Market demand for safe feed and food supply and consumer preferences for safe and healthy products are increasing. Control measures to counter threats to the feed supply need to be implemented as early as possible to prevent economic losses. Mycotoxins produced by certain groups of fungi are a problem that can disrupt the feed supply or pose a threat to the health of animals and humans. Biological control to detoxify contaminated feed ingredients can be carried out on a large scale economically. For example, lactic acid bacteria (LAB) can act as biological agents for eliminating mycotoxins. This study aimed to clarify the value of screening LAB to inhibit Aspergillus flavus> growth and detoxify aflatoxin B1 (AFB1). In this study, using a completely randomized design with three replications, five isolates of LAB (LA.1, LA.6, LA.8, LA.12, and LA.22) along with their supernatants were tested qualitatively and quantitatively for their ability to counter mycotoxins using A. flavus and corn kernels. The isolates with the best activity were identified by sequencing 16S rDNA. The results showed that the five LAB isolates can inhibit the growth of A. flavus and detoxify AFB1. Among these isolates, LA.12 showed the best performance, followed by LA.22, LA.8, LA.6, and then LA.1. The sequencing results confirmed that LA.12 was Lactobacillus harbinensis strain 487. All of the isolates in this study have the potential as biological agents for detoxifying AFB1, with isolate LA.12 appearing to be the most promising biodetoxification agent for feed (AFB1 in corn) based on its ability to inhibit pathogenic fungi. Keywords: biodetoxification, feedstuffs, lactic acid bacteria, pathogenic fungi.

Mastitis is an important disease that can reduce milk production and farmer income as well as negatively affect human health. This study aimed to summarize dairy mastitis in Indonesia, both subclinical mastitis (SCM) and clinical mastitis (CM), and its prevalence in different provinces, the diagnostic methods, and the animal species. Relevant studies on mastitis in dairy animals in Indonesia were obtained from PubMed, Scopus, ProQuest, Google Scholar, and Garuda. The title and abstract were screened for the eligibility of the studies. The full text of the selected studies was assessed and the data were extracted for analysis. To determine the pooled estimate of the prevalence of mastitis, a random-effects model was performed using the “Meta” and “Metaphor” packages in the R software version 4.2.2. The heterogeneity of several characteristics (mastitis type, provinces, animal species, and diagnostic methods) was evaluated through subgroup meta-analysis. Meta-regression analysis was conducted to assess the trend of mastitis prevalence reports over time. Publication bias was evaluated using Egger’s test and a funnel plot. A total of 735 studies were retrieved for the title and abstract screening, which resulted in the final selection of 37 studies with a total of 6050 samples for meta-analysis. The pooled estimate of mastitis prevalence in dairy animals in Indonesia was 59.44% (95% confidence interval [CI], 52.39%–66.49%). Based on mastitis type, SCM had a significantly higher prevalence than CM (58.24% [95% CI, 51.26%–65.23%] vs. 3.31% [95% CI, 1.42%–5.19%]). No significant difference was observed in the analysis of other subgroups. Among provinces, Central Java had the highest prevalence (66.62% [95% CI, 49.37%–83.87%]), whereas Yogyakarta had the lowest (41.77% [95% CI, 14.96%–68.58%]). Based on animal species, cow and goat had a prevalence of 63.42% (95% CI, 55.97%–70.86%) and 44.96% (95% CI, 28.26%–61.66%), respectively. Based on the diagnostic method, the California mastitis test resulted in 60.08% (95% CI, 52.11%–68.06%) and the Institut Pertanian Bogor test, 56.00% (95% CI, 41.20%–70.81%). No significant change in the prevalence of mastitis in Indonesia was observed from 2003 to 2022. This study demonstrates that the pooled estimate of mastitis prevalence in dairy animals in Indonesia is >50%. Based on subgroup analysis, SCM had a higher prevalence than CM; however, the prevalence between provinces, detection methods, and animal species in the 2003–2022 periods was not significantly different. A mastitis control strategy needs to be developed to reduce the prevalence of mastitis and further loss in milk production. Keywords: animals, cow, Indonesia, subclinical mastitis.

Brucellosis remains an endemic zoonotic disease in many developing countries, causing great harm to public health and devastating losses to livestock. One of the main reasons for the low effectiveness of anti-brucellosis measures is the lack of reliable methods for diagnosing infected animals throughout their lifespan. Classical serological tests, such as the tube agglutination test, rose Bengal plate test, and complement fixation test, as well as commercial enzyme-linked immunosorbent assay kits, are based on the detection of antibodies to the cell wall polysaccharide antigens of Brucella spp. smooth strains. As a result, they do not exclude cross-reactions with related bacteria and fail to differentiate between infected and vaccinated animals. Over the past decades, many attempts have been made to identify immunoreactive and pathogen-specific protein antigens. To date, several studies have investigated Brucella spp. recombinant proteins, including cell wall proteins, as the best antigens for diagnosing brucellosis in animals and humans. However, the available results on the specificity and sensitivity of serological tests based on cell wall proteins are ambiguous and sometimes contradictory. This review aims to provide an overview of the current state of knowledge of the diagnostic value of outer membrane and/or periplasmic proteins of Brucella spp. The goal is to identify future developments that may lead to reliable antigens for serological tests.

