Heterogenic colonization patterns by Leptospira interrogans in Rattus norvegicus from urban slums

Abstract We evaluated the renal colonization by Leptospira interrogans in Rattus norvegicus (rats), as it is the major natural reservoir of urban leptospirosis. We caught 72 R. norvegicus, out of which 32 were found to be positive for L. interrogans by immunofluorescence assay. From these rats, we selected 17 and divided them into six groups based on the mass-age/sex. We performed the immunohistochemistry test against L. interrogans in the kidney sections of the rats and systematically counted the colonized tubules (CTs) in 20 fields. The proportion of positive fields varied from 5% to 95%. The number of CTs in 20 fields varied from 0.5 to 85.5. These differences were not related to age or sex of the animals. The characterization of leptospiral colonization patterns in the natural reservoirs is important to better understand the host-pathogen interactions in leptospirosis.

Freqüência de aglutininas anti-Leptospira interrogans em eqüídeos, em Minas Gerais, 2003 a 2004 / Frequency of anti-Leptospira interrogans agglutinins in equidae in Minas Gerais, Brazil, from 2003 to 2004

The goal of this research was to find the frequency and spatial distribution of infection caused by Leptospira interrogans in equidae in Minas Gerais State from September 2003 to March 2004. Samples of blood serum (6,475) were analyzed by microscopic agglutination test. From the total, 381 samples were positive (5.9 percent), with title equal or superior to 1:200 for one or more serovars of leptospira. The most frequent serovars were Hardjo (Norma), Pomona, Bratislava, and Batavie. The higher frequency of equidae reagents were recorded at the North and Northeast region of Minas Gerais, followed by Triângulo Mineiro and Alto Paranaíba, then Central, West, Metropolitan area of Belo Horizonte, and South/Southwest.

Situación de la leptospirosis en el Urabá antioqueño colombiano: estudio seroepidemiológico y factores de riesgo en población general urbana / Leptospirosis in Uraba, Antioquia, Colombia: a seroepidemiological and risk factor survey in the urban population

Leptospirosis es una zoonosis de gran incidencia en regiones tropicales. Su prevalencia es desconocida en la región del Urabá colombiano. Entre marzo y octubre del año 2000 se realizó un estudio descriptivo de corte para determinar la seroprevalencia de anticuerpos contra Leptospira spp. y describir algunos factores de riesgo en nueve municipios del Urabá. La población incluida fue de 582 personas a las cuales se les tomó una muestra de sangre y se le aplicó una encuesta sobre factores de riesgo. La detección de anticuerpos contra Leptospira spp. fue realizada por inmunofluorescencia indirecta y por microaglutinación. La seroprevalencia general en la zona fue 12,5 por ciento (IC95 por ciento: 10,01-15,5). No hubo diferencias en cuanto al sexo, raza, oficio, edad, años de residencia en la zona y características de la vivienda. L. interrogans serovar Grippotyphosa fue la especie más prevalente, identificándose en 53 de los seropositivos. En 38 seropositivos los títulos detectados fueron iguales o mayores a 1:400. En conclusión, existe alta prevalencia de anticuerpos contra Leptospira spp. Es necesario orientar las medidas de control para disminuir el riesgo de exposición ambiental a leptospirosis por parte de los habitantes de la zona.

Leptospirosis is a widespread zoonosis in tropical regions. The prevalence is unknown in the Colombian region of Uraba. A cross sectional study was conducted from March to October 2000 in order to determine the prevalence of Leptospira spp. antibodies and describe risk factors in nine counties in the region. The sample consisted of 582 individuals, who answered a questionnaire and had blood samples drawn to determine risk factors. Detection of Leptospira spp. antibodies was based on indirect inmunofluorescence and microagglutination. Seroprevalence was 12.5 percent (95 percentCI: 10.01-15.5). No differences were observed according to race, gender, occupation, age, living conditions, or time of residence in the area. L .interrogans serovar Grippotyphosa was the most prevalent species, identified in 53 individuals. Titers were > 1:400 in 38 seropositive individuals. In conclusion, there is a high prevalence of Leptospira spp. antibodies in the area, where it is thus necessary to establish control measures to decrease the risk of environmental exposure to leptospirosis.

