Lima, C. Andrade

Magalhães, Marcelo

Rosa, C. A. Santa

Sulzer, Catherine R.

English

Cadernos Espinosanos (E-Journal)

Rev. Inst. Med. trop. São Paulo 15( 1) :38-42, janeiro-fevereiro, 1973 UDC 616. 986. 71 HUMAN LEPTOSPIROSIS CAUSED BY SEROTYPE ALEXI IN BRAZIL e. A. SANTA RosA (1), Marcelo MAGALHÃES (2 ), Catherine R. SuLZER (3 ) and C. Andrade LIMA ( 4) SUMMARY The results · of a serologic study of a patient serum and the isolation of leptospira . from hemo culture are reported. Cross-agglutination and agglutinin absorption tests revealed the isolated strain to be identical to serotype alexi. This is the first known isolation of this serotype in Latin America. This case was a very mild atypical case of leptospirosis. INTRODUCTION The first isolations of the members of the pifrogenes group in the W estern Hemisphere were made by YAGER & GocHENOUR 15 in 1952. Until a few years ago only serotype ictero-haemorrhagiae had been incriminated in hu-man leptospirosis in Bra'.fil. The importance of leptospirosis has been ~ore widely recogniz-ed in recent years in this country since other serotypes such as wolffi 5 and andamana 6 have been isolated from clinicai cases. Canicola 3 and grippotyphosa 4 have been incrimmated recently as the cause of human leptospirosis in the Bra~ilian area. Also sero-logic studies on humans indicated the pre-sence of a multiplicity of leptospiral serotypes, mainly in the State of São Paulo 2 • 7 , 8 , 9 , 11, 12, 13, 14 • The results of a 1968 serologic study of a patient serum and. the isolation and identifi-cation of leptospira from his blood are report-ed in this paper. MA TERIALS AND METHODS ln 1968, an adult male arrived from the hinterland of Brazil looking for a doctor in a private clinic in Recife, State of Pernambu-co, Brazil. His compli:tints were severe headache ( there was no evidente of meningeal irritation}, ele-vated temperature, abdominal pains, and a rash on the abdomen. Typhus fever was sus-pected at first. Stool and hemo cultures for Salmonellosis were carried out and blood was inoculated into tubes of Fletcher's semi-solid medium as a check for leptospires. The patient's serum was also tested for typhus fever and leptospi-rosis. The microscopic agglutination test em-ploying live antigens was the basic serological technique used with a selected battery of 22 different serotypes (Table I). The -serum di-lution titer was defined to be the highest di-lution in which 50 percent or more of the cells were agglutinated. The method of preparing antisera in rabbits and the; conduct of agglutinin absorption tests (1) Departamento de Microbiologia e Imunologia do Instituto de Ciências Biomédicas da U.S.P. (Av. Dr. Arnaldo, 455 - São Paulo) {2) Departamento de Microbiologia e Parasitologia da Universidade Federal de Pernambuco (3) Bacterial Immunology Unit, Center for Disease Control, Health Services and Mental Health Admi-nistration, Public Health Service, U. S. Department of Health, Education, and Welfare, Atlanta, Georgia, U.S.A. {4) Departamento de Clinica Médica da Universidade Federal de Pernambuco, Brasil 38 SANTA ROSA, C. A.; MAGALHÃES, M.; SULZER, C. R. & LIMA, C. A. - Human l-eptospirosis caused by serotype alexi in Brazil. Rev. Inst. Med. trop. São Paulo 15:38-42, 