Coronavirus disease-2019 (COVID-19) impacts increasing the use of disinfectants (benzalkonium chloride), which indirectly accumulate in water. The disinfectant accumulation in waters has been studied, but there has been no study of its impact on aquatic commodities, especially fish with a high sensitivity, one of which is goldfish (Carassius auratus). Benzalkonium chloride can potentially affect several body proteins, including the cytoskeleton, nervous and endocrine systems, and fish physiology. This study aimed to determine the impact of benzalkonium chloride as a disinfectant on the level of color brightness, growth, gill histopathology, and mortality in goldfish. This investigation provides input into the impact of using disinfectants to prevent COVID-19 on aquatic commodities. This study utilized goldfish specimens sourced from Tulungagung, East Java, Indonesia. The experiment involved different concentration levels of benzalkonium chloride: (T1) 0 mg/L, (T2) 0.03 mg/L, (T3) 0.06 mg/L, (T4) 0.09 mg/L, and (T5) 0.12 mg/L. The research data were subjected to an analysis of variance for analysis. In cases where significant differences were observed, Duncan’s test was conducted for color brightness, growth, and mortality data. Furthermore, if the gill histopathological data yielded significant differences, additional tests were applied (Kruskal–Wallis and Mann–Whitney test). The findings of this study demonstrated significant differences (p < 0.05) in the level of color brightness, growth, gill histopathology, and mortality in goldfish in response to varying concentrations of benzalkonium chloride. The relationship between the length and weight of the goldfish was analyzed using regression coefficients (b values), which were determined as 4.86, –0.04, –0.2, 0.8, and –0.07, respectively. Notably, the brightness level in the T2 group exhibited positive color results with a hue value of 11.55°, while optimal growth was observed in the T4 group, as evidenced by b value of 0.8. The gill histopathological data showed significant differences (p < 0.05). The scoring of histopathological damage in the goldfish gills ranged from 0 to 10, with higher scores indicating more severe damage. The highest total score of 10 was observed in the T5 group exposed to a concentration of 0.12 mg/L, resulting in an 85% mortality rate. This indicates that benzalkonium chloride, with its toxic compounds, can disrupt the respiratory system of fish and lead to death. The effects of benzalkonium chloride were evident even at a concentration of 0.03 mg/L. With increasing concentration, there was an increase in mortality rate, a decrease in growth, and a rise in histopathological damage to the gills. These findings highlight the negative impact of using conventional disinfectants on water and its organisms, emphasizing the need for further research on environmentally friendly alternatives. Keywords: color brightness, fisheries management, freshwater species, integrated multi-trophic aquaculture, mortality.

Colic is among the common health issues in equine health management. Gastrointestinal (GI) disorders are the most frequent causes of colic, but dysfunction of other organs and systems inside the abdominal cavity may also contribute. Therefore, it is crucial to identify risk factors for colic of specific etiologies. This study aimed to examine the incidence, risk factors, and best therapeutic management practices for horses with colic. A cohort of 256 horses living in Lamongan, East Java, Indonesia, was randomly recruited based on reports of colic symptoms by owners. Diagnosis and treatment were then conducted with the help of owners. Symptom profiles, risk factors, and therapeutic management strategies were analyzed by Chi-square tests. Of 256 horses enrolled, 217 (84%) were diagnosed with colic, of which 172 (79.3%) were cases of spasmodic colic, 33 (15.2%) of impaction colic, and 12 (5.5%) of intestinal obstruction/displacement. Male sex (χ2 = 16.27; p < 0.001), wheat bran feeding (χ2 = 15.49; p < 0.001), concentrate feed intake >5 kg/day (χ2 = 24.95; p < 0.001), no regular anthelmintic drug treatment (χ2 = 67.24; p < 0.001), GI parasite infection (χ2 = 65.11; p < 0.001), recurrent colic (χ2 = 91.09; p < 0.001), poor body condition score (χ2 = 71.81; p < 0.001), limited daily water access (χ2 = 127.92; p < 0.001), and indications of dental disease (χ2 = 9.03; p < 0.001) were identified as risk factors. The most effective therapies were gastric intubation (χ2 = 153.54; p < 0.001), Vitamin B complex injection (χ2 = 32.09; p < 0.001), fluid therapy (χ2 = 42.59; p < 0.001), and non-steroidal anti-inflammatory drug injection (NSAID). Colic is highly prevalent among horses in Lamongan, East Java, Indonesia. Proper diet, workload management, regular access to clean drinking water, and dental care can reduce colic risk. Recommended therapies include NSAID injection without other analgesics or spasmolytics, fluid therapy, Vitamin B complex, and gastric intubation. Keywords: colic, domesticated animals, lamongan, risk factors, therapeutic management.

The poultry industry faces an emerging muscular defect in chicken meat called white striping (WS). The biological processes associated with WS myopathy are immune system activation, angiogenesis, hypoxia, cell death, and striated muscle contraction. We examined the Troponin T3 (TNNT3), Toll-like receptor 2 (TLR2), and Toll-like receptor 4 (TLR4) genes based on their functions related to muscle contraction and the innate immune system. This study aimed to determine the muscle fiber characteristics (MFCs) and expression level of TNNT3, TLR2, and TLR4 genes in white striping chicken meat (WSCM). A total of 428 breast samples were randomly collected from a commercial poultry processing plant. The samples were classified into four levels: 0 (normal), 1 (moderate WS), 2 (severe WS), and 3 (extreme WS). Five samples per group were selected to evaluate MFCs, including total number of muscle fibers, muscle fiber diameter, cross-sectional area, endomysium thickness, and perimysium thickness. Five samples per group were selected for ribonucleic acid (RNA) isolation to evaluate the messenger RNA (mRNA) expression levels of TNNT3, TLR2, and TLR4 genes related to WS. Statistical analysis revealed that the total number of fibers, endomysium thickness, and perimysium thickness significantly differed between groups (p < 0.05). Muscle fiber diameter and cross-sectional area did not significantly differ (p > 0.05). The expression of the TNNT3 gene did not significantly differ among groups (p > 0.05). Toll-like receptor 2 and TLR4 mRNA expression significantly differed among groups (p < 0.05). These detailed MFCs will provide baseline information to observe WS in chicken meat. Toll-like receptor 2 and TLR4 genes may play a role in the occurrence of WS in chicken meat through non-specific immune reactions. Keywords: chicken, gene expression, myopathy, poultry, white striping.

