Natural feed additives are important for broiler sustainability during the post-antibiotic era. This study aimed to evaluate the possible effects of the dietary supplementation of Lactobacillus casei-fermented mixture of red rice and aromatic ginger (FERMIX) on growth, blood profiles, intestinal bacterial content, and morphological measurements of broilers. Two hundred broiler chicks were allotted to four groups with five replications, including CONTROL (chicks provided with basal feed), FERMIX025 (basal feed supplemented with 0.25% FERMIX), FERMIX050 (0.50% FERMIX), and FERMIX100 (1.00% FERMIX). FERMIX is an anaerobic fermentation product from a mixture of red rice flour and aromatic ginger using L. casei. Blood, organs, digesta, and carcass were collected on day 35. Final body weight, weight gain, feed intake, and feed conversion ratio did not differ (p>0.05) across treatments. FERMIX at 0.5% increased (p<0.05) spleen relative weight. Liver weight was lower (p<0.05) in broilers fed with 1% FERMIX. The liver weight linearly reduced (p<0.05) with the increased FERMIX levels. FERMIX at 0.25% elevated (p<0.05) broiler drumsticks' yield than that in other groups. Erythrocytes, hemoglobin, packed cell volume, and plasma total protein levels were quadratically higher (p<0.05) in FERMIX050 treated than in other groups. Increased FERMIX levels resulted in a linear decrease (p=0.08) in ileal lactose-negative Enterobacteriaceae counts. Villi height/crypt depth (CD) ratio was quadratically higher (p<0.05) in the duodenum of broiler supplemented with 0.5% FERMIX. The jejunal CD was quadratically lower (p<0.05) in FERMIX050 than in other groups. Although it did not influence growth, dietary FERMIX, especially at 0.5%, improved immune competencies, physiological conditions, and health of broilers' intestine. Keywords: broiler, fermentation, health, phytogenic, prebiotic, probiotic.

The World Health Organization considers multidrug-resistant (MDR) Klebsiella pneumoniae a major global threat. Horses harbor commensal isolates of this bacterial species and potentially serve as reservoirs for human MDR bacteria. This study investigated antimicrobial resistance in horses caused by extended-spectrum β-lactamase (ESBL)-producing K. pneumoniae. One hundred fifty-nine nasal swab samples were collected from horses with respiratory distress not treated with cefotaxime and erythromycin. Biochemical and serological identification was performed on all samples. Polymerase chain reaction (PCR) was used to detect 16S-23S ITS, mucoviscosity-associated gene (magA), uridine diphosphate galacturonate 4-epimerase gene (uge), and iron uptake system gene (kfu), blaTEM, blaSHV, and blaCTX genes. Sequence analysis and phylogenetic relatedness of randomly selected K. pneumoniae isolates carrying the blaTEM gene were performed. Ten isolates of Klebsiella spp. were obtained from 159 samples, with an incidence of 6.28% (10 of 159). Based on biochemical and serological identification, K. pneumoniae was detected in 4.4% (7 of 159) of the samples. Using PCR, all tested K. pneumoniae isolates (n=7) carried the 16S-23S ITS gene. By contrast, no isolates carried magA, uge, and kfu genes. The blaTEM gene was detected in all test isolates. Moreover, all isolates did not harbor the blaSHV or blaCTX gene. Sequence analysis and phylogenetic relatedness reported that the maximum likelihood unrooted tree generated indicated the clustering of the test isolate with the other Gram-negative isolate blaTEM. Finally, the sequence distance of the blaTEM gene of the test isolate (generated by Lasergene) showed an identity range of 98.4-100% with the blaTEM gene of the different test isolates. The misuse of antimicrobials and insufficient veterinary services might help generate a population of ESBL-producing K. pneumoniae in equines and humans, representing a public health risk. Keywords: extended-spectrum β-lactamase, horse, Klebsiella pneumoniae, multidrug-resistant, respiratory manifestation.

