Blood parasite infections in poultry, such as Plasmodium, are a serious threat to the poultry industry due to their potential to cause economic losses. To date, there has been inadequate research on the morphological and molecular detection of the different Plasmodium species that infect poultry in Indonesia. Therefore, this study aimed to analyze the morphological and molecular characteristics of Plasmodium spp. and the several predisposing factors for Plasmodium infection in layer chickens from three districts of Yogyakarta, Indonesia. One hundred and five blood samples from layer chickens were collected from 13 farms located in three districts of Yogyakarta (Sleman, Bantul, and Kulon Progo) between September and November 2022. Blood samples were subjected to microscopic and polymerase chain reaction (PCR) analyses. Sequencing was performed using basic local alignment search tools to identify the nucleotide structure of cytochrome b. Phylogenetic analysis of Plasmodium was performed using the MEGA-X software. Microscopic examination revealed that 17/105 positives (16.19%) were positive for blood parasite infection. Trophozoites, erythrocytic meronts, and microgametocytes of Plasmodium were found in blood samples. Based on the morphological examination, the species found in the samples was close to Plasmodium juxtanucleare. Polymerase chain reaction examination revealed that 21/60 samples were positive for Plasmodium (35%). The Plasmodium species identified from the sequenced samples were proven to be P. juxtanucleare. The P. juxtanucleare from Thailand was closely related to samples (99.64%–100%) with a genetic distance of 0%–1%. In addition, age, population, and cage type were not significantly associated with Plasmodium infection. Based on microscopic and PCR examinations, the Plasmodium species found in the three districts of Yogyakarta was P. juxtanucleare. The genetic distance between samples from the three districts of Yogyakarta was closely related (0%–1%) to P. juxtanucleare from Thailand and Japan. There was no correlation between Plasmodium infection and age, cage type, or population. Keywords: avian malaria, cytochrome b gene, layer chicken, polymerase chain reaction.

Improving the feed efficiency of beef cattle is necessary to increase the profitability of meat production. Implementing marker-assisted selection breeding systems can improve the genetic potential of beef cattle for increased productivity. This research aimed to study the effects of insulin-like growth factor (IGF)-1 C472T, growth hormone (GH) C2141G, and GH receptor (GHR) T914A polymorphisms on growth performance and feed efficiency in young Kazakh white-headed cattle. Young Kazakh white-headed cattle (n = 50) were grouped after weaning according to sex (28 bulls and 22 heifers) and they were genotyped according to the IGF-1 C472T, GH C2141G, and GHR T914A polymorphisms. The test period was conducted from 8 to 15 months of age. The experimental animals were evaluated for live weight (LW) at the beginning and end of the test period. They were also assessed for average daily gain, hip height, metabolic mid-weight (MMWT), actual dry matter intake (DMI), and residual feed intake (RFI). Significant differences in MMWT were found between the bulls with the IGF-1TT and IGF-1CT genotypes, which was a 2.2 kg increase in heterozygous cattle (p < 0.05). Heterozygous IGF-1CT bulls differed with a higher DMI of 0.087 kg/ day (p < 0.05) compared to IGF-1TT homozygotes. Carriers of the IGF-1TT genotype had the greatest feed efficiency at 0.068 kg/day (p < 0.05). Heifers with the GHCC genotype differed in their maximum DMI with an increase of 1.17%–1.57% (p < 0.05) relative to the other genotypes. The G allele in the GH C2141G polymorphism was associated with better (p < 0.05) feed efficiency in the Kazakh white-headed breed. The minimum DMI and RFI in GHR T914A heterozygous heifers were significantly inferior (p < 0.05) to the other genotypes. Association studies of the IGF-1 C472T, GH C2141G, and GHR T914A polymorphisms indicate a relationship between growth, development, and feed efficiency with the genetic characteristics of young Kazakh white-headed cattle. A significant (p < 0.05) dominant effect was found in the IGF-1 gene in bulls and in the GHR gene in heifers, which should be considered when breeding with heterogeneous parental pairs. The negative effect of the allele substitution in the IGF-1 C472T polymorphism was observed in the LW of heifers (−3.25 kg) at the age of 8 months and bulls (−6.05 kg) at 15 months. The substitution in the GH C2141G polymorphism was associated with a significant reduction in DMI by 0.036 kg (p < 0.05) and an increase in feed efficiency by 0.023 kg (p < 0.05) during the rearing of heifers. These results can improve the production efficiency of mature herds of Kazakh white-headed cattle. Keywords: feed efficiency, growth hormone gene, growth hormone receptor gene, insulin-like growth factor-1 gene, Kazakh white-headed breed, polymorphism.

Diabetes mellitus is a leading cause of mortality worldwide associated with hyperglycemia-induced hematological aberrations and thromboembolic complications. This study aimed to explore the modulatory effect of Terminalia catappa leaf aqueous crude extract (TCLE) on hematological and coagulation disturbances in a Type 2 diabetic rat model. High-fat diet streptozotocin-induced diabetic rats were treated orally with 400 and 800 mg/kg body weight TCLE daily for 28 days. Full blood count, coagulation parameters, plasma calcium (Ca), and erythrocyte glycogen (GLYC) levels were assessed using standard procedures. Terminalia catappa leaf aqueous crude extract treatment had a significant (p < 0.05) prolonging effect on clotting and bleeding times while increasing Ca, GLYC and mean corpuscular volume in diabetic rats. On the other hand, lymphocytes (LYM), platelet (PLT) count, mean PLT volume, neutrophil-LYM ratio (NLR), and PLT-LYM ratio (PLR) of TCLE-treated diabetic animals were significantly reduced (p < 0.05) compared with untreated diabetic animals. Lymphocyte, PLT count, NLR, and PLR correlated positively (p < 0.05) with plasma glucose, while a significant positive association was observed between Ca and GLYC. On the other hand, a strong negative association (p < 0.05) was observed between clotting time and fasting plasma glucose. These findings suggest that T. catappa leaf extract may be useful in reversing diabetic-mediated hematological anomalies due to its anticoagulant and anti-anemic activities. Keywords: coagulation, diabetes, hematology, high-fat diet, streptozotocin-induced, Terminalia catappa.

