Methicillin-resistant coagulase-positive staphylococci in new, middle-aged, and old veterinary hospitals in southern Thailand: A preliminary study.

Background and Aim: Methicillin-resistant coagulase-positive staphylococci (MRCoPS) cause pyoderma, dermatitis, and nosocomial infection. Numerous factors, including indiscriminate antimicrobial use (AMU) in veterinary medicine, cleaning practices, and AMU in hospitals, contribute to MRCoPS. However, the relationship between hospital age and MRCoPS has not yet been investigated. This study aimed to estimate the prevalence of MRCoPS in the treatment and operation rooms of new, middle-aged, and old veterinary hospitals. Materials and Methods: Samples were collected from small animal hospitals in Surat Thani, Nakhon Si Thammarat, and Songkhla in Thailand. Hospitals were defined as those that had been in operation for 5 years (new, n = 5), 5-15 years (middle-aged, n = 6), or >15 years (old, n = 3). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was used to identify 280 samples, and duplex polymerase chain reaction was used to identify resistance genes (mecA and blaZ). The VITEK2® automated system was then used to determine the minimum inhibitory concentration. Results: A total of 57 Staphylococcus species were identified and classified as coagulase-positive staphylococci (CoPS) (22/57, 38.60%) or coagulase-negative staphylococci (35/57, 61.40%), respectively. Nine of the 22 CoPS (40.90%) harbored the mecA gene, and 21 isolates (95.45%) harbored the blaZ gene. Interestingly, more MRCoPS was found in new hospitals (six isolates) than in middle-aged (one isolate) and old hospitals (two isolates), although there was no statistically significant difference in the presence of MRCoPS across new, middle-aged, and old veterinary hospitals (p = 0.095), Kruskal-Wallis test. There is a need for further detailed studies, including an increase in the number of hospitals in various locations. Conclusion: MRCoPS is a nosocomial pathogen that causes zoonotic and recurrent infections in veterinary hospitals. The prevalence of MRCoPS tended to be higher in new hospitals. Areas with heavy animal contact, such as hospital floors, are areas of particular concern, and cleaning/disinfection of these areas must be highlighted in hygiene regimens.

Seroprevalence of melioidosis and its association with blood profiles and pathogens in sheltered dogs in southern Thailand.

Background and Aims: Melioidosis is a notable zoonotic disease in Thailand that can affect both humans and animals. Although dogs are one of the most popular pets worldwide, there is a remarkable lack of information on the prevalence and knowledge of canine melioidosis. This study aimed to estimate the seroprevalence of melioidosis in sheltered dogs and its relationship with the blood profile and blood pathogens. Materials and Methods: Melioidosis in 156 dogs was analyzed using an indirect hemagglutination assay. Hematology and serum biochemistry tests were performed using an automated system. Blood pathogens (e.g., Ehrlichia, Anaplasma, Hepatozoon, and Babesia) were diagnosed using conventional polymerase chain reaction. Results: The seroprevalence rates of canine melioidosis and blood pathogen infection were 5.77% (9/156) and 50.64% (79/156), respectively. Seropositive dogs generally have higher lymphocyte counts and aspartate aminotransferase levels but lower total white blood cell, neutrophil, and platelet (PLT) counts than seronegative dogs. No statistically significant difference (p > 0.05) was observed between the seropositive and seronegative dogs' hematology and serum biochemistry findings. Neither the correlation between melioidosis and blood pathogen infection nor the association between melioidosis and thrombocytopenia was statistically significant (p > 0.05). Remarkably, dogs that had coinfections with both melioidosis and blood pathogens demonstrated a significantly reduced PLTcount (49,167 ± 7,167) compared with dogs that tested positive for melioidosis but negative for blood pathogens (139,333 ± 29,913) (p < 0.01). Conclusion: In southern Thailand, the prevalence of canine melioidosis was low but the prevalence of blood pathogens was high. Coinfection with blood pathogens can significantly reduce PLT counts, which may have a potentially serious impact. Future research should focus on conducting seroprevalence studies in the general dog population.

A systematic review and meta-analysis of the global prevalence and relationships among Burkholderia pseudomallei sequence types isolated from humans, animals, and the environment.

