In Thailand, domestic cats are the most common companion animal, and many are admitted to veterinary clinics for neutering surgery; however, such environment can induce stress. This is the first study to evaluate stress in hospitalized cats after neutering surgery using cat stress score (CSS) and salivary cortisol levels, including the impact of providing a hiding box (B) and/or administering a pheromone product to reduce stress. The study design was based on a randomized controlled clinical trial. A total of 80 domestic cats undergoing routine neutering surgery were assessed for their behavioral demeanor scoring system (DSS) as friendly (DSS1) and aggressive (DSS2) based on a DSS. During admission, the cats were randomly allocated to single standard cages with one of the following treatments: (B), feline facial pheromone (P), a combination of hiding box and the pheromone (BP), or no additional enrichment (C). Cat stress score, food intake, and hide-seeking behavior were recorded. The cortisol enzyme-linked immunosorbent assay kit was used to assess the salivary cortisol level. On the 1st day of admission, aggressive cats had a significantly higher CSS (4.16 ± 0.29) than friendly cats (3.27 ± 0.16). Both demeanor cat groups showed statistically significant reductions in stress levels earlier than the control group after providing the enrichments. Saliva cortisol measurements ranged from 0.24 to 0.66 ng/mL. No statistical differences in cortisol levels were observed between the 1st day and other days of admission. In contrast, no differences in food intake and hide-seeking behavior were seen within each group during the same period. Results suggested that stress and stress responses in cats depended on behavioral demeanor. The provision of enrichment, including hiding box and feline facial pheromone in singly housed caging reduced stress, especially in aggressive cats. However, salivary cortisol analysis, food intake, and hide-seeking behavior were ineffective for assessing stress in cats after neutering surgery. Keywords: cortisol, domestic cats, food intake, pheromone.

Slaughterhouses act as a significant public health hotspot in developing countries like Bangladesh. The study aimed to investigate small ruminants at slaughterhouses for pathological study and molecular detection of important zoonotic diseases. A total of 75 goats and 14 sheep were investigated from June 2019 to January 2020 at different slaughterhouses in Mymensingh division, Bangladesh. The targeted diseases were tuberculosis (TB), listeriosis, Q fever, brucellosis, anthrax, toxoplasmosis, hydatidosis, and linguatulosis. The tentative diagnosis was made based on gross and histopathological lesions. Polymerase chain reaction (PCR) was performed to confirm the causal agents of zoonotic diseases using disease-specific primers. Grossly, caseous nodule formation in the visceral organs; enlarged and calcifications of mesenteric lymph nodes (MLNs); hydatid cyst formation in the liver were the predominant lesions observed. Histopathologically, granuloma, caseous necrosis, and calcifications admixed with acid-fast bacteria in the MLNs, liver, spleen, and kidney were seen as suggestive of infectivity due to TB. Septic lymphadenitis mixed with rod-shaped bacteria, doughnut granuloma, fibroplasia accompanied by eosinophils and lymphocytic infiltration in MLNs, and portal granuloma were observed in listeriosis, Q fever, linguatulosis, and toxoplasmosis suspected cases, respectively. The PCR amplified Mycobacterium tuberculosis complex (372 bp), Mycobacterium bovis (600 bp), Listeria monocytogenes (517 bp), Toxoplasma gondii (512 bp), and Coxiella burnetii (687 bp) species-specific amplicons. In addition, linguatulosis and hydatidosis were identified in six and three goats, respectively. Brucellosis and anthrax were not detected in any cases. The slaughterhouse samples were also found to harbor the coexistence of different zoonotic pathogens. Deadly infectious zoonotic diseases in goats and sheep at slaughterhouses may cause widespread public health risks. As a result, more intensive monitoring and epidemiological surveys are required to successfully prevent and control zoonotic diseases. Keywords: pathology, polymerase chain reaction, small ruminants, zoonotic diseases.

The utilization of cassava leaves and peels, ceara rubber leaves, sweet potato leaves, Chinese Albizia leaves, and lophatheri leaves from Bojonegoro Regency has led to the poisoning of livestock due to antinutritional factors. Nevertheless, the plants are known to have bioactive components and potential antioxidant and antibacterial activity if appropriately processed. This study aimed to determine the antinutritional compounds as well as the antioxidant and antibacterial potential of these plants responsible for livestock poisoning in the Bojonegoro Regency. Extraction was performed by the maceration method using 70% (v/v) ethanol solvent. The samples were analyzed qualitatively to determine the presence of tannins, alkaloids, oxalates, cardiac glycosides, and cyanogenic glycosides. The antioxidant activity was determined using the 1,1-diphenyl-2-picrylhydrazyl method, while the antimicrobial activity was assessed by different testing concentrations (125, 250, and 500 mg/mL) against Staphylococcus epidermidis, Staphylococcus aureus, and Escherichia coli. The ethanolic extract of the plants was found to contain antinutritional tannins, alkaloids, cardiac glycosides, and cyanogenic glycosides suspected of causing livestock poisoning. Despite the presence of these antinutrients, all extracts also had antioxidant and antibacterial potential. Cassava peels and sweet potato leaves had the highest antioxidant activity, whereas Chinese Albizia leaves had the most potent antibacterial activity. Cassava leaves and peels, ceara rubber leaves, sweet potato leaves, Chinese Albizia leaves, and lophatheri leaves obtained from Bojonegoro Regency and used as agricultural waste contain antinutritional factors but also possess potentially effective antioxidant and antimicrobial components. Keywords: antimicrobial, antinutritional, antioxidant, Bojonegoro regency, livestock, plant poisoning.

Leptin (LEP) is an autocrine and paracrine factor produced by the fat pad and acinar epithelial cells of the breast. This study aimed to investigate the effects of LEP on yak mammary epithelial cells (YMECs) and the expression of STAT3. In addition, we evaluated the possible effects of prolactin (PRL) on the function of LEP. The YMECs were treated with 0, 50, 100, 200, 400, and 800 ng/mL LEP for 48 h in the absence of PRL and the presence of 500 ng/mL PRL. The growth activity of YMECs was measured using the cell counting kit-8 assay. The changes in the lactation signaling pathway-related factor STAT3 were detected at the mRNA, protein, and protein phosphorylation levels using the reverse transcriptase-quantitative polymerase chain reaction and Western blotting. To explore whether LEP affects the activation of STAT3 through JAK2/JAK3 in YMECs, the JAK2/3 signaling pathway inhibitor AG490 was used at a fixed concentration of LEP. Each concentration of LEP significantly promoted the expression of STAT3 mRNA (p < 0.05) in YMECs in the presence of PRL. In the absence of PRL, all concentrations of LEP were found to inhibit the expression of the STAT3 protein (p < 0.05). The expression of the STAT3 protein in YMECs was found to first increase followed by a decrease with an increase in the concentration of LEP. In addition, the phosphorylation level of STAT3 increased in all groups, except the 100 ng/mL concentration group. The STAT3 phosphorylation trend and protein expression were different, such that the level of protein phosphorylation was higher than that of the STAT3 protein (p < 0.05). The addition of AG490 reduced the expression of the STAT3 mRNA, STAT3 protein, and STAT3 phosphorylation in the LEP and LEP + PRL groups. Altogether, the results indicated that different concentrations of LEP exerted varying effects on the growth of YMECs and the expression of STAT3, and the activity of STAT3 was primarily activated by JAK2. The addition of LEP can effectively inhibit the downregulation of the JAK2/STAT3 signal pathway by AG490, mitigate its inhibitory effect on the proliferation of YMECs, and reduce apoptosis. We believe that these findings will provide a theoretical and experimental basis for future research in this field. Keywords: JAK2, leptin, mammary epithelial cells, STAT3, yak.