Seeds from Carica pubescens were observed to be abundant as waste. This waste contains active plant compounds whose utilization has not been studied in the livestock sector. This study was conducted to evaluate the potential of an aqueous seed extract of Carica pubescens (ASE) as an anthelmintic agent during rumen fermentation and methane reduction. Aqueous seed extract of Carica pubescens was prepared from C. pubescens cultivated in Wonosobo, Indonesia. Phytochemical analyses were performed to quantify the secondary metabolite content of ASE. In vitro adult worm mortality tests, scanning electron microscopy, and gas production tests were conducted to evaluate rumen characteristics, methane reduction, and the potential of ASE as an anthelmintic against Haemonchus contortus. Adult worms and ruminal fluid were collected from Indonesian thin-tailed sheep. Two-way analysis of variance followed by Tukey’s test was performed using the Statistical Package for the Social Sciences® 21.0 software to detect significant differences. In vitro study results showed that 1–5% ASE inhibited H. contortus after the 1st h of incubation, and 5% ASE suppressed 100% of adult H. contortus worms in the 5th h. Scanning electron microscopy analysis of ASE-treated worms ASE revealed damaged cuticle structures. ASE had no significant effect on pH, NH3, volatile fatty acid, acetate, propionate, butyrate, acetate: propionate, or microbial protein in rumen fluid (p > 0.05). The in vitro feed fermentation results showed that ASE significantly affected methane reduction. The inclusion of up to 5% ASE in sheep diets may serve as a potential alternative anthelmintic against H. contortus as well as a methane reduction agent, without deleterious effects on rumen fermentation. Keywords: Carica pubescens, fermentation, greenhouse, Haemonchus contortus, metabolite, ruminant.

Foot-and-mouth disease (FMD) virus causes continuous outbreaks, leading to serious economic consequences that affect animal productivity and restrict trade movement. The potential influence of the disease was due to the emergence of new strains or re-emergence of local strains with major antigenic variations due to genetic mutations. This study aims to evaluate circulating virus in samples collected from infected animals during an outbreak using antigenic characterization and identify whether there is an emergence of a new strain or mutation. Reverse-transcription polymerase chain reaction (RT-PCR) was used to screen 86 samples. Viral protein 1 (VP1) codon sequencing was performed. The virus was isolated from the samples inoculated on the baby-hamster kidney cell line and Enzyme-linked immunosorbent assay was performed for serotyping and antigen detection. Based on the RT-PCR screening results, 10 positive samples were selected for sequencing. The sequences belonged to the FMD serotype A African topotype originating from the ancestor prototype Sudan/77, with which it shared 98.48% ± 1.2% similarity. The divergence with local isolates from 2020 was 9.3%. In addition, the sequences were 96.84% ± 1.01% and 95.84% ± 0.79% related to Egyptian-Damietta type 2016 and Sudanese-2018, respectively. Divergence with vaccinal strains ranged from 10% to 17%. Amino acid sequence analysis revealed that the isolates had variation in the most prominent antigenic regions (residues 35–75) and the immunogenic determinants of the G-H loop of VP1 (residues 100–146 and 161–175). The current isolates should be included in the locally produced vaccine to provide broader immunogenic coverage against serotype A African topotypes. Keywords: Epicenters, foot and mouth disease virus, foot-and-mouth disease virus Africa type G-IV, mutations, outbreaks, viral protein 1.

Canine gastrointestinal tract (GIT) parasites are of public health and veterinary concern and are harmful to both humans and animals. The risk of transmitting GIT parasites can be minimized depending on dog owners’ knowledge and management practices. Therefore, this study aimed to assess dog owners’ general knowledge and practices regarding GIT parasites. A structured questionnaire containing 10-point Likert statements was administered to 130 respondents in Cebu, Philippines, to obtain information about their profile (age, sex, civil status, and educational attainment) and data regarding their home environment, number of dogs and other pets owned, and their knowledge and practices regarding canine GIT parasites. These respondents had previously provided canine stool samples for fecalysis. Most respondents were female (65.4%), 18–24 years old (55.4%), single (71.5%), and educated to college level (49.2%). Housing styles were mostly gated (76.9%), and >50% had a garden. The majority owned 1–3 dogs (56.9%) and did not own any other pets (73.8%). All knew about canine GIT parasites. There was a significant association between GIT parasite positivity in dogs and the owner’s sex (p = 0.005). Gastrointestinal parasite positivity was also associated with the owner’s knowledge about the transmission of parasites from the mother’s milk to her puppies (p = 0.001), canine intestinal parasites potentially causing weight loss, diarrhea, and vomiting (p = 0.001), and dogs potentially becoming infected with parasites from licking or sniffing the soil or grass where other animals have been (p = 0.005). Moreover, there was a significant association between GIT parasite positivity and the owner’s practice of daily cleaning of the areas where the dog defecated (p = 0.001), deworming every 6–12 months (p = 0.001), and unfailingly following the vaccination and deworming schedule (p = 0.006). Finally, the summative knowledge and practice level of the owners were significantly associated with GIT parasite positivity (p = 0.001). This study highlights the need for continuous education of dog owners to maintain good knowledge and practices to prevent GIT parasite infection. Frequent deworming (once a month) of adult dogs is recommended. Keywords: dogs, intestinal parasites, soil, vaccination.

Porcine circovirus 3 (PCV3) was recently reported in Malaysian commercial pig population in 2020 by conventional polymerase chain reaction (PCR), revealing a molecular prevalence of 17.02% in the sampled domestic pig population. This study aims to describe a chromogenic in situ hybridization (ISH) technique using digoxigenin (DIG)- labeled cloned PCV3 open reading frame 1 (ORF1) fragment DNA to detect and localize the PCV3 antigen in formalin-fixed, paraffin-embedded lung, and lymphoid tissue specimens. Since PCV3 was mainly detected in lung and lymphoid tissues, we obtained tissue specimens from these organs from the previous Malaysian PCV3 study. Digoxigenin-labeled ISH probes were designed to target a 69 bp region of PCV3 ORF1 spanning from the nucleotide positions (282–350). Light microscopy analysis revealed that chromogenic staining of PCV3 antigens was visualized within the cytoplasm of pneumocytes and lymphocytes, indicating positive ISH results. The results of molecular detection of PCV3 using PCR and ISH showed a high agreement of 90.91%, including for the negative PCV3 status for all samples. This study reports a chromogenic ISH technique using DIG-labeled probes targeting PCV3 ORF1 to detect PCV3 antigens in lung and lymphoid tissues. Despite the limited availability of PCV3 antibodies, ISH remains relevant for investigating PCV3 replication and pathogenesis and can be used complementarily with PCR for evaluating the localization of antigens in infected tissues. Keywords: chromogenic in situ hybridization, in situ hybridization, porcine circovirus type 3, porcine circoviruses, pigs.

