Lawsonia intracellularis is an obligatory intracellular bacterium associated with equine proliferative enteropathy (EPE), which significantly impacts equine health. Despite its clinical relevance, epidemiological and diagnostic approaches for this infection in horses have remained underexplored. This study aimed to evaluate the humoral immune response in horses immunized with an experimental vaccine for L. intracellularis and to determine the occurrence of anti-L. intracellularis antibodies in horses from southern Brazil using the flow cytometry antibody test (FCAT). A total of 12 horses were immunized with an experimental vaccine containing inactivated L. intracellularis and adjuvants. Blood samples were collected on day 0 (D0) and every 7 days until day 35 (D35) to monitor the antibody response using FCAT. In addition, serum samples from 79 horses from the North and Northwest regions of Rio Grande do Sul were tested to determine the occurrence of anti-L. intracellularis antibodies. The FCAT protocol was optimized for equine samples, and a cut-off fluorescence threshold was determined using Receiver Operating Characteristic curve analysis. FCAT demonstrated high accuracy, with a sensitivity of 100% and specificity of 92% at a fluorescence cut-off of ≥15%. Immunization triggered a robust humoral response, with a significant increase in fluorescence from day 7 to day 35. In the occurrence analysis, 26% of the horses tested positive for anti-L. intracellularis antibodies. The highest occurrence was observed in females (35.8%) and horses aged 16–22 years (50%). This study provided evidence that L. intracellularis infection is relatively common among horses in southern Brazil. FCAT was a sensitive and specific diagnostic tool for detecting anti-L. intracellularis antibodies in horses. The findings highlight the need for better diagnostic and preventive measures to control EPE in equine populations.

Electroejaculation (EE) is widely used for semen collection in bulls but raises concerns about animal welfare due to potential pain and stress. The physiological impact of EE on bulls remains a topic of debate, with previous studies yielding inconclusive results. This study aims to objectively evaluate pain and stress responses in bulls subjected to EE using electroencephalography (EEG) alongside hormonal, behavioral, and metabolite profiling. Eight bulls were subjected to EE in three replicates, with physiological and behavioral data collected before, during, and after the procedure. EEG parameters, including median frequency (MF) and total power (Ptot), were analyzed to assess cortical activity indicative of pain and stress. Blood samples were evaluated for stress-related hormones (adrenaline, noradrenaline, β-endorphin, and dopamine), while metabolomic analysis was conducted to identify biochemical alterations associated with stress. Behavioral indicators, including vocalization and muscle spasms, were recorded. EE induced significant increases (p < 0.05) in stress hormones at ejaculation, which gradually returned to baseline 20 min post-procedure. EEG metrics, such as MF and Ptot, significantly increased during EE (p < 0.05), indicating heightened cortical activity associated with nociception. Metabolomic analysis revealed distinct biochemical shifts, with variations in glucose, taurine, and norepinephrine profiles across baseline, stimulation, and recovery phases. Behavioral observations corroborated physiological findings, with bulls exhibiting signs of discomfort, such as struggling, arched back posture, and excessive salivation. The combined EEG, hormonal, and metabolomic findings confirm that EE is a stressful and painful procedure for bulls. The study provides robust evidence of neurophysiological and biochemical responses indicative of pain. These findings highlight the need for alternative semen collection methods to minimize animal distress and improve welfare standards.

Canine femoral fractures are prevalent in veterinary medicine, necessitating effective fixation methods to ensure stability and promote healing. Conventional bone plate fixation methods, including non-locking and locking plates, have inherent limitations, such as periosteal damage and mechanical failure. This study aims to evaluate the biomechanical performance of three fixation methods – non-locking bone plates, locking bone plates, and a novel double-rod clamp internal fixation system – using finite element analysis (FEA). A computed tomography-based canine femur model was created to simulate a midshaft commin-uted fracture with a 20 mm gap. Three fixation configurations were modeled: A non-locking bone plate, a locking bone plate, and a double-rod clamp system. FEA was performed to assess implant stress and proximal fragment displacement under physiological axial loading. Mesh refinement and multiple loading conditions were incorporated to enhance computational accuracy. The non-locking bone plate exhibited the highest implant stress (1160.22 MPa), surpassing the material yield strength and indicating a risk of mechanical failure. The double-rod clamp system demonstrated lower stress (628.34 MPa), whereas the locking bone plate had the lowest stress (446.63 MPa). Proximal fragment displacement was highest in the non-locking bone plate (2.37 mm), followed by the double-rod clamp system (0.99 mm), with the locking bone plate exhibiting the least displacement (0.34 mm), suggesting superior stability. The double-rod clamp system emerged as a promising alternative, offering a balance between stability and stress distribution while minimizing periosteal damage. While the locking bone plate provided the greatest stability, the double-rod clamp fixation demonstrated favorable mechanical properties and could serve as a cost-effective and minimally invasive alternative in veterinary orthopedics.

Water quality is a critical factor in duck farming, influencing growth performance, health, and productivity. Salinity in drinking water is a concern in regions affected by saline intrusion, such as the Mekong Delta, Vietnam. Notably, female ducks exhibit greater salt tolerance than males due to differences in kidney size and function. This study aimed to investigate the effects of saline drinking water on the growth performance, carcass traits, and blood biochemistry of crossbred Muscovy ducks (Cairina moschata). A completely randomized design was used, including five treatments with five replications per treatment. Each replication consisted of four crossbred Muscovy ducks, with an equal number of males and females. The treatments comprised freshwater (SW0.0) and diluted seawater with salinity levels of 0.15% (SW0.15), 0.3% (SW0.3), 0.45% (SW0.45), and 0.6% (SW0.6). Ducks were provided ad libitum access to feed and water throughout the experiment. Key parameters measured included dry matter intake (DMI), water intake, body weight (BW), carcass traits, meat quality, salt gland dimensions, and blood biochemical parameters. Saline drinking water significantly reduced DMI and BW gain, with male ducks more adversely affected than females. The final BW of males decreased with increasing salinity, while female ducks maintained stable BWs. Ducks consuming water with salinity levels of 0.45% and 0.6% had higher feed conversion ratios (4.83 and 4.68, respectively) compared to lower salinity groups (3.77–4.61). Carcass weight, breast weight, and abdominal fat percentage declined with increasing salinity, whereas thigh and drumstick weights remained unaffected. The crude protein content of thigh and drumstick meat decreased at higher salinity levels. Blood biochemical parameters, including sodium, chloride, urea, and creatinine levels, increased in ducks consuming highly saline water, indicating potential physiological stress. Ducks in higher salinity groups exhibited increased salt gland size, suggesting an adaptive response to saline conditions. Male crossbred Muscovy ducks can tolerate drinking water salinity levels up to 0.3%, whereas females can withstand salinity up to 0.6%. The greater salinity tolerance of female ducks makes them more suitable for farming in saline-affected areas. While increased salt gland size suggests an adaptive response, elevated blood biochemical markers indicate potential physiological stress. Future research should explore strategies to mitigate the negative effects of saline water on duck production, including dietary interventions with antioxidants such as vitamin E or selenium.