Bovine brucellosis is a disease of global socio-economic importance caused by Brucella abortus. Diagnosis is mainly based on bacterial culture and serology. However, these methods often lack sensitivity and specificity. A range of molecular diagnostic methods has been developed to address these challenges. Therefore, this study aims to investigate the diagnostic accuracy of molecular tools, in comparison to gold standard bacterial isolation and serological assays for the diagnosis of bovine brucellosis. The systematic review and meta-analysis were conducted based on analyses of peer-reviewed journal articles published between January 1, 1990, and June 6, 2020, in the PubMed, Science Direct, Scopus, and Springer Link databases. Data were extracted from studies reporting the use of molecular diagnostic methods for the detection of B. abortus infections in animals according to Preferred Reporting Items for Systematic Reviews and Meta-analyses (PRISMA) guidelines. The quality of included journal articles was assessed using the quality assessment of diagnostic-accuracy studies assessment tool and meta-analysis was carried out using Review Manager. From a total of 177 studies, only 26 articles met the inclusion criteria based on PRISMA guidelines. Data from 35 complete studies were included in the meta-analysis and used to construct 2 × 2 contingency tables. Improved diagnostic performance was observed when tissue (sensitivity 92.7% [95% confidence interval (CI) 82.0–98.0%]) and serum samples (sensitivity 91.3% [95% CI 86.0–95.0%]) were used, while the BruAb2_0168 locus was the gene of preference for optimal assay performance (sensitivity 92.3% [95% CI 87.0–96.0%] and specificity 99.3% [95% CI 98.0–100.0%]). Loop-mediated isothermal amplification (LAMP) had a higher diagnostic accuracy than polymerase chain reaction (PCR) and real-time quantitative PCR with sensitivity of 92.0% (95% CI 78.0–98.0%) and specificity of 100.0% (95% CI 97.0–100.0%). The findings of this study assign superior diagnostic performance in the detection of B. abortus to LAMP. However, due to limitations associated with decreased specificity and a limited number of published articles on LAMP, the alternative use of PCR-based assays including those reported in literature is recommended while the use of LAMP for the detection of bovine brucellosis gains traction and should be evaluated more comprehensively in future. Keywords: bovine brucellosis, Brucella abortus, meta-analysis, molecular diagnosis, systematic review.

Coccidian infection (coccidiosis) is one of the most important causes of illness and death in the fish population, including Asian sea bass. The fingerling developmental stage is sensitive to various infectious agents. Economic losses are sustained by the sea bass aquaculture industry due to coccidiosis annually. However, the related pathological changes in the Asian sea bass fingerlings' three-part intestine remain unknown. This study aimed to investigate the Asian sea bass fingerlings' infection rate, infection location and site, and specific pathological lesions in the small intestinal tissues in a marine cage farming operation. A cross-sectional study was conducted on 44 fingerling fishes. Major coccidia proportions were identified morphologically at both the macroscopic and microscopic levels. The infection number was determined based on coccidia presence at various intestinal locations and sites. All areas were assessed for pathological lesions using semi-quantitative grading. Analysis of variance was used to perform all data analyses using the SPSS software. Data were expressed as means ± standard deviation. p < 0.05 was considered statistically significant. All Asian sea bass fingerlings studied were infected with coccidia. Enteritis and mucosal necrosis were distinct lesions found in the anterior intestine, which had the highest infection rate (49.94%), followed by the mid intestine (35.63%), and the posterior intestine (22.43%). The most common coccidian infection site was extracellular (subepithelial), followed by intracytoplasmic, and epicellular sites. Histopathological lesion determination revealed that intestinal tissue inflammation and epithelial injuries were predominantly seen in the anterior gut (p < 0.05). There was a high coccidian infection rate in Asian sea bass fingerlings from marine cage farming operations. Infection and intestinal damage at the anterior intestine, a major site, led to fingerling death. Disease prevention in the nursery should be intensive from the fingerling period to decrease the fatality rate caused by coccidia. Keywords: Asian seabass, coccidian, fingerling, histopathology, Lates calcarifer, small intestine.

Health problems can be caused by consuming foods that have been processed in unsanitary conditions; hence, the study of the impact of contamination on food and its prevention has become critical. The disease caused by Klebsiella pneumoniae in food is increasing significantly every year across the world. The main factors that are essential for the virulence of K. pneumoniae are lipopolysaccharide and polysaccharide capsules. Furthermore, K. pneumoniae is capable of forming biofilms. Capsule polysaccharides, fimbriae types 1 and 3, are crucial virulence factors contributing to biofilm formation in K. pneumoniae. The food contamination by K. pneumoniae may not directly pose a public health risk; however, the presence of K. pneumoniae refers to unhygienic practices in food handling. This article aims to demonstrate that K. pneumoniae should be considered as a potential pathogen that spreads through the food chain and that necessary precautions should be taken in the future.

Toxoplasmosis is a disease caused by the protozoan parasite Toxoplasma gondii that affects both humans and animals, leading to abortions and significant clinical manifestations in pregnant and immunocompromised hosts, in addition to massive economic losses in animal industries. Data from Lebanon are scarce regarding the seroprevalence of T. gondii infection in livestock. This study aimed to estimate the seroprevalence and assess the associated risk factors of T. gondii infection in sheep and goats in Lebanon. A cross-sectional study was carried out from May 2020 to April 2021. Blood samples from 150 sheep and 145 goats (total 295) destined for human consumption were obtained from 20 Lebanese farms located in the North and Beqaa governorates. The anti-T. gondii immunoglobulin G antibodies were assayed through means of a modified agglutination test with a cutoff titer of 20. An overall seroprevalence of 48.5% (143/295) was reported: About 56.6% seroprevalence was found in sheep (85/150) and 40% (58/145) in goats. Adult age, female gender, and the wet season were significantly associated with an increased seropositivity rate of T. gondii infection (p < 0.001, p = 0.001, and p = 0.043, respectively). These results confirm the spread of T. gondii in sheep and goats destined for human consumption in various geographical regions in Lebanon. Therefore, continuous monitoring of T. gondii infection in livestock is warranted to control the spread of the infection and limit its potential transmission to humans through the consumption of raw or undercooked meat. Keywords: goats, Lebanon, risk factors, seroprevalence, sheep, Toxoplasma gondii.