The main purpose of a national bioresource center is to standardize, centralize, preserve, and ensure accessibility of microbial bioresources that accumulate there because of state research programs. The establishment of national bioresource centers for antibiotic-resistant microorganisms allows to solve practical problems in the field of veterinary service, as well as to develop effective chemotherapeutic and disinfectant drugs to overcome the mechanisms of resistance. This study aimed to outline the process of forming a national culture collection of antibiotic-resistant strains of zoonotic bacteria in the Russian Federation using two microbial strains. The object of research was isolates of Salmonella spp., Escherichia coli, Enterococcus spp., Campylobacter spp., Listeria monocytogenes, and Staphylococcus spp., all of which were obtained from biomaterials of farm animals, feed samples, bedding, water from livestock buildings, washouts from environmental objects, and food products. The resistance of bacterial isolates was determined using microbiological and molecular-genetic research methods. During monitoring studies, 1489 bacterial isolates were isolated. In total, 408 bacterial isolates were tested for sensitivity to antimicrobial agents, including E. coli (47.6%), Salmonella spp. (30.4%), Enterococcus spp. (11.3%), and Campylobacter spp. (10.8%). For genetic characterization, 95 isolates of Salmonella enterica, E. coli, Campylobacter spp., L. monocytogenes, Staphylococcus spp., Enterococcus spp. were chosen from the research collection, which was formed as part of the monitoring program for antibiotic resistance. Deposited isolates that underwent whole-genome analysis can be used as positive control samples both in the development and use of methods or test systems for the detection of various resistance genes in zoonotic bacteria. In addition, such isolates can also be used for microbiological studies related to determining the sensitivity of microorganisms to antibacterial drugs, for phenotypic studies in the diagnosis of various bacterial infections in animals and birds, and retrospective analysis of strains from numerous collections. Keywords: antibiotics, bioresource centers, genes, microorganisms, prevalence, resistance.

Antibiotics that increase growth have long been employed as a component of chicken growth. Long-term, unchecked usage may lead to microbial imbalance, resistance, and immune system suppression. Probiotics are a suitable and secure feed additive that may be provided as a solution. The objective of this research was to ascertain the effects of dietary multistrain probiotics (Lactobacillus acidophilus, Bifidobacterium spp., and Lactobacillus plantarum) on the morphology (length and weight) of reproductive organs and productivity performance of laying hens during the early stage of laying. One hundred ISA Brown commercial layer chicks of the same body weight (BW) that were 5 days old were divided into five treatments, each with four replicates and four chicks in each duplicate. There were five different dietary interventions: (T1) 100% base feed; (T2) base feed with 2.5 g of antibiotic growth promoter/kg feed; (T3) base feed plus probiotics; (T4) base feed at 1 mL/kg with probiotics; and (T5) base feed with probiotics, 3 mL/kg feed, 5 mL/kg of feed. The parameters observed were performance, internal and exterior egg quality, and the morphology (length and weight) of laying hens’ reproductive organs. Probiotic supplementation (L. acidophilus, Bifidobacterium, and L. plantarum) significantly affected the BW, feed intake, egg weight, yolk index, albumin index, Haugh unit, egg height, egg width, and morphology (length and weight) of laying hens’ reproductive organs compared to the control group (basic feed). In addition, there was no discernible difference between treatment groups in theeggshell weight and thickness variables across all treatment groups. When laying hens were between 17 and 21 weeks old, during the early laying period, microbiota inoculum supplements (L. acidophilus, Bifidobacterium, and L. plantarum) increased growth, the quality of the internal and external layers’ eggs, and the morphology of the laying hens’ reproductive organs. Keywords: external quality eggs, good health, growth performance, internal quality eggs, probiotics, reproductive organs.

To meet domestic demand for meat, Indonesia imports live cattle from Australia, which have non-bovine viral diarrhea (BVD)-free status. The consequence of importing live cattle from Australia is potentially introducing a novel BVD variant to Indonesia. Therefore, detecting BVD early and determining the agent’s characteristics and clinical symptoms are necessary. This study aimed to detect and characterize clinical symptoms of bovine viral diarrhea (BVD) and highlight the importance of farm management as a risk factor for the spread of BVD. This study aimed to provide information about the effectiveness of preventive measures against BVD in Australian-imported cattle at the Tanjung Priok Seaport Agricultural Quarantine. Bovine viral diarrhea is among the most common diarrheal diseases found in feedlots and is a severe health and economic problem in cattle. All cattle in a selected feedlot were examined for clinical symptoms on their first day of arrival. The sampling criteria included age, body weight, body temperature (BT), animal breath (AB), pulse (PL), conjunctivitis (CJ), hyperlacrimation (HL), hypersalivation (HS), DR, fever, limping leg (LL), emaciation, stomatitis (ST), weakness (WK), and coronitis (CR). In addition, 64 blood samples were taken from cattle that exhibited clinical symptoms of BVD. On the 3rd> day of arrival, a blood sample showing positive clinical symptoms was examined using antigen (Ag)-capture enzyme-linked immunosorbent assay (ELISA). The data from these clinical symptoms were analyzed alongside the laboratory results using multidimensional scale analysis, heatmap distribution, and principal component analysis (PCA). Furthermore, the positive serum samples obtained from the Ag-capture ELISA underwent a nested multiplex polymerase chain reaction and molecular detection and genetic characterization of BVDV based on the 5’ untranslated region of the viral genome, followed by sequence and phylogenetic tree analyses. Using PCA, 12 clinical symptom characteristics of BVD were determined from 13 clinical symptoms synergized with five cattle positive for Ag-capture ELISA. The clinical symptoms included internal factors such as physiological conditions of CJ, HL, HS, DR, BT, LL, loss of appetite, ST, WK, CR, AB, and PL. The screening test showed that five samples tested positive for the BVD Ag, while 59 tested negative. Phylogenetic tree analysis using a 360-nucleotide portion of the NS5B gene showed that Sample B23F5R had a distinct path compared to the other two samples in the phylogenetic diagram. The profile of sample B23F5R was closely related to BVDV reference subgenotype 1-a group (NCBI, access no. LC068605), with a homology percentage of 92.36%. Furthermore, this sample was similar to the BVDV reference 1-a, Strain 12, identified in Japan. The other two samples, B13F5R and A13F5R, showed close resemblance to the BVDV reference subgenotype 1-a that had been previously identified in Indonesia (NCBI, access no. MK411755), with homology percentages of 97.81% and 97.75%, respectively. The BVDV-1a strain is the main subtype present in beef cattle imported from Australia to West Java, Indonesia. The characteristics of clinical symptoms associated with BVD infection comprised 12 symptoms synergized with the positive sample in the PCA. The present results can facilitate the development of preventive and control measures for BVD circulation in Indonesia. Keywords: bovine viral diarrhea virus, clinical symptoms, detection, imported cattle, West Java.