The high salinity of drinking water has been a significant problem of the Mekong Rivers Delta. Animals drinking high salinity water altered feed and water intake (WI), urinary electrolytes excretion, and productivity. This study aimed to evaluate the effects of high salinity in drinking water on drinking and eating behaviors and kidney function in crossbred goats. The experiment was completely randomized with two treatments: freshwater (0%, seawater [SW0]) and water high in salinity (1.5%, SW1.5) from diluted SW, with five replicates (five animals per treatment). This experiment lasted 3 weeks: the 1st week for the pre-treatment period and the 2nd-3rd weeks for the post-treatment. Dry matter intake (DMI) and WI were recorded every day, while urine volume (UV) was determined from day 8 to day 21. Blood and urinary samples were collected on days 6, 14, and 21 of the study for electrolytes and creatinine analysis. The results demonstrated that both DMI and WI were affected by SW1.5 (p<0.05). Goats drinking from SW1.5 had lower DMI during D19–21, and the ratio of DMI/WI was significantly different during D16–21 (p<0.05). Interestingly, the UV from SW1.5 was higher than that from SW0 during D13–21 (p<0.05). Although the body weights (BW) of both groups were similar (p>0.05), the weight gain observed in the SW1.5 group tended to decrease (p=0.056) at the 2nd week. The concentration of electrolytes in blood did not differ between the groups (p>0.05). In contrast, the concentration and excretion of Na+ and Cl- in urine increased in SW1.5 goats at D14 (p<0.05), while creatinine levels in the blood remained normal (p>0.05). The study concluded that crossbred male goats can tolerate 1.5% saline water from diluted SW for 2 weeks. The high salinity in water influences drinking and eating behavior in growing goats. However, the adaptive mechanism by increasing urine output and reducing the reabsorption of Na+ and Cl- in the kidney is the key function and works faster than behavioral responses. The kidney apparently drives drinking behavior during high salinity water consumption. Keywords: electrolytes, drinking behavior, goat, kidney, saline water.

Stem cells are cells that can proliferate to form a new tissue, leading to its use in regenerative therapy. Stem cells will secrete biological factors, such as growth factors, cytokines, and other proteins to their surroundings and culture medium/conditioned medium (CM), altering tissue physiology. These factors can help wound healing, but their effect on third-degree burns is poorly understood. This research aimed to study the activity of mesenchymal stem cell-conditioned medium gel in healing and repairing third-degree burns on rats skin. Twenty-four Sprague–Dawley rats with burn wounds on the dorsal area were divided into four groups; the first group was treated with CM gel, with a concentration equivalent to 0.05% protein, the second group was treated with a placebo gel, the third group with silver sulfadiazine (SSD) cream (SSD-Burnazin contain 10 mg/g SSD), and the fourth group was not given any treatment, for 21 days, and on the final day, the rats were sacrificed, and the skins were taken. All topical treatments completely cover the wound area. Wound healing process indicators observed include wound diameter, scabs' formation, blister formation, and hair growth every day. The skins taken were processed with hematoxylin-eosin and Masson's trichrome staining. The indicators studied include neutrophil infiltration, mononuclear cell infiltration, neovascularization, collagen area, and re-epithelization ratio. CM shows better wound healing than other groups and faster hair growth. Keywords: conditioned medium, mesenchymal stem cell, third-degree burn.

