Molecular Detection and Characterization of Hemotropic Mycoplasma in Assamese Macaques (Macaca assamensis) of Northern Thailand.

Hemotropic mycoplasmas, also known as hemoplasmas, are parasitic bacteria that infect red blood cells, potentially leading to varying degrees of anemia across numerous mammalian species, including nonhuman primates. The present study aims to investigate the prevalence of hemoplasma infection and identify the species involved among free-ranging Assamese macaques (Macaca assamensis) inhabiting northern Thailand. A total of 133 blood samples were collected from Assamese macaques in Chiang Rai province, Thailand, and subjected to screening for hemoplasma infection utilizing nested PCR amplification targeting the 16S rRNA gene. Positive samples were subsequently analyzed through nucleotide sequencing and phylogenetic analysis for putative species identification. Current study results revealed that 17.3% (23/133; 95% CI 11.29-24.81) of Assamese macaques tested positive for hemoplasma infection using the nested PCR assay. Partial 16S rRNA sequences derived from hemoplasma isolates in Assamese macaques exhibited 99% homology, forming a cluster within the same phylogenetic clade as "Candidatus Mycoplasma haematomacacae," previously identified in long-tailed macaques, rhesus macaques, and Japanese macaques. These findings suggest the presence of "Ca. M. haematomacacae" not only in long-tailed macaques and rhesus macaques but also in Assamese macaques in Thailand. To our knowledge, this marks the first molecular detection of "Ca. M. haematomacacae" in Assamese macaques in Thailand. These results hold significance as they enhance our understanding of hemoplasma infection distribution among macaque populations in Thailand.

Isolation of Oral Bacteria, Measurement of the C-Reactive Protein, and Blood Clinical Parameters in Dogs with Oral Tumor.

Canine oral cancers have a poor prognosis and are related to chronic inflammation. This may pose a risk of secondary bacterial infection. This study aimed to compare the bacteria isolated from oral swab samples, values of C-reactive proteins (CRPs), and clinical blood profiles of dogs with and without oral mass. A total of 36 dogs were divided in three groups: no oral mass (n = 21), oral mass (n = 8), and metastasis groups (n = 7). Significantly, both the clinical groups (the oral mass group and metastasis group) showed anemia, a decrease in the albumin-to-globulin ratio (AGR), and an increase in the neutrophil-to-lymphocyte ratio (NLR), globulin-to-albumin ratio (GAR), CRP, and CRP-to-albumin ratio (CAR) compared to the normal group. CAR showed an increasing trend in the oral mass and metastasis groups (10 times and 100 times, respectively) compared to the no oral mass group (P < 0.001). Neisseria spp. (20.78%) was the main isolated bacteria in all groups. The main genera in the no oral mass group were Neisseria spp. (28.26%), Pasteurella spp. (19.57%), and Staphylococcus spp. (19.57%). Neisseria spp., Staphylococcus spp., Klebsiella spp., and Escherichia spp. were found equally (12.5%) in the oral mass group. Escherichia spp. (26.67%), Pseudomonas spp. (13.33%), and Staphylococcus spp. (13.33%) were the main genera in the metastasis group. Interestingly, Neisseria spp. decreased in the clinical groups (Fisher's exact = 6.39, P=0.048), and Escherichia spp. increased in the metastasis group (Fisher's exact = 14.00, P=0.002). The difference of oral bacteria in clinical dogs compared to healthy dogs may be related to microbiome alterations, and both the clinical groups showed the increment of inflammatory biomarkers. This suggested that further studies should be conducted on the correlation between the specific bacteria, CRP, blood clinical parameters, and type of canine oral mass.

First study on stress evaluation and reduction in hospitalized cats after neutering surgery.