Herbal plants have the potential to reduce the population of metagonic bacteria and protozoa due to the bioactive compound contained in herbal plants. This study aimed to evaluate the effect of herbal plant supplementation on rumen fermentation characteristics, methane (CH4) gas emissions, in vitro nutrient digestibility, and protozoan populations. This study consisted of two stages. Stage I involved determining the potential of herbal plants to increase total gas production (Orskov and McDonald methods) and reduce the protozoan population (Hristov method). Three potential herbs were selected at this stage and used in Stage II as supplements in the palm kernel cake (PKC)-based diet (30% herbal plants + 70% PKC). Proximate and Van Soest analyses were used to determine the chemical composition. In vitro dry matter digestibility (IVDMD), organic matter (IVOMD), and rumen fermentation characteristics were determined using Theodorous method. Conway microdiffusion was used to determine ammonia concentration (NH3). Gas chromatography was used to determine the total and partial volatile fatty acid production. The results of the first stage showed that seven herbal plants (Moringa oleifera, Rhodomyrtus tomentosa, Clerodendron serratum, Curcuma longa Linn., Urena lobata, Uncaria, and Parkia timoriana) significantly differed in terms of total gas production (p < 0.05). Herbal plants can increase gas production and reduce protozoan populations. The highest total gas production was observed using P. timoriana, M. oleifera, and C. longa Linn. Moringa oleifera plants were the most effective in lowering protozoa population. In Stage 2, the supplementation of herbal plants in PKC-based-diet significantly increased IVDMD, that was ranged from 56.72% to 65.77%, IVOMD that was ranged from 52.10% to 59.54%, and NH3, that was ranged from 13.20 mM to 17.91 mM. Volatile fatty acid partial and total gas production potential and CH4> gas emissions were also significantly different from those of the control (p < 0.05). Supplementation of M. oleifera, C. longa Linn., and P. timoriana in ruminant diet effectively increased total gas production, IVDMD percentage, and IVOMD, and reduced CH4 gas emissions and protozoa populations during rumen fermentation. Keywords: herbal plants, in vitro, methane, protozoa.

Bovine coccidiosis, caused by the protozoa Eimeria, is an important parasitic cattle disease that affects animal health and has economic impact worldwide. This study was conducted to report the first molecular prevalence and genetic diversity of Eimeria spp. in dairy cattle in Khon Kaen province, Thailand, and to identify the risk factors associated with Eimeria spp. infection. From July 2020 to October 2021, 296 fecal samples were collected from dairy cattle divided into three age groups, including <3-month-old calves, 3-month-old to 1-year-old calves, and >1-year-old cattle. Eimeria spp. were identified by multiplex polymerase chain reaction (PCR) amplifying 18S RNA gene and confirmed by DNA sequencing. Information regarding all associated risk factors was collected using questionnaires and analyzed using logistic regression tests in the Statistical Package for the Social Sciences program. Polymerase chain reaction results showed that 104 (35.13%) of 296 samples were positive for Eimeria spp. The <3-month-old calves (46.51%) had the highest infection rate. Moreover, multiplex PCR identified five species of Eimeria, namely, Eimeria bovis (32.69%), Eimeria zuernii (18.26%), Eimeria alabamensis (5.76%), Eimeria ellipsoidalis (3.84%), and Eimeria cylindrica (2.88%). An association was observed between risk factors and Eimeria spp. incidence (p < 0.05). DNA sequencing revealed the similarity of each Eimeria spp. with 91%–100% nucleotide identity. Phylogenetic tree analysis demonstrated the close relationships of clusters of E. bovis and E. zuernii, E. ellipsoidalis, and E. cylindrica and another cluster of E. alabamensis. The results confirm that Eimeria spp. are commonly found in dairy cattle, especially calves. The molecular test could be powerful for species identification. This study also provides epidemiological information for developing future strategies to control bovine coccidiosis. Keywords: bovine coccidiosis, dairy cattle, Eimeria spp., molecular prevalence, multiplex polymerase chain reaction, Thailand.