Leptospirosis: clinical presentation and correlation with serovars.

A total of 2400 patients with pyrexia of unknown origin and or suspected leptospirosis were included in this study. Dark field microscopy detected Leptospira in 690 cases, Leptospira serological Investigations proved positive in 570 out of these 690 patients. Among them 212 had the classical icteric and the other 358 had anicteric type of presentation. Notably eptospira interrogans serovar ictero haemorrhagiae infection was encountered in 212 patients. In 30 patients, who had multi organ dysfunction which included renal failure, hepatic dysfunction or meningitis was due to Leptospira interrogans Serovar cannicola. Coexsistense of leptospirosis and hepatitis B virus infection were noted in 15 patients. Antibody to Leptospira interrogans was demonstrated by Micro agglutination test (MAT) in addition to dark field microscopy positivity in these cases. Similarly HIV antibody was demonstrated in 30 of the 330 anicteric patients. 554 out of 570 cases responded to intra venous penicillin (216), and oral Doxycycline (182) and Augmentin (156), and the remaining 16 patients succumbed to death.

Molecular characterization of Leptospira sp. strains isolated from human subjects in São Paulo, Brazil using a polymerase chain reaction-based assay: a public health tool

A polymerase chain reaction (PCR)-based assay which amplifies repetitive DNA elements present within bacterial genomes was used to characterize and differentiate Leptospira sp. Thirty-five strains from a reference culture collection and 18 clinical isolates which had been previously analyzed by cross agglutinin absorption test (CAAT) were evaluated by this technique. PCR results from analysis of the reference culture collection showed no bands corresponding to serogroups Australis, Autumnalis, Bataviae, Celledoni, Cynopteri, Djasiman, Panama, Pomona, Pyrogenes, and Tarassovi. However, the PCR method was able to clearly discriminate the serogroups Andamana, Ballum, Canicola, Grippotyphosa, Hebdomadis, Icterohaemorrhagiae, Javanica, Sejroe, Semaranga, and Shermani. Clinical isolates previously characterized by CAAT as serovar Copenhageni, serovar Castellonis, and as serovar Canicola were in agreement with PCR results. The clinical isolate previously characterized as serovar Pomona was not differentiated by PCR. Forty additional clinical isolates from patients with leptospirosis obtained in São Paulo, Brazil were also evaluated by this PCR method. Thirty-nine of these were determined to belong to serogroup Icterohaemorrhagiae (97.5 percent) and one to serogroup Sejroe (2.5 percent). These results demonstrate that the PCR method described in this study has utility for rapid typing of Leptospira sp. at the serogroup level and can be used in epidemiological survey.

Presença de anticorpos da classe IgM de Leptospira interrogans em animais silvestres do Estado do Tocantins, 2002 / Presence of IgM antibodies for Leptospira interrogans in wild animals from Tocantins State, 2002

Quatrocentos e vinte e sete amostras de soro provenientes de animais silvestres foram testadas frente a 18 sorovariedades de Leptospira interrogans. De 286 amostras de Cebus apella, 46 (16,1 por cento) foram positivas para as sorovariedades pomona, brasiliensis, mini, swajizak, grippothyphosa, sarmin, fluminense, autumnalis, hebdomadis, guaratuba, javanica e icterohaemorhagiae. Das 82 de Alouatta caraya, 2 (2,4 por cento) foram positivas para as sorovariedades mangus e fluminense. Das 31 de Nasua nasua, 4 (12,9 por cento) foram positivas para as sorovariedades fluminense e javanica. Das 10 amostras de Cerdocyon thous, 2 (20 por cento) foram positivas para as sorovariedades fluminense e brasiliensis. Sete de Dasyprocta sp, 6 de Tamandua tetradactila e 5 de Euphractus sexcintus não apresentaram reatividade.