Scrub typhus and murine typhus are globally distributed zoonoses caused by the intracellular Gram-negative bacteria Orientia tsutsugamushi and Rickettsia typhi, respectively. Numerous studies have been undertaken on rickettsial illnesses in humans and animals, including arthropod vectors, in Thailand. However, the reports on the seroprevalence of antibodies to O. tsutsugamushi and R. typhi in buffaloes is extremely rare. Thus, this study aimed to estimate the seroprevalence of both rickettsial infections in water buffaloes (Bubalus bubalis) in Phatthalung Province, southern Thailand. From February to March 2023, a total of 156 serum samples were collected from 156 water buffaloes on 29 farms in Phatthalung province. The sera were screened for antibodies against O. tsutsugamushi and R. typhi using an indirect immunofluorescence assay. The seroprevalence of antibodies against O. tsutsugamushi and R. typhi in individual water buffaloes was 4.49% (95% confidence interval [CI]: 2.19%-8.97%) and 3.85% (95% CI: 1.77%-8.14%), respectively, whereas 31% (9/29) of the herds had buffaloes with antibodies. The number of buffaloes with scrub typhus infection and ectoparasite infestation was statistically significant (p < 0.05; odds ratio = 6.25 [95% CI: 1.19-33.33]). Intriguingly, the prevalence of scrub typhus antibodies in buffaloes that were not infested with ectoparasites was much higher than those that were. This is the first report of O. tsutsugamushi and R. typhi antibodies in water buffalo sera in Southern Thailand. Two serum samples showed a high antibody titer against O. tsutsugamushi. Seroprevalence mainly occurred in non-ectoparasite-infested buffaloes, especially for O. tsutsugamushi antibodies. At the herd level, one-third of the studied farms showed seroprevalence. Additional research on the occurrence of these pathogens in vectors and in other animal reservoirs is necessary. Keywords: indirect immunofluorescence assay, Orientia tsutsugamushi, Rickettsia typhi, seroprevalence, Thailand, water buffaloes.

To develop species-specific probiotics for poultry, it is ideal to obtain these probiotic microorganisms directly from the intestinal tract of broiler and egg-laying chicks in production environments to ensure adaptation to actual conditions. This study aimed to isolate lactic acid bacteria (LAB) from the intestinal tract of broiler and egg-laying chicks to determine their probiotic potential. Twenty-five Ross-308 broilers and 25 Isa Brown egg-laying chicks were raised until days 42 and 120, respectively; they were housed in an individual poultry building. Lactic acid bacteria were isolated and identified from the small intestine mucus of broiler and layer chicks and then evaluated based on resistance to acidic pH levels, bile salt concentration, and antagonistic activity against wild strains of Escherichia coli and Salmonella spp. selected strains with probiotic potential were identified by polymerase chain reaction and confirmed by rDNA sequencing. One hundred and fifty Gram-positive isolates were obtained; 28% (42) were catalase and oxidase negative and biochemical identification was made by crystal system: 76.2% (32) Enterococcus spp., 16.6% (7) Lactococcus spp., and 7.2% (3) Streptococcus spp.; and evaluated for hemolysin production; tolerance to low pH and bile salts, and antagonistic potential were carried out. Molecular characterization yielded 56% (24) Enterococcus faecium, and 44% (18) Enterococcus faecalis. About 81% (34) of strains were without vancomycin resistance genes criterion. This study isolated and characterized 36 strains of LAB with probiotic qualities, from the intestines of broiler and egg-laying chicks, selecting E. faecium, Enterococcus avium, and Enterococcus casseliflavus, Lactococcus garviae as promising strains for further in vitro and in vivo research. Keywords: Enterococcus avium, Enterococcus faecium, lactic acid bacteria, probiotics.