Swamp buffaloes play an important role in the rural economy of Indonesia. They consume various forages during their grazing time, including those with anti-parasitic potential. However, the information about the type and quality of forages and their potential as a natural anthelmintic for swamp buffalo is very limited. This study aimed to identify the diversity, quality, and anthelmintic potency of forages consumed by swamp buffaloes reared in Bantarkawung Subdistrict, Brebes District, Central Java Province, Indonesia. Samples of forages were obtained during three observation periods of the study, with a 12-week interval between each period. Forage diversity was evaluated by identifying its consumption by swamp buffaloes during their grazing activity in the field and feeding time in their shed. The quality of forages was analyzed using proximate analysis to measure their dry matter (DM), crude fiber (CF), crude protein (CP), crude fat (CFat), and ash contents. This is followed by the calculation of their total digestible nutrient based on the proximate analysis results. Botanical composition analysis was then conducted to measure the predominance of forages consumed by the livestock during their grazing activity. Literature reviews were carried out to explore forage’s anthelmintic activity. The results showed that swamp buffaloes consume nine species of forage in the shed and 47 in the grazing area, including nine legumes, 18 grass, and 20 others. Swamp buffaloes consumed forages of lower quality, which contained high CF contents and varying levels of other nutrients below their daily nutritional needs. The grazing activity allowed swamp buffaloes to consume a higher variety of forages with better nutritional quality, thereby enabling them to meet their nutritional needs. Legumes and other forages served as the major protein sources, providing CP of 20.03% DM and 11.53% DM, and CF levels of 17.01% DM and 20.35% DM, respectively. The results also showed that the consumption of these forages increased during the rainy season. The predominant species of legumes consumed were Leucaena leucocephala and Acacia spp., while Alternanthera sessilis and Merremia umbellata were the predominant species of other forages. A total of 13 of the 47 species could potentially be used as natural anthelmintic due to their secondary metabolites, namely, tannin, flavonoid, saponin, terpenoid, diterpenoid, and mimosine. These compounds exert anthelmintic effects by inhibiting egg-hatching and larval development, as well as damaging the surface structure of both larvae and adult worms, ultimately leading to the death of the parasites. Overall, swamp buffaloes consumed more variety of forages during grazing compared to when they were kept in sheds. While the low-variety and low-quality forages provisioned for swamp buffaloes in their shed resulted in a low nutrient intake below their daily requirement. Furthermore, daily grazing activities allowed swamp buffaloes to fulfill and supplement their need by consuming a variety of grasses, legumes, and other forages in their respective grazing areas. Some of these forages also have the potential to become natural anthelmintic because they contain secondary metabolites, such as tannins, flavonoids, saponins, terpenoids, diterpenoids, and mimosine. Keywords: forages, herbal anthelmintics, Indonesia, nutrition, pastoral, swamp buffalo.

We previously reported the mitigation effects of difructose anhydride III (DFA III) on mycotoxins, such as zearalenon and sterigmatocystin, based on the urinary concentrations of these molecules in calves. This study was aimed at evaluating the effects of dietary supplementation of DFA III and the fermented status of DFA III in the intestine by comparing serum levels of short-chain fatty acid (SCFAs) in DFA III-supplemented cattle with those in non-supplemented control cattle. Serum SCFA concentrations were measured in 30 Japanese Black heifers, aged 9–10 months, from two herds, using gas chromatography on days 0 (before DFA III supplementation), 9, and 14 after DFA III supplementation. A notably different trend was observed for isobutyric acid and enanthic acid, which may reflect the different fermentation status of supplementary DFA III in the intestine. Our results indicate the possibility that this trend observed in the intestinal tract following DFA III administration is associated with changes in the environment of intestinal bacterial flora, which may partially reflect the effects of DFA III supplementation on cattle. Difructose anhydride III supplementation for at least 2 weeks affects the trend of blood SCFA concentrations in cattle. Our results provide evidence supporting the effects of DFA III on the intestinal environment and intestinal barrier function. Keywords: cattle, difructose anhydride III, short-chain fatty acid.

Elemental analysis is a useful technique for predicting metabolic disorders in broiler chickens. Elemental imbalances are also important for the development of new methods to diagnose the health status of birds that can be implemented on a farm-wide scale. This study aimed to identify elemental markers related to pre-nosological diagnoses of metabolic disorders in broiler chickens. We compared birds given high-protein, high-carbohydrate, and high-fat diets. A control group received the standard diet recommended by the All-Russian Research and Technological Institute of Poultry, while experimental Group I received a diet with high-protein content, Group II received a diet with high-carbohydrate content, and Group III received a diet with high-fat content. At the end of the experiment, blood samples were taken for biochemical and elemental analysis. Biochemical analysis was carried out using an automated biochemical analyzer, and the levels of trace elements in the serum were assessed using inductively coupled plasma spectrometry. We found that the high-protein diet was accompanied by a decrease in chicken body weight, cholesterol, and several elements (i.e., P, Cr, Cu, Zn, and B) as well as an increase in the levels of Ca, Co, and Si. The high-carbohydrate diet led to a significant increase in glucose levels as well as a decrease in the levels of albumin, triglycerides, and Cr, Mn, Se, I, and Cu. Finally, the high-fat diet led to an increase in body weight, glucose, cholesterol, triglycerides, and the elements Cu, Zn, and Si as well as a decrease in the levels of Mg, Cr, and Fe. The determination of the levels of trace elements such as Co, Cr, Mn, Fe, and Cu in chicken blood serum may be an important indicator of the state of protein, carbohydrate, and lipid metabolism of poultry stock. Keywords: blood biochemistry, blood serum, broiler chickens, feeding, trace elements.

Healthy meat production is an important aspect of increasing sheep productivity. This study aimed to examine the influence of protected kapok seed oil (KSO) in combination with choline chloride (CC) on the feed utilization, lipid status, and performance of thin-tailed sheep. Thirty male thin-tailed sheep (approximately 6 months old, with an average body weight of 12.59 ± 1.48 kg) were divided into six treatment groups (five heads/treatment). Factor 1 consisted of two treatments: K1 (KSO supplementation at 10% supplementation and 75% protection level) and K0 (without KSO supplementation). Factor 2 consisted of three levels of CC: (C0: 0%; C1: 1.5% and C2: 3% feed dry matter (DM) basis supplementation levels). The variables measured were the DM consumption, DM digestibility, organic matter digestibility, nitrogen retention, daily body weight gain (DBWG), and blood and meat lipid status. The data were analyzed using analysis of variance in a completely randomized design in a factorial pattern of 2 × 3 × 5. Choline chloride supplementation (up to 3%) increased DM consumption in the K0C2 group. The CC and protected KSO (K1C2) supplementation combination resulted in the highest DM consumption level (p < 0.05). The protected KSO supplementation increased DBWG (the DBWG in the K1C0 group was higher than that in the K0C0 group, and the highest DBWG was found in the K1C2 group) (p < 0.05). Protected KSO and CC supplementation decreased cholesterol levels and increased the relative proportion of linoleic acid in meat (p < 0.05). Combined supplementation with protected KSO and CC improved the feed utilization and performance of male thin-tailed sheep. There were increases in DBWG, decreases in intramuscular fat and cholesterol levels, and increases in meat linoleic acid levels. Keywords: cholesterol, linoleic acid, male thin-tailed sheep, meat, performance.

