Milk, a nutritious food, is widely consumed in human diets; however, contamination by micro-organisms can negatively impact its quality and consumer health. Contamination by micro-organisms affects the quality of milk, which can affect the quality of the milk production chain. This study aimed to determine the changes in milk composition and antibiotic susceptibility related to bacteria isolated from dairy cow milk. Raw milk samples were collected from 72 dairy cows. All milk samples were subjected to the California Mastitis Test (CMT) for CMT score determination. We also investigated milk composition, bacterial culture (BC), and antibiotic susceptibility. About 47.22% and 30.56% of dairy cattle were positive for CMT + BC and automatic somatic cell count (ASCC) + BC, respectively. Fecal appearance and animal age were found to be risk factors for ASCC + BC positivity in dairy cattle. Bacteria were found in approximately 76% of milk samples, with the most common isolated species being α-hemolytic Streptococcus spp., coagulase-negative Staphylococcus spp., and Escherichia coli. Of these, 70% are resistant to at least one antibiotic. Variation in the multidrug resistance pattern was high in Klebsiella spp. Fecal appearance and animal age are risk factors for ASCC + BC positivity in dairy cattle. This study identified antibiotic and multidrug resistance patterns, which require comprehensive studies and effective surveillance systems. Remarkably, the use of antibiotic therapy in dairy cattle should be monitored.

Live-attenuated vaccines are the most successful type of vaccine and could be useful in controlling fowl adenovirus (FAdV) 8b infection. This study aimed to attenuate, molecularly characterize, and determine the immunogenicity, efficacy, and challenge virus shedding in broiler chickens. The FAdV 8b isolate (UPM08136) was passaged onto chicken embryo liver (CEL) cells until attenuation. We sequenced and analyzed the hexon and fiber genes of the passage isolates. The attenuated bioreactor-passage isolate was inoculated into 1-day-old broiler chickens with (attenuated and inactivated) and without booster groups and challenged. Body weight (BW), liver weight (LW), liver: body weight ratio (LBR), FAdV antibody titers, T-lymphocyte subpopulation in the liver, spleen, and thymus, and challenge virus load and shedding were measured. Typical cytopathic effects with novel genetic changes on CEL cells were observed. The uninoculated control-challenged (UCC) group had significantly lower BW and higher LW and LBR than the inoculated groups. A significantly higher FAdV antibody titer was observed in the challenged non-booster and attenuated booster groups than in the UCC group. T cells in the spleen and thymus of the liver of inoculated chickens were higher than uninoculated control group levels at all-time points and at different times. A significantly higher FAdV challenge virus load was observed in the liver and shedding in the cloaca of UCC chickens than in non-booster chickens. The FAdV 8b isolate was successfully attenuated, safe, and immunogenic. It reduces virus shedding and is effective and recommended as a vaccine against FAdV infection in broiler chickens.

Toxoplasma gondii and Trypanosoma evansi, the zoonotic protozoa responsible for toxoplasmosis and trypanosomiasis, are significant threats to the productivity and financial stability of livestock farming. T. gondii can be transmitted horizontally through ingestion of fecal oocysts and T. evansi through arthropod vectors. In addition, both species can be transmitted from mother to fetus through the placenta. This study aimed to assess the molecular prevalence of T. gondii and T. evansi transplacental-transmitted protozoans and to identify the epidemiological risk factors in recently calved female cattle across Phayao, Thailand. We collected 106 bovine placentas from beef and dairy cow full-term pregnancies in Phayao, Thailand. T. gondii and T. evansi DNA were detected using targeted B1 gene and expression site-associated gene (ESAG) species-specific polymerase chain reaction (PCR), respectively. Forty-three placentas were positive for T. gondii B1 PCR, whereas only one was positive for T. evansi ESAG PCR, resulting in an overall prevalence of transplacental-transmitted protozoan infection of 41.5% (44/106). The prevalence of T. gondii and T. evansi was 40.6% (43/106) and 0.9% (1/106), respectively. No significant correlation was found between T. gondii infection and various risk factors, including locality, age, and cattle type. The prevalence of transplacental-transmitted protozoan T. gondii infection was high among female cattle in Phayao, Thailand, whereas the prevalence of T. evansi infection was notably lower. Although the conventional modes of transmission differ between these two parasites, the transplacental transmission of T. evansi and especially T. gondii may play a crucial role in the persistence of these protozoan species in this area.