Anthelmintic resistance (AR) and acaricide resistance (ACR) pose great economic threat to communal livestock raised by rural communities, limiting sustainable production. This study was conducted to assess the occurrence of AR and ACR against nematodes and ticks that infest small ruminants (sheep and goats) from small-scale farming communities in the North West Province of South Africa, as well as document the associated risk factors. The study was conducted on small-scale farming locations in two districts of the North West Province, namely, Dr. Ruth Segomotsi Mompati district and Dr. Kenneth Kaunda district, from November 2019 to March 2020. A questionnaire survey based specifically on antiparasitic treatment and related management practices was administered to 86 small-scale farmers. A fecal egg count reduction test (FECRT) was used to determine in vivo AR in small ruminants against benzimidazole (BZD), levamisole, and macrocyclic lactone on nine ruminant farms. Then, deoxyribonucleic acid was extracted from L3 larvae and resistant nematodes were identified using a polymerase chain reaction, targeting the internal transcribed spacer 2 gene. An egg hatch assay (EHA) and a larval mortality assay (LMA) were used to determine in vitro AR against thiabendazole (TBZ and BZD) in the same farms. Acaricide resistance against fluazuron–flumethrin (Drastic Deadline eXtreme) pour-on was assessed using an adult immersion test (AIT) on Rhipicephalus evertsi. Questionnaire results indicated that most farmers (89%) relied solely on anthelmintics. Farmers used visual appraisal to estimate the dosage, which is the primary cause of resistance. The FECRT revealed AR in all the farms. Egg hatch assay results revealed AR development against TBZ in all districts, with >95% of the eggs hatching at variable doses. Larval mortality assay results revealed the development of resistance against BZD, with 50% of L3 larvae surviving at different doses in all farms. Adult immersion test results indicated that fluazuron-flumethrin (>99%) exhibited high acaricidal efficacy against R. evertsi by inhibiting tick oviposition. This investigation found that sheep and goats in the studied areas are developing AR to gastrointestinal parasites. The findings of in vivo tests showed resistance with fecal egg count reduction percentage of <95% or lower confidence limit of <90%. The results of EHA and LMA revealed no evidence of inhibition of egg development and larval mortality, indicating the development of resistance. Acaricide resistance was not detected against fluazuron–flumethrin, which is commonly used in the study areas. Thus, developing management methods for these economically significant livestock nematodes, including teaching small-scale farmers how to properly administer anthelmintics and acaricides to their livestock, is urgently needed. Keywords: acaricidal resistance, adult immersion test, anthelmintic resistance, egg hatch assay, gastrointestinal nematodes, larval mortality assay.

The gut microbiome plays an important role in the overall health and well-being of dogs, influencing various physiological processes such as metabolism, nutrient absorption, and immune function. Edible insects are a sustainable and nutritious alternative protein source attracting increasing attention as a potential component of animal feeds, including pet food. However, little is known about the effects of insect-based diets on the gut microbiota of dogs. This study aimed to examine the fecal microbiota of dogs fed a diet that substituted common protein sources (poultry meal) with the house cricket (Acheta domesticus [AD]) or mulberry silkworm pupae (Bombyx mori pupae [BMp]) at different levels. Fifteen healthy adult mixed-breed dogs were systemically randomized and assigned into each block under a completed randomized block design into the following five experimental dietary groups: control diet, 10% AD, 20% AD, 7% BMp, or 14% BMp for 29 days. The amounts fed to the dogs were based on the daily energy requirement. Fecal samples were collected on days 14 and 29 and analyzed for bacterial community structure using 16S ribosomal ribonucleic acid gene sequencing. At the phylum and genus levels, microbiota and their diversity were generally relatively similar among all treatments. The diets containing insects did not significantly alter the major phyla in the gut microbiome of dogs (p > 0.05). A few significant changes were found in the relative abundance of bacterial genera, with the levels of Allobaculum and Turicibacter being reduced in dogs fed a higher level of BMp. In contrast, only a decrease in Turicibacter was found in dogs fed the lower level of AD than the control diet (p < 0.05). Corynebacterium and Lactobacillus levels in the dogs fed 14% BMp were significantly increased compared with those in the control group (p < 0.05). These findings suggest that insect-based diets may slightly alter the gut microbiota of dogs. Further research is needed to fully understand the mechanisms by which insect-based diets influence the gut microbiota of dogs and the long-term potential health implications. Keywords: 16S ribosomal ribonucleic acid, canine, cricket, gut microbiota, insect, silkworm.

Several strains of Aspergillus fumigatus produce mycotoxins that affect the health and productivity of dairy cattle, and their presence in dairy cattle feed is a serious concern. This study aimed to determine the densities of A. fumigatus and gliotoxin in commercial dairy feed. More than 60 dairy feed samples were examined for fungal contamination, specifically for A. fumigatus, using phenotypic approaches and DNA sequencing of the internal transcribed spacer (ITS) and β-tubulin regions. Thin-layer chromatography and high-performance liquid chromatography (HPLC) were used to assess gliotoxin production in A. fumigatus. Real-time polymerase chain reaction (RT-PCR) was used to investigate the expression of gliZ, which was responsible for gliotoxin production. High-performance liquid chromatography was used to detect gliotoxin in feed samples. Aspergillus was the most commonly identified genus (68.3%). Aspergillus fumigatus was isolated from 18.3% of dairy feed samples. Only four of the 11 A. fumigatus isolates yielded detectable gliotoxins by HPLC. In total, 7/11 (43.7%) feed samples tested had gliotoxin contamination above the threshold known to induce immunosuppressive and apoptotic effects in vitro. The HPLC-based classification of isolates as high, moderate, or non-producers of gliotoxin was confirmed by RT-PCR, and the evaluation of gliZ expression levels corroborated this classification. The identification of A. fumigatus from animal feed greatly depended on ITS and β-tubulin sequencing. Significant concentrations of gliotoxin were found in dairy cattle feed, and its presence may affect dairy cow productivity and health. Furthermore, workers face contamination risks when handling and storing animal feed. Keywords: Aspergillus fumigatus, cattle feed, gliotoxin, gliZ, high-performance liquid chromatography, real-time polymerase chain reaction.