Background and Aim: Burkholderia pseudomallei, a highly pathogenic bacterium responsible for melioidosis, exhibits ecological ubiquity and thrives within soil and water reservoirs, posing significant infection risks to humans and animals through direct contact. The aim of this study was to elucidate the genetic diversity and prevalence patterns of B. pseudomallei sequence types (STs) across a global spectrum and to understand the relationships between strains isolated from different sources. Materials and Methods: We performed a systematic review and meta-analysis in this study. Extensive research was carried out across three comprehensive databases, including PubMed, Scopus, and ScienceDirect with data collected from 1924 to 2023. Results: A total of 40 carefully selected articles contributed 2737 B. pseudomallei isolates attributed to 729 distinct STs and were incorporated into the systematic review. Among these, ST46 emerged as the most prominent, featuring in 35% of the articles and demonstrating a dominant prevalence, particularly within Southeast Asia. Moreover, ST51 consistently appeared across human, animal, and environmental studies. Subsequently, we performed a meta-analysis, focusing on nine specific STs: ST46, ST51, ST54, ST70, ST84, ST109, ST289, ST325, and ST376. Surprisingly, no statistically significant differences in their pooled prevalence proportions were observed across these compartments for ST46, ST70, ST289, ST325, and ST376 (all p > 0.69). Conversely, the remaining STs, including ST51, ST54, ST84, and ST109, displayed notable variations in their prevalence among the three domains (all p < 0.04). Notably, the pooled prevalence of ST51 in animals and environmental samples surpassed that found in human isolates (p < 0.01). Conclusion: To the best of our knowledge, this study is the first systematic review and meta-analysis to investigate the intricate relationships between STs and their sources and contributes significantly to our understanding of B. pseudomallei diversity within the One Health framework.

Molecular characterization of Campylobacter spp. isolates obtained from commercial broilers and native chickens in Southern Thailand using whole genome sequencing.

Chickens are the primary reservoirs of Campylobacter spp., mainly C. jejuni and C. coli, that cause human bacterial gastrointestinal infections. However, genomic characteristics and antimicrobial resistance of Campylobacter spp. in low- to middle-income countries need more comprehensive exploration. This study aimed to characterize 21 C. jejuni and 5 C. coli isolates from commercial broilers and native chickens using whole genome sequencing and compare them to 28 reference Campylobacter sequences. Among the 26 isolates, 13 sequence types (ST) were identified in C. jejuni and 5 ST in C. coli. The prominent ST was ST 2274 (5 isolates, 19.2%), followed by ST 51, 460, 2409, and 6455 (2 isolates in each ST, 7.7%), while all remaining ST (464, 536, 595, 2083, 6736, 6964, 8096, 10437, 828, 872, 900, 8237, and 13540) had 1 isolate per ST (3.8%). Six types of antimicrobial resistance genes (ant(6)-Ia, aph(3')-III, blaOXA, cat, erm(B), and tet(O)) and one point mutations in the gyrA gene (Threonine-86-Isoleucine) and another in the rpsL gene (Lysine-43-Arginine) were detected. The blaOXA resistance gene was present in all isolates, the gyrA mutations was in 95.2% of C. jejuni and 80.0% of C. coli, and the tet(O) resistance gene in 76.2% of C. jejuni and 80.0% of C. coli. Additionally, 203 virulence-associated genes linked to 16 virulence factors were identified. In terms of phenotypic resistance, the C. jejuni isolates were all resistant to ciprofloxacin, enrofloxacin, and nalidixic acid, with lower levels of resistance to tetracycline (76.2%), tylosin (52.3%), erythromycin (23.8%), azithromycin (22.2%), and gentamicin (11.1%). Most C. coli isolates were resistant to all tested antimicrobials, while 1 C. coli was pan-susceptible except for tylosin. Single-nucleotide polymorphisms concordance varied widely, with differences of up to 13,375 single-nucleotide polymorphisms compared to the reference Campylobacter isolates, highlighting genetic divergence among comparative genomes. This study contributes to a deeper understanding of the molecular epidemiology of Campylobacter spp. in Thai chicken production systems.

Occurrence of multidrug resistance associated with extended-spectrum ß­lactamase and the biofilm forming ability of Escherichia coli in environmental swine husbandry.