In Kazakhstan, the study of Echinococcus infection among farm animals is crucial to monitor the invasion among livestock and map the data obtained. Unfortunately, there are only partial data on the study of Echinococcus among cattle's in Kazakhstan, which makes it difficult to conduct a comparative analysis of the epidemiological situation among livestock animals. The present study aimed to molecularly identify the species and haplotypes of the E. granulosus complex infecting cattle in Kazakhstan and investigate their genetic variation relative to mitochondrial (mt) targets. Individual cyst isolates (n = 700) were collected from infected cattle lungs and livers after slaughter from the slaughterhouse. Total DNA was extracted from the germinal layers of the cyst from each isolate. This DNA sequenced partial mt genes of cytochrome c oxidase 1 (450 bp) and NADH dehydrogenase 1 (1200 bp). We determined that all the sequences were detected as E. granulosus s.s., of which 69 (94.5%) samples belonged to G1, and only 4 (5.4%) samples belonged to the G3 genotype. After bioinformatic analysis, 38 haplotypes were identified. Our findings revealed that the G1 genotype of E. granulosus s.s. is the predominant cattle genotype in Kazakhstan. However, only one region showed the presence of two genotypes G1 and G3, in the sequence, which suggests that further research is needed to investigate the epidemiology of Echinococcus infection in cattle in Kazakhstan.

The high prevalence of QX-like variant among Thai isolates poses a significant threat to poultry production. In this study, we evaluated the protective efficacy of commercially available heterologous infectious bronchitis virus (IBV) vaccines against the local Thai QX-like strain in specific-pathogen-free (SPF) chicks from Thailand. The experiment involved 100 SPF chicks divided into 4 arms. Arms I and II received the TAbic IB VAR (233A) and Ibird (1/96) vaccines, respectively, on day 1. After 10 days, both arms received the H120 vaccine. Arms III and IV were non-vaccinated positive and negative controls. Challenge infection was local Thai QX-like virus on birds of Arms I, II, and III, and negative control of Arm IV. Clinical signs of infectious bronchitis (IB) and IBV detection using reverse transcription polymerase chain reaction were assessed at 2, 4, and 6 days post-challenge (dpc). At 6 dpc, the birds were humanely euthanized for post-mortem examination with the ciliostasis test and histopathological analysis of the tracheas, lungs, and kidneys. Virus shedding started at 4 dpc (33.3% positive) and reached 100% positivity at 6 dpc with obvious clinical respiratory symptoms in non-vaccinated-challenged birds. No detection of IBV in vaccinated-challenged arms. Ciliary activity scores were significantly lower in non-vaccinated-challenged birds at 23.64 (standard deviation [SD] ± 1.74) and 96.50 (SD ± 1.91) and 95.64 (SD ± 1.77), respectively (p = 0.05) than in vaccinated-challenged birds. The most remarkable histopathological changes were observed in non-vaccinated-challenged birds, with moderately severe changes in the trachea, lungs, and kidneys. On the other hand, birds in vaccinated-challenged arms showed no significant changes. This study demonstrated the efficacy of TAbic IB VAR (233A) or Ibird (1/96) vaccine combined with a Massachusetts serotype vaccine (H120) against the local Thai QX-like strain in SPF chicks, contributing valuable insights to the selection of suitable commercially available vaccines to combat the prevalent local QX-like strains in Thailand.

Halal restaurants participating in online food delivery services do not require halal certification. The Halal status of products through the Halal logo provides the consumer with information on the basis of which he decides to buy. Online transactions involve potential risks related to online processes, payment methods, and product quality. The aim of this study was to determine whether a declared Halal label is in accordance with the business processes implemented. Halal authentication of Gofood's meatball partner products in Yogyakarta and Solo Raya determined the incompatibility of meatball ingredients. Sixty meatball samples were collected from Yogyakarta and 30 samples from Solo Raya. Halal certification test was carried out using the thermal cycle polymerase chain reaction method at Universitas Gadjah Mada Animal Husbandry Laboratory and the results were used to identify pork contamination in meatballs. The addition of pork or pork meatballs was used as a control. Eight meatball restaurants in the Solo Raya and Yogyakarta areas were found to be contaminated with pig DNA. The results of the tracing materials and processes, i.e., the grinding stage, are critical because all samples were supposed to be made from beef. It is known from interviews that contamination with pig DNA at the milling stage was accidental. Restaurants that sell meatballs are committed to adhering to product labels that are 91.1% safe from pork contamination. The Halal and original beef labels were in accordance with their statements. This study highlights the concept of Halal authentication with traceability to overcome pork contamination in meat products.