The increasing human-wildlife interface due to urbanization and agricultural expansion has escalated the risk of zoonotic and interspecies disease transmission. Canine distemper virus (CDV), a highly contagious Morbillivirus, has been documented in various carnivorous and non-carnivorous species. In 2019, Malaysia reported its first case of CDV infection in a wild Malayan tiger (Panthera tigris malayensis) named Awang Besul in Kampung Besul Lama, Terengganu. However, the potential role of small mammals as intermediate hosts in CDV transmission remains poorly understood. This study aimed to investigate the role of small mammals as potential reservoir hosts for CDV and to provide molecular confirmation of CDV infection in these species, thereby assessing their role in enzootic viral maintenance and cross-species transmission to apex predators like the Malayan tiger. Wildlife sampling was conducted between July 2023 and May 2024 in Kampung Besul Lama, where CDV was previously detected in a Malayan tiger. A total of 77 small mammals from different species were captured using baited live traps. Species identification was performed based on morphological characteristics. Biological samples were collected through nasal and ocular swabs and analyzed using reverse transcription polymerase chain reaction to detect CDV RNA. Positive isolates were subjected to sequencing and Nucleotide Basic Local Alignment Search Tool analysis for molecular characterization. Molecular detection confirmed CDV RNA in three common tree shrews (Tupaia glis), marking the first documented case of CDV in this species. Phylogenetic analysis of the viral hemagglutinin (H) gene revealed a 99.50% nucleotide similarity to a previously reported Malaysian CDV strain (BesulMY/Malaysia/PP894823.1). These findings suggest that small mammals may act as overlooked reservoir hosts, facilitating viral maintenance and spillover between domestic animals and wildlife. This study provides the first molecular evidence of CDV infection in tree shrews, highlighting their potential role in sustaining CDV in an enzootic state and acting as a conduit for interspecies transmission. Given the critically endangered status of Malayan tigers, targeted CDV surveillance and One Health-based disease mitigation strategies are essential to prevent further spillover events that could accelerate species decline.

Infectious bursal disease (IBD) is an immunosuppressive disease caused by the IBD virus (IBDV), which adversely affects poultry vaccination programs. The novel variant IBDV (nvIBDV) has recently emerged in various regions, including Malaysia, raising concerns about its immunosuppressive potential and impact on Newcastle disease (ND) vaccination. This study aimed to investigate the immunosuppressive effects of the Malaysian nvIBDV strain (UPM1432/2019) in specific-pathogen-free (SPF) and broiler chickens and evaluate its influence on ND vaccine efficacy. SPF chickens were orally infected with nvIBDV across three passage levels to study virus-induced clinical signs, lesions, and atrophy. Broiler chickens were vaccinated with live and killed ND vaccines and subsequently challenged with nvIBDV to measure ND antibody titers. The genotype of nvIBDV was characterized using sequence analysis of segments A and B. Bursal histopathology and statistical analyses were conducted to evaluate the virus’s immunosuppressive effects. Infected SPF chickens displayed no clinical signs but showed significant bursal atrophy and lesions across all passages (p < 0.05). Broilers infected with nvIBDV exhibited no mortality or clinical signs; however, ND antibody titers significantly declined by 14 days post-challenge (1493.0 ± 746.1) compared with the unchallenged group (2975.7 ± 189.5; p < 0.01). Histopathological analysis revealed severe depletion of the bursal follicles, lymphoid cell aggregation, necrosis, and hemorrhage. Genotyping of nvIBDV identified as genotype A2dB1, consistent with strains from other regions. The Malaysian nvIBDV strain causes subclinical infections in SPF and broiler chickens, resulting in bursal atrophy and reduced ND vaccine-induced antibody titers. Silent spread and immunosuppressive effects present significant challenges to poultry health management and vaccination efficacy. Enhanced diagnostic and biosecurity measures are crucial for mitigating its impact.

Artificial insemination (AI) is a critical technique in sheep breeding programs, yet its success is influenced by factors such as semen quality, storage conditions, and insemination timing. This study examines the effects of different semen extenders, storage durations, and insemination schedules on semen motility and pregnancy rates in Palestinian Assaf sheep. The primary aim of this study was to assess and compare the efficacy of three semen extenders – Andromed, Indonesian, and Syrian – on the quality of stored semen and its subsequent effect on pregnancy rates following AI. In addition, the study investigated the impact of semen storage duration and the timing of insemination post-equine chorionic gonadotropin (eCG) injection on reproductive outcomes. Semen was collected from eight Assaf rams, diluted with one of three extenders, and stored at 4°C for up to 72 h. Sperm motility was analyzed using a computer-assisted sperm analysis system at different time intervals. AI was performed on 180 Assaf ewes across three farms in Palestine, with insemination conducted at either 48 h, 52 h, or both 48 and 52 h post-eCG injection. Pregnancy rates were determined through ultrasound 45 days post-insemination. Statistical analysis was conducted using the Statistical Package for the Social Sciences 22 (IBM® NY, USA), with p-values set at <0.05 for significance. Semen motility decreased significantly over time, with total motility (TM) declining from 0.85 ± 0.01 (fresh) to 0.63 ± 0.02 after 3 days of storage. The Syrian extender exhibited superior fast motility (FM) (0.35 ± 0.01) compared to the others. Strong positive correlations were observed between TM and progressive motility (0.90, p ≤ 0.01). Pregnancy rates did not significantly differ among extenders (Andromed: 0.58 ± 0.06, Indonesian: 0.54 ± 0.07, Syrian: 0.56 ± 0.08). However, insemination performed at both 48 and 52 h post-eCG injection resulted in the highest pregnancy rate (0.62 ± 0.07), while the second ejaculation showed a tendency for improved fertility outcomes (0.61 ± 0.07). Prolonged semen storage negatively impacts motility, though the Syrian extender preserves FM better than the others. Pregnancy rates were not significantly influenced by the extender type but were optimized by insemination at both 48 and 52 h post-eCG injection. These findings highlight the importance of refining semen preservation techniques and timing AI procedures to enhance breeding success in Assaf sheep.

Brucellosis is a zoonotic disease that remains underdiagnosed in humans due to its non-specific clinical presentations. Punjab, India, is particularly vulnerable to brucellosis due to its high-density livestock farming. This study aimed to estimate the seroprevalence of brucellosis in individuals presenting with non-specific clinical symptoms. A cross-sectional study was conducted at the Centre for One Health, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, from January 2021 to December 2021. A total of 137 serum samples were collected from individuals either self-referred or physician-referred for brucellosis testing. The samples were screened using the rose Bengal plate test (RBPT) and confirmed with the standard tube agglutination test (STAT). Data on demographics, symptoms, and occupational exposure were analyzed using SPSS version 26.0, with statistical significance set at p < 0.05. Overall, 17.52% (24/137) of individuals tested positive using STAT, with antibody titers ranging from 80 IU/mL to >2560 IU/mL. Seropositivity was higher in males (20.83%) than in females (9.77%). The highest seropositivity (42.9%) was observed in individuals aged 71–80 years. Among symptomatic individuals (n = 92), fever was the most common symptom (n = 79), followed by joint pain (n = 13). However, 15.6% of asymptomatic individuals also tested positive. No significant association was found between symptoms and seropositivity (p > 0.05). In addition, self-referred individuals (24.1%) had a higher seropositivity rate compared to physician-referred cases (12.7%). Among occupationally exposed individuals, veterinary officers showed the highest seropositivity. The study highlights a considerable seroprevalence of brucellosis among various symptomatic and among asymptomatic individuals. Given its non-specific clinical manifestations, routine serological screening is recommended, especially for high-risk groups. A One Health approach integrating human and animal health surveillance is crucial for effective disease control.