Extended-spectrum beta-lactamase (ESBL) production and biofilm formation are mechanisms employed by Escherichia coli to resist beta-lactam antibiotics. Thus, we aimed to examine antibiotic resistance associated with ESBL production and biofilm formation in E. coli isolates from swine farms in Southern Thailand. In total, 159 E. coli isolates were obtained, with 44 isolates identified as ESBL producers, originating from feces (18.87 %) and wastewater (8.80 %) samples. All ESBL-producing strains exhibited resistance to ampicillin (100 %), followed by the cephalosporin group (97.73 %) and tetracycline (84.09 %). Multidrug resistance was observed in 17 isolates (38.63 %). Among the isolates from feces samples, the blaGES gene was the most prevalent, detected in 90 % of the samples, followed by blaCTX-M9 (86.67 %) and blaCTX-M1 (66.67 %), respectively. In the bacteria isolated from wastewater, both blaGES and blaCTX-M9 genes were the predominant resistance genes, detected in 100 % of the isolates, followed by blaCTX-M1 (64.29 %) and blaTEM (50 %), respectively. Strong biofilm formation was observed in 11 isolates (36.67 %) from feces and 4 isolates (25.57 %) from wastewater samples. Notably, nearly 100 % of ESBL-producing strains isolated from feces tested positive for both pgaA and pgaC genes, which play a role in intracellular adhesion and biofilm production. These findings contribute to the understanding and potential control of ESBL-producing E. coli, and the dissemination of antibiotic resistance and biofilm-related genes in swine farms.

Effect of feeding toy and the presence of a dog owner during the feeding time on dog welfare.

Background and Aim: 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). Materials and Methods: 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. Results: 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. Conclusion: 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.

Factors associated with pet dog behavior in Thailand.

Background and Aim: Dog behavior problems pose serious public health and economic and animal welfare concerns. There are many factors influencing dog behavior. This study aims to explore factors associated with pet dog behavior in Thailand using the Canine Behavioral Assessment and Research Questionnaire (C-BARQ). Materials and Methods: The Thai version of the C-BARQ was validated in 2022. The online C-BARQ survey (Google Forms) was advertised and distributed on social media for 3 months. There were a total of 1827 respondents to the survey. The relationship among 12 subscales and variables was analyzed using Spearman's correlation. Results: "Excitability," "attachment- and attention-seeking," and "chasing" were the three most reported behavior problems. "Trainability," a favored dog behavior, also had a high score in the study. These behaviors were associated with the owner's age, house type, the dog's historical background, the number of dogs and the presence of other species in the household, the dog's training, diet types, and the daily number of meals. The lowest mean score was for "owner-directed aggression," and it was associated with the dog's gender and size, the number of dogs and the presence of other species in the household, the dog's training, and the number of daily meals. Conclusion: This is the first empirical study demonstrating factors related to dog behavior in Thailand. It provides an in-depth understanding of the prevalence and factors associated with Thai pet dog behavior and important knowledge for further studies to advocate for dog-human relationships and contribute to a reduction in dog abandonment in Thailand.

Hematological analysis of naturally infecting blood parasites in dogs.

Background and Aim: Blood parasite infections such as anaplasmosis, babesiosis, and ehrlichiosis are commonly found in domestic dogs, which adversely influence their health. Many dogs are infected with multiple blood parasites that cause more severe diseases than a single infection. This study aimed to investigate the effect of multiple blood parasite infections on the hematological profiles of dogs at a shelter in Southern Thailand. Materials and Methods: The blood samples from 122 dogs were collected to assess the hematology profiles of uninfected, single-infected, and multiple blood parasite-infected dogs. The results were compared using Kruskal-Wallis test and Dwass-Steel-Critchlow-Fligner pairwise comparisons. The infections were confirmed by polymerase chain reaction. Results: The results showed that all the infected dogs had significantly lower red blood cell (RBC) count, hemoglobin (HB), hematocrit (HCT), and platelet count (PLT) than the uninfected dogs. Although the dogs with triple infection had lower RBC, HB, HCT, and PLT than those with double and single infections, the difference was not statistically significant. Conclusion: We proposed that triple blood parasite infection with Anaplasma platys, Babesia vogeli, and Ehrlichia canis caused more severe disease than double and single infections. Evaluating the hematological profiles of dogs naturally infected with single, double, and multiple blood parasite infections without clinical signs can enhance their health and welfare.

Association of multilocus sequencing types and antimicrobial resistance profiles of methicillin-resistant Mammaliicoccus sciuri in animals in Southern Thailand.

