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. 2020 Mar 17;16(1):89.
doi: 10.1186/s12917-020-2230-0.

Leptospira infection and shedding in dogs in Thailand

Kerstin Altheimer  1 , Prapaporn Jongwattanapisan  2 , Supol Luengyosluechakul  2 , Rosama Pusoonthornthum  2 , Nuvee Prapasarakul  3 , Alongkorn Kurilung  3 , Els M Broens  4 , Jaap A Wagenaar  4   5 , Marga G A Goris  6 , Ahmed A Ahmed  6 , Nikola Pantchev  7 , Sven Reese  8 , Katrin Hartmann  9
Affiliations

Leptospira infection and shedding in dogs in Thailand

Kerstin Altheimer et al. BMC Vet Res. .

Abstract

Background: Leptospirosis is a widespread zoonosis and has been recognized as a re-emerging infectious disease in humans and dogs, but prevalence of Leptospira shedding in dogs in Thailand is unknown. The aim of this study was to determine urinary shedding of Leptospira in dogs in Thailand, to evaluate antibody prevalence by microscopic agglutination test (MAT) and enzyme-linked immunosorbent assay (ELISA), and to assess risk factors for Leptospira infection. In Northern, Northeastern, and Central Thailand, 273 stray (n = 119) or client-owned (n = 154) dogs from rural (n = 139) or urban (n = 134) areas were randomly included. Dogs that had received antibiotics within 4 weeks prior to sampling were excluded. No dog had received vaccination against Leptospira. Urine was evaluated by real-time polymerase chain reaction (PCR) specific for lipL32 gene of pathogenic Leptospira. Additionally, urine was cultured for 6 months in Ellinghausen-McCullough-Johnson-Harris (EMJH) medium. Antibodies were measured by ELISA and MAT against 24 serovars belonging to 15 serogroups and 1 undesignated serogroup. Risk factor analysis was performed with backwards stepwise selection based on Wald.

Results: Twelve of 273 (4.4%; 95% confidence interval (CI): 2.0-6.8%) urine samples were PCR-positive. In 1/273 dogs (0.4%; 95% CI: 0.01-1.1%) Leptospira could be cultured from urine. MAT detected antibodies in 33/273 dogs (12.1%; 95% CI: 8.2-16.0%) against 19 different serovars (Anhoa, Australis, Ballum, Bataviae, Bratislava, Broomi, Canicola, Copenhageni, Coxi, Grippotyphosa, Haemolytica, Icterohaemorrhagiae, Khorat, Paidjan, Patoc, Pyrogenes, Rachmati, Saxkoebing, Sejroe). In 111/252 dogs (44.0%; 95% CI: 37.9-50.2%) immunoglobulin M (IgM) and/or immunoglobulin G (IgG) antibodies were found by ELISA. Female dogs had a significantly higher risk for Leptospira infection (p = 0.023).

Conclusions: Leptospira shedding occurs in randomly sampled dogs in Thailand, with infection rates comparable to those of Europe and the USA. Therefore, the potential zoonotic risk should not be underestimated and use of Leptospira vaccines are recommended.

Keywords: Canine; Culture; Dogs; ELISA; Leptospira; MAT; PCR; Risk factors; Seroprevalence; Zoonosis.

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Conflict of interest statement

NPa is employed at IDEXX Laboratories, Ludwigsburg. This laboratory offers the IDEXX Leptospira RealPCR Test on a commercial basis and performed the testing in this study.

MGAG and AAA work at the OIE and National Collaborating Centre for Reference and Research on Leptospirosis and commercially performed MAT and phylogenetic analysis in the present study.

EMB and JAW work at the Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University and commercially performed IgM and IgG ELISA in this study.

None of these companies played a role in the study design, in the collection and interpretation of data, or in the decision to submit the manuscript for publication.

KH has given talks for MSD, Merial, Boehringer Ingelheim, and IDEXX. She participated in research funded by or using products from MSD, Merial, Boehringer, Zoetis, Megacor, Biogal, and Scil.

There is no commercial conflict of interest as the information generated here is solely for scientific dissemination. The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Evolutionary relationships of taxa. The evolutionary history was inferred using the Neighbor-Joining method. The optimal tree with the sum of branch length = 0.76583659 is shown. The percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (1000 replicates) are shown next to the branches. The tree is drawn to scale, with branch lengths in the same units as those of the evolutionary distances used to infer the phylogenetic tree. The evolutionary distances were computed using the Maximum Composite Likelihood method and are in the units of the number of base substitutions per site. The analysis involved 19 nucleotide sequences. Codon positions included were 1st + 2nd + 3rd + noncoding. All positions containing gaps and missing data were eliminated. There were a total of 245 positions in the final dataset. Evolutionary analyses were conducted in MEGA 7. The bar indicates 0.050 estimated substitution per sequence position. Dog D64 of the present study clusters within the genomospecies Leptospira interrogans
See this image and copyright information in PMC

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