Poultry meat is a popular food consumed globally. However, poultry farming is a source of Salmonella contamination which causes human salmonellosis. This study aimed to estimate the prevalence and antimicrobial resistance (AMR) of Salmonella among chickens and meat at poultry slaughterhouses in province study areas in Thailand and Japan. Chicken meat and feces samples were collected in Thailand and Japan. In Nakhon Ratchasima Province, Thailand, 558 samples were obtained from slaughterhouses from January 2021 to March 2022. In Gifu Prefecture, Japan, 140 samples (70 each of intestinal contents and meat) were purchased from slaughterhouses from June to October 2022. For Salmonella detection, the samples were cultivated according to the International Organization for Standardization 6579:2002/AMD 1:2007 method and confirmed using polymerase chain reaction (PCR) and agglutination tests for serotyping. Isolated Salmonella were tested for AMR to nine antibiotics using the disk diffusion method. Selected phenotypic multidrug-resistant (MDR) isolates were evaluated for AMR genes (AMRGs) using PCR. Salmonella prevalence from chickens and meat at slaughterhouses in Thailand and Japan was 41.2% and 40.7%, respectively. All the Salmonella isolates in Japan were serotyped as Schwarzengrund, and no Salmonella isolates were resistant to the nine antibiotics tested. In contrast, most of the Thai Salmonella isolates from chicken cloacal swabs and meat were resistant to doxycycline (78.3%) and colistin (63.5%). The prevalence of MDR Salmonella (MDRS) in chickens and meat from Thailand and Japan was 29.1% (67/230) and 0% (0/57), respectively. However, the most frequent AMRGs found in MDRS in Thailand were extended-spectrum beta-lactamase-Temoneira (ESBL-TEM) (45.1%). All isolated Salmonella from Japan revealed a class 1 integron gene (Int-1). Phenotypic MDRS isolates from Thailand showed the greatest correlation to AMRG and ESBL-TEM. Although there were no phenotypic AMR Salmonella isolates found in Japan, they can be found associated with Int-1, which may carry other AMRGs within the gene cassettes. Keywords: antimicrobial resistance, broiler chickens, meat, Salmonella, slaughterhouse.

The exchange of oxygen and carbon dioxide (CO2) in the incubator plays a key role in embryonic development and hatching. This study aimed to study the effect of non-ventilated (NV) intervals during the early stage of embryonic development on the hatching process. Hatching eggs (n = 7200) were equally divided into four treatment groups and incubated in four incubators. The first group was incubated in normal ventilated condition (V) during the setting phase of incubation. Ventilation holes of the three remaining incubators were closed for the first 3, 6, and 9 days and termed as NV groups (NV1, NV2, and NV3, respectively). A gradual increase in CO2 was allowed for NV groups, followed by opening the incubator holes to permit ventilation throughout the rest of the incubation periods. Obtained results demonstrated that CO2 concentration gradually increased up to 0.19% for the NV1 group, 0.41% for the NV2 group, and 0.90% for the NV3 group, while CO2 concentration remained at 0.08% during the first 9 days of incubation in the V group. Albumen pH was lowered for all NV groups. The highest hatchability percentage was recorded for NV3 followed by NV2 and NV1 groups. All NV groups represented earlier and narrower spread of hatch and higher hatched chick weight. Embryos and hatched chicks in the NV groups had higher hormonal levels of thyroxin and corticosterone. All non-ventilation periods had positive effects on narrowing the spread of hatch, increasing hatched chick weight and hatchability percentage compared to the normal V condition. Furthermore, the non-ventilation throughout the first 9 days of incubation yielded the best hatching results. Keywords: albumen pH, carbon dioxide, embryonic hormones, hatchability, non-ventilation.

Immune cells require toll-like receptor 4 (TLR4) to respond to lipopolysaccharides (LPS) by releasing pro-inflammatory cytokines. Peripheral blood mononuclear cells (PBMCs) are used to assess changes in cytokines released in response to diseases or pathogens. This study aimed to assess TLR4 gene expression in PBMCs from Leghorn chicken and the release of related cytokines. Peripheral blood mononuclear cells were isolated from blood samples obtained from Leghorn chicks. The PBMC cultures were stimulated with various concentrations of LPS (0.01-1 µg/ml). Polymerase chain reaction was used to detect TLR4 expression. The production of tumor necrosis factor-alpha (TNF-α) and interleukins (IL-1β and IL-6) was quantified using an enzyme-linked immunosorbent assay. We found that TLR4 was expressed in both non-stimulated and stimulated Leghorn chicken PBMCs. In addition, the release of TNF-α, IL-1β, and IL-6 levels in Leghorn chicken PBMCs increased significantly with an increase in LPS concentration (0.01–1 µg/mL) (p < 0.05). Although TLR4 was expressed in both non-stimulated and stimulated Leghorn chicken PBMCs, its expression was significantly higher in LPS-stimulated PBMCs Therefore, the chicken’s endotoxin response can be assessed by evaluating the pro-inflammatory cytokine production from PBMCs. Keywords: Leghorn chicken, peripheral blood mononuclear cell, pro-inflammatory cytokine, Toll-like receptor 4.