Background and Aim: In Thailand, domestic cats are the most common companion animal, and many are admitted to veterinary clinics for neutering surgery; however, such environment can induce stress. This is the first study to evaluate stress in hospitalized cats after neutering surgery using cat stress score (CSS) and salivary cortisol levels, including the impact of providing a hiding box (B) and/or administering a pheromone product to reduce stress. Materials and Methods: The study design was based on a randomized controlled clinical trial. A total of 80 domestic cats undergoing routine neutering surgery were assessed for their behavioral demeanor scoring system (DSS) as friendly (DSS1) and aggressive (DSS2) based on a DSS. During admission, the cats were randomly allocated to single standard cages with one of the following treatments: (B), feline facial pheromone (P), a combination of hiding box and the pheromone (BP), or no additional enrichment (C). Cat stress score, food intake, and hide-seeking behavior were recorded. The cortisol enzyme-linked immunosorbent assay kit was used to assess the salivary cortisol level. Results: On the 1st day of admission, aggressive cats had a significantly higher CSS (4.16 ± 0.29) than friendly cats (3.27 ± 0.16). Both demeanor cat groups showed statistically significant reductions in stress levels earlier than the control group after providing the enrichments. Saliva cortisol measurements ranged from 0.24 to 0.66 ng/mL. No statistical differences in cortisol levels were observed between the 1st day and other days of admission. In contrast, no differences in food intake and hide-seeking behavior were seen within each group during the same period. Conclusion: Results suggested that stress and stress responses in cats depended on behavioral demeanor. The provision of enrichment, including hiding box and feline facial pheromone in singly housed caging reduced stress, especially in aggressive cats. However, salivary cortisol analysis, food intake, and hide-seeking behavior were ineffective for assessing stress in cats after neutering surgery.

Risk factors and clinical and laboratory findings associated with feline immunodeficiency virus and feline leukemia virus infections in Bangkok, Thailand.

Background and Aim: Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) are retroviruses associated with chronic and neoplastic diseases in domestic and non-domestic cats. There has been increasing interest in the clinical importance of feline retroviruses in Thailand and the identification of associated risk factors in domestic cats. To prevent the spread of retroviral diseases and improve the management of retrovirus-infected cats, risk factors and associated clinical laboratory data must be clearly understood. This study aimed to identify the influence of household, lifestyle, health status, sterilization, clinical presentations, and laboratory findings on FIV- and FeLV-infected cats in Bangkok, Thailand. Materials and Methods: A total of 480 cats were evaluated for FeLV p27 antigen and FIV antibodies using Witness FeLV-FIV Rapid Test and SNAP FIV/FeLV Combo Test at a veterinary hospital service. Results: Of the 480 cats tested, 113 were positive for virus infection, including 60 for FeLV (12.5%), 40 for FIV (8.3%), and 13 for both FeLV and FIV (2.7%). The findings revealed that the risk factors for cats infected with FeLV, FIV, or both FeLV and FIV were significantly different compared with those for non-infected cats (p < 0.05). Multivariate analysis showed that multi-cat ownership is a risk factor for the high prevalence of feline retrovirus infection, as multi-cat households exhibited a higher prevalence of infection than single-cat households. Anemic and sick cats were also at a greater risk of testing positive for specific retrovirus infections. FeLV-infected cats had a higher risk of anemia and low erythrocyte and thrombocyte counts (p ≤ 0.0001), whereas FIV-infected cats were more likely to have anemia and leukocytopenia than controls. Conclusion: Knowledge of the risk factors for retroviral diseases and associated clinical and laboratory findings can be used to develop strategies to reduce FIV and FeLV infections in cats.

Using bioelectrical impedance analysis for modeling regression and predicting body fat accumulation in dogs in East Thailand.

Background and Aim: Obesity in dogs leads to several health problems, such as premature death, and contributes to other diseases. Recently, body fat percentage has been considered to represent the body condition of dogs, and bioelectrical impedance analysis (BIA) is the most effective method for accurately measuring body fat in dogs. In Thailand, information on the body condition of dogs is limited, and there is no standard body fat level for Thai or mongrel dogs. This study was designed to evaluate and analyze the body fat percentage in dogs through BIA using a handheld instrument. The results of this study can help enhance the quality of life and health of dogs and aid in setting a standard body fat level for Thai or mongrel dogs. Materials and Methods: The body fat percentage of 340 Thai and mongrel dogs in East Thailand was measured in the standing position, and the body condition score (BCS) (range, 1-5), sex, sterilization status, age, type of diet, and lifestyle were recorded. A linear regression model was developed to compare the variables and the predicted body fat percentage, and multiple linear regressions were used to analyze the factors for body fat increment. Results: The linear regression model used to estimate the percentage of body fat (y) for each BCS (x) was y = 0.84 + 8.36x (R2 = 0.7219; p < 0.0001); the average body fat percentage was 27.52% for all studied dogs; specifically, 24.83% for the Thai Bangkaew, 26.42% for the Thai Ridgeback, and 27.65% for mongrels. The median body fat percentage was significantly higher in female than in male dogs. We found that as age increases, body fat percentage also increases; this increasing trend begins at the age of 5 years. However, increasing the level of activity and decreasing meal frequency leads to an increase in body fat percentage in neutered male dogs. Conclusion: The average body fat percentage of dogs in East Thailand is 27.52% and this value is expected to increase when these dogs reach the age of 5 years. BIA is a valid and effective measurement tool for detecting the body fat percentage in dogs.

