Background and Aim: African horse sickness (AHS) has become a newly emerging disease after an outbreak in northeastern Thailand in March 2020. Mass vaccination in horses with live-attenuated AHS virus (AHSV) vaccine is essential for AHS control and prevention. This study aimed to monitor the longitudinal humoral immune response before and after a single vaccination using a live-attenuated vaccine against AHS in stallions, mares, and pregnant mares, including maternal immunity in foals born from pregnant mares during the outbreak in Thailand. Materials and Methods: A total of 13 stallions and 23 non-pregnant and 21 pregnant mares were vaccinated with live-attenuated AHSV vaccines. Serum samples from selected horses were collected on the day of vaccination and 1, 6, 8, 9, 10, and 12-months post-vaccination. Furthermore, seven serum samples of foals born from vaccinated pregnant mares were collected on parturition date and 1, 3, and 6-months old. The antibody titer against AHS in all collected serum samples was evaluated using a commercial enzyme-linked immunosorbent assay kit. All data were analyzed for mean and standard deviation for each group of samples using a spreadsheet program. Antibody titers between times were analyzed using a one-way analysis of variance as repeated measurement, and antibody titers between horse groups were analyzed using a general linear model for statistically significant differences when p < 0.05. Results: In stallion and non-pregnant mare groups, there were no statistically significant differences in antibody titers in all 6 time periods after vaccination. The antibody titer in the pregnant mare group showed a non-statistically significant difference between each gestation stage, except at 8 months post-vaccination. Furthermore, increasing antibody titers on days 1 and 3 after receiving colostrum in foals indicate the major role of transcolostral antibody transfer for AHS. Conclusion: This study demonstrated that a single AHS vaccination using a live-attenuated vaccine could stimulate high antibody titers sufficient for AHS control and prevention during the outbreak in Thailand. Similarly, the antibody response of vaccinated horses of both genders, including various stages of pregnant mares, was statistically not different. Keywords: African horse sickness, antibody, maternal immunity, stage of gestation, vaccination.

External hoof characteristics, balance, and conformation have been extensively studied in horses; however, mules remain understudied in these aspects. This study evaluated the size, shape, and symmetry of untrimmed and unshod forelimb hooves, compared the symmetry between forelimb hooves and stratified external forelimb hoof measurements based on the body condition score of mules raised in the foothill plains of northern Thailand. The forelimb hooves of 38 mules were photographed and 33 parameters, including angular and linear measurements, were analyzed. A multivariate analysis was used to explore the influence of sex, age, and body condition scores (BCS) on angular, linear, and area parameters. Additionally, one-way ANOVA followed by Tukey’s test was used to compare these parameters across different BCS groups. Despite the absence of shoeing and trimming, these mules exhibited optimal left–right forelimb hoof symmetry, with no significant (p < 0.05) differences in: outer wall length and inner wall length (OWL–IWL: Left 0.11 ± 0.66 cm; Right −0.12 ± 0.43 cm); sole length and sole width (SL^S–SW: Left 1.65 ± 0.76 cm; Right 1.46 ± 0.89 cm); dorsal hoof wall length and heel length (DHWL–HL: Left 4.00 ± 0.80 cm; Right 3.81 ± 0.72 cm); and frog length and frog width (FL–FW: Left 3.88 ± 1.13 cm; Right 3.82 ± 0.18 cm). However, significant (p < 0.05) differences were observed within each body condition score group for forelimb hoof measurements for DHWL, IWL, heel separation (HS), heel bulb distance (HBD), SW, FW, and FL, while sex and age had no significant differences across the study variables. These findings provide valuable insights into mule welfare and management, contributing to understanding of the interplay between overall health and hoof conformation in the study area.

External hoof characteristics, balance, and conformation have been extensively studied in horses; however, mules remain understudied in these aspects. This study evaluated the size, shape, and symmetry of untrimmed and unshod forelimb hooves, compared the symmetry between forelimb hooves and stratified external forelimb hoof measurements based on the body condition score of mules raised in the foothill plains of northern Thailand. The forelimb hooves of 38 mules were photographed and 33 parameters, including angular and linear measurements, were analyzed. Despite the absence of shoeing and trimming, these mules exhibited optimal left-right forelimb hoof symmetry, with no significant (p < 0.05) differences in: outer wall length and inner wall length (OWL - IWL); sole length and sole width (SLS - SW); dorsal hoof wall length and heel length (DHWL - HL); and frog length and frog width (FL - FW). However, significant (p < 0.05) differences were observed within each body condition score group for forelimb hoof measurements for DHWL, IWL, heel separation (HS), heel bulb distance (HBD), SW, FW, and FL. These findings provide valuable insights into mule welfare and management, contributing to understanding of the interplay between overall health and hoof conformation in the study area.

Trypanosomosis (Surra), caused by the hemoflagellate protozoan Trypanosoma evansi, has a negative impact on animal health and the livestock economy in many countries, including Thailand. Molecular epidemiological data and integrative control approaches for small-scale farms remain limited. This study determined the prevalence of Trypanozoon infection and its associated risk factors in Thai livestock under an integrative control approach. Molecular tools were used to characterize T. evansi typing. A total of 947 blood samples were collected from cattle and buffalo across 123 small-scale farms in 16 provinces covering four regions of Thailand, with diagnostic results and control recommendations communicated to farmers within 14 days. The ITS2-positive samples were then subjected to further molecular characterization, together with two dogs, seven horses, and an in vitro T. evansi isolate, comprising 40 Trypanozoon-positive samples. These were characterized using PCR targeting the TBR primer, maxicircle NADH5, VSG RoTat 1.2, and minicircle B, followed by ITS2-based phylogenetic analysis. Risk factors were assessed using Chi-square and Fisher’s exact tests, logistic regression, a generalized linear mixed model, and Firth’s penalized logistic regression. The overall prevalence of Trypanozoon infection was 3.17% (30/947), detected exclusively in beef cattle in the Northern region (15.30%; 30/196), specifically in Lampang (46.15%, 12/26), Tak (23.08%, 3/13), and Phrae (17.24%, 15/87) Provinces. No infections were reported in the Northeastern, Central, or Southern regions. Male sex, age under one year, and residence in the Northern region were significant individual-level risk factors, while farm-level lacrimation and the presence of Stomoxys were significantly associated with infection. Molecular characterization identified T. evansi type A and type non-A/B, with no type B or maxicircle-positive samples detected. Phylogenetic analysis grouped the Thai isolates with previously reported isolates from Thai deer and cattle, Iranian camels, and Colombian dogs. On follow-up, no Trypanozoon infection was detected in revisited farms. These findings confirmed that T. evansi, comprising types A and non-A/B, continues to circulate in Thai livestock, particularly in beef cattle in Northern Thailand. Data from this study can be used for surveillance and future development of more effective diagnostic systems.