Arrhythmic sudden cardiac death in dogs and horses often results from ventricular arrhythmia secondary to myocardial damage. Despite this, limited data exist on the histomorphometric changes in cardiac conduction fibers (CCFs) and cardiac conduction cells (CCCs) following spontaneous myocardial infarction (MI). This study aimed to characterize morphometric and histological alterations in conduction fibers and their junctions with cardiomyocytes in infarcted hearts of horses and dogs. Ten hearts from horses and 10 from dogs that had died suddenly were examined. Histological and immunohistochemical analyses were performed using hematoxylin and eosin, Masson’s trichrome, and periodic acid–Schiff staining to identify conduction fibers and assess glycogen accumulation. The thickness and density of conduction fibers, as well as the diameter of conduction cells, were measured using image analysis software. Statistical comparisons were conducted using t-tests, analysis of variance, and Cohen’s d-test. In horses, the diameter of CCCs was significantly smaller in infarcted cases (55.74 μm) compared to normal hearts (79.08 μm) (p < 0.001). In dogs, slight hypertrophy of CCCs (31.21 μm) was observed in normal hearts, whereas infarcted hearts exhibited reduced diameters (26.83 μm) (p = 0.114). The density of CCFs was 9.06% in horses and 7.99% in dogs (p = 0.846), while fiber thickness was 30.06 μm in horses and 29.86 μm in dogs (p = 0.263). Horses exhibited extensive myocardial fibrosis, particularly in the middle third and posterior left ventricle, while dogs displayed milder lesions distributed across the ventricle. This study demonstrates a reduction in CCC size in horses and minor hypertrophy in dogs, coupled with fibrotic myocardial lesions of varying severity. The observed histomorphometric changes provide insight into the structural impact of MI on conduction cells, which may contribute to ventricular arrhythmias in these species. These findings have implications for veterinary cardiology and the management of MI-related arrhythmic conditions.

Trypanosomiasis caused by Trypanosoma vivax is a significant cause of reproductive inefficiency and economic losses in cattle farming. While its impact on reproduction is known, the molecular mechanisms underlying these effects remain poorly understood. This study aimed to investigate the molecular effects of T. vivax on reproductive parameters in cattle and evaluate epidemiological risk factors associated with trypanosomiasis in beef cattle in northern Minas Gerais, Brazil. A dual approach combining in silico bioinformatics and epidemiological analyses was employed. Proteins linked to T. vivax were identified using the UniProt database, and protein interaction networks were constructed using the String V.12 platform. The epidemiological study involved serological diagnosis of trypanosomiasis through indirect immunofluorescence in 383 serum samples collected from 14 herds. Risk factors such as farming system, animal replacement frequency, vector presence, vaccination practices, and reproductive history were assessed through statistical analysis. Bioinformatics analyses suggested that T. vivax may exert molecular effects on bovine reproduction through the expression of toll-like receptor 2, nuclear factor kappa B (NF-κB), and nuclear receptor coactivator 7 proteins. The overall prevalence of T. vivax was 6.79%, with no direct association found between infection and reproductive performance. However, 57.7% of seropositive cattle belonged to farms with lower pregnancy rates, and 96% were from farms reporting abortions in the past 12 months. The semi-intensive/intensive farming system and shared use of syringes and needles during vaccination were identified as significant risk factors for T. vivax infection. The study provides evidence of T. vivax spread in northern Minas Gerais and highlights the need for improved control strategies, including vector management and proper sanitary practices. Bioinformatic analysis suggests that T. vivax may influence reproductive outcomes through the NF-κB signaling pathway, warranting further experimental validation. Future studies should investigate the molecular mechanisms of T. vivax in high-prevalence herds to refine disease management and mitigation strategies.

Egg production is a critical indicator of reproductive efficiency in laying hens. High environmental temperatures and humidity expose laying hens to heat stress, adversely affecting egg production, egg quality, feed intake, blood chemistry, health, and behavior. Despite the global economic significance of egg production, research on the impact of heat stress in tropical climates, particularly in locally adapted breeds, remains limited. This study investigates the effects of heat stress on egg production in a new synthetic breed of laying hens raised under hot and humid conditions. This study utilized secondary data from the Ligor chicken development project, covering the period from January 1 to December 31, 2023. A total of 1293 daily records of egg production from 872 laying hens, aged between 20 and 70 weeks, were analyzed. Egg production parameters, including hen day production (HDP) and average egg weight (AEW), were recorded daily. Environmental data, including temperature and humidity, were collected at hourly intervals using a data recorder. The heat stress index (HSI) was calculated based on temperature and relative humidity values. Statistical analyses, including regression modeling, were performed to assess the relationship between HSI and egg production. The findings demonstrated that heat stress negatively impacted egg production. A statistically significant negative correlation was observed between HSI and both HDP and AEW (p < 0.05). For each unit increase in HSI, HDP decreased by 1.29%, and AEW declined by 0.18 g. These results indicate that heat stress adversely affects the productivity of laying hens in tropical climates. Heat stress significantly reduces egg production and egg weight in laying hens under hot and humid conditions. The findings highlight the need for adaptive breeding strategies and improved environmental management to mitigate the adverse effects of heat stress. Future research should focus on genetic selection for heat resistance and the integration of precision farming techniques to optimize poultry production in tropical regions.

Paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is a significant infectious disease affecting livestock, including camels in Saudi Arabia, leading to severe economic losses. Despite its impact, limited genomic studies have been conducted to characterize MAP strains in camels and their antimicrobial resistance (AMR) profiles. This study aimed to (1) characterize the clinical and pathological findings of MAP infections in camels; (2) determine the seroprevalence of MAP in the eastern region of Saudi Arabia; (3) differentiate between MAP strains using polymerase chain reaction (PCR) and bioinformatics tools; (4) conduct a comprehensive genomic analysis; and (5) identify genes associated with AMR, virulence, and immune response. A total of 345 blood samples were collected for seroprevalence analysis using enzyme-linked immunosorbent assay (ELISA), and 68 rectal scraping samples were analyzed using Ziehl–Neelsen staining and PCR for strain differentiation. Seventeen clinical cases underwent clinical, postmortem, and histopathological examinations. Whole-genome sequencing and bioinformatics analyses were performed using the Bacterial and Viral Bioinformatics Resource Center to identify genetic variations, AMR genes, and phylogenetic relationships among MAP isolates. Clinical findings revealed progressive emaciation and chronic diarrhea in affected camels. Gross examination showed intestinal wall thickening and mesenteric lymph node congestion. Histopathological analysis indicated hyperactivation of crypts of Lieberkühn and mononuclear cell infiltration. PCR analysis identified a higher prevalence of the sheep (S) strain (162 bp) compared to the cattle (C) strain (310 bp). The overall seroprevalence of MAP was 8.11% (ELISA). Whole-genome sequencing identified 34 AMR genes and 10 virulence genes, with annotation revealing 4.7 million base pairs, coding sequences, transfer RNA, ribosomal RNA, and pseudogenes. Phylogenetic analysis grouped MAP strains into four distinct clades, indicating potential cross-species transmission. This study provides critical insights into the genetic diversity and AMR mechanisms of MAP strains in camels, emphasizing the need for targeted control strategies. The findings highlight potential zoonotic risks and inform future vaccine development to mitigate MAP infections in livestock.

