Performance and Cost-Efficiency of Single Hormonal Treatment Protocols in Tropical Anestrous Dairy Cows.

This retrospective study aimed to evaluate the performance of hormone treatment protocols, determine the factors associated with pregnancy success after hormone treatment, and compare the cost-efficiencies of two types of hormone treatment among cyclic and noncyclic anestrous dairy cows. The clinical records of 279 anestrous cows that received hormone treatment for artificial insemination (AI) from 64 herds in the western region of Thailand were obtained from Kasetsart University Veterinary Teaching Hospital from January to August 2017. The performance of the hormone treatment protocols, fixed-time AI (TAI) and estrus detection before AI (EAI), showed that the pregnancy risk for the TAI protocol was higher than that for the EAI protocol, but pregnancy per AI did not differ significantly between the two protocols in cyclic and noncyclic cows. Multivariate logistic regression analysis showed that cows receiving the TAI protocol were more likely to be pregnant compared to those treated with the EAI protocol. Cows with a 3.00 body condition score (BCS) < 3.75 after treatment and loose-housed cows were more likely to become pregnant. Treatment during winter showed higher pregnancy success than that in the summer and rainy seasons. The cost-efficiency analysis showed that the TAI protocol was the most cost-efficient option for noncyclic cows, whereas the EAI protocol was the most cost-efficient option for cyclic cows.

Isolation of bacteriophages specific to bovine mastitis-causing bacteria and characterization of their lytic activity in pasteurized milk.

Background and Aim: Bovine mastitis is one of the most serious issues in dairy production. It is caused by contagious and coliform pathogens such as Staphylococcus spp., Escherichia coli, and Klebsiella pneumoniae. In addition, the emergence of drug-resistant bacteria raises urgent concerns in the field of drug treatment, thus requiring the exploration of alternative treatments. Bacteriophage therapy has been shown to be a promising alternative approach for the control of antibiotic-resistant pathogens. In this study, we aimed to isolate phages specific to contagious mastitis and coliform mastitis, characterize the isolated phages, and examine their ability to lyse bacteria in pasteurized milk samples. Materials and Methods: The Staphylococcus phage vB_Sau-RP15 isolated from raw milk in our previous study was used in this study. Other three phages, vB_Eco-RN12i1, vB_Kpn-RN14i1, and vB_Ssc-RN20i3, were isolated from wastewater using E. coli 5823, K. pneumoniae 194, and Staphylococcus sciuri MM01 as hosts, respectively. The host range and efficiency of plating (EOP) were determined following phage isolation. Moreover, the lysis activities of these phages against their hosts were investigated in pasteurized milk using a multiplicity of infections (MOIs) of 10 and 100 at 37°C. Phages were applied using individual and combination phages. Results: According to the EOP results, all phages showed high specificity to their respective hosts. They are tailed phages with distinct morphologies. Individual phage treatments in spiked pasteurized milk with their respective bacterial hosts significantly reduced the bacterial counts in both MOI conditions during the first 2 h of the treatment (approximately 1-8 log reduction compared to the control). Although these phages specifically infected only their hosts, the phage cocktail resulted in a better result compared to the use of individual phage. However, bacterial regrowth was observed in all experiments, which may be related to the development of phage-insensitive mutants. Conclusion: Our findings suggest that the application of phages could be used to treat bovine mastitis. Phage cocktail is suitable to promote the efficacy of phage treatment in pasteurized milk. However, when considering the use of phages in dairy cows, certain phage properties in raw milk and in vivo and ex vivo should be highlighted to ensure their effectiveness as biocontrol agents for bovine mastitis treatment.

The influence of subclinical mastitis on the protein composition and protease activities of raw milk from lactating Thai-crossbred dairy cows.

