Toxoplasma gondii prevalence and risk factors in owned domestic cats from Nakhon Pathom Province, Thailand

Domestic cats are a potential source of Toxoplasma gondii infection for humans. This study was conducted to determine the seroprevalence and risk factors for T. gondii infection in domestic cats. Cat sera (n = 182) were tested for T. gondii IgG antibodies using the latex agglutination test (LAT) and the GRA7 of T. gondii (TgGRA7)-indirect enzyme-linked immunosorbent assay (iELISA). Univariable and multivariable logistic regression analyses were used to identify the factors associated with T. gondii infection. The overall prevalence rates were 18.1% (33/182) according to LAT, 19.2% (35/182) according to the iELISA and 17.0% (31/182) according to LAT and iELISA. Univariable analyses identified, outdoor access (P = 0.006), being a former stray cat (P = 0.001) and successful hunting behaviors (P = 0.04) as risk factors for T. gondii infection. Outdoor access (OR 2.63, 95% confidence interval (CI) 1.03–6.72) and is a former stray cat (OR 3.69, 95% CI 1.52–8.96) remained significant risk factors in multivariable analyses. This study indicated a relatively high seroprevalence of T. gondii among domestic cats. Cat owners can reduce the risk for T. gondii infection by not allowing their cats to roam free. Furthermore, education about the transmission of the parasite should be provided to prevent infection to the owners

Toxoplasma gondii and Neospora caninum prevalence and risk factors on goat farms in Kanchanaburi province, Thailand

Toxoplasma gondii and Neospora caninum are apicomplexan protozoan parasites that have been associated with reproductive problems in ruminants. Despite the high seroprevalence of T. gondii and N. caninum reported in goats worldwide, little information about the seroprevalence of these parasites in goats in Thailand. In this study, we investigated the seroprevalence of T. gondii and N. caninum in 389 goat samples from five districts of Kanchanaburi province, Thailand. An indirect enzyme-linked immunosorbent assay (iELISA) using recombinant TgGRA7 and NcSAG1 antigens was used for the detection of anti-T.gondii and anti-N.caninum antibodies, respectively. Risk factors for the occurrence of these protozoan parasites on goat farms were also assessed. Specific IgG against T. gondii and N. caninum was detected in 28.5% and 16.7% samples, respectively. Co-infections were observed in 11.8% of samples. The risk factors significantly associated with T. gondii seroprevalence was the presence of cats (odds ratio [OR]= 2.55, 95% confidential interval [CI]=1.67- 3.89). The presence of other domestic animals on farm contributed to the risk of T. gondii and N. caninum infection on goat farms (OR=1.692, 95% CI= 1.08- 2.63). The present study provides data on the seroprevalence and risk of T. gondii and N. caninum infections among goats in Kanchanaburi province, Thailand. Integrated and efficient measures are required to prevent and control infections on goat farms

Immunoproteomics to identify species-specific antigens in Neospora caninum recognised by infected bovine sera

Bovine neosporosis is a disease of concern due to its global distribution and significant economic impact through massive losses in the dairy and meat industries. To date, there is no effective chemotherapeutic drug or vaccine to prevent neosporosis. Control of this disease is therefore dependent on efficient detection tests that may affect treatment management strategies. This study was conducted to identify the specific immunoreactive proteins of Neospora caninum tachyzoites recognised by sera from cattle infected with N. caninum, Toxoplasma gondii, Cryptosporidium parvum, Babesia bovis and B. bigemina, and by sera from uninfected cattle using two-DE dimensional gel electrophoresis (2-DE) combined with immunoblot and mass spectrometry (LC-MS/MS). Among 70 protein spots that reacted with all infected sera, 20 specific antigenic spots corresponding to 14 different antigenic proteins were recognised by N. caninum-positive sera. Of these immunoreactive antigens, proteins involved in cell proliferation and invasion process were highly immunogenic, including HSP90-like protein, putative microneme 4 (MIC4), actin, elongation factor 1-alpha and armadillo/beta-catenin-like repeat-containing protein. Interestingly, we discovered an unnamed protein product, rhoptry protein (ROP1), possessing strong immunoreactivity against N. caninum but with no data on function available. Moreover, we identified cross-reactive antigens among these apicomplexan parasites, especially N. caninum, T. gondii and C. parvum. Neospora caninum-specific immunodominant proteins were identified for immunodiagnosis and vaccine development. The cross-reactive antigens could be evaluated as potential common vaccine candidates or drug targets to control the diseases caused by these apicomplexan protozoan parasites

