contact infections. They represent one group of healthy resistant individuals (among them.were two babies in the first month of life.) Hermaeche, Surreco, Feluffo, and Aleppo (1943) reported that from.3,592 children, admitted to hospitals for all causes without regard to intestinal disorders, 29 different types of Salmonella were isolated from 489 patients. The most prevalent types were S. mhimurium, Salmonella newport, Salmonella montevideo, _S_. 222.929 S. schottmiilleri and Salmonella bredeney. Thirty—four (9.04 per cent) of the 489 persons infected with Salmnella were carriers. Posch, Stofer, and Hirscfield isolated Salmonella cholerae- 531.; as the specific factor in an indolent, subcutaneous abscess in an aged Negro laborer who had suffered for 40 years as a re- sult of a severe contusion a few inches above the wrist. Animal Carriers The role played by the Salmonella in animal disease is an important one as all species of domestic nnmmals are in varied degree susceptible to infection. The work of Edwards and Bruner (1943), showed that swine were by far the most important source of Salmonella organism in mammals. Salmonellae have also been isolated from healthy cattle, dogs, rats, mice and birds. Wolff (1947) stated that _S_. cholerawsuis was the most prevalent of more than 30 Salmonellae isolated from swine. The second most frequently occurring type was _S_. typhimurium. Rubin, Scherago, and Weaver (1942) traced many outbreaks of Salmonella food poisoning in nan to the eating of pork from swine which had undergone rigid antemortem and postmortem veterinary inspection. Hormaeche and Salsansndi (1936) isolated Salmonella from the mesenteric lymph nodes of apparently healthy hogs slaughtered for meat purposes. They examined materials from 36 lots of hogs consisting of 20 animls each, and were able to isolate _S_a_:_L_- monella organisms from 22 (47.9 per cent) of these lots. These organism were: _S_. typhimurium, §_. m, §_. d_er_bz, g. newport, §_. montevideo, Salmonella muenchen, _S_. schottlm'illeri, g. breden Salmonella m, Salmonella 935133, and Salmonella _c_e_r_1_.'_c_>_. Monteverde (1942) isolated SalmOnella bomriensis from the mesenteric gland of a normal pig in Buenos Aires. Rubin, Scherago and Weaver (1942) examined 40 lots of hogs, each lot consisting of 25 animals, and found the incidence of Salmonella organisms to be 47.5 per cent. 0f the 50 hogs examined individually, 5 (10 per cent) yielded Salmonellae. From these positive lots and individual animls, 242 straim yielded 13 types of Salmonella. Cherry, Scherago and Weaver (1943) investigated the incidence of Salmonella organism in retail mrket mats. They found a higher incidence rate in pork products than in other mats. 0f the 250 meat sunples exanined, 13 (5.2 per cent) yielded Salmonella and 10 (5 per cent) of these were isolated from 170 pork products. The Salmonellaefound in pork samples were: _S_. typhinmrium, g, newport, g. m, Salmonella 531:, S. bredeney, Salmonella newirgton, _S_. derby, and Salmonella senftenberg. Bruner and Edwards (1946) reported the isolation of 11 types of Salmonella from 60 bovine outbreaks. The most fre- quent types found were Salmonella ¢_i_u_b.l_i_1_1., _S. mhimim, _S_. choleraesuis and Salmonella enterilidis. Guerrero (1943) stated that carrier cows are usually the source of Salmonella infection on farms where the disease is not prevalent. These cows usually abort their calves fr°mn§2$f monella infection at successive pregnancies. Johnson and Graham.(l944) reported the isolation °f.§2lf monslla choleraesuis :55, kunzendorf from a bovine fetus origi- nating from.a brucellosis free hard. Wolff (1947) stated: "Meat from cattle has oftul been in- criminated in outbreaks of Salmonella infection, though less often than pork. In fact, the first recorded Salmonella food infection was traced to meat of a diseased cow, in which‘S. enteritidis was isolated from the patients and from.the sus- pected meat.” Cherry, Scherago and weaver (1943) examined 64 beef samples. g. typhimurium was found in one sample. and _S_. senftenberg in two of the samples. Salmonella infection of the horse is quite common, but there is insufficient information about Salmonella carriers among horses. Jordan and Burrows (1945) give'g. abortus gggi_as the cause of infectious abortion of mares. .