Bartonella and Borrelia are zoonotic vector-borne pathogens that can infect dogs and humans. Data on Bartonella and Borrelia in dogs in the Philippines are lacking. This study was conducted to validate previous reports and further investigate the occurrence of Bartonella and Borrelia spp. in cities of Metro Manila. A total of 182 canine blood samples were acquired with DNA using a commercial extraction kit from selected veterinary clinics in the cities of Metro Manila and Laguna, Philippines. The mammalian actin was amplified through polymerase chain reaction (PCR), followed by PCR assays targeting Bartonella gltA and Borrelia flaB. Further PCR assays targeting 16S of Borrelia and ospA and ospC of Borrelia burgdorferi were performed for those that showed flaB bands. A positive band for gltA of Bartonella was observed in 9 (4.95%) samples, whereas a positive band for flaB of Borrelia was observed in 15 (8.24%) samples. Subsequent PCR assays for other genes of Borrelia were negative. These results confirmed the presence of Bartonella and warranted further investigation for the possible presence of other Borrelia species. Keywords: Bartonella, Borrelia, dogs, polymerase chain reaction, tick-borne pathogens.

Mastitis is an essential issue in dairy cows. Post-milking teat dips can help reduce this problem, but they employ harsh disinfectants, and many bacteria are becoming increasingly tolerant. This study aimed to investigate the antibacterial activity of clove, citronella, and sweet basil essential oils against the common bovine mastitis causative agents Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli and to develop an antiseptic post-milking teat spray for use in dairy cows. The in vitro antimicrobial activity of the essential oils was determined by broth microdilution and time-kill assays. Essential oil-based post-milking teat sprays were developed. The bacterial eradication efficacy of the formulations was determined by time-kill assays and their stability was tested by repeated freeze-thaw cycles. The most effective formulation was tested in dairy cows. The minimum inhibitory concentrations and minimum bactericidal concentrations of the tested essential oils against S. aureus, S. agalactiae, and E. coli were in the range of 0.78–6.25 µL/mL. The time-kill tests indicated that the essential oils’ antibacterial activity depended on concentration and contact time. All three essential oil-based post-milking teat spray preparations showed good stability. The citronella spray formulation showed the highest antibacterial potency. In in vivo testing, the citronella spray eradicated aerobic bacteria on the teat skin of cows (99.9% or 3-log10 reduction) within 1 min, which was non-inferior to a standard 0.54% iodine solution teat dip. Clove, citronella, and sweet basil essential oils were effective against S. aureus, S. agalactiae, and E. coli in vitro. Of these, citronella essential oil is the most promising to be developed as a post-milking teat spray with high antibacterial activity and excellent bacterial eradication properties in vivo. Keywords: antibacterial activity, essential oils, post-milking disinfection, spray formulation.

Subclinical mastitis (SCM) caused by erythromycin-resistant Staphylococcus aureus is a significant disease in lactating animals. Therefore, it is crucial to understand the genetic factors contributing to erythromycin resistance in S. aureus. This study aimed to estimate the prevalence of S. aureus in milk from subclinical mastitic cattle and buffaloes and tank milk samples as identified by probe-based real-time polymerase chain reaction (PCR) and the genotypic assessment of macrolide and erythromycin resistance profiles, as well as to analyze the phylogenetic relatedness of our local isolates of S. aureus. In total, 285 milk samples were analyzed using the California mastitis test to detect SCM. Milk samples were cultured on different specific Staphylococcus media. The presence of S. aureus was confirmed by Gram staining, the catalase and coagulase tests, the detection of hemolytic activity, DNase agar testing, and biofilm activity in Congo red medium. The genotypic identification of S. aureus (nuc) was performed. The determinants of erythromycin (ermA, ermB, ermC, and ermT) and macrolide resistance (msrA) were screened in all isolates. DNA sequencing of our local isolates of S. aureus was used to analyze their phylogenetic relatedness. Moreover, histopathological examination of tissue specimens of mammary gland was performed. The S. aureus positivity rates were 36.4%, 48.8%, and 63.6% in cattle, buffalo, and bulk tank milk, respectively. Probe-based real-time PCR molecularly confirmed all 62 S. aureus isolates. Thirty-one isolates were subjected to PCR to create profiles of their genotypic erythromycin resistance. ermA, ermB, ermC, and ermT were present in 5 (8%), 26 (41.9%), 18 (29%), and 15 (24.1%) S. aureus isolates, respectively. Moreover, msrA was found in three (4.8%) strains. Eight PCR products were produced using standard PCR for DNA sequencing. Multiple sequence alignment, phylogenetic tree construction, and analysis of nuc in S. aureus revealed a high degree of homology (100%) with S. aureus strains isolated from milk in cases of bovine mastitis in India and Kenya. Histological analysis of udder tissues revealed extensive aggregation of mononuclear inflammatory cells in the interstitial connective tissue, primarily lymphocytes, and macrophages. This study showed a high prevalence of erythromycin resistance in S. aureus isolates. This information is vital for controlling mastitis and the spread of resistance genes between bacterial strains and hosts. Moreover, the probe-based real-time PCR approach is helpful for the rapid screening of S. aureus isolates and the consequent efficient treatment and control of S. aureus mastitis. Keywords: antimicrobial resistance, bovines, Egypt, erythromycin resistance, genetic diversity, Staphylococcus aureus, subclinical mastitis.

Wheat bran (WB) is used extensively in animal feed. Despite its nutritional value, its use is limited because of its high-fiber content. We evaluated the effect of soybean oil on laying hen performance with maize meal partly replaced by WB. Thirty-six ISA Brown laying hens, 40 weeks old, were used in a completely randomized design in which laying hens were distributed in individual cages, with three replications of four birds and assigned to three treatments: T1 (basal diet), T2 (60% basal diet + 20% maize meal + 20% WB), and T3 (60% basal diet + 20% maize meal + 17.5% WB + 2.5% soybean oil). Compared with the control group (T1), replacing 20% of yellow maize with WB (T2) did not affect average live weight, egg laying rate, soft-shelled egg production, egg mass, feed conversion per dozen eggs, or laying hen viability (p > 0.05). When 20% of the maize meal was replaced with WB, feed intake and feed conversion per egg mass increased (p < 0.05). Furthermore, adding 2.5% soybean oil to feed containing WB improved laying hen performance by significantly reducing feed conversion per egg mass (p < 0.05). Adding 2.5% soybean oil to diets containing WB instead of 17.5% yellow maize improved the feed conversion per egg mass performance of laying hens. Keywords: hens, laying, soybean oil, wheat bran.