Molecular Detection and Genetic Diversity of Tick-Borne Pathogens in Goats from the Southern Part of Thailand

Tick-borne hemoprotozoan and rickettsial diseases affect the health and productivity of small ruminants in tropical and subtropical regions. Despite the large population of goats in the southern part of Thailand, there is limited information on the prevalence of tick-borne pathogens. In this study, polymerase chain reaction was used to detect the presence of Theileria spp., T. ovis, T. orientalis, Babesia ovis, Anaplasma ovis, and A. marginale in 262 goats from three provinces in the southern part of Thailand. In this investigation, Theileria spp. and A. ovis were detected while T. ovis, B. ovis, and A. marginale were not detected. Overall infection rates of Theileria spp. and A. ovis were 10.3% and 1.5%, respectively. The co-infections of two parasites was observed in 1.5% of goats. Sequence analysis showed the presence of T. luwenshuni and T. orientalis in the goat samples. This study is the first to use the molecular detection of T. orientalis in Thai goats, and presents genetic characterization using the major piroplasm surface protein (MPSP) gene. In the phylogenetic analysis, the T. orientalis MPSP sequence was classified as type 7. The A. ovis major surface protein 4 (MSP4) gene sequences shared high identities and similarity with each other and clustered with isolates from other regions. This study provides information about the prevalence and genetic diversity of tick-borne pathogens in goats in the study area, and is expected to be valuable for the development of effective control measures to prevent disease in animals in Thailand

Exposure to Toxoplasma gondii in Asian Elephants (Elephas maximus indicus) in Thailand

Toxoplasma gondii is the causative agent of toxoplasmosis in humans and various animal species worldwide. In Thailand, seroprevalence studies on T. gondii have focused on domestic animals, and information on infections in Asian elephants (Elephas maximus indicus) is scarce. This study was conducted to determine the seroprevalence of T. gondii infection in archival sera collected from 268 elephants living in Thailand. The serum samples were analyzed for anti-T. gondii immunoglobulin G antibodies using the latex agglutination test (LAT) and indirect enzyme-linked immunosorbent assay (iELISA) based on T. gondii lysate antigen (TLA-iELISA) and recombinant T. gondii dense granular antigen 8 protein (TgGRA8-iELISA). The prevalence of antibodies against T. gondii was 45.1% (121/268), 40.7% (109/268), and 44.4% (119/268) using LAT, TLA-iELISA, and TgGRA8-iELISA, respectively. Young elephants had a higher seropositivity rate than elephants aged >40 years (odds ratio = 6.6; p < 0.001; 95% confidence interval: 2.9–15.4). When LAT was used as the reference, TLA-iELISA and TgGRA8-iELISA showed a substantial (κ = 0.69) and moderate (κ = 0.42) agreement, respectively. Although our findings suggest the widespread exposure of Asian elephants to T. gondii in Thailand, the source of infection was not investigated. Therefore, investigation of the predisposing factors associated with toxoplasmosis is necessary to identify the potential risk factors for infection

Immunoproteomics to identify species-specific antigens in

Bovine neosporosis is a disease of concern due to its global distribution and significant economic impact through massive losses in the dairy and meat industries. To date, there is no effective chemotherapeutic drug or vaccine to prevent neosporosis. Control of this disease is therefore dependent on efficient detection tests that may affect treatment management strategies. This study was conducted to identify the specific immunoreactive proteins of Neospora caninum tachyzoites recognised by sera from cattle infected with N. caninum, Toxoplasma gondii, Cryptosporidium parvum, Babesia bovis and B. bigemina, and by sera from uninfected cattle using two-DE dimensional gel electrophoresis (2-DE) combined with immunoblot and mass spectrometry (LC-MS/MS). Among 70 protein spots that reacted with all infected sera, 20 specific antigenic spots corresponding to 14 different antigenic proteins were recognised by N. caninum-positive sera. Of these immunoreactive antigens, proteins involved in cell proliferation and invasion process were highly immunogenic, including HSP90-like protein, putative microneme 4 (MIC4), actin, elongation factor 1-alpha and armadillo/beta-catenin-like repeat-containing protein. Interestingly, we discovered an unnamed protein product, rhoptry protein (ROP1), possessing strong immunoreactivity against N. caninum but with no data on function available. Moreover, we identified cross-reactive antigens among these apicomplexan parasites, especially N. caninum, T. gondii and C. parvum. Neospora caninum-specific immunodominant proteins were identified for immunodiagnosis and vaccine development. The cross-reactive antigens could be evaluated as potential common vaccine candidates or drug targets to control the diseases caused by these apicomplexan protozoan parasites