Enhancing the nutritional and biological value of meat obtained from young surplus replacement animals of dual-purpose breeds is a critical objective in the livestock industry. This study aimed to investigate the impact of thyroglobulin (TG5, c. −422C > T) and leptin (LEP, c. 239C > T) polymorphisms on the productivity, chemical composition, and fatty acid (FA) profile of meat from Simmental bulls. A total of 26 Simmental bulls were genotyped for TG5 (c. −422C > T) and LEP (c. 239C > T) polymorphisms and reared under the same fattening conditions. Controlled slaughter was conducted at 18 months of age. Subsequently, the experimental animals were evaluated to determine their slaughter traits and the chemical and FA composition of ground beef and the longissimus dorsi muscle. The results showed that the TG5 (c. −422C > T) polymorphism significantly (p < 0.05) affected the differentiation of bulls in terms of the synthesis of stearic acid, linolenic acid, and total polyunsaturated FAs, as well as the fat and dry matter content in the longissimus dorsi muscle. Conversely, the presence of the T allele in the LEP (c. 239C > T) polymorphism was associated with increased dry matter and fat in ground beef, carcass weight, and internal fat weight. The analysis of slaughter traits and the chemical and FA composition of meat from the Simmental bulls genotyped for the TG5 and LEP genes revealed a genetic basis for the quantitative and qualitative characteristics of meat productivity. Thus, the genetic variability of bulls regarding the LEP and TG5 genes can be used to improve the quantitative and qualitative indicators of meat productivity in Simmental cattle through marker-assisted selection. Keywords: chemical composition, fatty acid, leptin, meat productivity, polymorphism, Simmental, thyroglobulin.

Infectious coryza (IC) or snot, is caused by Avibacterium paragallinarum and leads to upper respiratory disease in poultry. Various diagnostic methods are available, including isolation and identification through bacterial culture and biochemical tests. However, the isolation and subsequent identification of A. paragallinarum are challenging because the bacteria are fastidious and require specific growth factors. This study aimed to detect A. paragallinarum in clinical samples taken from the exudate of the infraorbital sinus of layer hens showing clinical signs of IC. Samples were collected from 10 layer hens with IC symptoms. Following DNA extraction, HPG-2 polymerase chain reaction (PCR) assays were performed. The PCR amplicons underwent electrophoresis to determine those of the correct target size (511 bp), and these were sequenced. The resultant sequences were analyzed using the National Center for Biotechnology Information (NCBI) basic local alignment search tool. MEGA X was used for bioinformatics analysis. The presence of A. paragallinarum was confirmed by HPG-2 PCR in 4/10 samples. Bioinformatics analysis showed that the amino acid sequence of the samples and the A. paragallinarum reference sequences in the NCBI database were grouped within the same cluster. Furthermore, the nucleotide sequences showed 98.64%–100% of similarity with the reference sequences. The phylogenetic reconstruction of partial pyrG sequences from 55 A. paragallinarum strains/isolates deposited in GenBank confirmed that the four HPG-2 PCR-positive samples fell within the A. paragallinarum cluster, separate from the Pasteurella multocida, Avibacterium spp., and Rodentibacter pneumotropicus clusters. Avibacterium paragallinarum infection was molecularly confirmed in 4/10 (40%) samples by HPG-2 PCR amplicon detection. Clustering of the pyrG partial gene sequences revealed that the positive samples fell within the A. paragallinarum cluster. Keywords: Avibacterium paragallinarum, bioinformatics, nucleotide sequencesm, polymerase chain reaction.

Administration is the main strategy for controlling ectoparasites in dogs. Ivermectin and fipronil are most extensively used to prevent and treat Rhipicephalus sanguineus sensu lato infestation in dogs in Thailand. Several researchers have reported resistance to acaricides in R. sanguineus s.l. globally, but documentation of acaricide resistance in the tick population in Thailand is lacking. In this study, we investigated the status of resistance to commonly used acaricides in Thailand in R. sanguineus s.l. Engorged brown dog tick females (10 tick populations) were field-collected directly from parasitized dogs in Maha Sarakham, Thailand, for toxicological bioassays with ivermectin and fipronil. Bioassays were performed in three replicates at 25°C-27°C and 80%-85% relative humidity under a 12-h/12-h photoperiod. The 50% of lethal concentration and its confidence intervals and the slope were estimated for each tick population using probit analysis. Resistance ratios (RRs) of field ticks were characterized based on the relative susceptible population of each acaricide. Six tick populations (P1-6) were tested for resistance to ivermectin, three of which (P2-4) exhibited low-level resistance to ivermectin (RR = 2.115-2.176). Of four tick populations (P7-10) treated with fipronil, two exhibited moderate-to-severe resistance (P7 and P9, RR = 21.684 and 4.387, respectively). All tick populations deemed resistant to acaricides had a history of exposure. Based on RR values, four R. sanguineus s.l. tick populations from Maha Sarakham province were resistant to ivermectin and fipronil. To the best of our knowledge, this represents the first documentation of acaricide-resistant populations of R. sanguineus s.l. in Thailand, and recommendations on tick control programs must be formulated with veterinarians and pet owners to prevent the development of further resistance. Keywords: bioassay, dog, fipronil, ivermectin, resistance, Rhipicephalus sanguineus sensu lato, Thailand.