Medio EMJH modificado para el cultivo de Leptospira interrogans serogrupo Ballum / Modified EMJH medium for cultivation of Leptospira interrogans serogroup Ballum

El serogrupo Ballum agrupa cepas de crecimiento fastidioso, con requerimientos nutricionales más exigentes que otras cepas patógenas de Leptospira. Fue evaluada la influencia de 37 compuestos nutricionales sobre el crecimiento de Leptospira interrogans serogrupo Ballum, tomando como base para el estudio al medio sintético EMJH. El crecimiento microbiano fue estimado espectrofotométricamente y por conteo directo en cámara de Petroff-Hausser. La estabilidad de la virulencia fue evaluada en hamsters mediante el cálculo de la dosis letal media. La estabilidad de la antigenicidad fue evaluada mediante Western blotting con antisuero policlonal específico. Bajo condiciones de cultivo controladas se logró triplicar los rendimientos de biomasa comúnmente obtenidos en el medio EMJH sin afectación de la virulencia y antigenicidad tras el incremento de la concentración de Tween 80 y la incorporación de acetato de sodio y extracto de carne. El incremento de la concentración de al menos 6 componentes del EMJH o la incorporación de una variedad de nuevos nutrientes no estimularon apreciablemente los rendimientos de biomasa o la velocidad específica de crecimiento del microorganismo. Los resultados obtenidos permiten disponer de un medio de cultivo enriquecido capaz de sustentar elevados rendimientos de biomasa de este serogrupo exigente de mayor circulación en humanos en Cuba.

Strains within the Ballum serogroup of spirochete Leptospira show fastidious growth with more exigent nutritional requirements than those of other Leptospira pathogenic strains. The influence of 37 nutritional compounds on the growth of Leptospira interrogans serogroup Ballum was investigated employing the synthetic EMJH medium as the base for the study. Microbial growth was estimated spectrophotometrically and direct counts were performed with a Petroff-Hausser counting chamber. Virulence stability was evaluated by calculating the mean lethal dose in hamsters. Antigenicity stability was evaluated by Western blotting using a specific antiserum. Cell yields commonly obtained in EMJH were triplicated without virulence or antigenicity depletions after culturing in a modified EMJH medium with an increased concentration of Tween 80, and the incorporation of sodium acetate and beef extract. Neither the increased concentration of at least 6 components of EMJH nor the incorporation of a variety of new nutrients stimulated cell yields or the growth rate of the microorganism. The results allow us to make use of an enriched culture medium that promotes high cell yields of this fastidious serogroup most prevalent in humans in Cuba.

Comparison of leptospiral serovars identification by serology and cultivation in northeastern region, Thailand.

Blood from patients suspected of leptospirosis 148 specimens were cultured for leptospira. Twenty two specimens were positive (15%). The isolated leptospira were tested against the 24 serovars of standard antisera by Microscopic Agglutination Test (MAT). It was found that all 22 leptospira isolates reacted strongly against L. autumnalis, except 1 isolate that also reacted against serovar djasiman. The patient's sera were collected from only 14 cases. When the sera of the 14 patients were tested with the 24 reference serovars by MAT it was found that sera reacted the most against L. australis and in decreasing order against L. bratislava, L. autumnalis, L. rachmati, L. copenhageni, L. javanica. There had some cross reactions against several serovars in a single patient. The present study showed inconsistency between culture results and serum assays. Since sera showed cross reactivities against several serovars, it was not possible to determine which serovar was etiologic. Therefore, the isolation of leptospira though time consuming is specific in the identification of the serovar.