Molecular detection and phylogeny of Ehrlichia canis and Anaplasma platys in naturally infected dogs in Central and Northeast Thailand.

Background and Aim: Ehrlichia canis and Anaplasma platys are tick-borne, Gram-negative bacteria that cause canine monocytic ehrlichiosis and canine cyclic thrombocytopenia, respectively. These diseases are of great importance and are distributed globally. This study aimed to create new primers for the identification of E. canis and A. platys in naturally infected dogs using polymerase chain reaction (PCR), DNA sequencing, and phylogenetic analysis using the 16S rDNA and gltA genes. Materials and Methods: In total, 120 blood samples were collected from dogs in three different locations (Saraburi, Buriram, and Nakhon Ratchasima provinces) in Central and Northeast Thailand. The molecular prevalence of E. canis and A. platys was assessed using PCR targeting the 16S rDNA and gltA genes. All positive PCR amplicons were sequenced, and phylogenetic trees were constructed based on the maximum likelihood method. Results: Ehrlichia canis had an overall molecular prevalence of 15.8% based on the 16S rDNA gene, compared to 8.3% based on the gltA gene. In addition, the overall molecular prevalence of A. platys using the 16S rDNA gene was 10.8%, while the prevalence rate was 5.8% using the gltA gene. Coinfection was 0.8% in Saraburi province. The partial sequences of the 16S rDNA and gltA genes of E. canis and A. platys in dogs in Central and Northeast Thailand showed 96.75%-100% identity to reference sequences in GenBank. Phylogenetic analysis of the 16S rDNA and gltA genes revealed that E. canis and A. platys sequences were clearly grouped into their own clades. Conclusion: This study demonstrated the molecular prevalence of E. canis and A. platys in Central and Northeast Thailand. The 16S rDNA and gltA genes were useful for the diagnosis of E. canis and A. platys. Based on the phylogenetic analysis, the partial sequences of the 16S rDNA and gltA genes in E. canis and A. platys were related to prior Thai strains and those from other countries.

Molecular identification of Trichuris trichiura and Hymenolepis diminuta in long-tailed macaques (Macaca fascicularis) in Lopburi, Thailand.

BACKGROUND AND AIM: Trichuris trichiura and Hymenolepis diminuta are helminthic intestinal parasites that infect humans and other animals, including non-human primates. However, molecular detection of these parasites remains scarce in long-tailed macaques (Macaca fascicularis), which coexist with human communities in Thailand. Thus, this study aimed to molecularly confirm the occurrence of Trichuris spp. and Hymenolepis spp. infection and determine the species of both parasites that were found in long-tailed macaques. MATERIALS AND METHODS: A total of 200 fecal samples were randomly collected from long-tailed macaques living in Lopburi, Thailand, and tested based on polymerase chain reaction (PCR) assays for Trichuris spp. and Hymenolepis spp. infections. The PCR products were submitted for DNA purification and sequencing. Phylogenetic analysis was performed using the maximum likelihood method. RESULTS: Of 200 tested samples, three (1.5%) were positive for Trichuris spp. Sequence analysis of all positive samples revealed the presence of T. trichiura, while eight samples (8/200, 4%) positive for Hymenolepis spp. were classified as H. diminuta. No significant associations were found between parasite infection and sex of macaques. CONCLUSION: This study revealed that long-tailed macaques harbor T. trichiura and H. diminuta. These results suggested that local residents and tourists must pay attention to limiting contact with long-tailed macaques and take hygienic precautions to reduce the risk of zoonotic and anthroponotic transmission of these parasites between humans and long-tailed macaques.