Clinical strains of microorganisms, including pathogenic yeast-like fungi (YLF), are resistant to currently used antifungal agents. Thus, it is relevant to study the combinations of existing antimicrobial drugs and a medicinal extract of plant origin (farnesol). In previous studies, farnesol showed a relatively strong anti-biofilm effect against Candida albicans. This study aimed to determine how much the resistance profile of non-biofilm microorganisms can change. Six clinical isolates of C. albicans and one reference strain were used to study the interaction of farnesol with the most used antimycotics. To determine the sensitivity of YLF to antimycotic drugs, such as nystatin (50 μg), amphotericin B (10 μg), ketoconazole (10 μg), clotrimazole (10 μg), voriconazole (10 μg), fluconazole (25 μg), miconazole (10 μg), and intraconazole (10 μg), the classic disk diffusion method was used. In the second stage, one of the six strains was used to simulate candidiasis of the gastrointestinal tract in an in vivo quail model. As an unusual experimental design, this study investigated the effects of pretreated C. albicans in quails, not the in vivo pathogenicity of C. albicans, after treatment with farnesol. The resistance profiles of Candida strains did not improve with farnesol in all strains. All concentrations of farnesol (100, 50, and 25 μM) demonstrated a fungistatic effect (i.e., an increase in drug sensitivity) in 23 of 56 (7×8) cases (41%). The remaining 54% demonstrated no changes in the resistance to antifungal drugs or deterioration of the indicators in rare cases (5%). At 100 μM farnesol, sensitivity improved in 33 of 56 cases (59%). Candidiasis or the severity of clinical disease of the quail digestive tract developed to a lesser extent if fungi were treated with farnesol. Farnesol does not always show a positive result on single cells without biofilm in the laboratory. However, in a biofilm or an in vivo model with biofilms, farnesol can be considered a new antimycotic drug or an additive to existing antimycotics. Keywords: amphotericin B, antimycotics, Candida albicans, clotrimazole, farnesol, fluconazole, intraconazole, ketoconazole, miconazole, nystatin, quail model, quorum sensing, voriconazole.

Udder health management is essential for the further development of milk production systems and public health. This process depends on the generation of knowledge regarding control, prevention, and promotion of health. In scientific literature, it is impossible to find a synthesis of the categories that would allow comprehension of the complex phenomenon udder health. Different research approaches have allowed this polysemic concept, described by some researchers as multifactorial and by philosophical perspectives as a social phenomenon, to be further studied. Thus, the objective of this systematic review was to systematize the conceptual categories of udder health and the use of the term in the original articles published in the scientific literature from the period 1962 to 2019. A systematic review with a broad approach was designed by applying the phases of identification, screening, selection, and inclusion criteria described in the Preferred Reporting Items for Systematic Reviews and Meta- Analyzes guide. An exhaustive search of original articles by specificity was carried out in the Science Direct, PubMed, Scielo, LILACS, and Google Scholar databases. The investigation was carried out on November 22, 2019. According to the inclusion criteria established, articles needed to be original studies, to be publications on bovine livestock, written in English, Spanish and Portuguese. Furthermore, the articles considered needed to tackle the term udder health so that its conceptual categorization could be extracted. Google Scholar patents and citations and articles removed from databases or not available were excluded from the study and those that, based on the reading of the complete text, considered the farming of animal species other than bovine. A qualitative synthesis of the year of publication, continent, approach, type of study, and conceptual category of udder health was carried out by calculating frequencies (Statistical Package for the Social Sciences version 24). In total, 165 articles were included in the study. Eight conceptual categories, consolidated over time, were systematized, showing that udder health is not a static problem, and that science has been responding through the generation of new knowledge around conceptual categories as different udder health problems emerge. Culture and politics were two categories, related to all the others, that stood out in the results. These two categories were of great interest in countries advanced in milk production and in the implementation of udder health policies, which acknowledge the producer and other actors of the production chain as fundamental political actors for policies, decision-making processes, and public health care to be effective. The lack of synonyms for the term udder health (e.g., mastitis) may have led to the exclusion of important articles in each category. However, the constriction to the term udder health was intentional and aimed at constructing the concept. Udder health is hereby understood as a health-disease process, different from the term mastitis, which from its semantic origin, refers only to the disease process. According to this study, the concept can be understood through the categories of traditional epidemiology based on risk factors and disease; microbiology; genetics, resistance, and immunity; animal welfare; nutrition; organic production; culture; and politics. Keywords: conceptual categories, culture, livestock productivity, politics, public health, udder health.

