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Complement Fixation Tests for Murine Typhus

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Complement Fixation Tests for Murine Typhus ice under glass seal. The animals' brains were Complement Fixation Tests removed for tissue infectivity tests and were also stored on dry ice. Tests of the tissues of For Murine Typhus serologically reactive animals and pools of their fleas aided in evaluating the significance of low On Small Mammals titers. Blood samples were obtained by cardiac By RUTH KEATON, B.S. puncture soon after capture of the animals. BILLIE JO NASH, B.A. Serums were separated aseptically and were J. N. MURPHY, JR., M.A., M.S.P.H. kept at ice-box temperature until examination. J. V. IRONS, Sc.D. Serums were inactivated 30 minutes immedi- ately before testing at 56° C. Each serum was subjected to a quantitative complement tT HE IMPORTANCE of commensal rats fixation test employing endemic typhus rick- and their fleas in the epidemiology of en- ettsiae. A slight modification of the pro- demic typhus was well establislhed in 1931 (1,2). cedure described by Brigham and Bensgton Following Dyer's report (3) on the experi- (11) was used. The result was recorded as mental infection of the woodchuck, meadow reactive when a 3 + or greater reaction was ob- mouse, and whitefooted mouse with endemic tained with satisfactory controls at a 1: 20 or tvphus, Brigham (4, 5) indicated that many greater dilution of serum. species of rodents and other mammals were ap- When a serum was reactive, the correspond- parently susceptible to endemic typhus. Spar- ing brain suspension was emulsified and inocu- row (6) recovered a strain of endemic typhus lated individually into hamsters for evidence of rickettsiae in the house mouse (.Mus musodus), typhus. With negative serologic findings, the as did Brigham (7) with his experiments on a brains of animals of the same species, particu- field mouse (Peromryscus sp.). larly from the same trapping area, were simi- In view of the wide distribution of endemic larly tested in pools of varying number. Pools typhus in commensal rats in Lavaca County of fleas from the same host species taken in the reported by Irons and associates (8), inquiry same area were similarly tested. Each hamster into the possible occurrence of infection in other was bled twice, both before inoculation and 3 small mammals seemed worth while. Morlan or 4 weeks later, and the serums were run in the and co-workers (9) used the complement fixa- quantitative complement fixation test. With tion test as an indication of natural infection, positive findings, titers tended to be high on the but Rickard and Worth (10) considered the second bleeding. This was the procedure recommended by Plotz, Wertman, and Bennett findings on the wild-caught cotton rat (Sig- inodon hispidus) to be nonspecific. (12) for identifying rickettsial agents isolated in guinea pigs or mice. The utilization of specific complement fixation tests is much more Methods economical than cross immunity and other tests Live animals were captured in 1945 and 1946 in the guinea pig for identifying endemic near Hallettsville, in southeast Texas. Gen- typhus rickettsiae. A summary of findings is shown in the table. erally, the ectoparasites were identified while alive. The ectoparasites to be tested for typhus The brain tissues of 3 of 17 house mice (Mm musculus) taken from 3 of 44 places surveyed were appropriately pooled and stored on dry yielded typhus rickettsiae. Most of the house mice were trapped on premises in close associa- Msi.s Keaton and Miss Nash are bacterzologists; tion with commensal rats. Curiously enough, Mr. Murphy i associate director, and Dr. Iromn those trapped in the fields were uniformly ncaa- is director of the bureau of laboratories, Tewas tive. Native rats and mice were found strictly State Department of Health. in the fields. Tests of the brain tissues from the cotton rat (S'igmodon hispidu8), the pack rat 28 Public Health Reports Results of complement fxation tests for typhus fever on animal serums Titer percentage Total Percent- _ Mammal species number age re- tested active 1:20 1:40 Above 1:40 - 9 Didelphis virginiana (opossum) _-_-__--27 6. 9 6.9 - |- --- Neotoma floridana (pack rat) -14 14. 2 7. 1 7. 1 Baiomys taylori (field mouse) -101 0 _ _ Signodon hispidus (cotton rat) -62 8. 0 8. 0 Spilogale indianola (civet cat) -2 0 _ -_ lMfus musculus (house mouse)- -_-_-_-_- _- _-_ 216 7. 8 . 9 4. 6 2. 