Coagulase-negative staphylococci (CNS) are becoming the major cause of clinical and subclinical bovine mastitis around the world. This study aims to estimate the prevalence, antibiogram, and frequency of the methicillin-resistant (MR) (mecA) gene in CNS collected from cows with subclinical mastitis. Thirty-four milk samples were collected from 20 cows. Fifteen subclinical mastitis samples (∼44.12%) were identified as CNS isolates. The Vitek2 compact system method was employed for the identification of the species. Furthermore, antibiotic sensitivity tests were performed against 10 different antibiotics for CNS strains. The mecA gene from isolated CNS was detected by conventional polymerase chain reaction (PCR). Staphylococcus haemolyticus was the most predominant isolated species with an incidence of 33.3% (5/15 isolates), followed by 26.7% for Staphylococcus sciuri and Staphylococcus vitamins (4/15 isolates), and 13.3% for Staphylococcus vitulinus (2/15 isolates), respectively. The highest resistance rates were determined to be 40% (6/15 isolates) against penicillin and oxacillin (OX), 33.3% (5/15 isolates) against clindamycin, 13% (2/15 isolates) against chloramphenicol, amoxicillin, and erythromycin, and 5% (1/15 isolates) against ciprofloxacin, respectively. The results revealed that the isolates were resistant to one or more antimicrobial agents, with five isolates displaying multiple antimicrobial resistance. Furthermore, the results exhibit that all CNS isolates had the mecA gene at 310 bp with a 100% frequency. Moreover, for detecting MR isolates, there are significant discrepancies between phenotypic and genotypic approaches, and only 6/15 CNS isolates phenotypically demonstrated OX resistance. The results emphasize the necessity of frequent monitoring of phenotypic and genotypic profiles of CNS isolates to ensure effective control measures and the prevention of multidrug resistance strain evolution. Keywords: coagulase-negative staphylococci, cows, mecA gene, polymerase chain reaction, subclinical mastitis.

Global warming has grave consequences on livestock production systems and profound negative effects on animal production. This study aimed to carry out an in vitro thermal stress stimulation (TSS) of bovine peripheral blood mononuclear cells (PBMCs) using different thermal assault conditions (TACs), including normal to extreme temperatures and varying durations of thermal exposure (DTE) to understand how PBMCs of Indian Zebu–Jersey crossbreds respond to various levels and durations of heat shock. Ten milliliters of blood were collected from 70 Indian Zebu–Jersey crossbreds under aseptic conditions and were sampled for isolating PBMCs. Peripheral blood mononuclear cells were divided into seven groups, each comprising 10 PBMC samples isolated from 10 different animals. Aliquots of 500 μL of PBMCs were stressed by exposure to different TACs (37, 40, and 45°C) for DTEs of 3 or 6 h. Subsequently, the cells were harvested. The control unstressed samples (500 μL aliquots of PBMCs) were exposed to no TAC (0°C) and zero DTE (0 h). Total RNA from all the treatment groups of PBMCs were isolated and quantitated. We found a very strong association between TACs and RNA levels. In addition, PBMCs viability was negatively affected by heat shock. This led to an exponential reduction in PBMC count as TACs toughened. Only 3.59 × 105 ± 0.34 cells/mL were viable after exposure to 45°C for a 6 h DTE. This cell viability was lower than that measured in controls subjected to no stress and zero time DTE (2.56 × 107 ± 0.22 cells/mL). We also observed a reduction in the concentration of RNA isolated from thermally stressed PBMCs. In vitro TSS of PBMCs provided biological information on the response of cellular systems to heat shock after exposure to TACs. This will help to mitigate and manage the effects of thermal stress in bovine species. The association between the reduction in PBMC count after in vitro TSS and the expression of heat shock protein 70 gene will be investigated in the future to further understand how Indian Zebu–Jersey crossbreds respond to in vitro thermal conditions. This will be used to determine the in vivo response of Indian Jersey crossbreds to different environmental thermal conditions and will further enable the in vivo understanding of thermotolerance potentials of bovine species for better adaptation, survival, and production performance. Keywords: bovine, cell survival, climate change, peripheral blood mononuclear cells, RNA synthesis, thermal stress.

Foot-and-mouth disease (FMD) is an extremely contagious viral disease that affects domestic and wild cloven-hoofed animals. In Egypt, FMD has been enzootic since the 1950s and caused great economic losses in cattle and buffalos over the past few years. This study aimed to detect FMD virus (FMDV) in serum and raw milk samples collected from infected and adjacent cattle and buffalos from different localities in El Menofia Governorate, Egypt. Blood and milk samples were collected from apparently diseased and adjacent 100 cows and 100 buffalos. Serum samples were prepared and used for the detection of FMDV using a non-structural protein enzyme-linked immunosorbent assay, while real-time reverse transcription-polymerase chain reaction (rRT-PCR) was used for the detection of FMDV in milk samples. Reproductive hormones were estimated using radioimmunoassay kits. Milk constituents were determined by Lactoscan. Of the 200 examined serum samples (100 cows and 100 buffalos), 56% and 44% were seropositive for FMDV non-structural protein antibodies in cattle and buffalo, respectively. Real-time reverse transcription-polymerase chain reaction results confirmed that all examined milk samples collected from seropositive animals were positive for FMDV. Estrogen and progesterone levels in the serum of seropositive and seronegative animals were measured, and FMDV was proven to significantly elevate estrogen and reduce progesterone levels in both non-pregnant and pregnant animals during different stages of pregnancy. The effect of the virus on milk composition and somatic cell count (SCC) was also studied, revealing that FMDV infection significantly decreased the level of milk fat, protein, and lactose but did not significantly affected minerals, pH, and conductivity. Moreover, it significantly increased the SCC. Data recorded in this study indicates a widespread occurrence of FMDV in cattle and buffalo all over Menofia Governorate, Egypt. Infected raw milk is of poor quality and, if put for commercial sale, may have health risks for consumers and play a significant role in spreading the virus. Moreover, FMDV may disturb some reproductive hormones, which could adversely affect cattle and buffalo productivity. Therefore, preventive programs and accurate diagnosis are essential for successful disease control. Keywords: foot-and-mouth disease, physiochemical properties of milk, raw milk, reproductive hormones.