Various management practices may cause the occurrence of reproductive failure indicated by repeat breeding in beef cattle. This study aimed to estimate the prevalence and the risk factors of repeat breeding in beef cattle in Sleman Regency, Indonesia. Observational and cross-sectional studies were used to determine the prevalence and the risk factors of repeat breeding. Sampling was conducted using a multistage cluster design. The sample size was determined using a sampling formula (n=4 PQ/L2). Questionnaire and interview data were evaluated descriptively. Chi-square analysis and odds ratio (OR) test were conducted to determine the association and association strength with a confidence level of 95%. Univariate, bivariate, and multivariate analysis through multivariate logistic regression test was done using Statistical Package for the Social Sciences, version 21.0 software. The results indicated that the prevalence of repeat breeding in beef cattle in the Sleman Regency was 30.4%. Multivariate analysis indicated that risk factors that significantly affected the repeat breeding were breeding experience (p=0.000; OR=3.378), knowledge of estrus cycle (p=0.000; OR=5.263), feed type (p=0.001; OR=6.061), feeding frequency (p=0.003; OR=2.77), shed hygiene (p=0.33; OR=2,381), and drainage system (p=0.000; OR=4,484). Various husbandry management significantly influence the incidence of repeat breeding in beef cattle in Sleman Regency with the type of feed, which was the highest risk factor. Hay should not be used as the main feed source since it might increase the incidence of repeat breeding. However, the other environmental factors such as season and presence of infection or parasite also need to be investigated further. Keywords: breeder, cow, epidemiology, reproductive failure, statistical analysis.

Duck eggs have high cholesterol levels; inulin addition combined with probiotic is known in several studies to lower cholesterol, while maintaining egg production capacity and blood hematology. This study aimed to investigate the effect of the addition of synbiotic preparations on egg production, egg quality, and hematology of Pengging ducks. A total of 200 female Pengging ducks aged 75 weeks (late production phase) and weighing 1467±90.87 g were maintained in litter cages, each measuring 1×1 ducks. The treatment included the addition of synbiotics between the inulin of gembili tuber (Dioscorea esculenta L. and Lactobacillus plantarum Ina CC B76) as follows: T0=control feed ("farmer feed"), T1=control feed+synbiotics 1 mL/100 g, T2=control feed+synbiotics 1.5 mL/g, and T3=control feed+synbiotics 2 mL/100 g in the feed. A completely randomized design was used in this study. The production performance, physical and chemical qualities of eggs, and hematological parameters of Pengging ducks were evaluated. The addition of synbiotics had no significant impact on the production performance, physical and chemical qualities of eggs, and hematological parameters (p>0.05), except for the egg yolk cholesterol content. The cholesterol content decreased significantly (p<0.05) with T2 and T3 treatments, but they had no significant effect (p>0.05). A significant decrease (p<0.01) in cholesterol levels was observed when the synbiotic dose was given at 1.5 ml/100 g feed (T2). However, there was no further decrease in cholesterol level when the synbiotic dose was increased to 2 ml/100g fed (T3). The addition of synbiotics preparations at 1.5 mL/100 g reduced the cholesterol content but did not improve egg production, egg physical quality, and hematology of Pengging ducks. Keywords: cholesterol, egg production, egg quality, inulin, late phase, Pengging duck.

The principal cytokines released by the host on infection include pro-inflammatory cytokines such as interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF). These cytokines were regarded as regulators of the host's response to infection. This study aimed to determine the release of pro-inflammatory cytokines from chicken peripheral blood mononuclear cells (PBMCs) following lipopolysaccharide (LPS) stimulation. Blood samples were collected from six Betong chickens. To isolate PBMCs, density gradient centrifugation was utilized. PBMC culture in RPMI1640 with 10% fetal bovine serum was stimulated with various concentrations of LPS (0, 0.01, 0.1, and 1.0 μg/mL). The production of TNF-α, IL-1β, and IL-6 was determined using an enzyme-linked immunosorbent assay. When the PBMCs were cultured for 24 h with varying doses of LPS, there was no significant variation in cell viability. TNF-α, IL-1β, and IL-6 levels were measured in Betong chicken PBMC. The release of these cytokines increased considerably as LPS concentration (0.01-1 μg/mL) increased (p<0.05). In vitro studies of the chicken immune response, notably the release of pro-inflammatory cytokines, can be conducted using PBMCs obtained from chicken blood. Keywords: chicken, interleukin-1β, interleukin-6, peripheral blood mononuclear cell, tumor necrosis factor-α.

Extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae have become a serious public health hazard worldwide. This importance is derived from the increase of new variants, particularly blaTEM, blaSHV, and blaCTX-M genes. This study aimed to examine ESBL-producing Escherichia coli isolated from different governorates in Egypt from dairy cows infected with subclinical and clinical mastitis. This study examined 207 milk samples for the resistance of isolates against 14 different antibiotics and ran serological identification of ESBL-producing E. coli isolates with complete antibiotic resistance. Genotypic and sequencing analyses of several resistance genes were conducted using a polymerase chain reaction. E. coli was identified in cases with subclinical mastitis (80.5%) and clinical mastitis (85.7%). ESBL-producing E. coli was isolated from 38.2% of subclinical mastitic milk compared to 39.3% in clinical cases, where O26:k60, O125:k70, and O25:k11 were the serotypes with complete resistance to antibiotics. ESBL-producing E. coli isolates were resistant to cefotaxime, amoxicillin, cloxacillin, oxacillin, rifampicin, and penicillin in 100% but susceptible to amoxicillin and clavulanic acid in 82.5% of the cases. Results also revealed that 51.25%, 52.5%, 66.25%, 77.5% and 60% of ESBL-producing E. coli isolates were responsive to ciprofloxacin, ofloxacin, norfloxacin, levofloxacin, and gentamycin, respectively. The detected genes were registered in GenBank as MW345819.1 and MW345820.1 for the E. coli blaTEM gene and MW295407 for the E. coli blaSHV gene. This study found ESBL-producing E. coli in mastitic milk samples from Egyptian dairy farms and confirmed the occurrence and circulation of the main antibiotic genes (blaTEM and blaSHV) in the samples. Regular and thorough surveillance of ESBL-producing E. coli and subsequent preventive actions are essential for preventing the spread of these resistance genes in the future, which could pose serious and catastrophic health risks. Authorities should cling to the concept of One Health to minimize the risk of new varieties. Keywords: antibiotic resistance, Escherichia coli, extended-spectrum β-lactamases, mastitis.

The coronavirus disease-19 (COVID-19) pandemic caused global economic and health problems. The pandemic increased the number of infectious diseases categorized as neglected diseases, such as staphylococcosis, which is caused by methicillin-resistant Staphylococcus aureus (MRSA). Celery is an herb that consist of antioxidants that can potentially act as antimicrobial agents. This study aimed to analyze the efficacy of celery alcoholic extract against systemic MRSA infections in rat models. In this study, 36 male, 6-month-old Sprague-Dawley rats (average weight: 300 g) were used as models. The rats were divided into six groups: Group K– (negative control), Group K+ (infected with MRSA without therapy), Group V (infected with MRSA+100 mg vancomycin per kg body weight [BW]), Group P1 (infected with MRSA+1 mg celery extract per kg BW), Group P2 (infected with MRSA+2 mg celery extract per kg BW), and Group P4 (infected with MRSA+4 mg celery extract per kg BW). The therapy was given once daily for 7 days. Blood and organs were taken on day 7 for hematology, serology, immunohistochemistry, and histopathology. Results showed that 4 mg celery extract per kg BW promotes the healing of MRSA systemic infections in rat models (p≤0.05). The better prognosis was indicated by the normalization of red blood cell indices, white blood cell, neutrophil and lymphocyte counts, Cluster of differentiation 4+, Cluster of differentiation 8+, and Cyclooxygenase-2 expression and absence of severe tissue damage. Celery extracts inhibited MRSA growth in the blood samples. It can be concluded that celery alcoholic extract can potentially be used as an antimicrobial agent against systemic MRSA infections. A clinical study regarding the efficacy of celery extract must be conducted to ensure its potency against MRSA infections in humans. Keywords: celery, efficacy, healing, methicillin-resistant Staphylococcus aureus, systemic infection.

