Comparison of polymerase chain reaction assay, bacteriologic culture, and serologic testing in assessment of prevalence of urinary shedding of leptospires in dogs

Harkin, Kenneth R.; Roshto, Yvette M; Sullivan, Jennifer T; Purvis, Tanya; Chengappa, M. M.

doi:10.2460/javma.2003.222.1230

Cited by 99 publications

(103 citation statements)

References 18 publications

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“…2) demonstrated that out of 89 seropositive cows, 23 responded positively in PCR, whereas Leptospira were found only in 11 urine samples. Similar studies were conducted by many researchers (20), who observed negative reaction in bacteriological studies, but positive in 50% cases using PCR (13). In the studies of these authors, PCR demonstrated very good results as compared to traditional methods, which was also confirmed in our study.…”

Section: Discussionsupporting

confidence: 80%

Leptospirosis in Cattle From Markets of Almaty Province, Kazakhstan

Kirkimbayeva

Łozowicka

Biyashev

et al. 2015

Bulletin of the Veterinary Institute in Pulawy

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This paper is the first study of the prevalence of leptospirosis in the cattle at slaughter from a rural area of Kazakhstan. Five hundred and seventy three samples of serum, urine, and kidneys from cattle of Alatau, Kazakh white and Auliyekol breed, aged from 2 to 5 years (unknown vaccination status), from the province of Almaty in the South-Eastern region were collected during four years (March 2010 to October 2013). The serological, bacteriological, and molecular analyses were performed. Serum samples were tested with 14 reference Leptospira serovars by microscopic agglutination test (MAT). MAT results showed that 89 (15.53%) serum samples had detectable antibodies against seven serovars of L. interrogans at a dilution of ≥1:100. Serovars: Pomona (38.2%), Tarassovi (27.2%), and Kabula (18.8%) were the most prevalent and their titres ranged from 100 to 1200. The spirochetes were detected in 11 samples of urine and nine samples of kidneys under dark-field microscope observation. The pure cultures were obtained from three samples. PCR technique confirmed leptospirosis in 23 out of 89 urine samples from cows, which showed the presence of leptospiral antibodies in microagglutination test. The high disease prevalence in cows indicates the high Leptospira contamination in this area. It was concluded that the bovine leptospirosis is an endemic and locally widespread disease in Kazakhstan, and that it may play a role in zoonotic transmission to humans.

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Section: Discussionsupporting

confidence: 80%

Leptospirosis in Cattle From Markets of Almaty Province, Kazakhstan

Kirkimbayeva

Łozowicka

Biyashev

et al. 2015

Bulletin of the Veterinary Institute in Pulawy

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“…There were variations in the sensitivity and specificity in detecting leptospiral organisms by various PCR methods (Wagenaar et al 2000). In addition, some PCR methods exhibited false positives in detecting pathogenic leptospires in clinical samples which crossreacted with saprophytic leptospires (Merien et al 1992;Wagenaar et al 1994;Harkin et al 2003). Selection of PCR methods is highly critical for the detection of leptospires in various samples.…”

Section: Discussionmentioning

confidence: 99%

Neglected leptospirosis in raccoons (Procyon lotor) in Indiana, USA

Tan

Dharmarajan

Beasley

et al. 2014

Veterinary Quarterly

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Background: Leptospirosis is a globally important zoonotic disease occurring clinically and subclinically in humans and animals. Objectives: To determine whether raccoons in Indiana carried leptospires in their kidneys. Animals and methods: Thirty-four raccoons were live-trapped from two forest patches in central Indiana. Following euthanasia, a portion of kidney (2 cm 2 ) from each raccoon was homogenized and used for leptospiral culture. Leptospiral cultures were subjected to DNA extraction and various polymerase chain reaction (PCR) procedures reported previously. Serum sample from each raccoon was collected and antibody titers to leptospiral serovars were determined by microscopic agglutination test (MAT). Results: All leptospiral cultures were positive for Leptospira by various PCR procedures. The PCR with the primers targeting the conservative region of LipL32 gene was the most sensitive PCR in the detection of pathogenic leptospires. The variable LipL32 PCR amplicons were sequenced and compared to the reference strains available in GenBank. Twelve kidney cultures had Leptospira interrogans, eight had Leptospira kirschneri and two had Leptospira borgpetersenii. They were predominantly Grippotyphosa serogroup. Antileptospire antibodies were detected in 16 out of 34 raccoons (47.1%) by MAT. There were titers ! 1:80 in 16 raccoons (47.1%) and titers ! 1:400 in 3 raccoons (8.8%). The highest leptospiral serovar-specific seroreactivity among 34 raccoons was L. interrogans Bratislava (38.2%) and L. interrogans Grippotyphosa (32.4%). Conclusions: Raccoons in Indiana carry leptospiral organisms in kidneys and the leptospires are predominantly L. interrogans species and of the Grippotyphosa serogroup. Clinical importance: The raccoons serve as reservoir hosts that exposure sources to wildlife, livestock, pets and humans.