Mosaicism – the presence of both wild-type and mutant alleles – is a serious problem for zygotic gene modification through gene editing using the Clustered regularly interspaced short palindromic repeats-Cas9 (CRISPR/ Cas9) system. Different delivery methods, such as microinjection (MI), electroporation (EP), and transfection (TF), can be used to transfer CRISPR/Cas9 components into porcine zygotes. This study aimed to develop a method that combines MI, EP, and TF to improve mutation efficiency mediated through the CRISPR/Cas9 system for a triple-gene knockout in pigs. The study consisted of three groups: The MI group with three simultaneously microinjected guide RNAs (gRNAs) targeting α-1,3-galactosyltransferase (GGTA1), cytidine 32 monophosphate-N-acetylneuraminic acid hydroxylase (CMAH), and β-1,4-N-acetyl-galactosaminyltransferase 2 (B4GALNT2); the MI + EP group with two gRNAs targeting GGTA1 and B4GALNT2 genes delivered into zygotes through MI, followed by EP of gRNA targeting the CMAH 1 h later; and the MI + EP + TF group with MI of gRNA targeting GGTA1 gene into zygotes, followed by EP of gRNA targeting CMAH 1 h later, and then TF of gRNA targeting the B4GALNT2 gene into zona-free zygotes after another hour. The rate of blastocysts carrying mutations in one or two gene(s) was significantly higher in the MI + EP + TF group than in the MI group. However, the blastocyst formation rate of zygotes in the MI + EP + TF group was lower than that of the zygotes in the other treatment groups. The combination of CRISPR/Cas9 delivery methods may improve the mutation efficiency of triple-gene edited porcine blastocysts. Keywords: clustered regularly interspaced short palindromic repeats/Cas9, electroporation, microinjection, porcine zygotes, transfection.

Low concentrations of heavy metals are toxic and pose a serious threat to human health and the environment. Therefore, regular assessments of the toxic metal content in poultry feed are crucial for evaluating feed quality and customer safety. It is difficult to determine the heavy metals in the poultry feed at the trace amount. Therefore, this study aimed to validate this method through the detection of three heavy metals, chromium (Cr), cadmium (Cd), and lead (Pb), in poultry feed samples. Graphite furnace atomic absorption spectrometry (GF-AAS) method was used to analyze the heavy metals in poultry feed according to the guidelines given by the Council Directive 333/2007/EC, Commission Decision 657/2002/EC. In this study, various parameters such as linearity check, limit of detection (LOD), limit of quantification (LOQ), recovery percentage, precision checks, repeatability, reproducibility, and uncertainty measurement were considered to validate and assess the method following international guidelines. Heavy metals, such as Pb, Cr, and Cd, were analyzed from the feed samples in the laboratory using the GF-AAS method (Model: AA-7000 Shimadzu, Japan) with high purity argon as the inert gas, and the absorbance was read at wavelengths of 283.0, 357.9, and 228.8 nm, respectively. The coefficient of variation (CV%) for system suitability and precision data was <10% for all the metals (Pb, Cr, and Cd) detected in this study. The overall CV% of repeatability and reproducibility ranged from 8.70% to 8.76% and 8.65% to 9.96%, respectively. The linearity of the calibration curves was excellent (r2 > 0.999) at various concentration levels for the three different metals. The recovery (%) was found to be 94.53, 93.97, and 101.63% for Pb, Cr, and Cd, respectively. The LOD values in feed were 0.065, 0.01, and 0.11 mg/kg, and the LOQ values were 0.22, 0.03, and 0.38 mg/kg for Cr, Cd, and Pb, respectively. The values recorded for the measurement uncertainty (%) were 11.48, 4.43, and 12.42% for Cr, Cd, and Pb, respectively. The results show that these study criteria or parameters have met the validated or acceptable range. Therefore, it is a reliable technique that can be used undoubtedly for the routine analysis of heavy metals in poultry feed samples across the globe. Keywords: graphite furnace atomic absorption spectrometry, method validation, poultry feed, toxic metals (Pb, Cr, and Cd).

Brucellosis is a prevalent infectious zoonotic disease that affects humans, livestock, and wildlife in many parts of the world. A cross-sectional study was conducted to estimate the seroprevalence and risk factors of brucellosis among farmers and patients attending six health centers in Sidi Kacem province (northwestern Morocco). Blood samples (3-5 mL) were collected. Among 1283 participants, 351 were males and 932 were females and tested for Brucella antibodies using rose Bengal plate test and immunoglobulin (Ig)M/IgG enzyme-linked immunosorbent assay (ELISA) for confirmation. The seroprevalence of brucellosis was 33.20% (426/1283) with a higher risk among males and rural residents. The univariable analysis revealed that contacting cattle, handling abortion products and manure, and consuming undercooked beef and goat meat were all risk factors for brucellosis. Furthermore, raw milk and milk derivatives were risk factors strongly linked to brucellosis. Our findings indicate a high prevalence of brucellosis associated with the consumption of raw meat, raw dairy products, milk, and close contact with infected animals. However, there are some limitations to this study, such as we did not use the ELISA test on all sera collected and individuals under the age of 18 were not included in the study. Moreover, building a database on the occurrence of brucellosis and associated epidemiological factors is critical for providing informed advice to policymakers to improve control strategies against this disease in Morocco. Keywords: human brucellosis, risk factors, seroprevalence, Sidi Kacem (Morocco).

Disposable imported nylon bags used in an in situ digestibility measurement restrict the effort of scientists to obtain more accurate information about ruminant feed quality due to their low affordability and environmentally unfriendly characteristics. This study aimed to find reusable local nylon fabrics to substitute imports. Five local fabrics (B1 = Abutai, B2 = Taffeta, B3 = Organza, B4 = N57, and B5 = M100) were used to make nylon bags and compared with the imported bag (B6 = Ankom technology). The research consisted of three steps: (1) Observing the similarity of the local nylon bag's hole number to the imported bag. (2) Testing feed in situ degradation (F1 = Napier grass, F2 = Cornmeal, and F3 = Dairy cattle total mixed ration) using bags B1–B6. (3) The reusability of the bag was tested using different washing methods (under running water [R1], rinse [R2], and ultrasonic water bath [R3]). It was shown that the hole numbers of B1 (1223 hole numbers) and B2 (1245 hole numbers) were not significantly different from B6 (1248 hole numbers). It was shown with dry matter degradability measurement using the in situ method that there was no significant difference in feed solubility (a), potential degradability (b), and the effective degradation between local fabrics (B1–B5) and B6. According to the degradation rate (c), there were interactions between the feeds and fabrics. For F1, all local fabrics were similar to B6, while for F2, only B1 was different from B6. For F3, only B5 was different from B6. It was also shown by the organic matter degradability measurements that there was a similar trend. The exception was the solubility (a) component in F3, in which it was shown that B1 was also different from B6. It was shown in the reusable test that there was no difference in the weight of the bag before and after all washing methods. In contrast, the hole number increased due to the shrinking of the bag after drying in a 60°C oven. According to this in situ study, local nylon bag B2 can substitute imported bags. A lower drying temperature is suggested to prevent shrinking and make the bag reusable. Keywords: digestibility, feed quality, in situ, local fabrics, nylon bag.