The high price of conventional diet ingredients led livestock producers to search for alternative feed sources such as faba beans (Vicia faba L.; FB). This study aimed to evaluate the effect of feeding FB on the growth performance of lambs. A total of 24 male lambs were distributed randomly into two groups and fed a control diet (CON; n=12) and 200 g/kg FB (FB200; n=12) dietary dry matter (DM). The study lasted for 70 days. The first 7 days were used for acclimatization, followed by 63 days of data collection (i.e., nutrient intake and digestibility and growth performance). A complete randomized design was used for the statistical analysis. The average initial body weight (BW) (20.54±0.798 kg) was similar between the diet treatment groups. Lambs fed the FB200 diet demonstrated higher (p≤0.008) nutrient intake than lambs fed the CON diet. The FB200 diet tended to improve the digestibility of DM, crude protein, and acid detergent fiber more than the CON diet (p≤0.072). Neutral detergent fiber and ether extract digestibility were higher (p<0.05) in lambs fed the FB200 diet than those fed the CON diet. Next, nitrogen retention increased (p<0.05) in lambs fed the FB200 diet compared with the CON diet. Final BW did not differ (p=0.221) between the two groups. However, the average daily gain was higher (p=0.028) in lambs fed the FB200 diet than in lambs fed the CON diet. Furthermore, the cost of gain decreased more (p=0.04) with the FB200 diet than with the CON diet. The results obtained in this study demonstrate the feasibility of using FB in feeding growing lambs, as it was shown to improve growth performance and reduce the cost of diet and gain. Keywords: Awassi lambs, faba beans, growth performance, nutrient intake and digestibility.

As a new feed additive, bile acid (BA) can promote the absorption and transport of lipids and fat-soluble vitamins. In recent years, BAs have been widely used in animal feed to promote fat absorption. Therefore, this study aimed to investigate the effect of bile salt supplementation in the diet of yellow-feathered broilers on messenger RNA (mRNA) expression of sterol regulatory element-binding protein 1 (SREBF1), fatty acid synthase (FAS), acetyl-coenzyme A carboxylase (ACC), and fatty acid transport protein 4 (FATP4). Four hundred and twenty commercial male chicks were randomly divided into seven groups (with four replicates per group and 15 chickens per replicate). They were fed diets supplemented with bile salts at 0, 1.5, 2.5, 3.5, 4.5, 5.5 mg/kg, and 2 mg/kg tylosin for 30 days. Changes in SREBF1, fatty acid transporter 4, FAS, and acetyl- CoA carboxylase genes in intestinal mucosa and liver of yellow-feathered broilers were determined using a quantitative fluorescence polymerase chain reaction. mRNA expression of SREBF1, FAS, ACC, and FATP4 in the small intestine decreased in chicks fed diets supplemented with 3.5 and 4.5 mg/kg bile salts (p<0.05) compared with the control group on 7 days and 14 d. The mRNA expressions of SREBF1, FAS, ACC, and FATP4 in liver tissue decreased in chicks fed diets supplemented with 4.5 and 5.5 mg/kg bile salts (p<0.05) compared to the control group on 7 days. The mRNA expression of SREBF1, FAS, ACC, and FATP4 in the liver at 14 days and the small intestine on 21 days also decreased in chicks fed diets supplemented with 4.5 mg/kg bile salts (p<0.05) compared to the control group. When contrasted with the control group on day 21, the mRNA expression of SRWBF1, FAS, ACC, and FATP4 detected in the liver was lower in chicks fed diets supplemented with bile salts (p<0.05). The dietary supplementation of bile salts at 4.5 mg/kg effectively regulates the expression of fat metabolism genes, such as SREBF1, FAS, ACC, and FATP4 mRNA. At this concentration, bile salts promote fat catabolism, inhibit fat synthesis, and play an essential role in improving the fat deposition of broilers. Keywords: acetyl-CoA carboxylase, fat metabolism, fatty acid synthase, fatty acid transporter 4, sterol regulatory element-binding transcription factor 1, supplemental dietary bile salts.