Background and Aim: Mammaliicoccus sciuri, formerly known as Staphylococcus sciuri, is an opportunistic pathogen in the environment, human and animal mucosa, and skin. Although this pathogen is becoming more resistant to drugs and harmful to animals and humans, basic knowledge of this pathogen remains limited. This study aimed to investigate a new multilocus sequencing type (MLST) related to the antibiotic resistance pattern of M. sciuri from animals in southern Thailand. Materials and Methods: We used 11 methicillin-resistant M. sciuri (MRMS) isolates in this study which were obtained from six horses, four cows, and one chicken of the previous study. Antimicrobial resistance (AMR) was re-evaluated based on the minimum inhibitory concentration using the VITEK® 2 automated system. Three AMR genes were examined, namely mecA, mecC, and blaZ. Staphylococcal chromosomal cassette mec (SCCmec) gene detection was performed through the multiplex polymerase chain reaction (PCR). Internal segments of the seven housekeeping genes, ack, aroE, ftsZ, glpK, gmk, pta1, and tpiA, were used for multilocus sequence typing. The population of resistant bacteria and the types of multidrug-resistant, extensively drug-resistant, and pandemic drug-resistant bacteria were classified through descriptive analysis. Results: mecA and blaZ genes were detected in all isolates; however, the mecC gene was not observed in any isolate based on the PCR results. All MRMS isolates revealed a non-typable SCCmec. Seven MLSTs (71, 81, 120, 121, 122, 199, and 200) were identified in this study. Conclusion: The characteristics of MRMS in Southern Thailand were variable, particularly in cattle and horses. The antibiogram and SCCmec types of this pathogen remain concerns with regard to antibiotic-resistant gene transmission among Staphylococcus and Mammaliicoccus species. All MLSTs in Thailand revealed the distribution among clones in Asia, including the virulence of a zoonotic clone in Southern Thailand.

Interactions between marine megafauna and plastic pollution in Southeast Asia.

Southeast (SE) Asia is a highly biodiverse region, yet it is also estimated to cumulatively contribute a third of the total global marine plastic pollution. This threat is known to have adverse impacts on marine megafauna, however, understanding of its impacts has recently been highlighted as a priority for research in the region. To address this knowledge gap, a structured literature review was conducted for species of cartilaginous fishes, marine mammals, marine reptiles, and seabirds present in SE Asia, collating cases on a global scale to allow for comparison, coupled with a regional expert elicitation to gather additional published and grey literature cases which would have been omitted during the structured literature review. Of the 380 marine megafauna species present in SE Asia, but also studied elsewhere, we found that 9.1 % and 4.5 % of all publications documenting plastic entanglement (n = 55) and ingestion (n = 291) were conducted in SE Asian countries. At the species level, published cases of entanglement from SE Asian countries were available for 10 % or less of species within each taxonomic group. Additionally, published ingestion cases were available primarily for marine mammals and were lacking entirely for seabirds in the region. The regional expert elicitation led to entanglement and ingestion cases from SE Asian countries being documented in 10 and 15 additional species respectively, highlighting the utility of a broader approach to data synthesis. While the scale of the plastic pollution in SE Asia is of particular concern for marine ecosystems, knowledge of its interactions and impacts on marine megafauna lags behind other areas of the world, even after the inclusion of a regional expert elicitation. Additional funding to help collate baseline data are critically needed to inform policy and solutions towards limiting the interactions of marine megafauna and plastic pollution in SE Asia.

Knema retusa is antibacterial and antibiofilm against antibiotic resistant Staphylococcus aureus and S. haemolyticus isolated in bovine mastitis.

This study aimed to assess antibacterial activity of Knema retusa wood extract (KRe) against antibiotic resistant staphylococci which are causative agents of bovine mastitis. From 75 cases of intramammary infections in dairy cows, 66 staphylococcal isolates were collected, including 11 Staphylococcus aureus isolates (17%) and 55 coagulase-negative staphylococci (83%). Sixty isolates (91%) formed strong biofilms. KRe had minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC) against the isolates ranging 32-256 ug/mL and 64-512 ug/mL, respectively. Two-hour KRe exposures at 4×MIC, viabilities of S. aureus and S. haemolyticus decreased by 3 log10 compared to the control. Scanning EM (SEM) showed that KRe disrupted the bacterial cells of both species. KRe at 1/16×MIC significantly inhibited biofilm formation (P < 0.05) in both S. aureus and S. haemolyticus. At 1/2×MIC, S. aureus and S. haemolyticus biofilm inhibition ranged from 75 to 99%. Cells within established biofilms were disrupted 66-83% by KRe at 32×MIC. Moreover, 1/2×MIC KRe reduced bacterial adhesion to glass surfaces observed by SEM. According to GC-MS analysis, the major compound in KRe was endo-2-hydroxy-9,9-(ethylenedioxy)-1-carbethoxy bicyclo [3.3.1] nonane (E2N). Molecular docking analysis of E2N has a high affinity for staphylococcal accessory regulator A (SarA), binding free-energy - 6.40kcal/mol. The results suggested that KRe may have medicinal benefits by inhibiting the growth, biofilm, and adhesion of antibiotic resistant staphylococci isolated from bovine mastitis.