§: typhimurium.has also occas- ionally been reported in horses. -5- Wolff (1947) stated that there have been reports of horse meat as a source of infection with Salmonella abortus _e_gu_i_, _S_. typhimurium and other varieties. Newson and Cross (1924, 1930, 1935) investigated several outbreaks of gastroenteritis in lambs caused by S. typhimurium. They regarded the long railway journeys the lambs led to make and the long periods of fastilg as predisposing factors. Henning (1936) described a new type of Salmonella isolated from a sheep at 0nderstepoort, South Africa, and was nannd Salmonella 0nderstem>ort. Henning (1939) stated that Salmonella abortus 9_v_i_s_ is the most common pathogenic Salmonella for sheep. Wolff, Henderson and McCallum (1948) examined 100 dogs. Eighteen of these animls were found to be excreting different Salmonella types: Salmonella manl'a ttan, _S. newport, Salmonella minnesota (both monOphasic and diphasic varieties), _S_. oranienburg, _S_. typhirmlrium, _S_. bredeney, _S_. worthington, Salmonella 51-7—0, Salmonella M, _S_. 33133, Salmonella kentucg, Salmonella illinois and Salmonella meleagrides. Wolff (1947) stated that more than 20 Salmonella types, all of which are known human pathogens, may be isolated from various conditions in dogs. Wolff (1947) stated that _S_. choleraesuis and _S_. paratyphi have been isolated from cats. -7- Litch and Meyer (1921) described an epidemic infection among rabbits caused by S. schot tmdlleri. There is little information in the literature about Sil- monella carriers among minks, rabbits and goats. Coburn, Armstrong and Wetmore (1942) reported S. m- 911523311. infection in chinchillas (chinchilla laniger . The organism was isolated from 32 of the 44 animals examined. Carrier animals were not detected by repeated cultural tests of fecal material. Four rodents of the domestic species of mice were found to be infected. The authors were of the epin- ion that wild rodents were the source of the infection in the chinchillas. There are nany reports in the literature about Salmonella carriers among rats and mice because of the importance of these animals in the spread of Salmonellosis. Duthie and Mitchell (1931) reported that several strains of the genus Salmonella appeared capable of inciting the form- ation of pseudo tubercles in rodents. Many attempts to isolate organisms of the Salmonella group from rat and mouse excreta have been made because of food poisonilg outbreaks. Meyer and Matsumura (1927) examined 775 . rats in the city of San Francisco, California, 58 rodents were infected either with S. enteritidis (28 cases) or S. m- murium (30 cases) . Krumwiede and Salthe (1924) were successful in isolating S. typhimurium from rodent excreta obtained in a bakery where prepared cream filling was found to be the cause of a food poisoning outbreak. Welch, Ostrolenk, and Bartram.(194l) made a study of rodent feces collected from.a large area of the United States without regard to history of intestinal disease. They found that only a small percentage (1.2 per cent) of these animals were excret- 113 food poisoning organism of the Salmonella type. While performizg the same experiment, they found that excreta of rats naturally infected with S. enteritidis and held at room tempera- ture may contain viable organisms for at least 148 days. Bartram, Welch, and Ostrolenk (1940) found that the majority of infected animals ceased to pass the organisms by the 15th day. Approximately 20 per cent continued to excrete Salmonella for as long as 7 weeks after infection which was the duration of their experiment. They suggest this as indicating, as in the case of human enteric carriers, that a small percentage of infected animals become carriers. Rubenstein (1944) stated that about 2 per cent of all per- sons infected with.Sa1mone11a excrete these organisms for a period longer than one year, while the clinically manifested disease, if present at all, does not last over a few weeks or days. He also stated that some strains are much more likely to produce carrier status than are others. Five per cent of all persons infected with Salmonella schottmfielleri become carriers, while human carriers of'S. choleraesuis have never been observed. -10- EXPERIMENTAL WORK materials In this study, 170 specimens were collected from.feces, intestines, colons, and lymph nodes of humans and of animals as shown.in.Table I. TABLE I Sources of ngples Individuals Feces Intes- Lymph Colon Total tine Nedes Humans 20 20 Pigs 2 2 4 l 9 Cattle 20 l 21 Horses l4 1 15 Sheep 6 l 7 Costs 4 4 Dogs 30 6 36 Cats 14 14 Rabbits 6 6 Minks 11 ll Chinchilla l l Hues ll 11 Rate 6 8 14 The human samples were obtained from Michigan State College students. The animal materials were taken from the following sources: muchigan State College Veterinary Clinic, Ndchigan State College barns, and private farms around East lensing. All the rat and mice specimens were collected from.the Department of Chemistry and from the Department of Physiology and Pharmacology of Michigan State College. Procedure Fecal samples about as large as a pea were dropped into tetrathionate broth and incubated for 24 hours at 37°C. Some of the fecal specimens were diluted with enough sterile water to make the samples soft, and from each of these streak inocu- lations were made on one McConkey plats and on one 88 agar plate. The plates were incubated with the tetrathionate broth cultures. When intestines, colons, and lymph nodes were used, a piece of the sample was dropped into tetrathionate broth, and another piece of the original sample was streaked on one NbConkey plate and on one 53 plate. All were incubated for 24 hours at 37° 0. Up to the first 38 samples the tetrathionate broth cultures were streaked upon SS only, after that the rest of the tetrathiomte cultures were streaked on.McConksy and SS plates. After the 24 hour incubation, all the plates were examined and the nonslactose-fermenting colonies were selected and inocu- lated into Kligler's iron agar slants. The plates were returmd to the incubator, re-examinsd at 48 hours and kept two more days in the incubator for observation. Gram stains were nade from the Kliglsr slant cultures. Gram.nsgative organisms showing salmonella-like reactions in.Kliglsr's media (acid and gas in the butt, alkaline slant with or without H28) were inoculated into the following carbohydrates: dextrose, lactose, sucrose, maltose, mannite, sorbitol and salicin. Tryptom broth, urea and lactose motility nedia were also inoculated. After 48 hours incubation, the carbohydrate media were observed for the pro- duction of acid and gas. If acid and gas were not present the incubation was continued and observations were nade daily. The tryptone broth cultures were tested for indol reaction anl tin lactose motility tubes were observed for motility and lactose fernnntati on. Proteus organisms were detected on the basis of urease activity. If acid and gas were produced in dextrose, maltose, mannite, sorbitol and not in lactose, sucrose and salicin; indol was not produced; urea was not decomposed; and hydrogen sulphide was produced or not; and the organism was motile, it was tentatively identified as a member of the Salmonella group. For determimtion of the species further fermentation studies with the necessary rare carbohydrates were done. Cultures were submitted to the Sal- monella typing Station, Bureau of Laboratories, Michigan State Department of Health, for antigenic analysis. Results The results from the 170 samples are as shown in Table II. S. typhimurium was the only Salmonella species isolated. This culture was isolated from the feces of a 5 year old dog in the Veterinary Clinic of Michigan State College. The dog had a whip- worm infestation. as“ gangs afieooaasm... o 3 m o spam 0 d“ .2” soda 0 a a H «a 3.23830 0 a." .2” m m m o» .5 omega 0 m m 333m 0 3 v." n .3” case *H on .n H o 0 ma assume: 0 m on a ma n on on amen o e e assoc o a H o aeonm 0 we a m 0 «ans weaken 0 H a n n e c» on season 0 am 3” mu a mum edupeo o a o e n n o» ob eavuso o m a e m . a norm 0 3 S” 5 onnom confirm sauce on: M 5 dozen cadence ueamaom soaoo nag InepoH neoem Hem ewe Headed Idem Ho .02 .oz Hanan. gem Ho 356m HH g9 DI SCUSSI ON Assuming that the technic employed in this work was adequate for the detection of Salmonella, it is obvious that the incidence of Salmonella carriers in the Lansing area is not great.‘ Selective media are toxic in some degree to all organisms and the use of tetrathionate broth and SS in succession for 38 of the samples may have resulted in inhibition of growth of Salmonellas. In a thorough examination for Salmonellag, one ought to employ a variety of combinations of media for each sample. Time and facilities did not permit such a far reaching investi- gation but these studies should be supplemented by further work of more intense nature. It is also possible that not enough cultures were made. -15- SUEQSIARY 1. One hundred and seventy samples of feces, intestines, colons, and lymph nodes obtained from humus and animls were examined bacteriologically for Salmonella. 