Theileria annulata is the most prevalent piroplasm infecting cattle across Egypt. Theileriosis is transmitted by ixodid ticks of the genus Hyalomma. This study aimed to estimate the prevalence of theileriosis in cattle and their associated risk factors for infection. A total of 570 blood samples were collected from cattle from five governorates (administrative districts) in Egypt and examined using a polymerase chain reaction assay to estimate the prevalence of tropical theileriosis and assess the associated risk factors. The overall prevalence rate was 16.49%, with the Alexandria district having the highest prevalence. The results revealed that the risk of theileriosis was elevated in older cattle (odds ratio [OR]=8.9, 95% confidence interval [CI]: 3.6- 21.9), especially in summers (OR=3.07, 95% CI: 1.4-6.3). Cattle heavily infested with ticks were at particular risk (OR=3.05, 95% CI: 2.1-4.5), as were those to which acaricide had not been applied (OR=13.7, 95% CI: 5.6-33.6). Understanding the risk factors associated with T. annulata infection and regular infection monitoring could reduce infection rates and economic losses and is essential for the implementation of efficient control programs. Keywords: cattle, Egypt, polymerase chain reaction, risk factors, Theileria annulata.

Theileria annulata infection is a tick-borne disease affecting ruminants in the tropical and subtropical regions causing severe economic losses. This study aimed to characterize circulating T. annulata isolates from four governorates (administrative districts) north and south of Egypt using polymerase chain reaction (PCR)-restriction fragment length polymorphism. Fifty samples were collected from the four governorates of Egypt and were examined by a PCR assay based on the heat shock protein 70 gene. The amplified product was subsequently digested using two restriction enzymes, Taq I and Alu I, to determine which pattern of T. annulata strains was involved. The findings revealed that one distinct pattern was observed for T. annulata isolates in the northern governorates and another one in the southern governorates. The Taq I enzyme produced three fragments (100, 175, and 270 bp), and the Alu I enzyme produced four fragments (60, 90, 125, and 270 bp). This study determined the presence of two distinct circulating genotypes of T. annulata among cattle in Egypt based on PCR-RFLP using the HSP 70 gene. More studies are needed in different parts of the country to investigate the virulence and strain variance of T. annulata in cattle. Keywords: cattle, Egypt, genotype, polymerase chain reaction-restriction fragment length polymorphism, Theileria annulata.

The pathogenesis of non-alcoholic steatohepatitis involves non-alcoholic fatty liver, oxidative stress, inflammation, and fibrosis. Although the long-term use of cinnamon bark in larger doses can negatively affect good health, proper use of its extracts effectively and efficiently improves health. Therefore, this study aimed to determine the minimal dose of Cinnamomum Burmannii extract through its activity in inhibiting oxidative stress in rats' livers treated with a high-fat and cholesterol diet (HFCD). Forty-two Sprague–Dawley rats (Rattus norvegicus), weighing 200-250 g body weight (BW), were divided into seven treatment groups with six replications: Normal, HFCD, atorvastatin, quercetin, and C. burmannii ethanol extract group, after which they were administered different dosages (i.e., 100, 200, and 300 mg/kg BW). Except for the normal group, rats were concomitantly administered HFCD with each treatment for 21 days. Then, their malondialdehyde (MDA) levels and superoxide dismutase (SOD) activity were assessed using colorimetry. However, their steatosis levels were determined based on histological preparations with hematoxylin-eosin staining. Duncan's multiple range test (DMRT) results indicated that all treatments had a significantly lower MDA than HFCD and normal rats (α=0.01). DMRT results also showed that treating with the C. burmannii ethanol extract at all dosages resulted in a significantly higher SOD activity level in HFCD rats than those treated with quercetin and atorvastatin (α=0.01). Furthermore, results showed that treatment with C. burmannii extracts at a dosage of 300 mg/kg BW incredibly maintained SOD activity as effective as quercetin, atorvastatin, and normal rats. Besides, while steatohepatitis levels of C. burmannii ethanol extract at dosages of 200 and 300 mg/kg BW commensurated with normal rats, steatohepatitis levels were significantly lower than those administered other concentrations or treatments (α=0.05). Ethanolic C. burmannii extracts protected the liver by regulating oxidative stress. Therefore, a 200 mg/kg BW dose is proposed as the minimal hepatoprotection dose to prevent fatty liver formation. Keywords: Cinnamomum burmannii, hepatoprotection, malondialdehyde, steatohepatitis, superoxide dismutase.