Background and Aim: Mastitis in dairy cattle is associated with a high rate of morbidity and death, which has major implications for milk production and quality. This study aimed to investigate the protein component and the activity of matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) in raw milk samples with different testing scores determined using the California mastitis test (CMT). Materials and Methods: Thirty cows were employed in the study, and milk from each quarter was tested for subclinical mastitis (SCM). According to the results of CMT, raw milk samples were classified into five categories: Healthy (score 0), trace (score T), weakly positive (score 1), distinctly positive (score 2), and strongly positive (score 3) for somatic cell count (SCC). The total milk protein was analyzed using the Bio-Rad protein assay, and the milk protein composition was determined using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis technique. In addition, gelatin zymography was used to evaluate changes in proteolytic abilities. Results: Milk samples with CMT scores of 1 and 3 had the highest total milk protein levels (32.25 ± 12.60 g/L and 32.50 ± 7.67 g/L, respectively), while the samples from healthy cows (CMT score 0) were only 6.75 ± 1.64 g/L. Globulin and lactoferrin were significantly increased in samples with a CMT score of 3 compared with those with other CMT scores. The bovine serum albumin level in samples with a CMT score of 2 was significantly (p < 0.05) higher than those with other CMT scores. No significant differences in casein abundance were found among samples with different CMT scores. Results from analysis of proteolytic activities demonstrated that the level of MMP-9 in samples with a CMT score of 3 was significantly (p < 0.05) higher than those with other CMT scores. Conclusion: The protein content and gelatinolytic activity of milk were drastically altered by the number of SCC, mainly due to SCM.

Long-term monitoring of immune response to recombinant lumpy skin disease virus in dairy cattle from small-household farms in western Thailand.

Lumpy skin disease (LSD) was firstly reported in Thailand in 2021 which affected the cattle industry. However, there is limited information on the immune response of LSDV infection in Thailand where recombinant vaccine strain circulated. The aim of this research was to study the duration of LSD immune response of subclinical and clinical animals after natural infection in dairy cattle. Sixty-six dairy cattle from ten farms in central and western regions of Thailand were investigated. Antibody was detected by virus neutralization test and ELISA. Cell mediated immunity (CMI)-related cytokine gene expressions were evaluated. Antibody was detected until at least 15 months after the noticeable symptom. Cattle with subclinical disease had lower antibody levels compared to animals which had clinical disease. IFN-γ and TNF-α levels were increased, while IL-10 level was decreased in the infected animals compared to the controls. This study elucidated immune responses in dairy cattle herd affected by recombinant LSDV.

Effect of claw blocks on the healing duration and lesion severities of claw lesions in lame cows in Western Thailand.

Background and Aim: Lameness is a major complication in dairy cattle affecting health and milk production. Several factors are found to contribute to this condition and specific treatments are required, including the process of claw trimming. The elevation of the claw, such as with the application of a claw block, was reported to be beneficial in the more severe cases. This study aimed to determine the efficiency of a claw block on claw lesions of lame cows in dairy farms in Western Thailand. Materials and Methods: Locomotion scores of 376 dairy cows were determined by a veterinarian using a scale of 1-5 (1 = normal; 5 = severely lame) at the time of the visit. Cows with a score of 3 or greater were defined as clinically lame. In total, 134 clinically lame cows from 11 dairy farms in the Kanchanaburi and Ratchaburi provinces were included in the analysis. Claw lesions included a white line abscess, bruised sole, sole ulcer, sole abscess, white line separate, and double soles. Wooden or rubber claw blocks were applied to the unaffected claw of the same hoof as the injured claw of 116 cows, which were classified as the treatment cases, and 18 cows were left untreated and classified as the control cases. Each cow was checked on every week of the healing process for 2 months unless the cow was culled earlier. Survival analysis was based on the Kaplan-Meier estimator and Cox Proportional Hazard regression. Results: The median healing time for lame cows with and without claw blocks was 21 and 36 days, respectively. After adjusting for the lesion severity and type, the lame cows with and without a claw block had hazard ratios of 2.16 and 3.08, respectively. The healing times between the four lesion types in cows with a claw block were not significantly different. The healing time was longer in lame cows, with a severity score of 4. Conclusion: The results from this study reveal that the treatment of lame cows with claw blocks promoted the healing capacity of claw lesions after claw trimming.

Rapid Spread and Genetic Characterisation of a Recently Emerged Recombinant Lumpy Skin Disease Virus in Thailand.

The emergence of the lumpy skin disease virus (LSDV) was first detected in north-eastern Thailand in March 2021. Since then, the abrupt increase of LSD cases was observed throughout the country as outbreaks have spread rapidly to 64 out of a total of 77 provinces within four months. Blood, milk, and nodular skin samples collected from affected animals have been diagnosed by real-time PCR targeting the p32 gene. LSDV was isolated by primary lamb testis (PLT) cells, followed by Madin-Darby bovine kidney (MDBK) cells, and confirmed by immunoperoxidase monolayer assay (IPMA). Histopathology and immunohistochemistry (IHC) of a skin lesion showed inclusion bodies in keratinocytes and skin epithelial cells. Phylogenetic analyses of RPO30 and GPCR genes, and the whole genome revealed that Thai viruses were closely related to the vaccine-derived recombinant LSDV strains found previously in China and Vietnam. Recombination analysis confirmed that the Thai LSDV possesses a mosaic hybrid genome containing the vaccine virus DNA as the backbone and a field strain DNA as the minor donor. This is an inclusive report on the disease distributions, complete diagnoses, and genetic characterisation of LSDV during the first wave of LSD outbreaks in Thailand.