Salinomycin sodium, a licensed coccidiostat in rabbits, is used for fattening at a dose of 20–25 mg/kg. Salinomycin toxicity may arise from many risk factors (e.g., overdosage or use in non-target animal species). Silymarin extracted from milk thistle has antioxidant, anti-inflammatory, and antiviral properties. This study aimed to investigate the adverse impacts of oral administration of salinomycin for 28 consecutive days and how to reduce its risks and side effects by administering silymarin. Eighty-four male New Zealand White bucks (1.750–2.000 kg) were randomly divided into seven groups (12 each). Group one was the control. Groups two and three were administered salinomycin orally (doses of 20 and 40 mg/kg ration). Group four was administered salinomycin (20 mg/kg ration) and silymarin (6.5 mg/kg body weight [BW]). Group five received salinomycin (40 mg/kg ration) and silymarin (13 mg/kg BW). Groups six and seven were administered silymarin at doses of 6.5 and 13 mg/kg BW. Rabbits were euthanized and slaughtered on day 29 using the Halal method. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine, urea, total proteins, albumin, total cholesterol, and high- and low-density lipoprotein (HDL and LDL) were analyzed in serum. Glutathione (GSH), superoxide dismutase (SOD), catalase, and malondialdehyde (MDA) were estimated in the liver. A histopathological investigation was performed on the liver and kidney. The MDA activity, AST, ALT, total protein, albumin, total cholesterol, triglyceride, LDL, urea, and creatinine values were significantly elevated in groups two and three. The GSH, catalase, SOD, and HDL were significantly lower in these groups than in the control group. There were moderate pathologic changes in the liver and kidney of the third group . However, the results of the fourth and fifth groups improved more than those of the second and third groups. The results of the sixth and seventh groups were nearly the same as those of the control group. Salinomycin toxicity was caused by oxidative damage because of reactive oxygen species formation. Silymarin (6.5 or 13 mg/kg BW) tends to prevent and treat accidental toxicity. However, the high dose of silymarin (13 mg/kg BW) had more renal and hepatoprotective capacities. Keywords: anticoccidial drugs, feed additives, liver enzymes, oxidative biomarkers, rabbits, salinomycin, silymarin.

Trichinellosis is a neglected and emerging foodborne zoonosis in Africa. Trichinella infection occurs through the consumption of raw or undercooked infected meat and meat products. This study aimed to assess pigs' management practices and determine the exposure of pigs and warthogs to Trichinella spp. in the northern area of Senegal. Surveys and observations were carried out among 40 pig farmers to assess husbandry practices regarding Trichinella spp. life cycle. In addition, 201 pig meat juices and 83 warthog meat juices were extracted and tested for anti-Trichinella antibodies by indirect enzyme-linked immunosorbent assay. Most (97%) of farms practiced a traditional farming system with free-ranging of pigs in 85% of farms. Farms had local pig breed without housing and supplementary feeding. Some farmers (27.5%) used slaughter waste to feed pigs and farmers were not aware that free-range farming is a source of infection to Trichinella infection. They were also unaware that some pig diseases could be transmitted to humans. The seroprevalence of Trichinella infection was 10.9% (95% confidence interval [CI]: 6.6–15.2%) in pigs and 10.8% (95% CI: 4.16–17.52%) in warthogs with significantly higher seroprevalence in male (22.2%: 95% CI: 6.6–37.8%) compared to female (9.2%; 95% CI: 4.9–13.5%) (p < 0.05). The confirmation of exposure to Trichinella spp. in this area in pigs and warthogs shows a significant risk of transmission of this disease to humans if the farming conditions and the health surveillance system are not respected. However, control measures are needed to reduce any risk of transmission of Trichinella infection to humans. Keywords: epidemiological studies, pig management, Senegal, Trichinella infection, warthog.

It is known that during the early postpartum and lactation periods in dairy cows, metabolic disorders develop, that is, ketosis, which can lead to secondary damage to internal organs. Therefore, it is important to address the issues of changing the lactating cows' clinical, laboratory, and physiological parameters regarding the development of hepatocardial syndrome. This study aimed to provide clinical and diagnostic justification for developing hepatocardial syndrome in highly productive dairy cows. The study was conducted on 20 black and white cows in the early postpartum period (7–10 days after birth), with a milk production level of >4500 kg of milk during the previous lactation period, a positive result in the formol colloid sedimentary test, the presence of deafness and splitting of heart sounds, changes in the size, or increased pain sensitivity of the percussion field of the liver. Clinically healthy dairy cows in the early postpartum period were used as controls (n = 24). Clinical, electrocardiographic, echocardiographic, and biochemical parameters were also evaluated. Dairy cows with hepatocardial syndrome developed arterial hypertension and sinus tachycardia, which led to a significant decrease in PQ and QT intervals at ECG. A significant increase in the diastolic size of the interventricular septum, systolic size of the free wall of the left ventricle, and diastolic and systolic sizes of the left ventricle and a significant decrease in the shortening fraction of the left ventricular myocardium were observed in the cows due to the development of hepatocardial syndrome. The affected cows demonstrated a significant increase in serum activity of gamma-glutamyl transferase, alanine aminotransferase, lactate dehydrogenase, creatine phosphokinase, alkaline phosphatase, troponin, malondialdehyde, diene conjugates, and ceruloplasmin and a decrease in glucose concentration. In addition, they demonstrated decreased activity of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase. Hepatocardial syndrome in dairy cows occurs due to ketosis, characterized by arterial hypertension, sinus tachycardia, a moderate decrease in myocardial contractility, oxidative stress, and cytolysis of cardiomyocytes and hepatocytes. Therefore, the control and prevention of the development of hepatocardial syndrome will make it possible to maintain the productive health and longevity of dairy cows. Keywords: early lactation, heart dysfunction, hepatocardial syndrome, ketosis in cows, liver pathology, transitional period.