Evidência do cão como reservatório da leptospirose humana: isolamento de um sorovar, caracterização molecular e utilização em inquérito sorológico / Evidence of dog as a reservoir for human leptospirosis: a serovar isolation, molecular characterization and its use in a serological survey

A leptospirose canina é conhecida como enfermidade de Stuttgard desde 1898, sendo os cães, depois dos roedores, considerados como a segunda principal fonte de infecção para o homem. O isolamento de um sorovar patogênico da urina de um cão, laboratorial e clinicamente identificado como tendo leptospirose, e sua utilização para testar amostras de soro de casos de leptospirose humana e canina, evidenciou a sua importância no ecossistema da região sul do Brasil. Os resultados do teste de soroaglutinação microscópica indicaram que 100 por cento das amostras de soro humano de 12 pacientes do banco de soro de 2001 do Centro de Controle de Zoonoses, que haviam reagido com títulos que variaram de 25 a 3.200 para o sorovar canicola, e 72 por cento das amostras de 105 soros caninos do mesmo banco de soro, também reagiram contra o novo isolado. O título médio e mediana dos soros humanos testados com a bateria de antígenos recomendada pela OMS, foi respectivamente 630 e 100, ao passo que os testados com o isolado foi de 1.823 e 400. Nos soros caninos, os títulos foram respectivamente de 347 e 100 para a bateria e de 1.088 e 200 para o isolado.

Common infecting leptospiral serovars in and around Pune, Maharashtra.

Leptospirosis is a disease with protean manifestations. The present study was conducted in Pune to examine the possibility of leptospiral infection among a group of patients with fever of undetermined origin and to identify the common infecting serovars. Serological evidence of leptospirosis was found in 22 of the 118 (18.6%) patients with the help of microagglutination test (MAT) using a battery of 9 antigens. The serovars responsible for infection included autumnalis in eight cases, copenhageni in six, pomona in three, grippotyphosa in two and australis, batavia and canicola in one case each. Thus, there appears to be a focus of leptospirosis in and around Pune with autumnalis and copenhagni as the common infecting serovars.

Leptospirosis in Barbados: a clinical study

A 39-month clinical study of leptospirosis was undertaken at the Queen Elizabeth Hospital, Barbados, Eighty-eight patients had a confirmed diagnosis of the disease during the period. The major serogroups identified were autumnalis (including a new serovar bim), icterohaemorrhagiae, ballum and canicola. The majority of patients presented with jaundice (95%,) anorexia and headaches (85%), fever (76%) and conjunctival suffusion (54%). While abnormal creatinine levels were seen in 49% of patients on admission, only 16% were judged to have had renal failure. The urine to plasma urea ratio showed high sensitivity and specificity in the diagnosis of pre-renal azotemia. Cardiac arrhythmias and myocarditis occurred in 18% of patients and pericarditis in 6%. An elevated serum amylase was found in 65% of cases. The bilirubin level took 5.5 weeks to return to normal. Thrombocytopenia was shown not to be due to a disseminated intravascular coagulation, and a randomised trial of high dose penicillin did not reveal any benefit to jaundiced patients. The overall mortality during the study was 5.7%

Characterization of trypsin extractable antigens of Leptospira interrogans serovars pomona, bataviae & L. biflexa serovar patoc.

Leptospira strains after treatment with trypsin released an antigen (Tx) which consisted of protein (4.2 mg/ml), carbohydrate (0.39 mg/ml) and hexosamine (0.025 mg/ml). Immunodiffusion, immunoelectrophoresis and indirect haemagglutination tests revealed serological cross reactions among the three strains used. Inhibition of microscopic agglutination reaction by the Tx antigen was observed in homologous system only. The antigen was partially sensitive to heat (80 degrees C for 10 min) as detected by immunodiffusion and microscopic agglutination inhibition. These data revealed the presence of serovar-specific and genus-specific antigens on Leptospira strains. It is suggested that this antigen could be useful in developing a serodiagnostic test for leptospirosis.