Staphylococcus aureus is a zoonotic pathogen with significant public health and economic implications. Its ability to tolerate environmental stress and regulate virulence through quorum sensing contributes to its persistence and pathogenicity. Rhodomyrtus tomentosa and its bioactive compound rhodomyrtone have demonstrated antimicrobial properties against Gram-positive, multidrug-resistant bacteria. This study aimed to evaluate the effects of these agents on quorum sensing inhibition and stress tolerance in S. aureus and Pseudomonas aeruginosa, providing insights into their potential as alternative antimicrobial strategies. The anti-quorum sensing activity of R. tomentosa extracts was assessed using Chromobacterium violaceum as a bioindicator. In addition, the effects on P. aeruginosa swarming motility were evaluated. Stress tolerance in S. aureus was examined by subjecting treated cells to acidic (pH = 5.0), alkaline (pH = 9.0), osmotic (7.5% NaCl), heat (43°C), and oxidative (1 mM H₂O₂) stress conditions. The survival rates were determined through colony-forming unit (CFU) counts following treatment with rhodomyrtone and ethanol leaf extracts. The ethyl acetate fraction of R. tomentosa leaf extract exhibited the highest violacein inhibition, followed by the ethanol extract. At 256 μg/mL, these extracts permitted P. aeruginosa colony formation but inhibited its swarming motility. Regarding stress tolerance, no surviving S. aureus cells were detected under any stress condition after 3–6 h of treatment with 2 × minimum inhibitory concentration (MIC) (1 μg/mL) of rhodomyrtone. In addition, 4 × MIC (128 μg/mL) of the ethanol leaf extract inhibited pathogen survival under all tested stress conditions except for alkaline and oxidative stresses. The findings suggest that R. tomentosa extracts and rhodomyrtone effectively inhibit quorum sensing and stress tolerance, offering a promising alternative antimicrobial approach. These compounds could be utilized in veterinary medicine and food safety to mitigate zoonotic pathogen contamination and combat antibiotic-resistant infections.

Brucellosis, caused by Brucella spp., is a zoonotic disease of major public health and economic significance. In Indonesia, surveillance efforts have predominantly focused on bovine brucellosis, leaving limited data on small ruminants despite their critical role as disease reservoirs. This study aims to estimate the seroprevalence of brucellosis in goats and sheep and to identify potential risk factors associated with its transmission in the Jabodetabek region, Indonesia. A cross-sectional study was conducted from May 2023 to November 2023, involving 18 herds of goats and sheep across Jakarta, Bogor, Depok, Tangerang, and Bekasi. A total of 665 blood samples (355 from goats and 310 from sheep) and 112 milk samples were collected. The samples were analyzed using the Rose Bengal Test, complement fixation test, and enzyme-linked immunosorbent assay. Descriptive statistics were used to estimate seroprevalence, and a Chi-square test was employed to evaluate risk factors. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated to assess associations between risk factors and seropositivity. The herd-level seroprevalence was 66.67% (12/18), with a mean within-herd seroprevalence of 10.39% (95% CI: 7.21–13.57). The animal-level seroprevalence was 6.17% (41/665), with the highest rates observed in Bogor City (11.89%), followed by Bekasi (8.91%), East Jakarta (8.00%), and Tangerang (4.58%). Depok City had no positive cases in serum tests, though two cases were detected through milk ELISA. Mixed-species farms exhibited a significantly higher risk of infection than single-species farms (OR: 0.30, 95% CI: 0.14–0.66, p < 0.05). No significant difference was observed between goats and sheep (p > 0.05), nor between males and females (p = 0.84). This study highlights a high seroprevalence of brucellosis in small ruminants within Jabodetabek, with mixed-species farming identified as a major risk factor. The findings underscore the need for enhanced surveillance, control measures, and public health interventions to mitigate disease transmission.

The Indonesian red claw crayfish (Cherax quadricarinatus) represents a significant aquaculture commodity with substantial economic importance; however, disease outbreaks, particularly tail ulceration caused by Aeromonas hydrophila, pose challenges to sustainable production. This study evaluated the efficacy of fermented palm kernel meal (FPKM), rich in mannan oligosaccharides, as a prebiotic additive to enhance non-specific immune responses in C. quadricarinatus. A total of 120 crayfish (mean weight 10.3 ± 0.15 g and length 7.53 ± 0.26 cm) were allocated in a completely randomized design comprising four dietary treatments: a control group without FPKM and probiotics and three experimental groups receiving diets supplemented with FPKM at concentrations of 40, 80, and 120 g/kg along with probiotics. Hemolymph samples were collected at baseline (day 0) and on days 3, 5, 7, and 14 post-treatment. Non-specific immune responses evaluated included total hemocyte count (THC), differential hemocyte count (DHC), phagocytosis activity (PA), phagocytosis index (PI), phenol oxidase (PO) activity, superoxide dismutase (SOD) activity, total plasma protein (TPP), and immune gene expression (lipopolysaccharide and β-1,3-glucan-binding protein [LGBP], lectin). The inclusion of FPKM significantly increased THC, with peak enhancement observed on day 7 at the highest FPKM concentration (120 g/kg). DHC remained stable across treatments. Phagocytic parameters, including PA and PI, showed significant improvements (74.5 ± 12.5% and 2.8 ± 0.41, respectively) by day 7 in the 120 g/kg treatment. PO and SOD activities significantly increased on days 5 and 14, respectively, with optimal responses at the highest dietary FPKM inclusion. TPP levels did not exhibit significant variation among treatments. Molecular analyses revealed marked upregulation of immune-related genes, notably lectin and LGBP, with peak expression detected in the 40 g/kg FPKM group. Supplementing diets with FPKM substantially improved non-specific immune responses in C. quadricarinatus. Optimal immune enhancement was generally achieved with 120 g/kg FPKM inclusion, although significant molecular immune responses were evident at lower concentrations. The findings underscore the potential of FPKM as a natural prebiotic to sustainably enhance crayfish immunity, thereby reducing dependency on antibiotics and contributing to eco-friendly aquaculture practices.

The administration of hot chili pepper in cattle nutrition has been suggested to influence immune responses, antioxidant activities, and physiological parameters. This study aimed to evaluate the effects of microencapsulated hot chili pepper (MCP) supplementation on blood metabolites, antioxidant capacity, and physiological parameters in crossbred dairy cows, contributing novel insights into metabolic and physiological adaptations under tropical production systems. Twenty-four crossbred lactating cows (Bos taurus × Bos indicus); average body weight 447.8 ± 89.6 kg; 53.54 ± 11.8 days in milk; average daily milk production 6.34 ± 1.91 L/cow/day) were randomly divided into two groups: A control group without additives and a treatment group supplemented with MCP (1,000 mg/cow/day) for 42 days. Blood samples were collected weekly for hematological analysis, biochemical evaluations, and oxidative stress biomarkers (2,2-diphenyl-1-picrylhydrazyl [DPPH], thiobarbituric acid reactive substances [TBARS], and trolox equivalent antioxidant capacity [TEAC]). Physiological measurements, including body condition score, heart rate, respiratory frequency, and urine pH, were also assessed weekly. Data were statistically analyzed using mixed-model procedures with repeated measures over time. Significant interactions between treatment and time were observed for hematocrit, red blood cells, and hemoglobin (p < 0.05), indicating physiological adaptations potentially related to increased water intake. Serum albumin levels were significantly lower in MCP-supplemented cows compared to controls (p = 0.006), suggesting a modulation of lipid transport mechanisms. Temporal variations significantly affected 75% of hematological parameters and 83% of biochemical parameters. Although antioxidant parameters (DPPH, TBARS, TEAC) did not differ significantly between groups, notable temporal changes were observed (p < 0.05). Physiological parameters showed significant temporal variations but no consistent effects due to MCP supplementation. Supplementation of dairy cows with MCP significantly impacted hematological parameters and serum albumin levels, revealing potential metabolic adjustments involving lipid transport and hydration status. However, oxidative stress markers and physiological parameters remained largely unaffected by the supplementation. These findings support the potential regulatory role of MCP in dairy cow metabolism, emphasizing its relevance as a dietary additive in tropical livestock production systems.