Stingless bees are generally found in tropical countries, including Indonesia. In West Sumatra, stingless bees are known as Galo-galo, consist of several species with different characteristics; however, the properties of honey produced by stingless bees have not yet been explored. This study aimed to determine the physicochemical, antioxidant, and antimicrobial activities as well as the microbiota profile of stingless bee honey from the bee species Heterotrigona itama, Geniotrigona thoracica, Tetrigona melanoleuca, and Tetrigona binghami that are intensively developed in West Sumatra, Indonesia. Honey produced by the stingless bee species H. itama, G. thoracica, T. melanoleuca, and T. binghami originating in West Sumatra was examined in the present study. The physicochemical properties (Association of Official Analytical Chemists), antioxidant activity (2,2-diphenyl-1-picrylhydrazyl technique), total phenols (Folin- Ciocalteu method), antimicrobial activity (Agar-Well diffusion test), total lactic acid bacteria, and microbiota diversity were measured in stingless bee honey samples. Stingless bee species significantly affected the physicochemical properties, antioxidant activity, total phenolic content, antimicrobial activity, and total lactic acid bacteria (p = 0.05), except for the crude fiber content. The carbohydrate profiles of honey produced by H. itama and T. binghami were dominated by monosaccharides, whereas those of honey from T. melanoleuca and G. thoracica were dominated by disaccharides. In terms of antioxidant activity (half maximal inhibitory concentration [IC50] value), there were no significant differences (p > 0.05) between honey from H. itama, T. melanoleuca, and T. binghami, but there were significant differences (p > 0.05) between honey from G. thoracica. The honey of G. thoracica and T. melanoleuca had the highest total phenolic content (65.65 ± 14.00 and 69.78 ± 8.06, respectively). In addition, honey from the four stingless bee species showed antimicrobial activity against the pathogenic bacteria Escherichia coli, Salmonella, Staphylococcus aureus, and Listeria monocytogenes. From the principal co-ordinate analysis (PCoA) results, it can be concluded that the microbiota profiles of the four stingless bee honey samples differed. The results showed that honey from H. itama, G. thoracica, T. melanoleuca, and T. binghami has different physicochemical characteristics, antioxidant activity, antimicrobial activity, and microbiota diversity. By knowing the content of this stingless bee honey, the results of this study can be used as information that this stingless bee honey has the potential as a functional food that is beneficial for health.

Cardiogenic embolism (CE) is a common complication of feline hypertrophic cardiomyopathy (HCM), leading to severe clinical symptoms. This study compared the effects of rivaroxaban and enoxaparin combined with clopidogrel on cats. This was a single-center, prospective, randomized controlled trial. In this study, rivaroxaban or enoxaparin plus clopidogrel was prescribed to 23 cats for at least one of the following events: Abnormal movement of the anterior mitral leaflet during systole, enlargement of the left atrium, spontaneous echocardiographic contrast, or presence of arterial thromboembolism. Oral rivaroxaban (2.5 mg, q24 h) was prescribed to six cats. Subcutaneous injections of enoxaparin (1 mg/kg, q24 h) plus oral clopidogrel (3 mg/kg, PO q24 h) for 60 days were administered to 17 cats. Renal insufficiency and bleeding complications were observed. Plasma concentrations of D-dimer, prothrombin time (PT), partial thromboplastin time, and international normalized ratio (INR) were evaluated. We analyzed the relationship between echocardiography parameters and the effects of coagulation. Blood samples were collected from all cats at baseline and at 1 and 2 months post-treatment. Rivaroxaban alone and in combination with enoxaparin and clopidogrel significantly affected PT and INR. In cats treated with 2.5 mg/kg rivaroxaban for 60 days, no bleeding or recurrence of thrombus formation was observed. These data support the use of rivaroxaban for the treatment of HCM-associated thromboembolism in cats. Treatment of HCM-associated thromboembolism with rivaroxaban alone demonstrated clinical effectiveness with no clinical complications in cats.

Lipopolysaccharide (LPS) is a robust endotoxin known to activate the immune system in cattle. The objective of this study was to investigate the effect of LPS on the morphology, cell viability, malondialdehyde (MDA), nitric oxide (NO), and total antioxidant capacity (TAC) of peripheral blood mononuclear cells (PBMCs) in Brahman and Brahman x Thai native crossbreed cattle. PBMCs were isolated from Brahman and Brahman x Thai native crossbreed cattle and treated with 0, 0.1, 1, and 10 μg/mL Escherichia coli LPS, respectively. Morphological changes in PBMCs were assessed at 24 and 48 h. In addition, we measured PBMC cell viability, MDA, NO, and TAC. LPS stimulation caused cell deformation and partial PBMC area enlargement, but there were no differences between Brahman and Brahman x Thai native crossbreed cattle. Stimulation at all levels did not affect the viability of PBMCs (p > 0.05). MDA and NO levels were significantly higher in Brahman cattle than in Brahman Thai native crossbred cattle (p < 0.05). TAC was significantly higher in Brahman x Thai native crossbred cattle than in Brahman cattle (p < 0.05). Immune cells of crossbreed cattle have a higher activation response to LPS than those of purebred cattle, and native crossbreed beef cattle have a higher antioxidant capacity than purebred beef cattle. This result may explain why hybrid cattle of indigenous breeds are more resistant to disease than purebred cattle.