The adhesive progestin patch was investigated for estrus synchronization in mares because this method is convenient and safe in other species. This study aimed to compare the efficacy of a transdermal progestin patch versus oral altrenogest on estrus synchronization, preovulatory follicle development, and ovulation. Twenty-four broodmares were randomly divided into two groups. In Group 1, mares (n = 12) were fed 0.044 mg/kg altrenogest daily for 14 days. Group 2 (n = 12) mares were treated with adhesive transdermal progestin patches for 14 days. Mares were observed for estrus daily, from day 3 until day 7, after hormone withdrawal. Follicular development was determined by ultrasonography. The preovulatory follicle size and ovulation time after hormone treatments and ovulation rate were determined for both groups. Plasma progesterone levels were measured during the experimental periods and 2 days after hormone termination in both groups. The results revealed that the transdermal progestin patch efficiently controlled follicular growth and estrus synchronization in mares. The percentage of mares exhibiting estrus was similar in the altrenogest (100.00%) and transdermal patch (91.67%) groups. Ovulation rates were equivalent with either altrenogest or progestin patch protocols (91.67% [11/12] vs. 83.33% [10/12]). In addition, the preovulatory follicle size was similar in mares treated with altrenogest and progestin patches. This study revealed that the transdermal progestin patch provides effective estrus synchronization and ovulation, similar to altrenogest treatment. However, the transdermal patch was more convenient with a shorter predictable ovulation time after estrus synchronization and should be considered as an alternative method for mares. Keywords: altrenogest, estrus synchronization, mare, progestin, transdermal patch.

The concentration of the feline acute-phase protein serum amyloid A (SAA) increases in cats with acute inflammatory diseases. However, it is unclear whether SAA concentration increases in cats with azotemic kidney disease or whether it can aid in differentiating acute kidney injury (AKI) from chronic kidney disease (CKD). Similarly, whether SAA concentration can be used as a prognostic marker is also unclear. Therefore, this study aimed to evaluate the SAA concentrations in cats with azotemic kidney disease and determine whether SAA concentrations can be used to differentiate between AKI, CKD, and “acute on CKD” (AoC). In addition, we evaluated whether SAA concentration could serve as a prognostic parameter. Moreover, we determined the correlations between SAA concentration and temperature; creatinine, urea, and albumin concentrations; leukocyte count; and urine protein/creatinine (UP/C). Forty-eight client-owned azotemic cats (creatinine >250 μmol/L) were included in this prospective study. Cats with pre- and post-renal azotemia were excluded from the study. The causes of azotemia were differentiated into AKI, CKD, and AoC. The SAA concentrations were analyzed through turbidimetric immunoassay at the time of admission. Data were analyzed using the Mann-Whitney U, Kruskal-Wallis, Chi-Square, Fisher’s exact, and Spearman correlation tests. p ≤ 0.05 was considered statistically significant. Serum amyloid A concentration increased in 5/12 cats with AKI, 7/22 cats with CKD, and 9/14 cats with AoC (p = 0.234). The median SAA concentration in cats with AKI, CKD, and AoC whose SAA concentration was ≥5 mg/L was 174 mg/L (10-281 mg/L), 125 mg/L (6-269 mg/L), and 143 mg/L (7-316 mg/L), respectively (p = 0.697), with no significant differences observed between the groups. The median SAA concentration did not differ significantly between survivors (125 mg/L, 10-316 mg/L) and non-survivors (149 mg/L, 6-281 mg/L; p = 0.915) with SAA concentration ≥5 mg/L. Serum amyloid A concentration increased in 44% of the cats with azotemia. However, it cannot be used to differentiate AKI from CKD or as a prognostic marker. Serum amyloid A concentration was correlated with neutrophil count, albumin concentration, and UP/C, and the presence of comorbidities may influence SAA concentration. Keywords: acute kidney injury, acute-phase protein, chronic kidney disease, International Renal Interest Society grading, uremia.

Equine influenza (EI) is a highly contagious disease that causes fever and upper respiratory tract inflammation. It is caused by influenza virus A, belonging to the Orthomyxoviridae family, with subtypes H3N8 and H7N7. This study presents data on the development of a real-time polymerase chain reaction (RT-PCR) assay using TaqMan probes to detect the H3 subtype of EI virus (EIV). The evaluation of the developed RT-PCR assay involved five strains of EIV as positive controls and ten nasopharyngeal swab samples collected from horses. RNA was isolated using the GeneJet Viral DNA and RNA Purification Kit, and primers and probes were designed using the Integrated DNA Technology PrimerQuest Tool. The assay was optimized by investigating the annealing temperature, primer and probes concentrations, sensitivity, and specificity. Sequencing was performed using the Thermo Fisher 3130 Genetic Analyzer, and the evolutionary history was inferred using the Neighbor-Joining method. The designed primers and probes, targeting the H3 gene, were found to be specific to the EIV. The RT-PCR assay was capable of detecting as low as 50 femtogram (f) or 3 × 103 copies of genomic RNA. No cross-reactions were observed with other respiratory viral and bacterial pathogens, indicating the high specificity of the assay. To evaluate its effectiveness, ten nasopharyngeal swab samples collected from farms in North Kazakhstan regions during disease monitoring were analyzed. The accuracy of the analysis was confirmed by comparing the results with those obtained from a commercial RT-PCR assay for EI identification. The developed RT-PCR assay exhibited high sensitivity and specificity for detecting the EIV. The results demonstrate that the developed RT-PCR assay is suitable for diagnosing EI. This simple, highly sensitive, and specific assay for detecting H3 EIV can be a reliable tool for diagnosing and surveilling EI. Implementing this RT-PCR assay in veterinary practice will enhance and expedite the timely response to potential outbreaks of EI, thus positively impacting the overall epizootic well-being of EI in Kazakhstan. Keywords: equine influenza, hemagglutinin, horses, primers, probe, real-time polymerase chain reaction assay, virus.

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. 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. 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. 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.