The indiscriminate use of antimicrobials in livestock farming has contributed to the emergence of antimicrobial-resistant pathogens, posing a significant public health challenge. This study aimed to assess the prevalence and antimicrobial resistance (AMR) profiles of Salmonella spp. isolated from swine and poultry in small- and medium-scale farms in Cambodia. A total of 638 fecal samples (273 rectal swabs from swine and 365 cloacal swabs from poultry) were collected from five provinces in Cambodia from June to September 2021. Salmonella spp. isolation followed ISO 6579:2002 guidelines, and antimicrobial susceptibility testing was conducted using the disk diffusion method, adhering to Clinical and Laboratory Standards Institute 2020 standards. Resistance was assessed against nine antimicrobial agents across five major classes. Overall, Salmonella was detected in 6.58% (42/638) of samples, with 6.96% (19/273) from swine and 6.30% (23/365) from poultry. Swine-derived isolates exhibited the highest prevalence in the growing stage (13.93%), whereas poultry isolates were most common in avian broilers (14.55%). High resistance was observed against β-lactams (penicillin, amoxicillin, and ampicillin), tetracyclines, and sulfonamides, with resistance rates ranging from 73.81% to 100%. Fluoroquinolone resistance (ciprofloxacin and norfloxacin) was moderate (29.77%), while aminoglycoside resistance (gentamicin) was low (2.38%). Multidrug resistance (MDR) patterns were identified in 91.30% (21/23) of poultry isolates and 52.63% (10/19) of swine isolates, with resistance spanning three to five antimicrobial classes. The presence of Salmonella in Cambodian swine and poultry farms and its high level of MDR underscore the urgent need for improved antimicrobial stewardship. The study highlights the risk of MDR Salmonella transmission through livestock production chains, emphasizing the necessity for stringent regulatory interventions, biosecurity measures, and surveillance programs to mitigate AMR spread in animal agriculture and public health sectors.

Family goat farming typically involves small herds managed with minimal infrastructure, leading to products of lower hygienic quality. This study aimed to characterize family goat farms in three distinct regions of Mexico (Durango, Campeche, and Querétaro) and to evaluate hygienic-sanitary indicators and associated risk factors affecting the quality of raw milk and fresh cheese. Seven representative family goat farms were selected based on specific inclusion criteria: Absence of reproductive management, seasonal milk production, manual milking, and artisanal cheese production. Paired samples of bulk raw milk and fresh cheese were collected from each farm. Samples underwent microbiological analyses, including total plate count (TPC), total coliform count (TCC), somatic cell count (SCC), and mold and yeast counts. Surveys addressing animal management, milking, cheese manufacturing, and sales practices were administered. Statistical analyses encompassed descriptive statistics, analysis of variance, cluster analysis, Fisher’s exact tests, and logistic regression. Among raw milk samples, only two farms met acceptable standards for TPC, SCC, and yeast counts according to Mexican regulations, while none complied for TCC. Similarly, cheese samples from two farms met standards for TPC, yeast, and molds, though none met the standards for TCC. Risk factors significantly associated with poor hygienic quality included inadequate pen hygiene, improper teat cleaning, failure to apply post-dip treatments, deficient hand washing, unsuitable milking techniques, lack of milk pasteurization, and insufficient refrigeration practices. Cluster analysis identified two distinct farm groups differentiated by management practices and hygienic standards, correlating with substantial differences in microbial quality indicators. The study identified critical gaps in the implementation of good livestock and manufacturing practices among family goat farms in Mexico. Key risk factors contributing to elevated microbial contamination included poor infrastructure, insufficient hygiene during milking and cheese processing, and inadequate storage conditions. The findings emphasize the necessity of promoting standardized hygienic practices and infrastructure improvements to enhance the sanitary quality of milk and cheese products from family goat farming systems.

Eggs represent a vital dietary source globally; however, bacterial contamination poses a substantial public health risk. This systematic review and meta-analysis aimed to evaluate the global prevalence of bacterial contamination in table eggs and to characterize the antibiotic resistance profiles of these pathogens, emphasizing their implications for public health. A comprehensive bibliographic search of Web of Science, MEDLINE (PubMed), CAB Abstract, and Google Scholar databases was performed, identifying 136 studies published between 1979 and 2022. The systematic review utilized Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines and advanced bibliometric techniques for data collection. Microsoft Excel and R software (v5.0) were employed for data consolidation and statistical analysis. Heterogeneity among studies was assessed using Higgins’ I² index, and a random-effects model was adopted for prevalence estimation and subgroup analyses. Seventeen bacterial species were identified in eggs, primarily Salmonella spp., Escherichia coli, Staphylococcus aureus, Campylobacter spp., and Listeria monocytogenes. Overall, eggshell contamination rates exceeded those of egg contents. Salmonella spp. isolates exhibited complete resistance (100%) to nitrofurantoin, novobiocin, and polymyxin and substantial resistance (>50%) to commonly used antibiotics such as amoxicillin (74.5%), penicillin G (89.1%), and colistin (83.1%). E. coli isolates showed total resistance to penicillin G (100%) and high resistance to amoxicillin (72.2%) and ceftazidime (95.6%). Antibiotic resistance varied significantly across regions, notably higher in Asian and African isolates. Multidrug-resistant strains of E. coli and Campylobacter spp. were also identified. This study underscores the high global prevalence of pathogenic bacteria in poultry eggs and highlights concerning antibiotic resistance trends, particularly among Salmonella spp. and E. coli. The findings emphasize the urgent need for region-specific biosecurity protocols and antimicrobial stewardship strategies to reduce egg contamination and control antibiotic-resistant pathogens, ultimately safeguarding public health and food safety.

Despite the global focus on coronavirus disease 2019 (COVID-19), the avian influenza virus (AIV) continues to circulate in Indonesia, particularly in traditional live bird markets (LBMs), which serve as critical nodes for virus amplification and interspecies transmission. This study aimed to investigate the co-circulation and genetic features of AIV, particularly the H9N2 subtype, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in LBMs in East Java during the COVID-19 pandemic. Environmental surveillance was conducted in seven traditional markets across four districts in East Java Province in 2021. Surface swabs were collected from high-risk areas, including poultry display tables, knives, cutting boards, and napkins. Samples were tested using reverse transcriptase polymerase chain reaction for influenza A and SARS-CoV-2. Positive AIV samples were further subtyped, sequenced, and analyzed for genetic markers associated with virulence, reassortment, and mammalian adaptation. Of 156 samples tested, 17 (10.9%) were positive for influenza A, with 3 (1.9%) confirmed as the H9 subtype. These H9-positive samples were collected from a knife, cutting board, and napkin in the same market location in Lamongan Regency. Phylogenetic and molecular analyses revealed that two isolates (LSJ/Env/83 and LSJ/Env/84) were H9N2 reassortants, harboring key molecular markers such as Q226L, T160A, and S138A in the hemagglutinin protein, indicative of increased affinity for human-type receptors. Additional substitutions in PB2, MP, and NS1 proteins were associated with enhanced replication and virulence in mammalian and avian hosts. All samples tested negative for SARS-CoV-2. This study demonstrates the continued environmental circulation of reassortant H9N2 AIVs in traditional markets during the COVID-19 pandemic, with isolates displaying genetic features indicative of zoonotic potential. These findings underscore the necessity for sustained genomic surveillance and stricter biosecurity interventions in LBMs to prevent cross-species transmission and mitigate pandemic risk.