Seroprevalence of bovine coronavirus and factors associated with the serological status in dairy cattle in the western region of Thailand.

BACKGROUND AND AIMS: Bovine coronavirus (BCoV) is a pathogen affecting the productivities of dairy cattle worldwide. The present study aimed to determine the seroprevalence and factors associated with BCoV serological status using a commercial indirect enzyme-linked immunosorbent assay (ELISA). MATERIALS AND METHODS: A cross-sectional study was conducted in the western region of Thailand. Blood samples were collected from 30 dairy herds. In total, 617 blood serum samples were tested using a commercial indirect ELISA for BCoV-specific immunoglobulin G antibodies. A questionnaire was used to collect data on the factors which have been identified as risk factors for BCoV antibody detection. The age and history of diarrhea of each animal were recorded. Fisher's exact test was performed to univariately assess the association between BCoV serological status and possible risk factors. Variables with Fisher's exact test p<0.10 were then evaluated using multivariate logistic regression to identify factors associated with BCoV serological status. The Bonferroni adjustment was used for multiple comparisons of significant variables in the final multivariate logistic regression model. RESULTS: No herd was free from antibodies to BCoV. The individual seroprevalence of BCoV was 97.89% (604/617). The prevalence within herds was in the range of 45.45-100%. Cattle >3 years of age were more likely to be seropositive to BCoV compared to cattle <1 year of age (p=0.003), with the odds ratio being 81.96. Disinfecting diarrhea stools were a protective factor for being BCoV seropositive, with odds ratios of 0.08 and 0.06 compared to doing nothing (p=0.008) and to clean with water (p=0.002), respectively. CONCLUSION: BCoV seropositive dairy cattle were distributed throughout the western region of Thailand. The probability of being seropositive for BCoV increased with increasing animal age. Cleaning the contaminated stool with appropriate disinfectants should be recommended to farmers to minimize the spread of the virus.

Association between milk urea nitrogen and first service conception in smallholder dairy farms under heat and humidity stress.

AIM: The study was to evaluate the relationship between the first service conception (FSC) and milk urea nitrogen (MUN) in smallholder dairy farms under heat and humidity stress. MATERIALS AND METHODS: Dairy cows from 43 dairy farms giving birth between November 2014 and April 2015 (n=295) contributed to the study. All cows were sampled monthly to measure milk compositions, and we collected additional farm data from farmers through a questionnaire. The first service during the first 120 days of lactation was the outcome of interest in this study. Multivariable logistic regression determined significant associations with FSC. RESULTS: The overall FSC was 22% and the mean MUN concentration was 11.55 mg/dl. The final FSC model included MUN concentration, the season of breeding, and protein energy ratio (PE ratio) in the diet. The odds of FSC were reduced by approximately 10% for each mg/dl higher MUN on the day of the milk sample that was nearest to the artificial insemination (AI) day. The odds of FSC were nearly 3 times higher when the first insemination occurred in winter compared to summer first services. Taking into account the nutritional factors, the odds of FSC were nearly 70% higher with an increase in PE ratio of 10 g of crude protein/Mcal from the mean of 35.90 g. CONCLUSION: This study of smallholder dairy farmers in the hot and humid climate of Thailand confirmed that season, nutritional management, and MUN concentration were associated with FSC. MUN appears to be a useful indicator to monitor the effects of diet on reproductive performance from this study.

Molecular characterization of bovine ephemeral fever virus in Thailand between 2013 and 2017.