Porcine epidemic diarrhea (PED) is a severe infectious disease that causes very high mortality in newborn piglets up to 2-3 weeks age. The main cause of repeated outbreaks of PED in infected farms is the continuing circulation of the PED virus (PEDV). Improper gilt management, including inappropriate gut feedback, commingling, and inadequate immunization, causes a prolonged virus circulation in breeding herds. Moreover, insufficient transfer of passive immunity through the colostrum to newborn piglets can also increase infection risk. Therefore, a gilt management program that controls infection should focus on infection monitoring and acclimatization. We investigated the source of recurrent PEDV outbreaks and examined how the effect of immunization methods, specifically using gut feedback mechanism and vaccination, can reduce PEDV circulation and improve immune responses in replacement gilts. The study site was a segregated commercial production farm with endemic PEDV. The acclimatization methods included gut feedback and vaccination. This longitudinal study evaluated two strategies of gilt acclimatization against PEDV: Program 1 (routine farm management) and Program 2 (early feedback program and all-in-all-out system). Levels of PED RNA in fecal samples were measured using quantitative reverse transcription-polymerase chain reaction, and the PEDV S gene was sequenced. Porcine epidemic diarrhea-specific immune responses were assessed using enzyme-linked immunosorbent assay and the serum neutralization test. Porcine epidemic diarrhea outbreaks occurred in the farrowing, nursery, and finishing units and farrowed litters 5–10 days old were symptomatic of PED. Phylogenetic analyses of the S gene showed PEDV sequence divergence between PEDV field strains and vaccine strain, which may contribute to periodic outbreaks and continued persistence of PEDV in the farm. After gut feedback and acclimatization, replacement gilts from Program 1 continued to shed PEDV before being introduced to sow herds, while those from Program 2 did not shed PEDV before being introduced to sow herds. However, the components of the immune response against PEDV in serum samples, including specific immunoglobulin (Ig)G, specific IgA, and neutralizing antibodies were lower in gilts of Program 2 than those in Program 1. We speculate that implementing the appropriate gilt acclimatization program can control PEDV circulation in farm. However, the acclimatization methods in Program 2 did not induce a strong and adequate immune response in replacement gilts. Therefore, maternal immunity levels and the degree of protection against PEDV require further study. Keywords: acclimatization, gilt, immune response, phylogenetic tree, porcine epidemic diarrhea.

Anaplasma spp. are common rickettsia species described in ruminant hosts, including cattle. The clinical signs of anaplasmosis range from asymptomatic to mortality. However, there are insufficient studies on epidemiology surveys of this blood pathogen. This study aimed to estimate the prevalence and risk factors of anaplasmosis in beef and dairy cattle in Northeast, Thailand. A total of 187 blood samples of beef and dairy cattle were collected from five provinces in Northeast Thailand. Anaplasma spp. infections were screened by microscopic examination and polymerase chain reaction targeting specific genes (msp4 gene for Anaplasma marginale and 16S rRNA gene for Anaplasma platys and Anaplasma bovis). Moreover, the associated risk factors for the infections were evaluated. Overall, blood samples from cattle revealed that 17.6% (33/187) were positive for Anaplasma spp. by microscopic examination and 20.8% (39/187) were positive by DNA amplification. Of these 20.8%, 17.6% were A. marginale and 3.2% were A. platys. However, A. bovis infection was not detected. Infection with Anaplasma spp. and A. marginale showed a significant association with breed and gender (p < 0.05) while age and packed cell volume levels showed no significant statistical relationship between Anaplasma spp. infected and uninfected animals. This study indicated that anaplasmosis is distributed in beef and dairy cattle in Thailand; therefore, prevention and control strategies for these pathogens should be improved. This information will benefit veterinarians and cowherds by avoiding vector exposure and eliminating tick breeding sites. Keywords: anaplasmosis, beef cattle, blood parasite, dairy cattle, molecular detection.

Feline chronic gingivostomatitis (FCGS) is a frequent chronic inflammatory condition in the oral cavity with an etiopathogenesis not completely identified. This study aimed to contribute to the knowledge of FCGS by identifying the presence of feline calicivirus (FCV) antigens and natural killer (NK) cells and comparing them. Forty biopsies from the oral mucosa of cats diagnosed with chronic gingivostomatitis were subjected to immunohistochemical techniques to evaluate cells with FCV antigens and NK cells positive for CD56. NK cells were identified in all samples, with an average of 725.3 ± 409.1 cells. Regarding FCV, it was identified in 18 out of 30 samples (60%), with a different number of cells with virus in between the analyzed cases. In all cases, the number of cells infected with FCV was lower than the number of NK cells present in the same samples, but there was no statistical association between them. This preliminary study shows that NK cells are present in gingivostomatitis lesions not exclusively caused by FCV-stimulus, as only 60% of all cases were positive for this virus, but other antigens should be considered in the etiology of FCGS. Keywords: feline calicivirus, feline chronic gingivostomatitis, immunohistochemistry, natural killer cells.

Climate change challenges with incremental sea level cause saltwater intrusion, which has affected the quality of freshwater and groundwater in coastal provinces, particularly the Mekong River Delta provinces of Vietnam. Interestingly, Bach Thao goats are predominant in this area and well adapted to saline water under tropical conditions. Therefore, this study investigated the salt tolerance threshold of Bach Thao goats drinking diluted seawater (DSW). The experiment was performed using seven Bach Thao male goats (20.60 ± 1.12 kg) and divided into two phases. In Phase 1 (control, C), all goats were provided fresh water (FW) for 7 days from two identical buckets, and daily water intake (WI) was recorded from both buckets. In Phase 2 (preference test, PT), each goat was provided FW from the first bucket and DSW from the second bucket or vice versa. The concentration of DSW for the preference test was 0.5%-2.0%. Body weight and dry matter intake showed no differences according to DSW consumption; however, WI was significantly lower during Phase 2 (p < 0.05), which was due to the lower WI with 1.5% and 2% of DSW (p < 0.05). Goats showed a similar preference for fluid intake between FW and DSW at 0.0%–1.0% levels and began avoiding DSW at 1.5% and rejected at 2.0% of DSW. Goats consuming 1.5% of DSW showed increased respiration rate from 13:00 to 19:00 h and rectal temperature at 13:00 h (p < 0.05). Goats can tolerate up to 1.0% of DSW. Shifting to FW activated aversive drinking to 1.5% and 2.0% of DSW. This behavioral response was prominent at 0.5% DSW. Moreover, goats that drank 1.5% of DSW had decreased thermoregulation. Keywords: body weight, climate change, freshwater, salt tolerance.