Brucellosis is a zoonotic disease that causes enormous losses in livestock production worldwide and has a significant public health impact. None of the brucellosis-free countries is currently able to guarantee their ability to prevent the introduction of the pathogen due to the increase in tourism and the expansion of migration. The timely identification of infected animals is an effective means of preventing brucellosis and minimizing the epidemiological risk. The tube agglutination test, Rose Bengal plate test, complement fixation test, and enzyme-linked immunosorbent assay, which are routinely used to identify seropositive productive animals, have limitations and results that do not always correlate. The indirect hemagglutination assay (IHA) stands out among non-traditional methods because it is affordable, has a simple protocol, and is more reliable than classical serological tests, especially in cases of questionable and/or false-negative results. The diagnostic value of the IHA has long been studied by laboratories in several countries, but mostly by post-soviet research teams; therefore, the results continue to be published in Russian-language journals, ensuring that the local scientific community can access the results. In addition, the efficacy of this test for the diagnosis of brucellosis and other infectious diseases has not yet been reviewed. The purpose of this review was to summarize the results of studies on the development and use of IHA for the diagnosis of brucellosis and to determine the prospects for further improvement

The quality of canine sperm can be influenced by many factors, such as breed, body weight, age, ejaculatory frequency, nutrition, and environment. In the UK, it is common practice for standard Bull Terriers (SBT) and miniature Bull Terriers (MBT) to require male donors during a short breeding period. The aim of this study was to evaluate the effect of semen collection frequency on ejaculate volume and nine sperm parameters in SBT and MBT males, considering age and body condition score (BCS). Ejaculates from six adult SBTs and four MBTs were collected 5 times at two consecutive intervals (Time Series [TS]1, 24 h vs. TS2, 48 h), 1 week apart. Ejaculate volume, concentration, total output, viability (live sperm), subjective total motility, vigor, and total morphological defects, including head, midpiece, and tail defects of sperm, were evaluated. A multivariable mixed linear model for repeated measures was used to analyze the effects of semen collection frequency, age, breed, and BCS on ejaculate volume and sperm parameters. Semen collection frequency, age, and, to a lesser extent, breed, and BCS significantly affected sperm parameters. Semen collection frequency affected all sperm parameters (p < 0.05) but not ejaculate volume (p > 0.05). Total sperm output, sperm vigor, total motility, and tail defects decreased (p < 0.05) at the end of TS1. However, sperm parameters remained relatively constant (p > 0.05) in TS2 between semen collection sessions. Overall, poorer sperm parameters were observed in older dogs (aged 5-8 years) than in younger dogs (aged 4 years). MBT produced less (p < 0.001) ejaculate volume (3.2 ± 0.2 mL vs. 4.3 ± 0.2 mL: Least Squares Mean ± Standard Error of Mean), lower total sperm output (221.8 ± 19.2 x 10⁶ vs. 348.6 ± 19.2 x 10⁶) and lower total morphological defects (25.0 ± 1.1% vs. 31.3 ± 0.9%), and a higher percentage of live sperm (77.0 ± 1.4% vs. 71.7 ± 1.1%) than SBT. In addition, a BCS of 4 positively influenced (p < 0.05) viability, vigor, and total sperm motility. Despite differences in age, breed, and BCS, better sperm parameter values were observed in all semen collection sessions. However, intensive semen collection (TS1) appears to be less effective in maintaining good sperm quality. For breeding or artificial insemination purposes, a 48-h interval between collection sessions is recommended for both breeds. The results of this study could be used to further optimize assisted reproductive technologies in both breeds.

Donkey and mare milk have high nutritional and functional values, but their lactic acid bacteria (LAB) content remains poorly studied and undervalued in the Algerian dairy industry. This study aimed to isolate and select LAB strains that produce antimicrobial substances during fermentation and to characterize the probiotic profiles of each extracted strain to indicate their potential for antioxidant and proteolytic activity. This study focuses on isolating and identifying lactic acid bacterial strains from 10 Equid-fermented milk samples collected in two regions of El Bayed Wilaya (Algeria). Identification of LAB strains was obtained by 16S rRNA sequencing. The probiotic properties of important strains and their aromatic productivity power are assessed. To evaluate their antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, Chryseobacterium joostei, Pseudomonas aeruginosa, and Escherichia coli, we selected 21 strains. Different induction methods have been used to amplify the antibacterial effects against these pathogenic strains. Among a total of 60 identified strains, 31 had a probiotic profile, and most were catalase-negative. Aromatic productivity power was observed in eight strains: Lactiplantibacillus plantarum, Lactobacillus casei, Lactobacillus paracasei, Weissella confusa, Weissella cibaria, Leuconostoc mesenteroides, Leuconostoc lactis, and Lactobacillus sp1. Our results provide insight into the considerable diversity of LAB present in fermented donkey and mare milk. To meet the expectations of the Algerian dairy industry, it is important that the probiotic skills of the nine selected strains are met. In addition, a significant number of these strains may have important probiotic activity and biotechnological potential.