2. ‘§, typhimmrium.was isolated from the feces of a 5 year old male dog which was infested with whipworms. All other samples failed to yield Salmonellas. -15- BIB‘LI OGRAPHY Bartram, NE. T., Welsh, H., and Ostrolenk, M. 1940 Incidence of members of the Salmonella group in rats. 3’. Infectious Diseases, ‘§_6_, 222-226. Bornstein, S., Saphra, I., and Straws, L. 1941 Frequency of occurrence of Salmonella species. J. Infectious Diseases, §_9_, 59-64. Bruner, D. W., Edwards, P. R. 1946 Salmonellosis of domestic animls. Proc. 50th. Ann. Meet. U. S. Livestock Sanitary Association ps 194-198. Cherry, W. B., Scherago, M., and Weaver, R. H. 1943 The occur- rence of Salmonella in retail meat products. Am. J. Hyg., §§_, 173-177. Coburn, D. H., Armstrong, W. H., and Wetmore, P. W. 1942 Obser- vations on bacteria treatment of Salmonella m-murium infection in chinchillas. American J'. Vet. Research, a, 96-99e Duthie, R. C., and Mitchell, C. A. 1931 Salmonella enteritidis infection in guinea pigs and rabbits. J. Am. Vet. M. Assoc., 7—8., 27-41. Edwards, P. H., and Bruner, D. W. 1943 The occurrence and dis- tribution of Salmonella types in the United States. J. Infectious Diseases, 12, 58-67. Galton, M. M., and Quan, A. L. 1943 Varieties of Salmonella isolated in Florida durilg 1942. Am. J'. Hyg., 519, 173-177. Guerrero, R. P. 1943 Salmonellosis of calves in trepical coun- tries. J’. Am. Vet. M. Assoc., 103, 152. Henning, M. W. 1936 Salmonella onderstepoort: A new type of Salmonella from a sheep. J. Hyg; 33, 525-530. Henning, M. W. 1939 The antigenic structure of Salmonella ob- tained from domestic animls an! birds in South Africa. Onderstepoort 3., 121, 79-189. Hormaeche, E., Surraco, W. L., Feluffo, C. A., and Aleppo, P. L. 1943 Causes of infantile summer diarrhea. Am. J. Dis. Children, fl, 539-551. -17- Hormaeche, E., and Salsamendi, H., 1936 Sobre la presencia de Salmonellas en los ganglios mesentericos de cerdoe normales. Arch. Urug. Med. Cir. y Espec., 3, 665. Johnson, L. E., and Graham, R. 1944 Results of bacterio- logical examination of aborted bovine fetuses from brucellosis-free herds Cornell Veterinarian, 32, 36-40. Jordan, 1!. 0., and Burrows, W. 1945 Textbook of bacteriology 14th ed. W. B. Saunders Co., Philadelphia _and London. p. 395. Krumwiede, C. 1924 Stuiies on the Paratyphoid-Enteritidis group, VIII, An epidemic of food infection due to a para- tnhoid Bacillus of rodent origin. Am. J. Hyg., 11:, 23-32. Litch, V. M., Meyer, K. F. 1921 A spontaneous epidemic among laboratory rabbits caused by a paratyphoid _B. Bacillus related to the rodent group. .7. Infectious Diseases, fig, 27’42e Meyer, K. 1"., Matsumnra, K. 1927 The incidence of carriers of B. aertgzcke and B. enteritidis in wild rats of San Francisco. J. Infectious Diseases, .4_l_, 395-404. Monteverde, .T. I. 1942 A new Salmonella type. Nature, 149, 472. Newson, I. E., and Cross, 1". 1924 An outbreak of paratyphoid dysentery in lambs. J. Am. Vet. Med. Assoc., N.S.V. 12, 289-500. Newson, I. E. and CrossJ’. 1930 Paratyphoid Dysentery in lambs. J. Am. Vet. Med. Assoc., N.S.V. 29, 91-92. Newson, I. E. and Cross, 1'. 1935 Third outbreak of paratyphoid dysentery in lamh.e Jo A111. Vet. Med. Asam e, NeSeVe 29-, 534—536. Posch, .T. I... Stofer, B. E., and Hirshfield, J’. W. 1944 Salmon- ella choleraesuis as a cause of indolent subcutaneous abscess (Human; J. Am. Vet. Med. Assoc., 104, 85. Rubin, H. I... Scherago, M., and Weaver, R. H. 1942 The occur- rence of Salmonella in the lymph glams of normal hogs. Am. J“. Hyg., _3_6_, 43-47. -18- Rubenstein, A. D., Feemster, R. E., and Smith, H. N. 1944 Salmonellosis as a public health problem in wartime. Am. J. Pub. Health, _34, 841. Schiff, F., and Saphra, I. 1941 A new Salmonella type: Salmonella havens. J. Infectious Diseases, _6_8_, 125-127. Seligmnn, E., Saphra, I. and Wassermnn, M. 1943 An analysis of 1,000 cases bacteriologically identified by the New York Salmonella Center. Am. J. Hyg., {_58, 226-247. Stone, W. S. 1943 Food handlers in the army and their rela- tionship to Salmonella food poisoning. Am. J. Pub. Health, §_3_, 706-708. Welch, S. 91., Dehler, S. A., and Havens, L. C. 1925 The prevalence of typhoid carriers in a general papulation. Je Ame MOde ASSOCe, _82, 1056-1039e Welch, H., Ostrolenk, M., and Bartram, M. T. 1941 Role of rats in the spread of food poisoning bacteria of tin Salmonella mUPe Anie «Te PUbe Health, a, 332-340e Wolff, A. H., 1947 The public health significance of animal Salmonella infections. J. Am. Vet. Med. Assoc., 111, 474-480. Wolff, A. H., Hendermn, N. D., McCullam, G. L. 1948 Salmonella from dogs and the possible relationship to Salmonellosis in man. Am. J. Pub. Health, _3_8_, 403-408. -19- IBRARIES lellilllll I 9 608 M'Tlrl‘l'filmflllrlllllillllzll “W W 3 119 3 0 3 01 2