1973. TABLE r Serotypes used as antigens in microagglutination test Serotype 1 - icterohaemorrhagia-e 2 - canicola 3 - pomona 4 - grippotyphosa 5 - taras,sovi 6 - sejroe 7 - australis 8 - bataviae 9 -mini 10 - ballum 11 - wolffi 12 - panama 13 - pyrogenes 14 - javanica 15 - autumnalis 16 - butembo 17 - celledoni 18 - poi 19 - andamana 20 - saxkoebing 21 - brasiliensi's 22 - patoc are described in detail elsewhere 10• Antigen and antisera from recognized serotypes were used in the cross-agglutination and aggluti-nins absorption tests. The isolated strain was designated as LT 22-69, it was tested preliminarly by tbe micros-copie agglutination test and then by aggluti-nin-absorption tests. The illness lasted only one week. There was no jaundice and the patient recovered with no specific therapy. Strain RGA Hond Utrecht IV Pomona Moskwa V Perep•elicin M 84 Ballico Van Tienen Sari Mus 127 3705 CZ 214 K Salinem Veldrat Bat. 46 AKyiami A Butembo Celledoni Poi CH 11 Mus 24 LT 966 Patoc I RESULTS Stool and hemo cultures were negative for salmonellae, as were the serological results from the Widal test. Leptospires were seen in Fletcher's semi-solid medium five days after inoculation. Serologic studies for leptospirosis revealed agglutinating antibodies against serotypes ta-rassovi 1 :3200, pyrogenes 1: 1600, pai 1 :400 and saxkoebing 1 :200. ,39 SANTA ROSA C. A.; MAGALHÃES, M.; SULZER, C. R. & LIMA, C. A. - Human }eptospirosis caused by seroty;e alexi in Brazil. Rev. Inst. M ed. trop. São Paulo 15: 38-42, 1973. Results of preliminary tests (Table II) ir~di-cate that the isolated serotype was serologi-cally identical with serotype a/,exi. This was later confirmed by the agglutinin-absorption test performed at the Center for Disease Con-trol in Atlanta, Georgia, U. S. A. (Table III). 40 TABLE II Cross-agglutination reactions of leptospiral strain LT 22-69 with antisera against 30 Jeptospiral serotypes and pyrogenes serogroup Titer against LT 22-69 antisera Antiserum Homologous antigen 22-69 against antigens in column 1 ballum 6,400 - -canicola 12,800 - -copenhageni 25,600 - -bataviae 25,600 - -grippotyphosa 25,600 - -autumnalis 51,200 - -pomona 25,600 - -wolffi 12,800 - -australis 6,400 - -tarassovi 6,400 - -georgia 12,800 - -javanica 6,400 - -celledoni 12,800 - -fort-bragg 12,800 - -sentot 25,600 - -djasiman 12,800 - -borincana 12,800 - -cynopteri 12,800 - -Pyrogenes serogroup pyrogenes 25,600 25,600 800 -~--··-zanoni 25,600 400 ~ . .-~----400 abramis 12,800 1.600 3,200 biggis 12,800 25,600 400 hamptoni 12,800 1,600 800 alexi 12,800 25,600 6,400 robinsoni 6,400 1,600 400 manilae 6,400 - 200 myocastoris 6,400 100 800 varela 12,800 - 400 cam-Jo 25,600 6,400 6,400 guara tuba 25,600 800 400 (-) Negative at 1:100 final dilution SANTA ROSA, C. A.; MAGALHÃES, M.; SULZER, C. R. & LIMA, C. A. - Human 1-eptospirosis caused by serotype alex~ in Brazil. Rev. Inst. Med. trop. São Paulo 15:38-42, 1973. TABLE III Results of cross agglutinin-absorption t-ests on leptospiral strain LT 22-69 with members of the Pyrogenes serogrour:> Antiserum Reciprocal of tit-er against antigen Homo!ogous Absorbing strain 1 Serotype Absorbed with Before After Before After pyrogenes 22-69 25,600 12,800 25,600 100 abramis 22-69 12,800 51,200 800 -hamptoni 