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Leptospirosis is an important zoonotic disease that is often associated with animal carriers and contamination of the environment via infected urine. This study aimed to assess pathogenic leptospiral carriage in Nan province, a rural area of Thailand where leptospirosis is endemic. Samples from 20 villages were obtained during the period 2013 to 2016, comprising urine samples collected from asymptomatic people (n=37) and domestic animals (n=342), and environmental water samples (n=14). Leptospira were cultured in Ellinghauson McCullough Johnson and Harris (EMJH) media. An rrs nested PCR identified 9.92% (95% confidence interval (CI) 6.96-12.88) of the urine and water samples as being positive for Leptospira spp., and phylogenetic analysis was conducted on the 443bp amplicons. Leptospira weilii, which has not previously been identified in Thailand, was recovered from 13 cattle, 9 pigs, 2 dogs, 2 water samples and 1 goat. L. interrogans was found in 4 dogs, 3 pigs, 3 cattle, 1 human and 1 water sample. Four leptospiral strains were isolated and multilocus sequence typing (MLST) analysis was performed on these. Three novel sequence types were identified, including two singletons of L. interrogans in ST26 and ST33, and one of L. weilii in ST94, with this having a close relationship to previous isolates from cases of human leptospirosis in Laos and China. Our results revealed that pathogenic Leptospira occur commonly in asymptomatic domestic animals, humans and environmental water samples in Nan Province, and emphasize the high potential for zoonotic transmission in the province.

Aims: This study was to investigate and to characterize methicillin-resistant coagulase-positive staphylococci (MRCoPS) harboring in dogs and people associated with dogs in Thailand. Methods and Results: Staphylococci were collected from 100 dogs, 100 dog owners, 200 small animal veterinarians and 100 people without pet association. Species of MRCoPS were identified phenotypically and genotypically. Molecular characteristics were determined by multilocus sequence typing (MLST), pulsedfield gel electrophoresis (PFGE) and SCCmec typing, and antimicrobial susceptibility was assayed by broth microdilution and by microarray analysis for resistance genes. Methicillin-resistant Staphylococcus pseudintermedius (MRSP), methicillin-resistant Staphylococcus schleiferi subsp. coagulans (MRSSc) and methicillin-resistant Staphylococcus aureus (MRSA) were isolated from dogs (45, 17 and 1%, respectively), veterinarians (8, 2 and 1Á5%, respectively) and dog owners (3, 2 and 0%, respectively). Seventeen sequence types (STs) were identified among 83 MRSP isolates which specifically carried SCCmec V, II-III, ΨSCCmec 57395 and three uncharacterized SCCmec types. MRSP ST 45, 68 and novel STs including 169, 178, 181 and 183 were shared among canine and human isolates. Most of MRSA ST398 and MRSSc carried SCCmec type V. The MRCoPS commonly displayed multiple resistances to tested antimicrobials and carried various resistance genes. Conclusion: Variety of MRCoPS, especially new MRSP clones, distributed in dogs and people in Thailand. Significance and Impact of the Study: The existence of MRCoPS circulating between dogs and humans in Thailand provides indirect evidence of interspecies transmission and represents a potential public health hazard.

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In Thailand, leptospirosis is considered an emerging disease in humans and animals. Many species can shed pathogenic Leptospira, including domestic cats (felis catus), which might be able to pose a risk to humans. There are no studies on Leptospira infections in cats in Thailand, but in other countries, it was demonstrated that cats can shed pathogenic Leptospira with high prevalences. The aims of this study were to evaluate whether outdoor cats in Thailand shed pathogenic Leptospira in their urine, and to determine antibody prevalence and risk factors associated with Leptospira infection. Two hundred and sixty outdoor cats were prospectively recruited. Urine samples were tested by real‐time PCR targeting the lipL32 gene of pathogenic Leptospira. Urine was additionally cultured for 6 months in Ellinghausen‐McCullough‐Johnson‐Harris medium to grow Leptospira. Antibodies against 24 serovars (Anhoa, Australis, Autumnalis, Ballum, Bataviae, Bratislava, Broomi, Canicola, Celledoni, Copenhageni, Coxi, Cynopteri, Djasiman, Grippotyphosa, Haemolytica, Icterohaemorrhagiae, Khorat, Paidja, Patoc, Pomona, Pyrogenes, Rachmati, Saxkoebing, Sejroe) belonging to 16 serogroups were determined using microscopic agglutination tests. Risk factors were analysed by Fisher's exact test. Urine samples of 2/260 cats (0.8%; 95% confidence interval (CI): 0.1%–2.8%) were PCR‐positive, but none of the 260 urine samples were culture positive. Leptospira antibodies were detected in 14/260 cats (5.4%; 95% CI: 3.0%–8.6%) with titers ranging from 1:20 to 1:160 (serovars: Anhoa, Autumnalis, Celledoni, Copenhageni, Djasiman, Icterohaemorrhagiae, Patoc). Cats aged ≥4 years were significantly more often infected with Leptospira than younger cats. No other significant risk factors were found. In conclusion, outdoor cats in Thailand can shed DNA and, possibly, viable, pathogenic Leptospira in their urine, although at a much lower prevalence than expected when compared to countries with similar climate. Thus, cats can be a potential source of infection for people. Further studies are needed to determine the role of cats in transmitting this zoonotic disease in Thailand.

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Porcine epidemic diarrhea virus (PEDV) is a coronavirus which causes severe diarrhea and fatal dehydration in piglets. In general, probiotic supplements could enhance recovery and protect piglets against enteric pathogens. Seven local lactic acid bacteria (LAB), (Ent. faecium 79N and 40N, Lact. plantarum 22F, 25F and 31F, Ped. acidilactici 72N and Ped. pentosaceus 77F) from pig feces were well-characterized as high potential probiotics. Cell-free supernatants (CFS) and live LAB were evaluated for antiviral activities by co-incubation on Vero cells and challenged with a pandemic strain of PEDV isolated from pigs in Thailand. Cell survival and viral inhibition were determined by cytopathic effect (CPE) reduction assay and confirmed by immunofluorescence. At 1:16, CFS dilution (pH 6.3-6.8) showed no cytotoxicity in Vero cells and was therefore used as the dilution for antiviral assays. The diluted CFS of all Lact. plantarum showed the antiviral effect against PEDV; however, the same antiviral effect could not be observed in Ent. faecium and Pediococcus strains. In competitive experiment, only live Lact. plantarum 25F and Ped. pentosaceus 77F showed CPE reduction in the viral infected cells to <50% observed field area. This study concluded that the CFS of all tested lactobacilli, and live Lact. plantarum (22F and 25F) and Pediococcus strains 72N and 77F could reduce infectivity of the pandemic strain of PEDV from pigs in Thailand on the target Vero cells.

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