A systematic review and meta-analysis of integrated studies on antimicrobial resistance in Vietnam, with a focus on Enterobacteriaceae, from a One Health perspective.

Vietnam is a low- and middle-income country (LMIC), a primary food producer, and an antimicrobial resistance (AMR) hotspot. AMR is recognized as a One Health challenge since it may transfer between humans, animals and the environment. This study aimed to apply systematic review and meta-analysis to investigate the phenotypic profiles and correlations of antimicrobial-resistant Enterobacteriaceae across three compartments: humans, animals and the environment in Vietnam. A total of 89 articles found in PubMed, Science Direct, and Google Scholar databases were retrieved for qualitative synthesis. E. coli and non-typhoidal Salmonella (NTS) were the most common bacterial species in studies of all compartments (60/89 studies). Among antimicrobials classified as critically important, the resistance levels were observed to be highest to quinolones, 3rd generation of cephalosporins, penicillins, and aminoglycosides. Of 89 studies, 55 articles reported the resistance prevalence of E. coli and NTS in healthy humans, animals and the environment against ciprofloxacin, ceftazidime, ampicillin, gentamicin, sulfamethoxazole-trimethoprim, chloramphenicol was used for meta-analysis. The pooled prevalence was found highest in E. coli against ampicillin 84.0% (95% CI 73.0-91.0%) and sulfamethoxazole-trimethoprim 66.0% (95% CI 56.0-75.0%) while in NTS they were 34.0% (95% CI 24.0-46.0%), 33.0% (95% CI 25.0-42.0%), respectively. There were no significant differences in the pooled prevalence of E. coli and NTS to these antimicrobials across healthy humans, animals and the environment, except for ceftazidime-resistant E. coli (χ2 = 8.29, p = 0.02), chloramphenicol-resistant E.coli (χ2 = 9.65, p < 0.01) and chloramphenicol-resistant NTS (χ2 = 7.51, p = 0.02). Findings from the multiple meta-regression models indicated that the AMR levels in E. coli (ß = 1.887, p < 0.001) and the North (ß = 0.798, p = 0.047) had a higher fraction of AMR than NTS and other regions of Vietnam. The outcomes of this study play an important role as the baseline information for further investigation and follow-up intervention strategies to tackle AMR in Vietnam, and more generally, can be adapted to other LMICs.

First isolation of verocytotoxin-producing Escherichia coli O157:H7 from sports animals in Southern Thailand.

Background and Aim: Escherichia coli O157:H7 is enterohemorrhagic E. coli, which produces verocytotoxin or Shiga toxin. It is a well-known cause of severe diseases in humans worldwide. Cattle and other ruminants are the main reservoirs of this organism. Sports animals, such as fighting bulls, riding horses, and fighting cocks, are economic animals in Southern Thailand. This study aimed to identify E. coli O157:H7 from the rectal swabs of these sports animals and determine the antimicrobial susceptibility patterns of isolated bacteria. Materials and Methods: The rectal swabs were collected from 34 fighting bulls, 32 riding horses, and 31 fighting cocks. The swabs were cultured on MacConkey (MAC) Agar; the suspected colonies were then identified by VITEK® 2 GN card, and the antimicrobial susceptibility was tested by VITEK® 2 AST N194 in VITEK® 2 Compact automation. Escherichia coli O157:H7 was confirmed by culturing on sorbitol MAC agar, the ability to grow at 44°C, and the presence of H7 antigen. In addition, the eaeA (E. coli attaching and effacing), along with stx1 and stx2 (Shiga cytotoxins) genes, were determined using polymerase chain reaction. Finally, the cytotoxicity of Shiga toxin was confirmed using the Vero cytotoxicity test. Results: Fifty-five suspected isolates (56.70%), which were collected from 19 fighting bulls (55.88%), 13 riding horses (40.63%), and 23 fighting cocks (71.13%), were identified as E. coli. However, one sample (Bull H9/1) from fighting bulls had an equal confidence level (50%) for E. coli and E. coli O157. The confirmation of this isolate demonstrated that it was sorbitol non-fermenter, could assimilate L-lactate, was unable to grow well at 44°C, and reacted with anti-serum to H7 antigen. In addition, it was positive with stx2 and eaeA genes, and the toxin affected Vero cells by a dose-dependent response. The antimicrobial susceptibility test revealed that five out of 55 (9.09%) E. coli isolates were resistant to antimicrobial agents. All five isolates (21.74%) were collected from fighting cocks. Escherichia coli Cock H4/3 was only one of the five isolates resistant to three antimicrobial agents (ciprofloxacin, moxifloxacin, and trimethoprim/sulfamethoxazole). Fortunately, it was not multidrug-resistant bacteria. Conclusion: This is the first report on detection of E. coli O157:H7 in fighting bulls and antibiotic-resistant characteristic of E. coli in fighting cocks in Southern Thailand. This research is beneficial in preventing the dissemination of E. coli O157:H7 or antimicrobial agent-resistant E. coli in sports animals and humans.