Detection and genetic characterization of "Candidatus Mycoplasma haemomacaque" infection among long-tailed macaques (Macaca fascicularis) in Thailand using broad-range nested polymerase chain reaction assay.

BACKGROUND AND AIM: Hemoplasmas are defined as small, epicellular parasitic bacteria that can infect the red blood cells of several mammalian species. Diseases caused by these bacteria range from asymptomatic infections to acute hemolytic anemia. However, data on hemoplasmas in non-human primates in Thailand remain to be limited. Therefore, this study aims to determine the occurrence and genetic diversity of hemoplasmas among long-tailed macaques in Thailand. MATERIALS AND METHODS: Blood samples were collected from 339 long-tailed macaques in three provinces of Thailand. DNA was then extracted from the blood samples and tested for hemoplasma using broad-range nested polymerase chain reaction (PCR) based on the 16S rRNA gene. PCR-positive samples were sequenced, and phylogenetic analysis for species identification was conducted. RESULTS: In total, 38 (11.2%) out of the 339 samples were found to be positive for hemoplasmas, based on the broad-range nested PCR assay of the 16S rRNA gene. The 16S rRNA sequences of Mycoplasma spp. were highly similar (98-99% identity) to "Candidatus Mycoplasma haemomacaque." Furthermore, phylogenetic analysis using maximum likelihood demonstrated that the sequences were located in the same cluster of "Ca. M. haemomacaque." CONCLUSION: The detection of hemoplasmas among long-tailed macaques in Thailand is reported. Genetic characterization confirmed that these hemoplasmas are closely related to "Ca. M. haemomacaque." These results indicate that long-tailed macaques in several locations in Thailand may be infected and serve as reservoirs for this parasite.

The effectiveness of marine based fatty acid compound (PCSO-524) and firocoxib in the treatment of canine osteoarthritis.

BACKGROUND: NSAIDs are accepted as the most predictably efficacious medical treatment of the clinical signs of osteoarthritis (OA). The marine-based fatty-acid compound PCSO-524 has been proposed as an adjunctive treatment for canine OA, however benefits of this agent is still controversial. The purpose of this study was to evaluate and compare the effectiveness of PCSO-524 combined with the NSAID firocoxib using force plate gait analysis, orthopedic assessment score (OAS) and canine brief pain inventory score (CBPI) in dogs with OA. A prospective, randomized, double-blinded study was conducted. Seventy-nine dogs that had hip and/or stifle OA were assigned randomly into three treatment groups: firocoxib, PCSO-524 and combination of firocoxib and PCSO-524, orally for 4 weeks. Peak vertical force (PVF, expressed as a percentage of bodyweight), OAS, CBPI, serum prostaglandin E2 concentration, hematology and blood chemistry values were evaluated before treatment (Day0), as well as at the second (Day14) and fourth week (Day28) during treatment. RESULTS: Within group analysis revealed significant increases in PVF over the 4-week treatment period for firocoxib, PCSO-524 and the combination (p < 0.05). Mean increases in PVF were 3.25 ± 4.13, 2.01 ± 3.86, 4.11 ± 4.69%BW (mean ± SD) respectively. The OAS showed non-significant change in all treatment groups. There were significant decreases in CBPI pain severity score (PSS) and CBPI pain interference scores (PIS) within some groups over time, however no significant differences were found between the groups. Significantly decreased serum PGE2 concentration (p < 0.05) was found in the combination group. Significant increases in BUN and creatinine (p < 0.05) compared to pre-treatment values were found in the firocoxib and combination groups but not in the PCSO-524 group at day28, but all values in all dogs remained within the normal ranges. CONCLUSIONS: The results of this study suggested combination of both PCSO-524 and firocoxib is more effective in alleviation of inflammation and improvement of weight bearing ability when compared to the uses of either PCSO-524 or firocoxib alone. Further clinical studies are needed to confirm this, and to determine if there is any benefit of PCSO-524 over placebo.