The high cost and limited availability of maize in poultry production necessitate the exploration of alternative feed sources. Bread waste (BW), a nutrient-rich by-product, offers a sustainable substitute. However, concerns regarding its protein degradation during processing and limited research in laying hens restrict its broader application. Probiotics such as Bacillus amyloliquefaciens CECT 5940 may enhance nutrient utilization and economic viability when combined with BW. This study aimed to evaluate the effects of partially replacing maize meal with BW, with or without the inclusion of B. amyloliquefaciens CECT 5940, on the productive performance and economic outcomes in laying hens. Sixty 52-week-old Lohmann Brown laying hens were randomly assigned to one of three treatments (n = 20 per treatment; 4 replicates of 5 hens each): (T1) basal diet; (T2) basal diet with 20% BW replacing maize; and (T3) basal diet with 20% BW and 0.8% B. amyloliquefaciens CECT 5940. Performance parameters and economic indicators, including feed intake, egg production, feed conversion ratios, gross revenue, contribution margin, and break-even point, were assessed over 45 days. No significant differences (p > 0.05) were observed in laying rate, egg mass, feed conversion per dozen eggs, or live weight across treatments. Economically, substituting 20% of maize with BW significantly reduced feed cost per kilogram and egg production cost per unit and per dozen (p < 0.05). The inclusion of B. amyloliquefaciens further enhanced the profitability index, gross value added, and reduced the break-even point compared to T1 and T2 (p < 0.05). Replacing 20% of maize with BW, with or without B. amyloliquefaciens CECT 5940, maintains productive performance in laying hens while significantly improving economic efficiency. This strategy offers a viable and sustainable alternative to conventional feed formulations in poultry production.

Optimizing feed strategies is critical in livestock production to enhance animal performance, nutrient utilization, and meat quality. Feed form, such as pelleted, mash, or blended forms, significantly influences these parameters. Investigating the optimal feed form for fat-tailed sheep production can improve economic outcomes and meat quality. This study aimed to evaluate the impact of dietary feed forms – pelleted (P10), mash (M10), and a blended form consisting of 50% pellet and 50% mash (M5P5) – on nutrient digestibility, animal performance, carcass characteristics, and meat quality attributes in fat-tailed sheep. Fifteen fat-tailed lambs were randomly allocated to three experimental groups receiving either100% mash feed (M10), a 50:50 mixture of mash and pelleted feed (M5P5), or 100% pelleted feed (P10) for a 90-day feedingtrial. Feed intake parameters (dry matter, organic matter, crude protein, crude fat, crude fiber, and nitrogen-free extract)were recorded. Nutrient digestibility was assessed, and production performance measures, including body weight gain,average daily gain (ADG), feed efficiency ratio (FER), and feeding cost per gain were determined. Post-slaughter carcasstraits, proximate meat composition, cholesterol content, pH, water-holding capacity (WHC), cooking loss, and shear forcevalues were evaluated. Dietary feed forms had no significant impact (p > 0.05) on feed intake, nutrient digestibility, ADG, or FER.However, significant differences (p < 0.05) emerged in carcass traits, notably with increased hot carcass weights observedin M5P5 (19.57 kg) and P10 (19.40 kg) compared to M10 (17.10 kg). Feed form significantly influenced meat-to-boneratio, with M5P5 and P10 groups exhibiting superior ratios relative to the mash-fed group. Meat quality analysis indicatedsignificant variations (p < 0.05) in WHC and cooking loss; the M5P5 group demonstrated enhanced WHC (63.2%) andreduced cooking loss (18.4%) compared to other treatments. Proximate composition, cholesterol content, pH, and shearforce were unaffected by feed form (p > 0.05). The blended mash-pellet diet (M5P5) effectively enhanced specific meat quality parameters, notably WHC and cooking loss, without compromising growth performance or nutrient utilization efficiency. These findings indicate potential for the strategic use of blended feeds in fat-tailed sheep production to optimize meat quality attributes, although further studies examining long-term economic and metabolic impacts are recommended.

Hepatobiliary disorders represent a significant clinical concern in canine medicine, contributing to substantial morbidity and mortality. However, comprehensive data on clinical presentation and hematological and biochemical alterations associated with these disorders in Vietnam remain limited. This study aimed to characterize the clinical manifestations and hematobiochemical profiles of dogs diagnosed with hepatobiliary disorders in Ho Chi Minh City, Vietnam. Eighty client-owned dogs diagnosed with hepatobiliary disorders through histopathological confirmation were retrospectively evaluated. Clinical signs, age, breed, and sex were recorded. Hematological indices – including red and white blood cell counts, hemoglobin concentration, hematocrit, and platelet (PLT) count – and biochemical parameters – such as total protein (TP), albumin (ALB), globulin, bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), blood urea nitrogen (BUN), creatinine (CREA), and uric acid – were assessed. Data were analyzed using one-way analysis of variance and Pearson’s Chi-square test, with statistical significance set at p < 0.05. Hepatitis was the most prevalent disorder (38.8%), followed by biliary disorders, hepatic lipidosis, neoplastic disorders (each 16.3%), and cirrhosis (12.5%). Vomiting (60.0%), jaundice (57.5%), and diarrhea (48.8%) were among the most frequent clinical signs, with significant variability across disorder types (p < 0.05). A significant variation in PLT counts was observed, with the highest values in hepatic lipidosis cases (p = 0.04). Biochemical analysis revealed marked elevations in total bilirubin, ALT, AST, ALP, and GGT across disorders (p < 0.05), particularly in hepatitis and neoplastic cases. Dogs with cirrhosis exhibited significantly reduced TP and ALB concentrations. Elevated BUN and CREA were also noted in dogs with neoplastic conditions, suggesting concurrent renal involvement. This study is the first to delineate the clinical and hematobiochemical characteristics of canine hepatobiliary disorders in Vietnam. The findings underscore the diagnostic value of integrating clinical signs with laboratory indices, particularly elevated liver enzymes and hypoalbuminemia, in the identification and differentiation of hepatobiliary conditions. These insights may enhance clinical decision-making and contribute to improved diagnostic accuracy and therapeutic outcomes in veterinary hepatology.