Bovine ephemeral fever (BEF) is an arthropod-borne disease caused by bovine ephemeral fever virus (BEFV), a negative sense, single-stranded RNA virus. BEFV is endemic in tropical and sub-tropical regions including Thailand, a country in mainland Southeast Asia. However, there are few studies on BEFV and no available information regarding molecular characteristics of BEFV in Thailand. Therefore, the aims of this study were to genetically characterize Thai BEFVs and reveal their evolutions by phylogenetic analysis of G gene ectodomain sequences. From 2013 to 2017, blood samples were collected from bovine that matched with BEF case definition from three regions of Thailand. Thai BEFV G genes and a whole genome of an isolate, East Asia/TH/LRI0045/2016, were sequenced and characterized. Additionally, their phylogenies were constructed. This is the first report on genetics of BEFV in Southeast Asia. G ectodomain encoding region of Thai BEFV found during 2013-2017 are closely related to the second and third sub-clades of East Asia lineage. In addition, we observed mutation in the putative P' ORF of all Thai BEFVs which generated a premature stop codon. Thai G gene sequences are closely related to those of mainland Chinese and Taiwanese isolates. The whole genomic sequences of Thai BEFV and East Asia/China/JT02 L/2002 possess common characteristics, suggesting shared evolutionary relationship between East and Southeast Asian strains. Further studies on relationship between animal translocation, circulation of BEFV in Greater Mekong subregion and acquisition of more G gene sequences may improve understanding of BEFV epidemiology in mainland Southeast Asia.

Exploring factors associated with bulk tank milk urea nitrogen in Central Thailand.

AIM: The study was to determine seasonal fluctuations and non-nutritional factors associated with bulk tank milk urea nitrogen(BTMUN). MATERIALS AND METHODS: A total of 58,364 BTM testing records were collected from 2364 farms in Central Thailand during September 2014-August 2015. Using square root BTMUN as the outcome, other milk components, farm effect, and sampling time were analyzed by univariable repeated measures linear regression, and significant variables were included in multivariable repeated measures linear regression. RESULTS: The average BTMUN (standard deviation) was 4.71 (±1.16) mmol/L. In the final model, BTM fat and protein percentages were associated with BTMUN as quadratic and cubic polynomials, respectively. BTM lactose percentage and the natural logarithm of somatic cell counts were negatively linearly associated with BTMUN. At the farm level, the BTM lactose association was negatively linear; herd BTMUN decreased following an increase of herd lactose average, and BTM lactose slopes were quite different among farms as well. Sampling time had the highest potency for the estimation of BTMUN over time, with lows and highs occurring in August and October, respectively. The variation in test level BTMUN was decreased by 18.6% compared to the null model, and 6% of the variance could be explained at the farm level. CONCLUSION: The results clarify seasonal variation in BTMUN and the relationships among other BTM constituents and BTMUN, which may be useful for understanding how to manage lactating dairy cattle better to keep BTM constituents within normal ranges.

Molecular and phylogenetic characterization of bovine coronavirus virus isolated from dairy cattle in Central Region, Thailand.

Bovine coronavirus (BCoV) is involved mainly in enteric infections in cattle. This study reports the first molecular detection of BCoV in a diarrhea outbreak in dairy cows in the Central Region, Thailand. BCoV was molecularly detected from bloody diarrheic cattle feces by using nested PCR. Agarose gel electrophoresis of three diarrheic fecal samples yielded from the 25 samples desired amplicons that were 488 base pairs and sequencing substantiated that have BCoV. The sequence alignment indicated that nucleotide and amino acid sequences, the three TWD isolated in Thailand, were more quite homologous to each other (amino acid at position 39 of TWD1, TWD3 was proline, but TWD2 was serine) and closely related to OK-0514-3strain (virulent respiratory strain; RBCoV).The amino acid sequencing identities among TWD1, TWD2,TWD3, and OK-0514-3 strain were 96.0 to 96.6%, those at which T3I, H65N, D87G, H127Y, andQ136R were changed. In addition, the phylogenetic tree of the hypervariable region S1subunit spike glycoprotein BCoV gene was composed of three major clades by using the 54 sequences generated and showed that the evolutionally distance, TWD1, TWD2, and TWD3 were the isolated group together and most similar to OK-0514-3 strain (98.2 to 98.5% similarity). Further study will develop ELISA assay for serologic detection of winter dysentery disease.

Influence of colostral quality on serum proteins in dairy calves raised in smallholder farms in Thailand.

