Oral vaccination of dogs against rabies has the potential to achieve mass coverage and thus deplete the virus of its most important reservoir host species. There is, however, no established non-invasive method to evaluate vaccine release in the oral cavity, following bait ingestion. In this study, two pre-selected marker methods in conjunction with their acceptance were assessed in local Thai dogs. Shelter dogs (n = 47) were offered one of four randomized bait formulations; bait type A-, containing Green S (E142) in a fructose solution; type B-, containing Patent Blue V (E131) in a fructose solution; type C-, containing the medium used for delivery of oral rabies vaccine in baits commercially produced; and type D-, containing denatonium benzoate, which was to serve as the negative control, due to its perceived bitterness. Patent Blue V was found to possess overall stronger dyeing capacities compared to Green S. Furthermore, there was no significant difference in the acceptance or bait handling of Patent Blue V baits compared to those containing the oral rabies vaccine medium alone, suggesting the potential use of this dye as a surrogate for rabies vaccine when testing newly developed bait formats.

In total, 901 dairy cow sera and data were collected from 51 farms in Nakhon Pathom, Ratchaburi and Kanchanaburi provinces (Western Region of Thailand). Serum samples were processed via the multispecies ELISA method to detect IgG antibodies against Toxoplasma gondii infection. The results demonstrated that the calculated true prevalence was 1.48% (95% CI, 0.64–2.75%) for the individual-level and 29.41% (95% CI, 18.71–43%) for the farm-level. The univariate risk factor analysis showed that the number of total owned cats, the presence of stray cats, and the frequency of cleaning per day were significant factors (p < 0.2). These three factors were subjected to logistic regression analysis, and the results revealed that the frequency of cleaning farms per day was a potential risk factor for T. gondii-seropositive farms (OR = 2.745, 95% CI, 1.15–8.69, p = 0.02). The frequency of cleaning might increase the T. gondii oocyst distribution within the barn area, thus increasing the possibility of infection. Our findings show that T. gondii continues to circulate in the dairy cow population in the western part of Thailand. The presence of cats on farms was not found to be associated with T. gondii infection, but the high frequency of cleaning the floor was, and contributed to the potential risk of infection.

The objective was to detect T. gondii and various protozoan oocysts in the feces of sheltered cats in Thailand using two molecular approaches, Sanger sequencing and un-targeted Next-generation sequencing (NGS). A total of 166 fecal samples from shelter cats, in 26 samples, Toxoplasma gondii oocyst–like structures were detected. The harvested oocysts were grouped into nine pooled samples. DNA was extracted from all pooled samples and tested using quantitative PCR (qPCR), employing coc1 and coc2 primers, which are commonly used to amplify Apicomplexan DNA. The sequenced of the qPCR products were analyzed by Sanger sequencing. Sequence from all pooled samples had similarity to Cystoisospora spp. To further characterize the oocyst species, NGS was performed. Bioinformatic analysis was conducted using a de novo assembler to generate scaffolds, which were then aligned against a custom database of coccidian whole-genome references. This analysis revealed the presence of T. gondii DNA in three pooled samples. In addition, DNA from other protozoan parasites—including Eimeria spp., Cystoisospora spp., Besnoitia besnoiti, Hammondia hammondi, and Cryptosporidium parvum—was also detected. These findings indicate that T. gondii is circulating among shelter cats, many of which were formerly stray. Moreover, NGS sequencing provided more comprehensive information on the diversity of coccidian species in cat feces compared with Sanger sequencing of PCR-amplified targets.