Weaning is a critical period in calf development, particularly under tropical conditions where nutritional stress and parasitic infestations can precipitate anemia. Anemia compromises growth and survival; yet, few studies have evaluated the effectiveness of hematinic therapy in anemic calves before weaning in tropical systems. This study aimed to assess hematological responses in anemic dairy calves treated with a hematinic complex and to establish anemia thresholds based on hematological indices in healthy calves raised under tropical conditions. A total of 22 Holstein × Zebu calves were studied from January to April 2024 in Tabasco, Mexico. Calves were grouped as healthy (n = 10; hematocrit [HCT] >24%) or anemic (n = 12; HCT <24%). The anemic group received an intramuscular hematinic complex for 5 consecutive days and three additional doses at day 42. Hematological parameters were measured every 21 days using an automated analyzer. Anemia thresholds were defined from the healthy group using mean ± 2 standard deviations. Data were analyzed using a repeated-measures design over time. Anemia thresholds were determined as HCT <21.3%, hemoglobin (HGB) <7.5 g/dL, and red blood cell (RBC) <5.4 × 106/μL. At baseline, anemic calves exhibited significantly lower HCT (21.4%), HGB (6.9 g/dL), and RBC (5.9 × 106/μL) compared to healthy controls (p < 0.01). Three calves presented with microcytic hypochromic anemia and one with macrocytic anemia. By day 42, hematinic-treated calves surpassed the anemia threshold, reaching an HCT of 25%, and further increased to 30% following the second treatment. HGB and RBC levels also improved, showing no significant differences from healthy calves at study end. Males responded more robustly to treatment than females. The administration of a hematinic complex effectively restored hematological parameters in anemic calves within 42 days, with sustained improvement following a second application. This intervention is recommended as part of pre-weaning health protocols to mitigate anemia-related growth setbacks in tropical cattle systems.

Fermented black soldier fly larvae (BSFL) have emerged as a sustainable and economically viable protein source in aquaculture. However, their potential as a replacement for marine fish in the diets of Asian swamp eels (Monopterus albus, ASEs) remains underexplored. This study assessed the effects of partially substituting marine fish with fermented BSFL on ASE growth performance, intestinal development, and hepatic health. A total of 480 ASEs were randomly assigned to four dietary groups: control (40% marine fish), BSFL34 (13.4% BSFL), BSFL61 (24.1% BSFL), and BSFL82 (32.8% BSFL), replacing marine fish on a dry matter basis. All diets were isonitrogenous and isoenergetic. Fish were reared in net cages for over 90 days, and parameters including survival rate, growth metrics, muscle and liver histology, intestinal morphology, gene expression (quantitative real-time polymerase chain reaction), and inflammatory protein levels (Western blotting) were assessed. Survival rate was significantly higher in the BSFL61 group (p < 0.05). Growth performance was not impaired across BSFL-fed groups, although BSFL61 showed reduced body weight compared to BSFL82 (p < 0.05). Muscle fiber size, satellite cell number, and muscle triglyceride (TG) content remained unchanged. BSFL82 showed increased hepatic TG accumulation (p < 0.05) and reduced liver fibrosis, while BSFL61 exhibited a significantly lower hepatosomatic index and increased fibrosis. Intestinal villus height was reduced in BSFL34 and BSFL61, while goblet cell density increased in all BSFL groups. Notch1 expression was upregulated in BSFL61 and BSFL82, whereas ctnnb1 and wnt5a were downregulated. Inflammatory markers nuclear factor-kappa B and interleukin-1 beta were elevated in BSFL-fed groups, indicating an activated mucosal immune response. Partial replacement of marine fish with fermented BSFL enhanced ASE survival, modulated intestinal immunity, and improved mucosal barrier function, without compromising overall growth performance. However, excessive inclusion may induce hepatic lipid accumulation and affect intestinal morphology. These findings support the use of fermented BSFL as a sustainable aquafeed ingredient, though inclusion levels should be carefully optimized to balance health benefits and growth efficiency.

Mastitis remains a major health challenge in dairy cattle, often caused by Gram-positive pathogens. Toll-like receptors (TLRs) and chemokine receptors (CXCRs) play essential roles in the innate immune response of mammary epithelial cells (MECs). However, the differential expression of these genes in response to specific mastitis-causing Bacillus spp. has not been comprehensively evaluated. This study aimed to characterize the temporal gene expression patterns of TLR and CXCR family members in murine mammary epithelial HC11 cells exposed to Bacillus cereus and Bacillus subtilis, thereby providing insights into their immunological roles in mastitis pathogenesis. HC11 cells were cultured and infected with B. cereus and B. subtilis (5 × 10⁷ colony-forming units/mL) and incubated at 37°C with 95% O2 and 5% CO2 for 48 h in RPMI 1640 medium supplemented with serum and antibiotics. Gene expression of interleukin (IL)-6, IL-8, TLR2, TLR4, IL-1 alpha (IL-1α), and CXCR1 was evaluated by quantitative real-time polymerase chain reaction at 0, 6, 12, 24, 48, and 72 h post-infection. Expression levels were normalized to glyceraldehyde-3-phosphate dehydrogenase and analyzed using ΔCt methods and Spearman correlation. TLR2 exhibited a biphasic expression pattern, with early upregulation followed by suppression, while TLR4 showed higher expression in response to B. subtilis than B. cereus. IL-6 displayed prolonged expression under B. subtilis challenge but was transient under B. cereus exposure. IL-1α showed consistent expression across both bacterial challenges, suggesting its potential as a stable biomarker for mastitis susceptibility. CXCR1 exhibited delayed but sustained expression, indicative of its role in secondary neutrophil recruitment. IL-8 expression correlated with early immune activation and chemotactic signaling. The immune response of HC11 MECs to Gram-positive bacterial infection is gene- and pathogen-specific. TLR and CXCR genes show distinct temporal profiles, underscoring their utility in understanding epithelial-driven immune defense. These findings provide molecular insights into mastitis pathogenesis and identify IL-1α, IL-6, and CXCR1 as promising targets for genetic selection and therapeutic intervention.

Staphylococcus pseudintermedius is an opportunistic zoonotic pathogen frequently implicated in skin and wound infections in companion animals. Its ability to form biofilms complicates treatment by increasing antibiotic resistance. Rhodomyrtone, a potent acylphloroglucinol isolated from Rhodomyrtus tomentosa, exhibits promising antibacterial activity against Gram-positive bacteria. This study aimed to develop rhodomyrtone-rich fractions and evaluate their antibacterial and antibiofilm activities against S. pseudintermedius. Ethanolic extracts of R. tomentosa leaves were subjected to acetone partitioning followed by quick column chromatography, yielding fractions F1–F15. Fractions F3–F7 were selected based on thin-layer chromatography and 1H nuclear magnetic resonance for rhodomyrtone content and quantified by high-performance liquid chromatography. Antibacterial activity against Staphylococcus aureus American Type Culture Collection (ATCC) 25923, S. pseudintermedius ATCC 49444, and 10 clinical S. pseudintermedius isolates was assessed using broth microdilution to determine minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values. Fraction F4, with the highest rhodomyrtone content, was further investigated using time-kill kinetics, scanning electron microscopy (SEM), and a crystal violet assay for biofilm inhibition. Fraction F4 contained the highest rhodomyrtone concentration (489.08 mg/g) and demonstrated the most potent antibacterial activity, with MIC and MBC values ranging from 0.5 to 2 μg/mL and 2 to 8 μg/mL, respectively, against clinical isolates. The time-kill study revealed a 4-log reduction (99.99%) in bacterial load within 8 h at 2× MIC. Biofilm formation by all tested S. pseudintermedius isolates was significantly inhibited at sub-MIC concentrations of F4 (p < 0.05). SEM analysis showed notable morphological disruptions in bacterial cells treated with F4, suggesting membrane damage as a possible mechanism of action. Fraction F4 from R. tomentosa leaf extract exhibited strong antibacterial and antibiofilm activity against S. pseudintermedius, comparable to that of pure rhodomyrtone and superior to doxycycline. These findings support the potential use of rhodomyrtone-rich fractions as standardized herbal antibacterial agents in veterinary medicine, providing an effective alternative for treating drug-resistant staphylococcal infections.