A conventional feeding bowl is the primary method that dog owners use to feed their dogs, but this may not encourage natural behaviors and may even exacerbate unwanted behaviors. This study aimed to compare a conventional feeding bowl to a feeding toy in relation to behavior, cortisol levels, and heart rate variability (HRV). The behaviors of four dogs were recorded and analyzed while being fed using either a stainless bowl (B) or a feeding toy (T) and either alone (A) or accompanied by a dog owner (O) for 30 min with each treatment (BA, BO, TA, and TO treatments). The dogs that were fed alone with the stainless bowl (BC) or the feeding toy (TC) were fed for 15 min/day for 7 days with their treatment, and serum cortisol levels measured on the first and last days of treatment. The dogs fed by the stainless bowl (BH) or the feeding toy (TH) with the owner present for 15 min for each treatment had their heart rate (HR) and HRV recorded by Polar® H10 during feedings The results were compared using a one-way analysis of variance (ANOVA), repeated measure ANOVA, and Student’s t-test. The dogs spent more time eating and interacting with the feeding toys than stainless bowls. The activity of the dogs was higher when using feeding toys, particularly with the TO treatment. Cortisol levels were significantly lower on day 7 than on day 1 of the TC treatment. The dogs’ HR was higher during TH treatment than during BH treatment. All HRV parameters were decreased significantly when feeding the dog with the toys. The results of this study support the idea that feeding enrichment supports the natural feeding behaviors of dogs as they mimic hunting and playing behaviors. This reduced unwanted behavior, cortisol levels, and HRV, and increased food consumption, eating duration, and active behaviors. The presence of the dog’s owner is important because it can enhance feeding and active behaviors, and feeding enrichment can improve the dog’s welfare and the dog-human relationship. Keywords: cortisol, dog behavior, dog welfare, dog-human relationship, feeding toy, heart rate variability.

The risk factors for cholangiocarcinoma (CCA) are opisthorchiasis and the intake of a combination of nitroso compounds through the consumption of traditionally fermented fish, which is very popular in areas where liver flukes are endemic. The incidence of CCA remains high because this cultural habit of rural people has been altered. Therefore, decreasing nitrate and nitrite concentrations in fermented fish are an alternative approach to reducing the risk of CCA. Thus, this study aimed to reduce nitrate and nitrite concentrations in fermented foods by heating and investigated its effect on CCA development in a hamster model. We used Association of Official Analytical Chemists method 973.31 to measure the nitrate and nitrite concentrations in both fermented fish (pla-ra [PR]) and pickled fish (pla-som [PS]) before and after boiling for 5 and 30 min, respectively. The same samples were fed to Opisthorchis viverrini (OV)-infected or -uninfected hamsters for 3 months. Thereafter, the hamsters’ liver and blood were collected for analysis. The levels of nitrates and nitrites in PS and PR significantly decreased following boiling for 5 and 30 min. The OV-PR and OV-PS groups showed dramatically increased numbers of inflammatory cells, fibrosis surrounding the bile duct, and focal fibrotic areas. However, after boiling the fermented dishes for 5 and 30 min, the extent of inflammatory cell infiltration and intensity of fibrosis in these groups were decreased. Our findings suggest that boiling reduces nitrate and nitrite toxicity in fermented dishes, as evidenced by reduced hepatic inflammation. However, regardless of heating, kidney tissues are adversely affected when fermented meals are consumed daily. Keywords: cholangiocarcinoma, fish, liver flukes, opisthorchiasis.

Antibiotic resistance is a component of antimicrobial resistance and is often referred to as the silent pandemic. It is one of the causes of global health problems that must be addressed. Resistance occurs due to frequent misuse and overuse of antibiotics by dairy farmers. Therefore, this study aimed to examine the influence of the characteristics of dairy farmers and analyze the variables that directly and indirectly affected the knowledge, attitudes, and practices (KAP) of dairy farmers regarding antibiotic resistance in a dairy farmers' cooperative in North Cianjur. A cross-sectional design was used with a structured questionnaire validated by Pearson correlation and reliability tested with Alpha Cronbach. Data were obtained from interviews with 75 dairy farmers in KPSCU, Cianjur District, West Java, Indonesia. The outcome variables were analyzed using descriptive and pathway analyses. The results showed that dairy farmers had good and sufficient knowledge (42.7%), the attitude level was positive (81.3%), and all farmers had sufficient practices (100%). The results also showed that most dairy farmers were over 40 years of age, with 54.7% having elementary school education. Approximately 50.7% of farmers had been engaged in breeding for 10–20 years, 80% owned their livestock, and 76% had never attended training. There was a significant relationship between age, education, training, and knowledge level. Knowledge was the primary factor influencing the overall attitude. In addition, age, education, type of business, knowledge, and attitude were factors that influenced the practice of antibiotic resistance. Training and education really influenced KAP of farmers. Therefore, the best way to reduce antibiotic resistance is by increasing farmers’ knowledge and understanding of antibiotic resistance and monitoring the use of antibiotics. Keywords: antibiotic resistance, knowledge, pathways analysis, questionnaire.