Bovine mastitis has long been considered the most important cause of economic losses in the dairy industry. Staphylococcus aureus is the most frequently isolated pathogen from bovine mastitis cases worldwide. Capsular polysaccharides (CPs) of serotype 5 (CP5) or serotype 8 (CP8) are the most prevalent capsule genotypes related to infections associated with S. aureus in humans. However, a variety of CPs has been reported in ruminants and other hosts. Information regarding the relationship between genotypic and phenotypic capsule variation and bovine mastitis in Jordan is scarce. Thus, we aimed to determine the prevalence of S. aureus capsule genotypes CP5 and CP8 in milk from bovine mastitis cases and the antimicrobial susceptibility profile of the recovered isolates in 27 dairy farms in Jordan. Staphylococcus aureus strains were isolated from bovine mastitis cases in two districts of Jordan. All S. aureus isolates were initially identified using conventional biochemical and microbiological methods. Subsequently, confirmation of the identity of S. aureus was performed by polymerase chain reaction (PCR) targeting nuc gene. Capsule polysaccharide typing was performed by PCR specific for CP5 and CP8. In addition, we assessed the antibiotic susceptibility profile of S. aureus isolates against commonly used antimicrobials by the disk diffusion method according to Clinical and Laboratory Standards Institute guidelines. We collected 148 clinical isolates of S. aureus from bovine mastitis cases in the Zarqa (67.6%, n = 100) and Irbid (32.4%, n = 48) districts. Most isolates possessed capsule genotypes (91.3%), predominantly CP8 (88.6%). Only 8.7% of the isolates were nontypeable by PCR. In addition, we found statistically significant differences between the geographical region and the status of methicillin-resistant capsule genotypes (p < 0.05). The rates of resistance to β-lactam, macrolide, and fluoroquinolone antibiotics were very low, but resistance to tetracyclines was considerably high (22.3%). Significantly, mastitis isolates from Irbid showed a higher rate of resistance to ciprofloxacin (8.3% vs. 0%), while isolates from Zarqa showed a significantly higher rate of resistance to gentamicin (12.0% vs. 6.2%). We established associations between capsule genotypes and antimicrobial resistance and the pathogenic behavior of S. aureus isolated from bovine mastitis cases. Further studies are necessary to fully elucidate the role and mechanisms of capsular expression in the epidemiological and molecular variability of S. aureus in bovine mastitis. Keywords: antibiotic susceptibility, bovine mastitis, capsule genotypes, polymerase chain reaction, Staphylococcus aureus.

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

Trichinellosis remains a dangerous disease for humans and animals, which can lead to a lethal outcome. The study of specific body reactions in response to invasion by different types of Trichinella can help in the early diagnosis of the disease. This study aimed to investigate the hematological, biochemical, and serological characteristics of rabbits experimentally infected with trichinellosis, as well as the possibility of using changes in these parameters at various disease stages for early hematological, biochemical, and serological diagnosis of trichinellosis. Three groups of rabbits were orally infected with Trichinella nativa and Trichinella spiralis derived from encysted T. spirtalis larvae in pork muscle samples. The first and second groups were infected with T. nativa and T. spiralis, respectively, while the third group served as control by receiving a physiological solution. An ADVIA 2120i automatic hematology analyzer with a blood smear staining module was used to determine the hematological parameters of rabbits. Antigens were used in an enzyme-linked immunosorbent assay (ELISA) to detect antibodies in the sera of infected rabbits that were supernatants containing excretory-secretory antigens (ES-Ag) and somatic antigen (S-Ag). The detection of biochemical responses to the invasion of T. nativa and T. spiralis isolates was detected and hematological parameters were featured in two cases. Trichinella nativa increased the number of erythrocytes, neutrophils, eosinophils, monocytes, basophils, and thrombocytes on day 7 in rabbits. Creatine kinase (CK) is regarded as the most important indicator for the early detection of parasite invasion. Blood biochemistry showed no active response to T. spiralis infection. However, counts of erythrocytes, neutrophils, lymphocytes, and CK rose significantly. In both color indicators, the number of thrombocytes decreased. Enzyme-linked immunosorbent assay with ES-Ag and S-Ag of these isolates demonstrated the ability to detect antibodies as early as 7 days after infection, with a significant increase in the marker up to 70 days. On the 7th day after infection, blood tests of infected animals revealed CK-N-acetyl-cysteine (18.2%) and neutrophils (43%) when infected with T. nativa and neutrophils (26.7%) and lymphocytes (20%) when infected with T. spiralis. These indicators may serve as specific parameters for the early detection of Trichinella spp. invasion. Keywords: diagnostics, experimental infection, rabbits, Trichinella nativa, Trichinella spiralis, trichinellosis.

Antimicrobial resistance (AMR) is a global problem that affects human and animal health, and eggs can act as a vehicle for pathogenic and non-pathogenic resistant bacteria in the food chain. Escherichia coli is an indicator of food contamination with fecal materials as well as the occurrence and levels of AMR. This study aimed to investigate the presence of AMR, integrons, and virulence genes in E. coli isolated from eggshell samples of three egg production systems, from supermarkets in Thailand. A total of 750 hen's egg samples were purchased from supermarkets in Phayao Province: Cage eggs (250), free-range eggs (250), and organic eggs (250). Each sample was soaked in buffered peptone water (BPW), and the BPW samples were incubated at 37°C for 18–24 h. All samples were tested for E. coli by the standard conventional culture method. Then, all identified E. coli were tested for antimicrobial susceptibility to 15 antimicrobial agents by the agar disk diffusion method. All E. coli strains were subsequently found to have virulence genes and Classes 1 and 2 integrons by polymerase chain reaction. Among the eggshell samples, 91 samples were identified as having E. coli (cage eggs, 24 strains; free-range eggs, 27 strains; and organic eggs, 40 strains). Then, among the E. coli strains, 47 (51.6%) were positive for at least one virulence gene. The proportion of AMR in the eggshell samples was 91.2% (83/91), and streptomycin (STR), ampicillin (AMP), and tetracycline (TET) had a high degree of resistance. Among the E. coli strains, 27 (29.7%) strains were positive for class 1 or 2 integrons, and integron-positive strains were commonly found in STR-, AMP-, and TET-resistant strains. Multidrug resistance (MDR) was detected in 57.1% (52/91) of the E. coli strains, with STR-AMP-TET (5.5%) as the most frequent pattern. The proportion of MDR in cage eggs was 75.0% (18/24), which was higher than in both free-range and organic eggs. On the other hand, 53.2% (25/47) of E. coli carrying virulence genes had MDR, distributed across the production systems as follows: Cage eggs, 76.9% (10/13); free-range eggs, 63.6% (7/11); and organic eggs, 34.8% (8/23). Escherichia coli was detected in eggshell samples from all three egg production systems. The high level of virulence genes, AMR, and integrons indicated the possibility of dissemination of AMR among pathogenic and commensal E. coli through eggshells. These findings could be a major concern to farmers, food handlers, and consumers, especially regarding raw egg consumption. Keywords: antimicrobial resistance, eggshells, Escherichia coli, integrons, virulence.