Query fever (Q fever) is an endemic zoonotic disease and ruminants are considered to be the primary source of infection in humans. It is caused by Coxiella burnetii which is an obligate intracellular bacterial pathogen with a worldwide distribution. This study estimated the prevalence of Q fever in livestock with a history of abortion in Makkah Province, Saudi Arabia. Sera from 341 camels, 326 sheep, and 121 goats of either sex from various locations (Makkah, Jeddah, AL-Taif, AL-Qunfudah, AL-Laith, and AL-Kamil) were examined using a Q fever indirect enzyme-linked immunosorbent assay. Among the 788 serum samples, 356 animals had anti-Coxiella burnetii immunoglobulin G antibodies with an overall seroprevalence of 45.4%. Significant differences were observed in seroprevalence between species and locations. Camels had the highest percentage of Q fever-positive sera, with a prevalence of 50.4%, followed by goats (44.6%) and sheep (36.8%), with a high significant difference between animals (p = 0.000). The prevalence was significantly higher in Makkah (65.4%) than in Jeddah (28.8%). C. burnetii infection is prevalent in agricultural animals, especially camels maintained at livestock farms in Makkah province. Therefore, these animals considered as the main source of Q fever infections in Saudi Arabia, which is also a reason for the abortion in these animals. Therefore, there is an urgent need for further studies on Q fever infection with interventional approaches for prevention and control.

Keratitis is a serious ocular infection often caused by pathogenic micro-organisms such as Acanthamoeba spp. Among other harmful microbes, Acanthamoeba keratitis presents a particular challenge due to its resistance to conventional antimicrobial agents. Piper betle Linn., commonly known as betel leaf, has been traditionally used for its medicinal properties. This study aimed to assess the potential of the leaf ethanol extract of P. betle Linn. in the treatment of Acanthamoeba triangularis in monoculture and co-culture with two prevalent pathogenic bacteria, Staphylococcus aureus and Pseudomonas aeruginosa, associated with keratitis. Minimum inhibitory concentrations (MICs) of A. triangularis, S. aureus, and P. aeruginosa extracts in monoculture and coinfected conditions were examined. In addition, this study explored the potential of the extract in preventing Acanthamoeba adherence in both monoculture and co-culture environments. Scanning electron microscopy (SEM) analysis confirmed the impact of the extract on Acanthamoeba cell membranes, including acanthopodia. Furthermore, a time-kill kinetic assay was used to validate the amoebicidal activity of the extract against A. triangularis and the tested bacteria. MICs for trophozoites, cysts, P. aeruginosa, and S. aureus in the monoculture were 0.25, 0.25, 0.51, and 0.128 mg/mL, respectively, whereas the MICs for Acanthamoeba coinfected with bacteria were higher than those in the monoculture. This extract inhibited the growth of A. triangularis trophozoites and cysts for up to 72 h. Moreover, P. betle extract effectively prevented the adherence of Acanthamoeba to contact lenses under monoculture conditions. SEM analysis confirmed that P. betle extract affects the cell membrane of Acanthamoeba, including Acanthopodia. In addition, the time-kill kinetic assay confirmed that the extract contained amoebicidal activity against A. triangularis, including the tested bacteria. Notably, S. aureus was more susceptible than A. triangularis and P. aeruginosa to P. betle extract treatment. Unexpectedly, our study revealed that S. aureus negatively affected A. triangularis in the co-culture after 3 days of incubation, whereas P. aeruginosa facilitated the growth of A. triangularis in the presence of the extract. This study provides compelling evidence of the anti-adhesive and anti-Acanthamoeba properties of P. betle leaf extract against A. triangularis under monoculture and co-culture conditions. The observed impact on Acanthamoeba cell membranes, coupled with the time-kill kinetic assay results, underscores the potential of P. betle leaf extract as a promising agent for combating Acanthamoeba-related infections in humans and animals.

Semen storage is an important reproductive method used in artificial livestock breeding. However, oxidative stress during storage reduces the quality of sperm. Melatonin supplementation in semen storage medium has not been well studied, but it has been shown to protect cells from oxidative stress. Therefore, this study aimed to determine the effect of melatonin supplementation on sperm quality parameters and antioxidant gene expression levels in semen extenders during cold storage. Semen extenders with melatonin concentrations of 0 (control), 0.1, 0.2, and 0.3 mM were added as treatment. The treated semen was then stored at 5&degC for 72 h using a cold storage method, and quality parameters, including percentage of progressive motility, membrane integrity, intact acrosome, and DNA integrity, were measured every 24 h. In addition, messenger ribonucleic acid abundance levels of glutathione peroxidase (GPx) and superoxide dismutase (SOD) genes were sampled after 0 and 72 h of cold storage. All observed sperm quality parameters decreased with increasing cold storage time; however, 0.2 mM melatonin demonstrated superior protection of sperm quality during cold storage. Gene expression analysis showed that GPx levels decreased significantly (p < 0.05) after 72 h in semen without melatonin but not in the melatonin-treated groups. A similar trend was also observed in SOD, indicating that exogenous antioxidants effectively protected the sperms. Melatonin supplementation at 0.2 mM in semen extenders during cold storage maintains sperm quality parameters for up to 72 h because melatonin protects sperm from oxidative stress. These findings can be used to improve the semen storage protocol by combining semen extender and antioxidant.