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“…Nonspecific vaccine titers can be detected by a microscopic agglutination test (MAT) up to three months after immunization (MILLER et al, 2011). Harkin et al (2003) and Minke et al (2009) showed that the vaccines prevent renal colonization, leptospiruria over long periods and death after extreme challenges in experimental infections.…”

Section: Introductionmentioning

confidence: 99%

Seroepidemiology of leptospirosis in pet dogs in the urban area of the municipality of Jataizinho, Paraná

et al. 2012

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The study of the prevalence of anti-leptospiral antibodies in pet dogs in urban areas and the variables associated with the risk of infection can aid in the identification of the serovars present in the region and indicated the reservoirs involved in this important zoonosis. The aim of the present study was to determine the prevalence of anti-leptospiral antibodies in 653 dogs from 369 households distributed throughout the urban area of the municipality of Jataizinho, Paraná (PR), and to investigate the variables associated with the risk of leptospiral infection in these animals. Serum samples were subjected to microscopic agglutination test (MAT) against 23 serovars of pathogenic leptospires. Among the 132 (20.21%) reactive samples, 15 (11.36%) were reactive for more than one serovar, and the most likely serovar could not be determined. Among the 117 (88.73%) samples that reacted to a single serovar, the serovar Canicola was considered to be the most likely in 89 cases (76.07%). Variables associated with the risk of disease were investigated using an epidemiological questionnaire administered to each owner, and the data obtained were analyzed using the statistical software EpiInfo. The significant variables (p ≤ 0.05) associated with seropositivity for leptospirosis in pet dogs in the urban area of the municipality of Jataizinho, PR, were the presence of rats in the household, adult or elderly dogs, free access of the dog to the street and contact with stray dogs. The results indicate a high prevalence of pet dogs in the urban area of Jataizinho, PR, with anti-leptospiral antibodies and suggest the need to educate the population of the municipality about the vaccination of dogs, as well as, the epidemiological role of the dog in urban leptospirosis. Key words: Epidemiology, microscopic agglutination, test, Leptospira spp., serovar Canicola, diagnosis, risk variables ResumoO estudo da prevalência de cães domiciliados em áreas urbanas com anticorpos antileptospiras e de variáveis associadas ao risco de infecção, pode auxiliar na identificação dos sorovares presentes na região e sugerir os reservatórios envolvidos nesta importante zoonose. O objetivo deste trabalho foi determinar a prevalência de anticorpos antileptospiras em 653 cães de 369 domicílios distribuídos na área urbana do município de Jataizinho, PR, e estudar variáveis associadas ao risco de infecção por leptospiras nesses animais. As amostras de soro foram submetidas a prova de soroaglutinação microscópica (SAM) frente a 23 sorovares de leptospiras patogênicas. Das 132 (20,21%) amostras de cães reagentes, 15 (11,36%) reagiram para mais de um sorovar, não sendo possível determinar o sorovar mais provável, e das 117 (88,73%) que reagiram para um sorovar somente, o sorovar Canicola foi considerado o mais provável em 89 (76,07%). Variáveis associadas ao risco da doença foram investigadas através de um questionário epidemiológico aplicado a cada proprietário e os dados obtidos foram analisados pelo programa estatístico EpiInfo. As variáveis signifi...

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Comparison of polymerase chain reaction assay, bacteriologic culture, and serologic testing in assessment of prevalence of urinary shedding of leptospires in dogs

Abstract: Irrespective of health status, 8.2% of dogs were shedding pathogenic leptospires. Serologic testing was a poor predictor of urinary shedding. Clinically normal dogs that shed leptospires may pose a zoonotic risk to their owners.

Cited by 99 publications

References 18 publications

Leptospirosis in Cattle From Markets of Almaty Province, Kazakhstan

Leptospirosis in Cattle From Markets of Almaty Province, Kazakhstan

Neglected leptospirosis in raccoons (Procyon lotor) in Indiana, USA

Seroepidemiology of leptospirosis in pet dogs in the urban area of the municipality of Jataizinho, Paraná

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