Probiotic species have been proven to be beneficial on broiler performance; however, most studies have focused on industrial chickens with fast growth, whereas little information concerning the use of these species on native chickens is available. This study aimed to investigate the effects of probiotics Lactobacillus plantarum (LP) and Bacillus subtilis (BS) on the mortality, growth rate, and carcass characteristics in native Noi chickens challenged with Salmonella Typhimurium. We divided 420 1-day-old Noi chicks into seven different treatment groups (n = 60): negative control (no S. Typhimurium, no probiotics or antibiotics); positive control (PC, S. Typhimurium infection, no probiotics or antibiotics); and S. Typhimurium infection and supplementation with LP, BS, LP + BS, enrofloxacin, and commercial probiotics, respectively. Treatment was for 96 days, and the chicks were orally challenged with S. Typhimurium at 22 days old. No deaths occurred during the 4 weeks post-infection in the negative control, LP, or LP+BS groups. The PC group had the highest mortality rate (20%). Re-isolation of S. Typhimurium from the liver, spleen, and heart showed reduced bacterial counts at 1 week post-infection in the LP, BS, and LP + BS groups. The lowest body weight gain was observed in the PC group (949 g/bird), and chicks in the LP group gained 1148 g/bird. An improved feed conversion ratio was noted in the groups receiving probiotic supplementation (3.42–3.50 kg feed/kg gain). There was little evidence that probiotics affected carcass percentage and related parameters, such as breast, thigh and drumstick, and wings. Lactobacillus plantarum or BS dietary supplementation to native Noi broilers resulted in a lower mortality rate and improved body weight gain but did not affect carcass characteristics. Keywords: Bacillus subtilis, growth, Lactobacillus plantarum, mortality, Noi chicken.

Globally, toxoplasmosis is an important zoonotic parasite infection of many warm-blooded animals (including humans). Toxoplasma gondii oocysts are widespread, and their contamination can be primarily attributed to the members of the Felidae family. This study aimed to estimate the prevalence and determine the dense granule antigen 6 (GRA6) genotype of T. gondii among domestic cats in the Phayao Province, Thailand. A total of 124 fecal samples were collected from owned cats in the Muang district, Phayao, Thailand, from January to December 2020. Fecal samples were tested for the presence of T. gondii DNA using targeted B1 gene polymerase chain reaction (PCR) amplification, and positive samples were subsequently analyzed for their T. gondii genotype through PCR-restriction fragment length polymorphism (RFLP) analysis and sequencing of the GRA6 gene. Among the 124 samples, 46 (37.1%) were tested positive for T. gondii. Only 10 positive DNA samples were successfully amplified for the GRA6 marker. Subsequent PCR-RFLP and sequence analyses indicated that all T. gondii isolates from cats in Phayao belonged to GRA6 genotype I. Data revealed that toxoplasmosis is remarkably distributed among (studied) domestic cats in Phayao, Thailand. Moreover, the virulent GRA6 allele was found to be circulated among domestic cats in this area. However, no significant correlation was observed between infection rates and different risk factors, which indicated that pet cats of any age, gender, or breed have similar risks of being infected with T. gondii. Our results further suggested that infective oocysts of T. gondii are widely distributed and that environmental contamination with these oocysts will introduce more risks of disease transmission to humans and other animals. Keywords: domestic cats, genotype, granule antigen 6, Toxoplasma gondii, toxoplasmosis.

Bovine mastitis has a negative impact on animals, and improper antibiotic use has caused an increase in bacterial resistance. Therefore, medicinal plants could serve as an alternative treatment for this condition. Polyphenols have potential as antibiotic agents. Oak bark has long been used as a medicine and has shown antibacterial effects. Moreover, research on heather plant demonstrated that it has antibacterial properties. This study aimed to assess the antibacterial effects of oak (Quercus robur) bark and heather (Calluna vulgaris L.) herb extracts against common bovine mastitis pathogens. Dried oak bark and heather herb were used to prepare extracts using 30%, 50%, and 70% ethanol and acetone as solvents. Their polyphenol content was determined using the Folin–Ciocalteu method. Bovine mastitis-inducing clinical isolates of Escherichia coli, Streptococcus agalactiae, Streptococcus uberis, Serratia liquefaciens, Staphylococcus aureus, and reference cultures of S. aureus and E. coli were used for antibacterial tests. All extracts were screened through a disk diffusion test to ascertain their antibacterial effects, and the minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined for the most effective extracts. Oak bark extracts had variable antibacterial effects against S. aureus and Streptococcus strains, but no statistically significant difference was observed in activity against E. coli. The disk diffusion test showed that the oak bark extracts obtained using acetone and ethanol at 30% yielded the best results. However, the 70% acetone oak extract alone affected all types of bacteria. Further antibacterial tests of 70% acetone and 30% ethanol oak extracts revealed that the lowest MIC and MBC scores were against S. aureus strains and E. coli reference cultures. Conversely, the heather herb extracts exhibited broader activity against all types of bacteria, although better results were observed against Gram-positive bacteria. There was also a negative correlation between solvent concentration and antibacterial effect (p < 0.05, r = –0.507). The highest inhibition zone scores and broadest spectrum were observed in samples prepared in 30% ethanol. There was no statistically significant correlation between the phenolic content of plants and their antibacterial effects. Oak bark and heather extracts could be used as potential antibacterial agents against bovine mastitis pathogens. Keywords: antibacterial, bovine mastitis, Escherichia coli, heather herb, in vitro, oak bark, Staphylococcus aureus, Streptococcus.

Developing curcumin into nanosized particles is one of the approaches to overcome the limited use of curcumin. This study aimed to prepare curcumin into nanosized particles to increase the curcumin level in the rat's liver and hepatoprotective effect in rats. Curcumin into nanosized particles formulated using ionic gelation method. Rats were divided into four groups (n = 6): Normal, negative, curcumin, and curcumin modified into nanosized particles were treated with 100 mg/ kg body weight orally for 14 days. Hepatic curcumin level was investigated using liquid chromatography with tandem mass spectrometry, antioxidant activity by malondialdehyde (MDA), and hepatoprotective effect by aspartate transaminase (AST), alanine transaminase (ALT), and histopathology. The curcumin level in the rat's liver in the curcumin group was 12.19 ng/mL, and that in those receiving modified into nanosized curcumin was 209.36 ng/mL. The MDA levels in the normal, negative, curcumin, and curcumin modified into nanosized particles groups were 1.88, 4.87, 3.38, and 1.04 nmol/L, respectively. The AST levels in these groups were 57.12, 130.00, 102.13, and 74.28 IU/L, and the ALT levels were 21.63, 61.97, 39.38, and 28.55 IU/L. The liver histopathology scoring showed that curcumin in nanosized particles was better than curcumin in degeneration of fat, lymphocyte infiltration, and necrosis. There was a 17 times increase in curcumin level in the liver of rats treated with curcumin modified into nanosized particles. Curcumin modified into nanosized particles showed more significant improvement as antioxidant and hepatoprotector than curcumin. Keywords: antioxidant, curcumin modified into nanosized particles, hepatoprotective.

