Isolation of Leptospira spp from dogs, bovine and swine naturally infected

Freitas, Júlio César de; Silva, Francielle Gibson da; Oliveira, Renata Cristina Gobbi de; Delbem, Alexandre Cláudio Botazzo; Müller, Ernst Eckehardt; Alves, Lucimara Aparecida; Teles, Paulo Sérgio

doi:10.1590/s0103-84782004000300030

Cited by 35 publications

(31 citation statements)

References 7 publications

(12 reference statements)

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“…The antibiotics, at the concentrations used in the EMJH medium, were found to be able in inhibiting contaminants but not leptospires. Myers and Varela-Diaz (12) found that subcultures were needed further on into an antibiotic containing medium, for total elimination of surviving contaminants after 24h, differently from what was described by Freitas et al, Turner, and Schönberg et al (6,16,19), that sub cultured after two days.…”

Section: Resultsmentioning

confidence: 94%

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EMJH medium with 5-fluorouracil and nalidixic acid associated with serial dilution technique used to recover Leptospira spp from experimentally contaminated bovine semen

Miraglia¹,

Moraes²,

Melville³

et al. 2009

Braz. J. Microbiol.

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Bovine semen experimentally contaminated with Leptospira santarosai serovar Guaricura was submitted to the modified EMJH medium with 5-fluorouracil (300mg/L) and nalidixic acid (20mg/L), named as "selective medium" and using the serial dilution technique, in order to evaluate the percentage of recovery of the added microorganism. The selective EMJH medium was found with higher percentage of recovery of leptospiras and minor losses of samples due to contamination with opportunistic microorganisms than the non-selective EMJH medium: 151/376 (40.0%) of positive growth; and 38/376 (10.0%) contamination and 58/376 (15%) and 129/376 (34.0%), respectively. These results were statistically significant (p<0. 0001; Fisher). Differences were found when the frequencies of positive leptospires recovery have been compared in the serial dilution technique (10 -1 to 10 -4 ) between the selective and non-selective media at different dilution factors. At 1/10 th dilution the percentages found were (0%, 0/80) and (38%, 30/80), at 1/100 th dilution, (3%, 2/80) and (49%, 39/ 80) and at 1/1,000 th dilution, (25%, 20/80) and (50%, 40/80), respectively. The percentage of recovery of leptospires was found to be directly proportional to the dilution used. The methodology of the serial dilution technique (setting at least three dilutions) and the use of selective EMJH medium have been found to be efficient for the isolation of leptospires from the bovine semen samples.

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Section: Resultsmentioning

confidence: 94%

“…Freitas et al (6) described the leptospires isolation of dogs, bovine and swine naturally infected by leptospires. The urine samples from 14 dogs and three bovines and also kidney, liver, ovary and uterus body samples from 36 slaughtered sows with unknown health records were used.…”

Section: Introductionmentioning

confidence: 99%

EMJH medium with 5-fluorouracil and nalidixic acid associated with serial dilution technique used to recover Leptospira spp from experimentally contaminated bovine semen

Miraglia¹,

Moraes²,

Melville³

et al. 2009

Braz. J. Microbiol.

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“…De fato, como o procedimento de tipificação das estirpes isoladas empregado só identificou a espécie das quatro estirpes e o sorogrupo de duas, não foram identificados os sorovares das mesmas, sendo necessário o emprego dos procedimentos clás-sicos para a identificação dos antígenos de superfície de natureza lipopolissacarídica, tais como a prova de absorção cruzada de aglutininas ou conjuntos de anticorpos monoclonais como já foi realizado em diversas localidades do Brasil, identificando em algumas ocasiões sorovares autóctones, só registrados até o presente unicamente no Brasil (SANTA ROSA; SULZER; CASTRO, 1972;VASCONCELLOS et al, 2001;DEL-BEM et al, 2002;MARVULO et al, 2002;CORRÊA et al, 2004;FREITAS et al, 2004;OLIVEIRA et al, 2005;AHMED et al, 2006;FAGUNDES et al, 2008;MIRAGLIA et al, 2008;ZACARIAS et al, 2008).…”

Section: Metodologiaunclassified

“…infectando animais no Brasil, Santa Rosa, Sulzer e Castro (1972) (AHMED et al, 2006;FAGUNDES et al, 2008). Freitas et al (2004) isolaram a estirpe LO-1 da urina de um cão da cidade de Londrina, PR, tipificada como sorogrupo Canicola sorovariedade Canicola (OLIVEIRA et al, 2005;AHMED et al, 2006;FAGUNDES et al, 2008). Delbem et al (2002) isolaram estirpe L04 do fígado de um suíno, tipificada como pertencente ao sorogrupo Canicola sorovariedade Canicola (FREITAS et al, 2004;FAGUNDES et al, 2008).…”

Section: Introductionunclassified

“…Freitas et al (2004) isolaram a estirpe LO-1 da urina de um cão da cidade de Londrina, PR, tipificada como sorogrupo Canicola sorovariedade Canicola (OLIVEIRA et al, 2005;AHMED et al, 2006;FAGUNDES et al, 2008). Delbem et al (2002) isolaram estirpe L04 do fígado de um suíno, tipificada como pertencente ao sorogrupo Canicola sorovariedade Canicola (FREITAS et al, 2004;FAGUNDES et al, 2008). Zacarias et al (2008) isolaram a estirpe L014 da urina de um bovino abatido no estado do Paraná, tipificada como pertencente ao sorogrupo Canicola sorovariedade Canicola (AHMED et al, 2006).…”

Section: Introductionunclassified

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Pesquisa de leptospiras e de anticorpos contra leptospiras em animais e humanos de propriedades rurais nos biomas brasileiros Pantanal e Caatinga

Silva

Santos

Silva

et al. 2015

Braz. J. Vet. Res. Anim. Sci.