Microscopic agglutination test (MAT) for the diagnosis of leptospirosis requires live cultures and is serovar-specific, while polymerase chain reaction (PCR) requires expensive equipment and sample preparation. The rLipL32 protein is conserved and can be used for the production of immunoglobulin G (IgG) anti-rLipL32 antibody, which can be used as a biomarker for leptospirosis diagnosis. This study aimed to produce and characterize an IgG anti-rLipL32 antibody as a biomarker for leptospirosis diagnosis. Escherichia coli rLipL32 was cultured and analyzed by PCR and sequencing. Cultures were used for rLipL32 protein expression and purification and the rLipL32 protein was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The rLipL32 protein was used to produce anti-rLipL32 serum and was analyzed by enzyme-linked immunosorbent assay (ELISA). Serum was purified to obtain IgG anti-rLipL32 antibody and characterized by SDS-PAGE and western blotting. PCR was able to amplify the LipL32 gene from E. coli rLipL32, and sequencing analysis showed 99.19% similarity with pathogenic Leptospira. SDS-PAGE analysis showed a 32-kDa band. ELISA results showed an increase in OD in anti-rLipL32 serum compared to preimmune serum. Western blotting results showed that the IgG anti-rLipL32 antibody was able to bind and cross-reacts with pathogenic Leptospira serovar but not with E. coli or Staphylococcus aureus. IgG anti-rLipL32 antibody has high specificity and sensitivity against Leptospira pathogens. These findings suggest that IgG anti-rLipL32 antibody is a promising biomarker for the diagnosis of leptospirosis.

Established antimicrobial resistance (AMR) surveillance in companion animals is lacking, particularly in low-middle-income countries. The aim of this study was to analyze AMR and its risk factors in Escherichia coli isolated from dogs at two veterinary centers in Lima (Peru). Ninety dogs were included in the study. Antimicrobial susceptibility was established by disk diffusion, whereas microdilution was used to determine colistin susceptibility. Mechanisms related to extended-spectrum β-lactamases (ESBL) and colistin resistance were determined by polymerase chain reaction. Clonal relationships of colistin-resistant isolates were assessed by XbaI-pulsed-field gel electrophoresis. Thirty-five E. coli strains were isolated. High levels of resistance to ampicillin (57.1%), nalidixic acid (54.3%), tetracycline (48.6%), and azithromycin (25.7%) were detected. Cephalosporin resistance levels were ≥20% and those for colistin were 14.3%. Twelve (34.2%) isolates were ESBL producers; of these, six bla_CTX-M-55 (50.0%), 2 (16.6%) bla_CTX-M-15, and 2 (16.6%) bla_CTX-M-8-like genes were found. The five colistin-resistant isolates were clonally unrelated, with four of them presenting amino acid codon substitutions in the mgrB gene (V8A) or mutations in the mgrB promoter (a12g, g98t, and c89t). Furthermore, dog age, <6 years (p = 0.027) and raw diet (p = 0.054) were associated with resistance to a greater number of antibiotic families. Despite small number of samples included, the study found that dogs studied were carriers of multidrug-resistant E. coli, including last-resort antimicrobials, representing a public health problem due to close contact between dogs and humans. This issue suggests the need for larger studies addressed to design strategies to prevent the spread of resistant micro-organisms in small animal clinics and domestic settings.

Filarial nematode typically produces a larval stage (microfilariae) in the bloodstream of vertebrate hosts, where microfilariae reside in the blood or subcutaneous tissues. Filarial nematodes cause human diseases, such as river blindness and elephantiasis, which are widely studied. However, in avian species, they are overlooked because they are nonpathogenic. In Thailand, microfilaria can be found in wild birds and domestic chickens. Recently, an increase in the number of blood samples submitted to veterinary diagnostic laboratories may have increased the number of microfilariae. Therefore, knowledge about filarial species and reliable detection methods are important. Therefore, this study aimed to investigate the efficacy of buffy coat smear and polymerase chain reaction (PCR)-based methods for the detection of microfilaria in domestic chickens. In addition, parasites were identified using the sequence of the cytochrome c oxidase subunit 1 (COX1) gene. Giemsa-stained buffy coat smears from a previous study were reanalyzed. These available buffy coat smears were prepared from 55 domestic chickens raised as backyard free-ranging in Southern Thailand. Fifty-seven frozen genomic DNA extracted from chicken blood were used to detect the presence of the COX1 gene in Onchocercidae nematodes. The nested PCR protocol for amplification of the OnchoCOI_ R2-OnchoCOI_ R2 fragment of the COX1 gene was applied from a previous report. Sequences of COX1 were analyzed to identify Onchocercidae nematodes and if they were single or mixed infections. We constructed Bayesian phylogenetics to identify parasites and assessment of the relationship between filarial nematodes in avian species and other vertebrate hosts. Buffy coat smears from 15 samples revealed microfilaria. Of these 15 samples, only eight were positive for COX1 nested-PCR amplification. The other two buffy coat-negative samples were also positive for nested-PCR. Sequencing of these 11 nested PCR-positive samples revealed that almost all of them were Onchocercidae nematodes. Bayesian phylogenetic analysis showed that chicken Onchocercidae spp. were grouped with other avian filarial nematodes. However, all chickens Onchocercidae spp. showed a double peak in the sequencing chromatogram, indicating mixed filarial infection (species or haplotypes). Therefore, no chicken Onchocercidae sequence was deposited on National Center for Biotechnology Information, GenBank. Giemsa-stained buffy coat smear was a reliable method for the detection of chicken microfilaria in routine veterinary diagnostic laboratories. Development of a new PCR-based method is necessary. This method may provide greater sensitivity and specificity of detection. In addition, the PCR method allowed us to access the genetic characteristics of nematodes, which helped us maximize our knowledge of nematodes. Further investigations, such as the pathogenicity of filarial nematodes in chickens and their potential vectors, are required.