Sangyod rice bran extract enhances Lacticaseibacillus paracasei growth during the exponential phase and antibacterial activity of L. paracasei supernatant against zoonotic and foodborne pathogens.

Background and Aim: Prebiotics are a group of nutrients or compounds that are degraded by the gut microbiota, including Lacticaseibacillus paracasei. The probiotic plays an important role in adhesion to the gut and is able to produce antimicrobial substances to inhibit pathogens. This study aimed to investigate the effects of Sangyod rice bran extract on the growth promotion of L. paracasei. Furthermore, antibacterial activity of the extract and L. paracasei supernatants cultured in De Man, Rogosa and Sharpe (MRS) medium plus the extract against zoonotic and foodborne pathogens was investigated. Materials and Methods: Antibacterial activity of the crude extract and the oil from Sangyod rice bran against the pathogens, including Bacillus cereus, Staphylococcus aureus, Escherichia coli, Avian pathogenic E. coli, and Pseudomonas aeruginosa was investigated using broth microdilution assay. The effects of the crude extract and the oil on the growth and adhesion of L. paracasei were further determined. The antibacterial activity of L. paracasei supernatant cultured in the medium supplemented with the extract and the oil against the pathogens was determined by agar well diffusion assay, followed by the broth microdilution assay. Finally, the chemical constituents and antioxidant activity of the crude extract and the oil from Sangyod rice bran were investigated. Results: The crude extract and the oil from Sangyod rice bran enhanced L. paracasei growth during the exponential phase. Furthermore, the crude extract at 0.25 mg/mL significantly enhanced the adhesion of L. paracasei to the surface compared with the control. Both minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values of the crude extract against B. cereus and S. aureus were 0.5 and 1.0 mg/mL, respectively. All pathogens were sensitive to the supernatant of L. paracasei with similar MIC and MBC ranging from 12.5% v/v to 50% v/v. However, the MIC and MBC values of L. paracasei supernatant grown in MRS medium plus the crude extract and oil were not significantly different compared to the supernatant obtained from MRS alone. The crude extract had free radical scavenging activities with IC50 values at 0.61 mg/mL. Conclusion: The results suggested the potential benefits of the crude extract from Sangyod rice bran for inducing the growth and the adhesion of L. paracasei and inhibiting zoonotic and foodborne pathogens.

Association of synovial expression of growth and differentiation factor 5 (GDF5) with radiographic severity of knee osteoarthritis.

Objectives: The growth and differentiation factor 5 (GDF5) gene plays a significant role in facilitating chondrogenesis, and GDF5 polymorphism is a genetic factor contributing to osteoarthritis (OA). However, the role of GDF5 expression in the synovial membrane remains unclear. The aim of this study was to determine the expression of GDF5 in the synovium in patients with primary knee OA. Materials and methods: Thirty patients scheduled for total knee arthroplasty were enrolled. Patients were grouped according to the Kellgren and Lawrence classification (KL) as grade 3 (15 patients) and grade 4 (15 patients). Synovial tissue was collected, and the GDF5 expression level was determined by real-time PCR. Ten synovial samples were randomly selected to evaluate the degree of synovitis. Results: Baseline characteristics did not differ between the two groups. The expression of GDF5 was significantly higher in the KL4 group (median expression 3.50, range 1.45-13.62) than in the KL3 group (median expression 1.81, range 0-9.46) (p value = 0.02). Histological staining of the synovium indicated low-grade synovitis in both groups. Conclusions: GDF5 expression in the synovium was positively associated with the radiographic severity of knee OA. The difference in GDF5 expression between the KL3 and KL4 groups supports the hypothesis that, through GDF5, the synovium may have important roles in cartilage maintenance and homeostasis in primary knee OA.

First study on molecular detection of hemopathogens in tabanid flies (Diptera: Tabanidae) and cattle in Southern Thailand.