22-69 25,600 12,800 800 -robinsoni 22-69 3,200 3,200 1,600 -biggis 22-69 12,800 25,600 6,400 -alexi 22-69 25,600 800 25,600 100 cam-lo 22-69 25,600 25,600 12,800 -guara tuba 22-69 3,200 3,200 800 -LT 22-69 pyrogenes 3,200 1,600 800 -LT 22-69 abramis 3,200 1,600 1,600 200 LT 22-69 hamptoni 3,200 1,600 6,400 200 LT 22-69 alexi 12,800 - 12,800 -LT 22-69 myocastoris 3,200 1,600 800 -LT 22-69 cam-lo 3,200 1,600 6,400 200 LT 22-69 guara tuba 3,200 3,200 400 200 LT 22-69 zanoni 3,200 3,200 400 -LT 22-69 LT 22-69 1 3,200 o 100 ') (-) Negative at 1:100 dilution (final dilution) (O) Was not done (not needed) DISCUSSION This case was a very mild atypical case of leptospirosis caused by serotype alexi; this is the first known isolation of this serotype in Latin America. The dinical features of human leptospirosis vary considerably; the illness can be severe as fulminating hepatitis and as mild as in-fluenza. The classic manifestations that sug-gest W eil's disease are fever, severe prostra-tion, icterus, renal decompensation, hemorrha-gic phenomena, and vascular collapse. All these are found in Brazilian leptospirosis. Milder forms of leptospirosis are more diffi-cult to recognize, but a leptospiral etiology should be considered in any febrile patient when severe headaches, vomiting, myalgia, conjuntiva! injection, proteinuria and neutro-philia are noted. Leptospirosis is to be sus-pected even more whenever there is a history of exposure to those environmental conditions that favor infection. Leptospirosis can easily be mistaken for typhoid fever, acute sinusitis, and influenza among other · infections. However, there are some clinica! manifestations in the early phase that should suggest leptospirosis, namely, an explosive onset with chills, a rapidly rising fever, severe conjuntivitis, and marked myal-gia accompanied by nausea and vomiting. The illnesses attributed to infection by serotype alexi can range from mild to severe 1. This case also exemplifies the value of blood culture in the laboratory diagnosis of leptos-pirosis. When the patient was first seen, ty-phoid fever was considered a diagnostic possi-bility. Stool specimens were cultured for ente-ric pathogens and blood specimens were cul-tured in Fletcher's medium to check for lep-tospires. Five days after the blood culture was made leptospires were found. The immunologic response of the patient is very interesting in that high serum titers were obtained against serotype tarassovi 1 :3200 and 41 SANTA ROSA, C. A.; MAGALHÃES, M.; SULZER, C. R. & LIMA, C. A. - Human leptospirosis caused by serotype alexi in Brazil. Rev. Jnst. Med. trop. São Paulo 15 :38-42, 1973. pyrogenes 1: 1600 with cross-agglutination with poi and saxkoebing. . The serological results also corroborated the Cultural resuit-RESUMO Leptospirose humana causada peÚJ sorotipo alexi, em Recife Os Autores relatam um caso de leptospirose humana, benigna e atípica. Através de hemo-cultura, em meio de Fletcher, foi isolada uma amostra de leptospira. Pela tipagem sorológi-ca completa (testes de aglutinação cruzada e absorção de aglutininas) a amostra foi das-sificada como sorotipo alexi. Com o soro do paciente foi feito o tes.te de aglutinação mi-croscópica obtendo-se o seguinte resultado: tarassovi 1 :3200, pyrogenes 1: 1600, poi 1 :400 e saxkoebing 1 :200. Este é o primeiro isola-mento do sorotipo alexi na América Latina. 