Indigenous beef cattle engaged in bullfighting in Southern Thailand represent a distinctive and valuable breed. Gastrointestinal (GI) parasites, which are recognized as important pathogens, have a negative impact on the overall health and physical performance of these fighting bulls. This study aimed to estimate the prevalence of GI parasitic infections and identify factors associated with these infections in a fighting bull population in Southern Thailand.  Fecal samples (n = 4,244) from fighting bulls were submitted to the Laboratory and Diagnostic Centre of the Teaching Animal Hospital, Faculty of Veterinary Science, Rajamangala University of Technology Srivijaya. We examined the samples using simple flotation and centrifugal sedimentation methods. Individual animal profiles and demographic data were collected.  The overall prevalence of GI parasitic infections was 93.2%. Nine GI parasites were identified as Paramphistome spp. [PP]. being the most prevalent (93.2%), and the highest annual prevalence occurred in 2019 (97.9%). The infection rates of various parasite species were significantly related to the years of study, geographic area, season, and age group (p < 0.05). The prevalence of parasitic infection was higher on the west coast (98.6%) than on the east coast (98.0%). PP, Eurytrema spp., Strongyles spp., and Buxtonella spp. infections differed significantly among the seven provinces of Southern Thailand (p < 0.05). The prevalence of GI parasitic infections was higher during the rainy season (98.5%) than during the summer (97.7%). Bulls aged 7.0–7.9 years and 8.0–8.9 years had the highest parasite infection rate (99.2%) compared with those aged 8.0–8.9.  GI parasitic infections continue to be a significant health concern among fighting bulls in Southern Thailand. Regular epidemiological investigations are crucial for developing effective preventive programs and control strategies and providing basic knowledge for bull farmers. Keywords: bullfighting, gastrointestinal parasites, geographical areas, prevalence, Thailand.

Anaplasma marginale is an obligate intraerythrocytic rickettsial parasite that infects cattle in tropical and subtropical regions. There is no evidence that A. marginale inoculation can be used to culture Rhipicephalus annulatus in rabbits. This study aimed to determine the molting of R. annulatus larvae, nymphs, and adults on rabbits as well as nymphs and adults of R. annulatus on calves with or without A. marginale. Transstadial, horizontal, and transovarial transmissions of A. marginale in R. annulatus reared on rabbits and calves were evaluated. Engorged female ticks were collected from field samples of A. marginale-infected and non-infected cattle. We divided the eight rabbits into two groups: A and B. Group A rabbits were infected with A. marginale through parenteral inoculation, whereas Group B rabbits were kept as a control. The “clean rabbits” in Group B were observed for tick rearing without A. marginale. Polymerase chain reaction was used to screen A. marginale in rabbits and stages of tick. The complete life cycle of R. annulatus with or without A. marginale was observed on rabbits.  A 6.5-day longer life cycle was observed in ticks harboring A. marginale than in ticks without A. marginale. To observe transstadial transmission, transstadial, horizontal, and transovarial transmissions of A. marginale in R. annulatus ticks were experimentally observed in one clean calf fed separately with infected nymphs and female adult ticks.  We experimentally observed transovarian, transstadial, and transovarial transmission of A. marginale in R. annulatus ticks as a biological vector reared on calves and rabbits. We used rabbits as a model animal for rearing R. annulatus ticks and culture of A. marginale. <br>Keywords: Anaplasma marginale, horizontal, rabbits, Rhipicephalus annulatus, transovarial, transstadial.

The impact of exogenous melatonin on the sperm quality of small ruminants is controversial. Therefore, this study aimed to synthesize previous findings on the influence of melatonin injection on sperm quality, steroid hormones, and testicular blood flow in small ruminants. Thirty studies were analyzed by computing the raw mean difference (RMD) as the effect size between the control and melatonin treatment groups, using the inverse of the variance for the random-effect model of the method of moments by DerSimonian and Laird. We assessed heterogeneity among studies using Q test. I2 statistic was used to classify the observed heterogeneity. We used Egger’s regression method to indicate publication bias.  Melatonin injection (p &lt; 0.05) affected sperm concentration (RMD = 0.42 × 109/mL), morphology (RMD = 2.82%), viability (RMD = 2.83%), acrosome integrity (RMD = 4.26%), and DNA integrity (RMD = 1.09%). Total motility (RMD = 5.62%), progressive motility (RMD = 7.90%), acrosome integrity (RMD = 8.68%), and DNA integrity (RMD = 2.01%) of post-thawed semen in the melatonin-treated group were also increased (p &lt; 0.05). Similarly, treatment with melatonin (p &lt; 0.05) enhanced total motility (RMD = 5.78%), progressive motility (RMD = 5.28%), curvilinear velocity (RMD = 4.09 μm/s), straight-line velocity (RMD = 5.61 μm/s), and average path velocity (RMD = 4.94 μm/s). Testosterone (RMD = 1.02 ng/mL) and estradiol 17-ß levels (RMD = 0.84 pg/mL) were elevated (p &lt; 0.05) in the melatonin-injected group. Melatonin implantation ameliorated testicular blood flow, as indicated by a significant reduction (p &lt; 0.05) in the resistive index (RMD = 0.11) and pulsatility index (RMD = –0.15).  Melatonin administration can increase the reproductive performance of small male ruminants.