The objective of this study was to analyze the influence of colostral quality on serum proteins in calves. Samples were collected from visited farms in Kasetsart University Veterinary Teaching Hospital at Kamphaeng Saen and Nong Pho Animal Hospital. In total, 35 dairy farms contributed 80 dams and calves' samples. Colostrum samples from 80 dairy cows and blood samples from their calves were taken to evaluate colostral immunoglobulins (Ig) and immunoglobulin G (IgG), and calf serum protein and IgG. Total colostral Ig, colostral and serum IgG, and serum protein were measured by a colostrometer, single radial immunodiffusion, and refractrometer, respectively. Immunoglobulin G and serum protein concentrations increased in the 1st day after birth, and maximum concentrations were seen in the 2nd day and then decreased in the 7th and 14th days. Average ± SD total colostral IgG concentrations at calving date and at 1 and 2 days after calving were 93.85 ± 33.89, 37.11 ± 23.51, and 17.23 ± 9.4 mg/mL, respectively. The profile of total Ig and IgG concentrations in colostrum had a similar pattern, with the maximum concentrations obtained in calving date and rapidly decreased thereafter. Low IgG concentrations were seen in the 7th and 14th day after calving. The calves that were fed with high quality colostrum had higher serum protein at 1 day of age, 7.49 ± 1.01 g/dL, than calves fed with low quality colostrum, 6.40 ± 0.86 g/dL (P < 0.01). The increase in serum protein after first colostrum feeding of high and low quality colostrum was 1.55 ± 1.07 and 0.81 ± 0.69 g/dL, respectively (P = 0.02).

Prevalence and risk factors for intestinal parasite infection in goats raised in Nakhon Pathom Province, Thailand.

The objectives of this study were to determine the prevalence and risk factors for intestinal parasite infection in goats raised in Nakhon Pathom Province. Fecal samples from 190 goats in 12 herds were collected per rectum. Questionnaires focusing on the general information about the farm and farmers, and management practices on farms were completed. Modified McMaster counting technique was performed to detect eggs of intestinal parasites. Each egg found was classified into three groups of parasites based on its characteristics. Individual and herd prevalence were calculated. Multiple logistic regressions were performed to analyze risk factors of infection. Herd prevalence of intestinal parasite infection was 100%, while individual prevalence was 79.47%. Strongyle group was the most common intestinal parasite found in this study. Moniezia spp. (8/190) and Trichuris spp. (1/190) were also found. The average number of eggs found was 1,176 eggs per gram of feces. Risk factors for intestinal parasite infection were housing system, deworming interval, and type of goat in herd. Goats housed in groups had higher infection rates compared with goats housed individually (odds ratio (OR) = 6.34; P value = 0.009). Goats in herds in which anthelmintic drugs were administered in intervals of greater than 3 months were more likely to become infected with intestinal parasites (OR = 33.07; P value <0.001). Goats in herds that kept only dairy goats were less likely to become infected than herd that kept only meat goats or kept both dairy and meat goats (OR = 46.20 and 8.75; P value <0.001 and 0.011, respectively).

Risk factors of Neospora caninum infection in dogs and cats in dairy farms in Western Thailand.

The objectives of this study were to investigate the risk factors of Neospora caninum infection in dogs and cats found in dairy herds in western Thailand. A case-control study was conducted in dairy herds in three western provinces including Nakhon Pathom, Ratchaburi and Kanchanaburi. Blood samples of pets from 14 positive dairy herds and 26 herds from negative neighbouring farms which were randomly selected, in total blood samples from dogs and cats from 40 herds were collected and examined for antibodies against N. caninum infections using competitive ELISA and indirect fluorescent antibody test (IFAT). No seropositive cats were found. Of the 38 positive dogs, four (10.5%) were seropositive to N. caninum, which was higher than the proportion of seropositive in the negative herd population (one of 76, 1.3%). The higher proportion of seropositive farm dogs as compared with neighbouring dogs tended to be significant (P = 0.055).

Relationships among milk urea-nitrogen, dietary parameters, and fecal nitrogen in commercial dairy herds.

Our study objective was to determine the ability of milk urea-nitrogen concentrations ([MUN]) to predict fecal nitrogen concentrations ([Fecal N]) in commercial dairy herds. A total of 83 dairy herds were each visited 3 times within 48 h after a monthly herd milk test. For each farm visit, forages were sampled for nutrient analyses, which were entered into a computerized ration evaluator, and fecal samples were taken per rectum from each of 6 cows (2 early-, 2 mid-, and 2 late-lactation). Fecal samples were pooled, mixed, and analyzed for nitrogen content. Fecal nitrogen concentrations were compared with the routinely measured nutritional parameters from the ration evaluation, and the herd average [MUN] for the previous milk test date using mixed linear regression analyses. Total protein supplied in the ration was significantly positively associated with [Fecal N], but herd average [MUN] was not associated (P > 0.10) with [Fecal N].