Background and Aim: Dog blood parasites are important tick-borne diseases causing morbidity and mortality in dogs worldwide. Four dog blood parasites species are commonly found in Thailand: Babesia canis, Hepatozoon canis, Ehrlichia canis, and Anaplasma platys. They are transmitted easily by tick species. However, there is little prevalence data available in Thailand. Diseases presentation of blood parasites infection is similar, but the treatment of each species is different. Current diagnosis mainly relies on microscopic examination of a stained blood smear, which has low sensitivity. Therefore, accurate diagnosis is important. This study aims to evaluate the efficacy of the conventional polymerase chain reaction (PCR) method and routine blood smears in the detection of four blood parasites species in dogs from Buriram Province, Thailand. Materials and Methods: In total, 49 EDTA-blood samples were collected from dogs in Buriram Province, Thailand. Blood parasite infection was compared using the Giemsa-stained blood smear technique to identify the parasite under a 100× oil immersion with PCR amplification of the 18S rDNA gene of B. canis and H. canis and the 16S rDNA gene of E. canis and A. platys. Results: Only one dog out of 49 was positive for H. canis based on microscopic examination whereas the PCR results showed that 2.04% (1/49), 4.08% (2/49), 36.73% (18/49), and 30.61% (15/49) of dogs were positive for B. canis, H. canis, E. canis, and A. platys, respectively. Moreover, coinfection was found in 16.33% (8/49) of dogs. Conclusion: This study is the first report to demonstrate the molecular prevalence of blood parasites in domestic dogs in Buriram Province. The results indicated that the PCR method exhibited much higher sensitivity and reliability for blood parasites diagnosis in dogs. Therefore, our data support serious concern regarding the diagnostic technique used in routine blood testing and also provide prevalence data for the management and control of blood parasites in this area.

The purpose of the present study was to establish normal electroretinogram (ERG) parameters using 56 normal eyes of four dog breeds common in Thailand: poodle, Labrador retriever, Thai ridgeback, and Thai Bangkaew. Standard ERG findings were bilaterally recorded using a handheld multi-species ERG unit with an ERG-jet lens electrode for 28 dogs under preanesthesia with diazepam, anesthesia with propofol, and anesthesia maintenance with isoflurane. There were significant differences in the mean values of ERG amplitudes and implicit times among the four dog breeds (p < 0.05) except for the b-wave implicit time of the photopic 30 Hz flicker response with 3 cd.s/m2 (p = 0.610). Out of the four breeds, Thai Bangkaew had the longest implicit time (p < 0.001) of scotopic low intensity responses, b-wave of scotopic standard intensity responses (3 cd.s/m2), a-wave of the higher intensity response (10 cd.s/m2), and a-wave of the photopic single flash response (3 cd.s/m2). For the b/a ratio, only the ratio of the Cone response was significantly different among the different breeds. In this summary, normal ERG parameters for four dog breeds were reported. Data from the investigation supported the hypothesis that determination of breed-specific limits of normality for ERG responses is necessary for individual clinics and laboratories.

Background Tear proteomic analysis has become an important tool in medical and veterinary research. The tear collection method could influence the tear protein profile. This study aims to evaluate the protein profiles of dog tears collected using microcapillary tubes (MT), Schirmer tear strips (ST), and ophthalmic sponges (OS). Methods The tear samples were collected using MT, ST, and OS. Tear protein profiles were analyzed using two-dimensional electrophoresis (2-DE) and the different protein spots’ expression was compared. Fourteen protein spots were identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Results Tear protein concentrations ranged from 2.80 to 4.03 μg/μL, with no statistically significant differences among collection methods. Protein expression in each collection method differed in terms of both the number and intensity of the spots. There were 249, 327, and 330 protein spots found from tears collected with MT, ST, and OS, respectively. The proteins albumin, haptoglobin, and lactoferrin identified from OS were found to have higher spot intensities than other methods of collection. The use of MT demonstrated the downregulation of nine proteins. Conclusions The recent study supported that tear protein analysis is affected by different tear collection methods. Although ST is commonly used for tear collection, it provides insufficient information to study particular tear proteins.

Background and Aim: Toxoplasma gondii is recognized as a zoonosis causing toxoplasmosis in animals globally. Cat is a definitive host of T. gondii and sheds oocyst through feces, which can infect human beings and animals through contaminated food ingestion. A precise diagnostic test is essential to prevent T. gondii infection in both humans and animals. This study aimed to develop and evaluate the pETite- dense granule antigen 7(GRA7)-based indirect enzyme-linked immunosorbent assay (ELISA) to detect T. gondii infection in cats. Materials and Methods: T. gondii-GRA7 was cloned and expressed in the Expresso® small ubiquitin-related modifier (SUMO) T7 Cloning and Expression System. The recombinant pETite-GRA7 was purified using HisTrap affinity chromatography and confirmed using Western blot analysis. The recombinant protein was used to develop and evaluate the indirect ELISA for T. gondii infection detection. In total, 200 cat sera were tested using pETite-GRA7-based indirect ELISA and indirect fluorescent antibody test (IFAT). The statistical analysis based on Kappa value, sensitivity, specificity, positive predictive value, negative predictive value, χ2 test, and receiver operating characteristic (ROC) curve was used to evaluate the performance of the test. Results: A 606 bp GRA7 polymerase chain reaction (PCR) product was obtained from T. gondii RH strain genomic DNA. The gene was cloned into the pETite™ vector and transformed to HI-Control Escherichia coli BL21 (DE3) for protein expression. Approximately 35 kDa of recombinant pETite-GRA7 was observed and Western blot analysis showed positive bands against anti-6-His antibody and positive-T. gondii cat serum. A sample of 0.5 μg/mL of pETite-GRA7 was subjected to indirect ELISA to detect T. gondii infection in the cat sera. The results showed sensitivity and specificity of pETite- GRA7-based indirect ELISA at 72% and 96%, respectively. An acceptable diagnostic performance was characterized by high concordant results (94%) and substantial agreement (Kappa value=0.65) with IFAT. The seroprevalence levels of ELISA and IFAT were 10% and 9%, respectively, and were not significantly (p>0.05) different. The expected performance of ELISA at different cutoff points using the ROC curve analysis revealed 89% sensitivity and 92% specificity at the cutoff value of 0.146, with a high overall assay accuracy (area under the curve=0.94). Conclusion: In this study, the pETite™ vector, N-terminal 6xHis SUMO fusion tag, was used to improve the solubility and expression level of GRA7. The recombinant pETite-GRA7 showed enhanced protein solubility and purification without special condition requirements. This pETite-GRA7-based indirect ELISA showed high concordant results and substantial agreement with IFAT. ELISA revealed an acceptable sensitivity and specificity. These initial data obtained from cats' sera demonstrated that pETite-GRA7-based indirect ELISA could be a useful method for local serological diagnosis of T. gondii infection in cats in Thailand.

Data from this report provides reference values for scotopic ERG measurements in these two wild cat species. It showed that the normal scotopic ERG responses have some differences between the two species which might be due to the skull conformation, eye size or physiology of the retina.