3 Perognathus hispidus (pocket mouse) -3 0 Peromyscus leucopus (white-footed mouse) -5 0 Geomys breviceps (gopher) -28 0 Procyon lotor (raccoon)- 13 0 Reithrodontomys fulvescens (harvest mouse) - 9 0 ------------------------------ (Neotoma foridana), and the opossum (Didel- other pools of fleas collected from nonmurine phis virginiana) were uniformly negative. hosts (13, 14) was perhaps fortuitous, as it is A pool of 20 fleas (Ctenocephalide8 felis), possible that the fleas had acquired the infection collected from two opossums at Hallettsville from rats. At any rate, these findings do not in in the summer of 1946 and not previously re- any way detract from the primary role of com- ported, gave an unequivocal positive test for mensal rats in the epidemiology of endemic typhus. These opossums were trapped in an typhus fever. oil mill which had not been dusted with DDT. The opossums showed negative complement fix- Summary ation tests. Eight pools of fleas collected from negative rats and mice and tested for harborage Endemic typhus rickettsiae were recovered of typhus rickettsiae gave negative results. from brains of three house mice taken from human habitations and from a pool of 20 fleas Discussion collected from two opossums. House mice trapped in fields were uniformly Although occasional serums from cotton rats, negative. wood rats, and from an opossum were reactive Unequivocal evidence of typhus in small in the complement fixation test for endemic mammals other than commensal rats and mice typhus, the titers were relatively low. How- was not obtained. ever, similar findings on commensal rats prob- on the cotton ably would have been considered as evidence of Serologic findings pack rat, rat, infection or immunity to typhus. Low titers and opossum were of doubtful significance. and failure to demonstrate the infectivity of brain tissues of cotton rats, pack rats, or opos- ACKNOWLEDGMENT sums cast doubt on the specificity of serologic findings on these animals. The percentage of The Lederle Laboratories, Pearl River, N. Y., sup- house mice showing positive serologic findings plied the endemic typhus antigen used in the study. was also relatively low, but the titers were not invariably low, and endemic typhus rickettsiae REFERENCES were obtained from the brains of three house (1) Dyer, R. E., Rumreich, A., and Badger, L. F.: A mice. The relatively few places harboring house virus of the typhus type derived from fleas col- mice with evidence of past infection contrasted lected from wild rats. Pub. Health Rep. 46: sharply with findings for the domestic rat (8). 334-338 (1931). (2) Mooser, H., Castaneda, M. Ruiz, and Zinsser, The finding of typhus rickettsiae in a pool of Hans: Rats as carriers of Mexican typhus fleas collected from the two opossums, and in fever. J. A. M. A. 97: 231-232 (1931). Vol. 68, No. 1, January 1953 29 232500-53 43 (3) Dyer, R. E.: Endemic typhus fever: Susceptibility (10) Rickard, E. R., and Worth, C. B.: Complement of woodchucks, house mice, meadow mice, and fixation tests for murine typhus on sera of wild- white-footed mice. Pub. Health Rep. 49: caught cotton rats in Florida. Am. J. Hyg. 723-724 (1934). 53: 332-336 (1951). (4) Brigham, G. D.: Susceptibility of the opossum (11) Brigham, G. D., and Bensgton, I. A.: A study of (Didelphis virginiana) to the endemic typhus. the complement flxation and Weil-Felix reac- Pub. Health Rep. 51: 332-337 (1936). tions in wild rats as related to the isolation of (5) Brigham, G. D.: Susceptibility of animals to the virus of endemic typhus. Pub. Health Rep. endemic typhus fever. Pub. Health Rep. 60: 29-46 (1945). 52: 660-62 (1937). (12) Plotz, H., Wertman, K., and Bennett, B. L.: (6) Sparrow, H.: Enqufte sur la presence du virus Identification of rickettsial agents Isolated in typhique chez les souris de Tunis. Arch. Inst. guinea pigs by means of specific complement Pasteur de Tunis 24: 435-460 (1935). fixation. Proc. Soc. Exper. Biol. & Med. 61: (7) Brigham, G. D.: A strain of endemic typhus fever 76-81 (1946). a field mouse. Pub. Health Rep. isolated from (13) Irons, J. V., Bohls, S. W., Thurman, D. C., Jr., 52: 659-660 (1937). and McGregor, T.: Probable role of the cat (8) Irons, J. V., Murphy, J. N., Jr., and David, D. E.: The distribution of endemic typhus in rats in flea, Ctenooephalides felis, in transmission of Lavaca County, Texas. Pub. Health Rep. 63: murine typhus. Am. J. Trop. Med. 24: 359- 692-694 (1948). 362 (1944). (9) Morlan, H. B., Hill, E. L., and Schubert, J. H.: (14) Irons, J. V., Beck, Oleto, and Murphy, J. N., Jr.: Serological survey for murine typhus infection Fleas carrying endemic typhus rickettsiae in southwest Georgia animals. Pub. Health found on nonmurine hosts. Am. J. Bact. 51: Rep. 65: 57-63 (1950). 609-610 (1946). Public Health Service Staff Announcements A new post has been created in the Bu- May 1948. Before that he was for 5 years reau of State Services of the Public Health the venereal disease control officer of the Service to give over-all direction to field Chicago Board of Health and medical offi- research. Dr. Justin M. Andrews will fill cer in charge of the Chicago Intensive the post as an Assistant Surgeon General Treatment Center for venereal disease.
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