Nosocomial infections caused by Salmonella spp. are common in veterinary facilities. The early identification of high-risk patients and sources of infection is important for mitigating the spread of infections to animal patients and humans. This study investigated the occurrence of Salmonella spp. among patients at a veterinary academic hospital in South Africa. In addition, this study describes the environmental factors that contribute to the spread of Salmonella spp. in the veterinary facility. This study used a dataset of Salmonella-positive animals and environmental samples submitted to the bacteriology laboratory between 2012 and 2019. The occurrence of Salmonella isolates at the veterinary hospital was described based on source, month, season, year, and location. Proportions and 95% confidence intervals were calculated for each variable. A total of 715 Salmonella isolates were recorded, of which 67.6% (483/715) came from animals and the remainder (32.4%, 232/715) came from environmental samples. The highest proportion (29.2%) of Salmonella isolates was recorded in 2016 and most isolates were reported in November (17.4%). The winter season had the lowest (14.6%) proportion of isolates reported compared to spring (31.3%), summer (27.8%), and autumn (26.4%). Salmonella Typhimurium (20.0%) was the most frequently reported serotype among the samples tested, followed by Salmonella Anatum (11.2%). Among the positive animal cases, most (86.3%) came from equine clinics. Most reported isolates differed based on animal species with S. Typhimurium being common in equines and S. Anatum in bovines. In this study, S. Typhimurium emerged as the predominant strain in animal and environmental samples. Equines were the most affected animals; however, Salmonella serotypes were also detected in the production animals. Environmental contamination was also a major source of Salmonella species in this study. To reduce the risk of transmission, strict infection prevention and control measures (biosecurity) must be implemented.

The high prevalence of diabetes mellitus in Indonesia indirectly reflects the high risk of developing chronic wounds that are susceptible to infection. Methicillin-resistant Staphylococcus aureus (MRSA) is an infectious pathogen that is resistant to various antibiotics. Therefore, antibiotic therapy is ineffective enough to treat chronic hyperglycemic wounds caused by MRSA infection. Curcuminoids have anti-inflammatory and antibacterial effects by inhibiting the enzymatic pathways involved in the pathogenesis of inflammation. Collagen is a tissue regeneration inducer. The combination of these two ingredients is expected to be an alternative therapy for MRSA-infected hyperglycemic chronic wounds without the risk of antibiotic resistance. The aim of this study was to investigate the efficacy of hydrolacin-gel in wound healing and inhibiting the growth of MRSA bacteria, as well as to determine the optimal concentration of curcuminoids combined with collagen marine peptides (CMPs). Hydrolacin-gels were prepared by homogenizing curcuminoid nanoemulsions and CMPs. The evaluation of preparation includes stability tests and antibacterial activity tests. Wound diabetic mice were treated with various combinations of curcuminoid and CMPs. Wound healing was observed based on malondialdehyde levels as a marker of oxidative stress and histopathological changes in the skin wound. Hydrolacin-gel was formulated by combining curcuminoid nanoemulsion (more water soluble) and CMPs, with the ratio of formula 1 (1:2, curcuminoid 43.3 mg and CMPs 5.58 mg), formula 2 (1:1, curcuminoid 86.8 mg and CMPs 3.72), and formula 3 (2:1, curcuminoid 130.2 mg and CMPs 1.87 mg) calculated based on the effective dose of curcuminoid 200 mg/kg body weight (BW) and CMPs 0.9 g/kg BW. Hydrolacin-gel had a potential antibacterial activity against MRSA. Hydrolacin-gel induced wound tissue repair and reduced oxidative stress caused by inflammation in diabetic-infected MRSA. Hydrolacin-gel could be used for healing MRSA-infected diabetic wounds, especially formula 3 with the ratio of curcuminoid: CMPs = 2:1. Hydrolacin-gel combining curcuminoid nanoemulsion and CMPs effectively inhibited the inflammatory process and increased re-epithelialization in MRSA-infected diabetic wound healing. Hydrolacin-gel with curcuminoid (130.2 mg) and CMPs (1.87 mg) at a concentration ratio of 2:1 appeared to be the best formula against MRSA infection in diabetic wounds.