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The occurrence of Leptospira and of seroreactivity against Leptospira was investigated in animals and humans from six farms located in two Brazilian biomes that have different geoclimatic conditions: Pantanal -municipalities of Miranda (MS), Itiquira (MT) and Poconé (MT) and Caatinga -municipalities of Sobradinho (BA), Garanhuns (PE) and Sobral (BA). Blood and urine samples of wildlife, domestic animals and humans were collected at each property. The samples were collected from February to April 2012 in Caatinga and from July to September 2012 in Pantanal. The serological reactivity against Leptospira spp. was verified by microscopic agglutination technique (MAT) made with a collection consisting by 24 antigens of Leptospira spp. The leptospires research was carried out by urine samples crop sown in Fletcher resources and Ellinghausen -McCullough -Johnson -Harris (EMJH). Crops with growth of leptospires were referred to the Leptospirosis Laboratory of the Institute of Pathobiology, National Institute of Agricultural Technology, Buenos Aires, Argentina and isolated Leptospira strains were genotyped with the technique of Multiple Locus Variable Number Tandem Repeat Analysis (MLVA). The classification procedure employed the VNTR 4, 7, 9, 10, 19, 23, 31, LB4 and LB5, which discriminate strains of L. interrogans and L. borgpetersenii. In Pantanal, 17 wildlife, 65 domestic animals and two humans were examined. In Caatinga, seven wild animals were examined, along with 100 domestic animals and 26 humans. Of 84 blood samples tested in Pantanal, 47 (55.95%) were positive and, of 133 in Caatinga, 59 (44.36%) were reactant. By Fisher's exact test, considering a 0.05 significance level, there was no difference between the proportions of serum reagent animals against Leptospira spp. in two biome reviews (p = 0.063). The predominant serovars in SAM reactions were: 1) Pantanal -Bratislava (wildlife, dogs and humans), Grippotyphosa (horses and cattle); 2) Caatinga -Copenhageni (humans and dogs), Patoc (horses and cattle), Panama (sheep and goats), Patoc, Copenhageni and Australis (wildlife). Four strains of Leptospira were isolated: two in Sobradinho, BA, L. interrogans serogroup Pomona in Cavea aperea and L. interrogans in Euphractus sexcinctus; and two in Sobral, CE, L. interrogans in Cerdocyon thous and L. interrogans serogroup Pomona in Euphractus sexcinctus.

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A microbiological and serological study of leptospirosis among pigs in the Morogoro municipality, Tanzania

Kessy¹,

Machang’u

Swai³

2009

Trop Anim Health Prod

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Serological and microbiological studies on leptospirosis in pigs from Morogoro municipality, Tanzania were carried out between October 2007 and May 2008. Serum samples (n = 385) from apparently healthy pigs were tested by microscopic agglutination test for antibodies against live cultures of six known Leptospira interrogans serovars: Pomona, Icterohaemorrhagiae, Ballum, Tarassovi, Grippotyphosa and Hardjo. Significant positive titres were detected in 4.42% (17/385) of all the tested serum samples. Asceptically collected samples, urine (n = 236) and kidney tissues (n = 214), were cultured in enriched Fletcher's and Ellinghausen McCullough-Johnson and Harris media and assessed, at weekly intervals for 24 weeks, for growth by dark-field microscopy. Two leptospiral organisms were isolated from the urine samples. There was a statistical association between seroposivity and location that the subjects reside in (P < 0.05), whereas it was not significantly associated with sex nor age (P > 0.05). The evidence of pig exposure to different serovars and the isolation of the leptospiral organisms confirm that the infection is present in pigs although with an overall low prevalence. Apart from its economic importance on to the pig industry, this disease is a potential zoonotic public health risk in Tanzania, especially because of the lack of studies on leptospirosis among persons who handle pigs and pork products.

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Isolation of Leptospira spp from dogs, bovine and swine naturally infected

Abstract: Isolamento de Leptospira spp de cães, bovinos e suínos naturalmente infectados ABSTRACT Leptospira isolation allows definitive diagnosis of the infection. Contamination by microorganisms is one of the

Cited by 35 publications

References 7 publications

EMJH medium with 5-fluorouracil and nalidixic acid associated with serial dilution technique used to recover Leptospira spp from experimentally contaminated bovine semen

EMJH medium with 5-fluorouracil and nalidixic acid associated with serial dilution technique used to recover Leptospira spp from experimentally contaminated bovine semen

Pesquisa de leptospiras e de anticorpos contra leptospiras em animais e humanos de propriedades rurais nos biomas brasileiros Pantanal e Caatinga

A microbiological and serological study of leptospirosis among pigs in the Morogoro municipality, Tanzania

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