Indonesia is home to a diverse array of native cattle breeds - such as Aceh, Bali, Madura, and Sumba Ongole - that are uniquely adapted to their regional environments and integral to the country’s agricultural and cultural heritage. This review synthesizes current research on the genetic diversity and phylogenetic relationships of Indonesian native cattle using microsatellite markers, a robust molecular tool for assessing genetic variation. Analysis of multiple studies reveals that geographical isolation, selective breeding, and human-mediated practices have shaped the genetic structure of these populations. Notably, Bali and Madura cattle exhibit distinctive genetic clusters reflecting island isolation and long-standing breeding traditions. Findings also underscore concerns regarding genetic erosion caused by uncontrolled crossbreeding with exotic breeds, which threatens local adaptability and increases the risk of inbreeding. Key genetic parameters - including allele richness, heterozygosity, and inbreeding coefficients - suggest varying degrees of genetic integrity among the breeds, with several populations showing signs of inbreeding depression. Microsatellite data further demonstrate clear phylogenetic separation among breeds, offering valuable insights for breed identification and conservation planning. The primary strength of this review lies in its comprehensive integration of genetic studies across diverse breeds and islands, providing a national-scale perspective. However, limitations include the underrepresentation of certain breeds and reliance on microsatellite data without integration of high-resolution genomic tools. Future research should incorporate advanced molecular techniques e.g., sngle-nucleotide polymorphism arrays and whole-genome sequencing) and longitudinal monitoring to inform targeted conservation strategies. This review advocates for the incorporation of molecular genetic data into national breeding and conservation programs. Strengthening such initiatives is essential for preserving Indonesia’s indigenous cattle as valuable genetic resources for climate-resilient, sustainable livestock production.

Pacific white shrimp (Penaeus vannamei) is a principal species in global aquaculture. However, outbreaks of Vibrio parahaemolyticus, the etiological agent of acute hepatopancreatic necrosis disease (AHPND), cause substantial economic losses. β-glucans derived from marine diatoms, Chaetoceros muelleri (CH) and Thalassiosira weissflogii (TH), have shown potential as immunostimulants to enhance shrimp resistance to pathogenic infections. This study aimed to evaluate the effects of β-glucans derived from CH, TH, and their combination on growth performance, immune responses, and survival of P. vannamei and to elucidate the underlying molecular mechanisms through transcriptomic and gene silencing approaches. Juvenile shrimp were assigned to four dietary groups for 30 days: Control (β-glucan-free), β-glucan from CH, TH, and a mixture of both (CH and TH) (CHTH). Growth performance, total hemocyte count (THC), and survival rate were evaluated. RNA-seq was performed on hepatopancreas samples after 14 days to identify differentially expressed genes (DEGs). Key immune-related DEGs were validated by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Functional analysis of the lysozyme (Lyz) gene was conducted through RNA interference (RNAi), followed by V. parahaemolyticus challenge. The CHTH diet group exhibited significantly enhanced growth metrics and the highest survival rate. Transcriptomic analysis revealed 1,902 DEGs in the CHTH group compared to control, with 915 upregulated and 987 downregulated genes. qRT-PCR validated the expression trends of selected immune-related genes, notably Lyz, which showed robust upregulation. RNAi-mediated Lyz knockdown reduced survival upon bacterial challenge, confirming its role in β-glucan-induced immunity. β-glucans derived from CH and TH, particularly in combination, significantly enhance growth performance and immunocompetence in P. vannamei. These findings underscore the potential of marine diatom-derived β-glucans as viable immunostimulants to mitigate AHPND in shrimp aquaculture, offering a sustainable alternative to antibiotic use.

Heat stress (HS) significantly compromises broiler performance, intestinal health, and immune responses, posing a growing threat under climate change. Probiotics (PROs) have been proposed as a nutritional intervention to mitigate HS effects, yet their efficacy through drinking water remains understudied. This study aimed to evaluate the effects of water-supplemented PROs on growth performance, physiological responses, intestinal morphology, and inflammatory biomarkers in broiler chickens under thermo-neutral (TN) and HS conditions. A total of 192 one-day-old Ross 308 broiler chicks were randomly allocated to four treatment groups (6 replicates/group): TN with control (CON) water, TN with PRO-supplemented water, HS with CON water, and HS with PRO-supplemented water. PROs (Bacillus subtilis and Bacillus pumilus) were administered in drinking water at 50 mg/L from day 1 to 35. Birds in the HS groups were subjected to 3 h daily heat exposure (33°C ± 2°C) from days 31 to 35. Growth performance, physiological indicators, intestinal histomorphology, and blood biomarkers were assessed. HS significantly increased rectal temperature (Tr) (1.6°C; p < 0.01) and respiratory rate (57 breaths/min; p < 0.01). PRO supplementation reduced Tr by 0.17°C (p = 0.01) but did not affect performance metrics (feed intake, body weight gain, feed conversion ratio). HS reduced villus height (VH) and surface area in the jejunum and ileum (p < 0.05), while PROs partially ameliorated VH under HS. No significant effects of PROs were observed on serum amyloid A or tumor necrosis factor-alpha levels. However, PROs tended to reduce diamine oxidase levels (24%; p = 0.09). HS also decreased albumin and glucose levels (p ≤ 0.02). HS adversely affected intestinal integrity and selected blood metabolites. Although PRO supplementation had limited effects on performance and inflammatory biomarkers, it conferred modest thermoregulatory benefits and tended to improve intestinal permeability. Further research is warranted to optimize PRO formulation and assess synergistic strategies for HS mitigation in poultry.

Nanotechnology offers innovative strategies to enhance livestock productivity and sustainability. Silicon-containing ultrafine particles (UFPs) have shown potential benefits in animal nutrition, yet their effects on gastrointestinal microbial composition and ruminal digestion in cattle remain poorly understood. This study was to evaluate the impact of dietary supplementation with silicon-containing UFPs on ruminal digestibility, bacterial taxonomic structure, and predicted metabolic functions in the gastrointestinal microbiota of cattle. A 42-day controlled feeding experiment was conducted on 20 Kazakh white-headed bulls (12 months old, 305 ± 10.4 kg), divided into control and experimental groups (n = 10 each). The experimental group received a diet supplemented with SiO₂ UFPs (2 mg/kg feed). Digestibility coefficients were measured using standard methods, and ruminal fluid samples were subjected to 16S ribosomal RNA sequencing and Kyoto encyclopedia of genes and genomes -based functional profiling. UFP supplementation significantly increased the digestibility of dry matter (3.5%), crude fiber (3.5%), crude protein (5.2%), and organic matter (8.11%) compared to the control group. The experimental group exhibited elevated relative abundances of Prevotellaceae, Lachnospiraceae, Oscillospiraceae, and genera Prevotella, Ruminococcus, and Selenomonas. Functional prediction analysis revealed higher proportions of genes involved in carbohydrate metabolism (e.g., starch, galactose, and amino sugar pathways), lipid metabolism, oxidative phosphorylation, and the biosynthesis of key vitamins and cofactors. Microbial diversity metrics (Chao1, Shannon) indicated significant changes in alpha diversity, with moderate shifts in beta diversity. Dietary inclusion of silicon-containing UFPs enhances nutrient digestibility and induces favorable modifications in the ruminal microbiota, including functional pathways linked to energy and macronutrient metabolism. These findings support the integration of nanotechnology-based feed additives in cattle nutrition to improve feed efficiency, productivity, and potentially reduce environmental impacts such as methane emissions.