Background and Aim: Female tabanids play a key role in disease transmission as mechanical vectors for various hemopathogens, but only a limited number of studies have been conducted on them. This study aimed to investigate the occurrence of hemopathogens in tabanid flies compared to those found in nearby cattle hosts. Materials and Methods: Tabanids were collected using a Nzi trap for three consecutive days per month during the dry season (February-May 2021). Furthermore, blood samples were collected from 20 beef cattle (Bos taurus) raised in the same area where the flies were captured. Conventional polymerase chain reaction (PCR) was used to detect hemopathogenic DNA in flies and beef cattle. Results: In total, 279 female tabanids belonging to five species were collected: Tabanus megalops, Tabanus rubidus, Tabanus mesogaeus, Chrysops dispar, and Chrysops fuscomarginalis. Notably, T. megalops was the most abundant species, accounting for 89.2% of the flies collected (n = 249). PCR technique revealed that 76.6% of T. megalops carried at least one pathogen (Anaplasma, Ehrlichia, Babesia, or Theileria). In addition, all beef cattle had multiple hemopathogenic infections (Anaplasma marginale, Ehrlichia spp., Babesia bigemina, Babesia bovis, and Theileria spp.). Conclusion: Although T. megalops could carry many hemopathogens, it might not be an important vector due to the limited number of flies and parasitic load. Furthermore, T. megalops could be utilized to monitor the presence of hemopathogens in the study area, but not the disease occurrence in the individual host species. Knowing the presence of hemopathogens in flies could help manage the disease in this area.

Effect of feeding different types of ß-glucans derived from two marine diatoms (Chaetoceros muelleri and Thalassiosira weissflogii) on growth performance and immunity of banana shrimp (Penaeus merguiensis).

ß-glucans are produced by many organisms and could be used as supplementary feed to enhance immunity and growth in some aquatic animals. This study aimed to compare the effectiveness of ß-glucans derived from two marine diatoms (Chaetoceros muelleri and Thalassiosira weissflogii) as growth promoters and immunity enhancers in banana shrimp (Penaeus merguiensis). Shrimp were divided into 3 groups: the control group was fed without ß-glucan; the second and the third group were fed with 2 g kg-1 of ß-glucan derived from C. muelleri and T. weissflogii, respectively. Shrimp were fed over a 30-day period to determine growth performance (final weight, weight gain, average daily gain (ADG), and feed conversion ratio (FCR)) at day 15 and day 30, respectively. The immune parameters determined were total hemocyte count (THC), phenoloxidase activity (PO) and immune gene expression. Survival rates were measured after 14 days of the feeding trial and Vibrio parahaemolyticus infection (6, 24, 48 h post infection). There was no significant difference (P > 0.05) for growth stimulation of shrimps between the two types of ß-glucans (C. muelleri or T. weissflogii). Notably, shrimps fed with ß-glucans had a higher final weight, weight gain, and ADG (P < 0.05) than shrimps fed with the control diet, while FCR of shrimps fed with both ß-glucans was lower when compared to the control diet. Immune parameters, THC, PO, and gene expression of anti-lipopolysaccharide factor (ALF) and crustin were significantly higher (P < 0.05) in shrimps fed with ß-glucans, especially with ß-glucans from C. muelleri than the control group both before and after V. parahaemolyticus infection. Expression of penaeidin 3 and peroxiredoxin genes was significantly higher in shrimps fed with ß-glucans after bacterial infection. Histopathology of hepatopancreas revealed an increase in blasenzellen hepatopancreatic epithelial cells (B cells) after 14 days of feeding which remained higher following infection with V. parahaemolyticus. The survival rate of shrimps fed with the diet containing ß-glucan derived from either C. muelleri (82.2%) or T. weissflogii (77.8%) after V. parahaemolyticus infection was significantly higher than for the control group (51.1%) (P < 0.05). In conclusion, we propose that feeding banana shrimps with ß-glucans derived from marine diatoms either C. muelleri or T. weissflogii at a 2 g kg-1 diet can significantly improve their growth performance and immunity.

Multiple blood pathogen infections in apparently healthy sheltered dogs in southern Thailand.