Antibiotic residues in livestock farming have been identified as a potential cause of antimicrobial resistance in humans and animals. This study aimed to determine whether antibiotic residues were present in the chicken meat, eggs, feces, and feed collected from all four districts in the Mymensingh division of Bangladesh. To detect antibiotic residues in the collected samples, qualitative thin-layer chromatography (TLC) and quantitative high-performance liquid chromatography (HPLC) were used. A total of 230 samples were analyzed for commonly used 11 antibiotics residue. Out of these, 40 meats and 40 feces samples were collected from broilers and layers, 30 egg samples from duck and layer, and 120 feed samples from both broilers and layers from the study area. Thin-layer chromatography was used for screening the presence of antibiotic residues; TLC-positive samples were then subjected to further HPLC analysis to determine the residue concentrations. Thin-layer chromatography analysis revealed that 23.5% of the tested samples contained residues from six different antibiotic classes (tetracyclines, quinolones, beta-lactams, sulfonamides, aminoglycosides, and macrolides). Thin-layer chromatography analysis showed that 35% and 25% of the meat samples were positive for residues from the broiler and layer, respectively. About 15% and 30% of layer and duck egg samples had positive residues, respectively. Out of 120 feed samples analyzed, about 15.8% had various antibiotic residues. In addition, feces samples from broilers and layers had 50% and 35% antibiotic residues, respectively. A total of 2.5% meat and 3.3% egg samples had antibiotic residues above the maximum residue limit (MRL). Based on the findings of this study, the highest percentage of oxytetracycline, followed by doxycycline and ciprofloxacin, were detected in feed samples, and oxytetracycline was detected in meat and egg samples. This study clearly showed the misuse of antibiotics in the poultry sector in Bangladesh. Although antibiotic residues below the MRL level are suitable for human consumption, they may result in antimicrobial drug resistance to pathogens. Keywords: antibiotic residues, broiler, high-performance liquid chromatography, layer, maximum residue limit, thin-layer chromatography.

Social isolation stress (SIS) and individual housing have been shown to cause abnormal cognitive insufficiencies, altered anxiety levels, and signs of psychiatric diseases. Acorus calamus (AC), commonly known as Sweet Flag, has been widely used in India to treat neurological, metabolic, and respiratory disorders, indicating its potential therapeutic value. This study aimed to determine the antidepressant and antioxidative effects of AC on rats subjected to long-term, social-isolation-induced stress. This study involved 2-month-old male rats (24) weighing approximately 180200 g bred in-house. The rats were divided into four groups (n = 6): Group-1 received saline, Group-2 received SIS, Group-3 received only 50mg/kg AC, and Group-4 received 50mg/kg AC and SIS for 6 weeks. After this, behavioral, biochemical, and neuronal assay was conducted. Behavioral experiments showed significantly higher activity levels (p < 0.001) in AC-treated rats than in the SIS group. In addition, rats subjected to SIS with AC treatment exhibited enhanced total antioxidants, superoxide dismutase, and neuronal assays compared to rats subjected to SIS alone. Acorus calamus treatment improved the antidepressant and antioxidant potential against SIS in rat brain tissue. Moreover, we proved that AC can effectively reverse the neurotoxicity induced by SIS in animal models. As we battle against the coronavirus disease 2019 pandemic and social isolation, AC could be considered a supplementary treatment to alleviate depressive-like symptoms in our present-day lifestyle. Keywords: antidepressant, antioxidative effects, brain tissue, coronavirus disease 2019 pandemic.

Pregnancy affects maternal hemodynamics. The changes in autonomic nervous system activity for hemodynamics adaptation in pregnant horses are still unclear. Thus, this study aimed to examine the effect of pregnancy on electrocardiographic, vasovagal tonus index, and echocardiographic variables in horses. A total of 23 Thai native crossbred mares without any cardiac abnormalities were included in this study. The animals were assigned into two groups, a non-pregnant mare group (n =12) and a pregnant mare group (n = 11). Electrocardiogram recordings (paper speed = 25 mm/s and calibration = 10 mm/mV) were performed to obtain six limb leads (leads I, II, III, aVR, aVL, and aVF). The vasovagal tonus index (VVTI) was calculated to assess variability in heart rate over short periods using just 20 consecutive beats. Cardiac structure and function were evaluated by echocardiography. Heart rate, P wave duration, PR interval, QRS duration, QT interval, and T wave duration were significantly different between non-pregnant and pregnant horses (p < 0.05). Pregnant horses had significantly lower VVTI than non-pregnant (p < 0.05). There were no significant differences in cardiac structures including % interventricular septum (IVS), % left ventricular posterior wall (LVPW), IVS in diastole, left ventricular internal diameter at end-diastole, LVPW thickness at end-diastole, IVS in systole, left ventricular internal diameter at end-systole, LVPW thickness at end-systole, and left atrium/aortic roots ratio between the two groups. However, the pregnant horses had a significantly higher cardiac output and % ejection fraction than non-pregnant horses (p < 0.05). This study provided the first evidence that hemodynamic adaptations during pregnancy modified cardiac conduction, vasovagal tonus index, and echocardiographic variables in horses. Keywords: echocardiography, electrocardiogram, horses, pregnancy, vasovagal tonus index.