1. 2. 3. REFERENCES ALEXANDER, A. D.; BENENSON, A. S.; BYRNE, R. J.; DIAZ RIVERA, R. S.; EVANS, L. B.; GOCHENOUR, W. S.; HALL, H. E.; HIGHTOWER, J. A.; JEFFRIES, Helen; de JESUS, J.; MARTINEZ, E.; PANIAGUA, M.; PONS, J. A.; RAMOS-MORALES, F.; RODRI-GUEZ-MOLINA, R.; SWISHER, K. Y.; WOO-DWARD Jr., T. E. & YAGER, R. H. - Lep-tospirosis in Puerto Rico. Zoonos. Res. 2:125-227, 1963. CORRÊA, M. O. A. - Leptospiroses em São Paulo. Rev. Inst. Adolfo Lutz 29/30:29-37, 1969/1970. CORRÊA, M. O. A.; AMATO NETO, V.; PE-DRO, R. de J.; KONICHI, S. R. & FLEURY, G. C. - Considerações sobre caso humano de leptospirose canicola, com isolamento e iden-tificação do agente etiológico, pela primeira vez no Brasil. Rev. Soe. Brasil. Med. Trop. 5: 55-58, 1971. 4. CORRÊA, M. O. A. - Comunicação pessoal. 5. CORRÊA, M. O. A.; HYAKUTAKE, S.; NA-TALE, V.; GALVÃO, P. A. A. & AGUIAR, H. C. - .Estudos sobre a Leptospira wolffi em São Paulo. Rev. Inst. Adolfo Littz 25;27 :11-26, 1965/1967. 42 6. CORRÊA, M. O. A.; HYAKUTAKE, S.; NA-TALE, V.; TIRIBA, A. C.; MARTIRANI, I.; GALVÃO, P. A. ,A.; ALBANO, A.; FILIPPI, J. de; FARHAT, C. K. & AMATO NETO, v. - Leptospirose humana por Leptospira an-damana. Rev. Inst. Med. trop. São Paulo 13: 137-143, 1971. 7. COSTA, E. A.; REZENDE, M. & LINS, Z. -Leptospiroses no Estado do Pará e Territó-rio Federal do Amapá. Rev. Inst. Adolfo Lutz 29/30::J.-4, 1969/1970. S. COSTA, E. A.; CORRÊA, M. O. A.; NATALE, V. & SADATSUNE, T. - Leptospirose com soro-aglutinação positiva para Leptospira ja-vanica em Boca do Acre, Amazonas. Rev. Inst. Adolfo Lutz 29/30:13-18, 1969/1970. 9. EDELWEISS, E. L. - Leptospiroses no Rio Grande do Sul. Rev. Inst. Adolfo Lutz 29/30: 5-11, 1969/1970. 10. GALTON, M. M.; MENGES, R. W.; SHOTS, E. B.; NAHAMIAS, A. J. & HEAT, C. W. -Leptospirosis: epidemiology, clinica] manifes-tations in man and animals, and methods in laboratory diagnosis. Public Health Serv. Publ., n.o 951. United states Government Printing Office. Washington, 1962. Jl. 12. 1 "' "· MAGALDI, C. - Contribuição à Epidemiolo-gia das Leptospiroses. Investigação em Tra-balhadores da Rede de Esgotos da Cidade de São Paulo. Tese. São Paulo, 1962. MEIRA, D. A. & SANTA ROSA, C. A. - Es-tudo epidemiológico e clinico da leptospirose em detentos de São Paulo. Rev. Hosp. Clin. Fac. Med. Univ. São Pau,lo 19 :163-172, 1964. PESTANA DE CASTRO, A. F.; SANTA RO-SA, e. A.; ALMEIDA, w. F. de & TROISE, C. - Pesquisa de Aglutininas anti-leptospira entre magarefes em alguns municípios do Estado de São Paulo. Rev. Inst. Med. trop. São Paulo 8:287-290, 1966. 14. SANTA ROSA, C. A.; COSCINA, A. L.; CAS-TRO, A. F. P.; SILVA, A. S. da & QUEIROZ, J. c. - Pesquisa de aglutininas anti-leptos-pira em soros de trabalhadores de ·diversas profissões. Rev. Microbial. 1: 19-24, 1970. 15. YAGER, R. H. & GOCHENOUR Jr., W. S. -Leptospirosis in North America. Amer. J. Trop. Meà. & Hyg. 1:457-461, 1952. Recebido para publicação em 17/7/1972. 

Human leptospirosis caused by serotype alexi in Brazil

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Human leptospirosis caused by serotype alexi in Brazil

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