In developing countries such as Thailand, free-ranging dogs are frequently involved in road accidents and contribute to the cost of public healthcare. Shelters play a vital role in communities because they help to control the population of unwanted and free-ranging dogs. This study aimed to investigate blood pathogen infection in sheltered dogs, as it is one of the factors contributing to animal welfare. Blood samples were randomly collected from 141 dogs from the largest shelter (approximately 400-500 dogs in total) in southern Thailand. Blood pathogens were detected using both PCR and light microscopy. Four blood pathogens were identified: Anaplasma platys, Ehrlichia canis, Babesia canis vogeli, and Hepatozoon canis. No trypanosomes were detected. The incidence of blood parasite infection was 56.7% (80/141) by PCR, and 28.4% (40/141) by microscopy. E. canis was the most prevalent pathogen, accounting for 46.1% (65/141) of the cases, while multiple infections accounted for 22% (31/141) of the cases. A triple infection with E. canis, A. platys, and B. canis vogeli was observed in 5.7% (8/141) of the cases. Although PCR is far more sensitive than microscopy, it appears to have equivalent specificity. In conclusion, this study reported a high occurrence of blood pathogen infections in clinically healthy sheltered dogs. Many of them were infected with multiple pathogens and may have been infected before entering the shelter. These findings suggest that a blood test is necessary to screen dogs prior to their admission to the shelter to prevent disease transmission and enhance animal welfare.

The combination of BMP12 and KY02111 enhances tendon differentiation in bone marrow-derived equine mesenchymal stromal cells (BM-eMSCs).

The Wingless and Int-1 (WNT) and bone morphogenic protein/growth differentiation factor (BMP/GDF) signalling pathways contribute significantly to the development of the musculoskeletal system. The mechanism by which they contribute is as follows: BMP/GDF signalling usually promotes tendon differentiation, whereas WNT signalling inhibits it. We hypothesised that inhibiting WNT and subsequently stimulating BMP signalling may enhance the tenogenic differentiation of stem cells. The objective of this study was to determine whether a combination of WNT inhibitor (KY02111) and BMP12/GDF7 protein could enhance the differentiation of bone marrow-derived equine mesenchymal stromal cells (BM-eMSCs) into tenocytes. Cells were cultured in five treatments: control, BMP12, and three different combinations of BMP12 and KY02111. The results indicated that a 1-day treatment with KY02111 followed by a 13-day treatment with BMP12 resulted in the highest tenogenic differentiation score in this experiment. The effect of KY02111 is dependent on the incubation time, with 1 day being better than 3 or 5 days. This combination increased tenogenic gene marker expression, including SCX, TNMD, DCN, and TNC, as well as COL1 protein expression. In conclusion, we propose that a combination of BMP12 and KY02111 can enhance the in vitro tenogenic differentiation of BM-eMSCs more than BMP12 alone. The findings of this study might be useful for improving tendon differentiation protocols for stem cell transplantation and application to tendon regeneration.

Antimicrobial resistance in Staphylococcus pseudintermedius on the environmental surfaces of a recently constructed veterinary hospital in Southern Thailand.

Background and Aim: Staphylococcus pseudintermedius is a zoonotic bacterium commonly found in animals, especially dogs. These bacteria can survive on environmental surfaces for several months. The infection of S. pseudintermedius from the environment is possible, but properly cleaning surface objects can prevent it. This study aimed to investigate the prevalence of methicillin-resistant S. pseudintermedius (MRSP) in the environment of a recently constructed veterinary hospital in Southern Thailand, where we hypothesized that the prevalence of MRSP might be very low. Materials and Methods: At three different time points, 150 samples were collected from different environmental surfaces and wastewater across the veterinary hospital. The collection was done after the hospital's cleaning. Bacteria were purified in the culture before being identified as species by biochemical tests and polymerase chain reaction (PCR). Next, the antimicrobial-resistant profile was performed using an automated system (Vitek 2). Finally, the antimicrobial resistance genes were identified using PCR. Results: Fifteen colonies of S. pseudintermedius were isolated from the surfaces of eight floors, four tables, two chairs, and one rebreathing tube. Fourteen colonies (93.3%) were multidrug-resistant (MDR) and carried the blaZ gene (93.3%). The majority of colonies were resistant to benzylpenicillin (93.3%), cefovecin (93.3%), ceftiofur (93.3%), kanamycin (93.3%), and neomycin (93.3%). Notably, only four colonies (26.7%) were methicillin-susceptible S. pseudintermedius, whereas 11 colonies (73.3%) were MRSP and carried both the mecA and blaZ genes. Five MRSP (45.5%) were resistant to at least 14 antimicrobial drugs, represented as extensively drug-resistant (XDR) bacteria. Ten of eleven MRSP (90.9%) were Staphylococcal chromosomal mec type V, while another displayed untypeable. Despite the routine and extensive cleaning with detergent and disinfectant, MRSP isolates were still detectable. Conclusion: Many isolates of MRSP were found in this veterinary hospital. Almost all of them were MDR, and nearly half were XDR, posing a threat to animals and humans. In addition, the current hospital cleaning procedure proved ineffective. Future research should be conducted to determine the bacterial biofilm properties and bacterial sensitivity to certain detergents and disinfectants.