Perennial allergic rhinitis (AR) is a chronic upper respiratory disease, with inflammation mediated by immunoglobulin E in the nasal mucosa caused by house dust mites. Recently, allergen immunotherapy showed promising allergic healing in patients with a definite history of sensitization. Based on this finding, a product was developed using Indonesian house dust mite (IHDM). This study aimed to optimize the allergenic rhinitis mouse model that was generated using IHDM to test the in vivo sensitivity and safety of this product. Seven groups of mice were used for effectiveness testing – normal, negative control with IHDM challenge, positive control with 0.1% histamine challenge, and AR group by both IHDM-induced sensitization at 12.5, 50, 250, or 500 μg and IHDM challenge. Mice were sensitized by intraperitoneal administration of IHDM once a week for 3 consecutive weeks. Thereafter, the challenge was given intranasally 5 times on alternate days. The number of nose rubbing and sneezing was noted. Eosinophil infiltration was assessed histologically using hematoxylin and eosin staining. The expression of interleukin-5 (IL-5) mRNA in the nasal mucosa was determined using semi-quantitative reverse transcription-polymerase chain reaction. The induction of AR with IHDM significantly increased the number of nose rubbing and sneezing in the mouse model. Eosinophil infiltration was observed in the nasal mucosa; however, no significant change occurred in the expression of IL-5 mRNA. Overall, these data indicate that IHDM allergenic extract could be an effective sensitizing agent in a mouse model of AR. Although the use of IHDM is a limitation of this study because other sources of house dust mites might have different effects, this study provides a proper model for immunotherapy effectivity testing for in vivo pre-clinical studies. Keywords: allergen immunotherapy, allergic healing, allergic rhinitis, Indonesian house dust mites, neglected disease.

Coccidiosis is an enteric infection caused by a protozoon (Eimeria tenella). Coccidiosis is known to have a negative impact on the economy. Coccidiosis is controlled using anticoccidial drugs, antibiotics, and vaccines. Various coccidial vaccines differ in application technique, attenuation method, and the species used. Coccidial vaccines can be spray or gel-based (Form). This study aimed to compare the effect of application and approaches between spray and gel vaccines for coccidiosis. Specific pathogen-free chicks were vaccinated with different vaccines. Fecal samples were taken on 21 days post-vaccination for vaccine take, and then a challenge test was done on day 21. Post-vaccination oocyst counts in gel vaccinated groups were more than the spray vaccinated ones as it recorded (1400 and 2200) oocyst/g, but the gel vaccines resulted in lower post vaccinal titer which was (10000 and 12500) oocyst/g. Results of quantitative real-time polymerase chain reaction test post-vaccination were (23.72, 20.29) cycle threshold (CT) for spray vaccines and (18.75, 17.62) CT for gel vaccinated group. By challenging all the experimental groups, the microscopic and macroscopic lesion of gel vaccines resulted in score 1, while spray vaccines groups recorded score 2 and the control non-vaccinated challenged chickens showed score 4. The non-vaccinated/non-challenged group recorded a score of zero. These results can help poultry producers to decide which delivery system will provide the best results for their production system. The gel vaccines showed a better protection rate and lower shedding, which means more protection of birds and public health. Keywords: coccidia, Eimeria tenella, anticoccidial drugs and vaccinations, spray.

Ruminant slaughterhouse is one of the food-producing units to meet the protein demand of the people in Central Java. This study aimed to evaluate the implementation of sanitation and hygiene in ruminant slaughterhouses in Central Java based on their veterinary control number (NKV) certification and the microbiological quality of the meat produced. This study was conducted from September 2021 to December 2021. Thirty-three priority slaughterhouses, representing 33 districts/cities in Central Java Province, were assessed for their hygiene and sanitation practices according to the NKV criteria mandated by The Minister of Agriculture Regulation No. 11/2020 on NKV Certification for Animal Production Unit. Sixty-six meat samples from these slaughterhouses were obtained for microbiological analysis. The total plate count (TPC), counts of Escherichia coli and Staphylococcus aureus, and the presence of Salmonella spp. were determined. The microbiological tests followed the standard national testing procedure according to the Indonesian National Standard 2897:2008 on Method of Analysis for Microbiological Contaminants in Meat, Eggs, Milk, and its derived products. The sanitation hygiene assessment of the 33 slaughterhouses showed that seven (21.2%) met the NKV criteria level 3, while the others did not. The average TPC of the meat samples was 1.57 × 105 CFU/g (4.93 log10), the S. aureus count was 7.6 CFU/g, and the E. coli count was 9.2 most probable number/g. Only one sample (1.50%) tested positive for Salmonella spp. A comprehensive assessment comparing the NKV criteria with the level of meat contamination showed that the ruminant slaughterhouses that satisfied the NKV criteria had more meat samples (85.71%), on average, that complied with the Indonesian National Standard for microbial contamination compared with those that did not satisfy the NKV criteria (69.23%). The odds ratio was 2.67. Most of the priority ruminant slaughterhouses in Central Java did not meet the NKV standards. The research only looks at the level of hygiene sanitation according to NKV standards in slaughterhouses, the level of contamination produced does not reflect the level of the consumer; therefore, the level of contamination should continue to be investigated at the post-production stage. Keywords: bacterial contamination, hygiene sanitation, meat, slaughterhouse.

Hyperglycemia associated with hyper- or hypo-insulinemia is a hallmark of type 2 diabetes mellitus, which is firmly linked to decreased male infertility. Recently, bee venom (BV) has shown potential health prosperities, including antidiabetic; however, no study focuses on the effect of BV on male fertility in diabetic conditions. This study aimed to detect the effect of BV on histological and hormonal alteration of the testis in diabetic mice. Twenty adult male mice were selected and assigned to four groups: Control, diabetic (150 mg/kg alloxan), BV1 (diabetic + 0.5 mg/kg BV), and BV2 (diabetic + 1 mg/kg BV). After 35 days, the serum levels of glucose, insulin, testosterone, follicular-stimulating hormone, luteinizing hormone, and prolactin were estimated. The histological structure of the testes was also evaluated. Alloxan-induced hyperglycemia and decreased insulin concentrations were reversed significantly by BV. Furthermore, diabetic mice exhibited various alterations in fertility hormones, while these disturbances were improved considerably to normal concentrations by BV. Similarly, alloxan-induced changes in sperm and testis histological parameters such as motility, viability, abnormality, sperm count, the number and diameter of seminiferous tubules, and the number of Leydig and Sertoli cells were significantly ameliorated to the normal condition by BV. Changes in the number, size, and shape of seminiferous tubules, the number of Leydig and Sertoli cells, and initial degeneration and vacuolization in interstitial cells and spermatogonia and spermatocyte were seen in diabetic mice. All these changes were shifted almost to normal structure by BV. The BV could be used as an alternative therapeutic agent that manages the markers related to diabetic conditions concomitant with the improved histological structure of the testes and hormone production to accelerate male fertility. Keywords: diabetic mice, hyperglycemia, Sertoli cells, spermatogonia.