Toxoplasma Serostatus in Thai Free- Range Chickens: Prevalence and Two Diagnostic Methods

Toxoplasma seroprevalence in free-range chicken, which directly feed from ground, is a good indicator for detecting an environment contamination by T. gondii oocysts. Many serological methods have been used for diagnosing Toxoplasma infection in animals including chicken. Selection of appropriate methods primarily depend on their sensitivity and specificity as well as laboratory techniques, special reagent or tool requirements and regional prevalence status. The aims of this study were to determine the prevalence of T.gondii antibody in free-range chickens in Thailand and to compare two serodiagnostic methods, indirect fluorescent antibody test (IFAT) and Modified Agglutination Test (MAT). Six hundred free-range chicken sera were collected from 2 villages in Kanchanaburi Province, Thailand, 300 in 2014 and 300 in 2015. The age of chickens was between 3-6 mo, of which chickens of 2015 were older than of 2014. Collected sera were test for T. gondii IgG antibody by both methods. Over all Toxoplasma seroprevalence were 33.0% and 17.7% by IFAT and MAT, respectively. There was poor agreement between the two serological methods (kappa=0.09). IFAT in this study revealed higher positive percentage than MAT. Even though the way of living and environment in the present study was suitable for Toxoplasma transmission, the seroprevalence in Thai free-range chickens was low compared to other tropical regions such as Africa or South America. A low Toxoplasma prevalence was also found in humans, cats and other intermediate hosts in Thailand.Toxoplasmosis in Thailand causes low burden compare to other infectious diseases, however, consume well-cooked free-range chicken meat and proper practice during handling raw meat when cooking should be advocated to prevent and reduce this infection.</p

Evaluation of Immunodiagnostic Performances of <i>Neospora caninum</i> Peroxiredoxin 2 (NcPrx2), Microneme 4 (NcMIC4), and Surface Antigen 1 (NcSAG1) Recombinant Proteins for Bovine Neosporosis

Bovine neosporosis is among the main causes of abortion in cattle worldwide, causing serious economic losses in the beef and dairy industries. A highly sensitive and specific diagnostic method for the assessment of the epidemiology of the disease, as well as it surveillance and management, is imperative, due to the absence of an effective treatment or vaccine against neosporosis. In the present study, the immunodiagnostic performance of Neospora caninum peroxiredoxin 2 (NcPrx2), microneme 4 (NcMIC4), and surface antigen 1 (NcSAG1) to detect IgG antibodies against N. caninum in cattle were evaluated and compared with that of the indirect fluorescent antibody test (IFAT). The results revealed that NcSAG1 had the highest sensitivity and specificity, with values of 88.4% and 80.7%, respectively, followed by NcPrx2, with a high sensitivity of 87.0% but a low specificity of 67.0%, whereas NcMIC4 showed sensitivity and specificity of 84.1% and 78.9%, respectively, when compared with IFAT. A high degree of agreement was observed for NcSAG1 (k = 0.713) recombinant protein, showing the highest diagnostic capability, followed by NcMIC4 (k = 0.64) and NcPrx2 (k = 0.558). The present study demonstrates that NcSAG1 is helpful as an antigen marker and also demonstrates the potential immunodiagnostic capabilities of NcPrx2 and NcMIC4, which could serve as alternative diagnostic markers for detecting N. caninum infection in cattle. These markers may find utility in future treatment management, surveillance, and risk assessment of neosporosis in livestock or other animal host species. Further research should be directed toward understanding the in vivo immune response differences resulting from immunization with both recombinant proteins

Additional file 1: Figure S1. of Development of a loop-mediated isothermal amplification technique and comparison with quantitative real-time PCR for the rapid visual detection of canine neosporosis

Map showing the sampling areas and the types of samples collected in this study. (TIFF 2909 kb