Snail meat and digestive tract hydrolyzate fermented with a consortium of preserved rice water microorganisms could serve as new sources of probiotics. Microorganisms from the examined feed supplement were isolated, identified, and characterized for resistance at low pH and with bile salts. The study aimed to determine the potential hydrolysate of the snail meat and digestive tract as a novel probiotic supplement for animal feed at various pH values and Oxgall. The submerged fermentation method was conducted for 21 days to examine the novel probiotic that originated from snail microorganisms in the hydrolyzed liquid fermented by finely ground snail meat and the digestive tract. The microorganisms in the hydrolyzate were isolated by a spread plate method, while the potential of the probiotic hydrolyzate was tested for resistance to pH values of 2, 2.5, 3, and 4, as well as resistance to bile salts at Oxgall concentrations of 0.2%, 0.3%, 0.5%, 1%, and 2%. The hydrolyzate profile of snail meat and digestive tract contained five isolates of lactic acid bacteria that could serve as potential probiotics. The application of fermentation technology using a consortium of preserved rice water microorganisms can convert snail meat and the digestive tract into novel probiotic products that could be utilized in feed supplements. Keywords: bile, pH, probiotics, snails, supplements.

Micronutrient imbalances pose a severe threat to the health and productivity of livestock and poultry. In this regard, a further stage in feeding science development will control and optimize the intake of mineral substances, including determining the elemental composition in some biosubstrates. One of these biosubstrates can be a feather. However, the amount of available information on the content of trace elements in laying hens is limited, complicating the laboratory data interpretation. Therefore, this study established reference intervals for the concentrations of the main essential and toxic elements in laying hens in different periods of ontogenesis. The study was conducted on clinically healthy Hisex Brown laying hens at the age of 10 (n=150), 30 (n=150), 120 (n=150), 150 (n=150), and 210 (n=150) days. All examined birds were born and raised on the territory of the South Ural biogeochemical province of Russia. The sampling of feathers was carried out by plucking the flight feathers of the wing. Inductively coupled plasma dynamic reaction cell mass spectrometry determined the elemental composition of the feather according to 25 parameters. The results showed that at the minimum age (10 days), the highest concentrations of chemical elements were observed in laying hens. Subsequently, as they grew older, in the period from the 30th to the 120th day, there was a significant decrease in these indicators. Later, from the 120th to the 150th day, a statistically significant increase in concentrations was replaced. Little growth and relative concentration stability were observed in the last part of the experiment (150-210 days). Chemical element concentrations in feathers were computed in reference ranges for each age group. The calculated ranges of chemical element concentrations in laying hens can be used to compile norms for their content in the body; however, it is worth noting that these ranges can vary depending on the biogeochemical province of breeding and the bird's age. Keywords: age, essential and toxic elements, feathers, laying hens, reference intervals.

Anthrax is one of the endemic strategic diseases in East Indonesia, particularly in the provinces of South Sulawesi, West Sulawesi, Gorontalo, East Nusa Tenggara, and West Nusa Tenggara. Anthrax is an important disease due to its zoonotic and economic impact on the livestock industry. This study aimed to identify the molecular characteristics of Bacillus anthracis in East Indonesia using multilocus variable-number tandem repeat (VNTR) analysis (MLVA). Isolates were obtained from an investigation of anthrax outbreaks in five provinces of East Indonesia from 1982 to 2020. Conventional polymerase chain reaction for B. anthracis was used to identify MLVA-8. Deoxyribonucleic acid sequencing analysis was based on MLVA-8 primers for VNTR identification of the phylogenetic relationship among 24 isolates of B. anthracis obtained from 17 distinct districts/cities in East Indonesia. Tandem Repeats Finder was used for VNTR identification, and Molecular Evolutionary Genetics Analysis X was used to construct phylogenetic analysis. In this study, 24 isolates were classified as genotype or lineage A. There were four subgroups of B. anthracis circulating in East Indonesia based on eight molecular marker loci sequence results. The findings of this study show that MLVA-8 typing might be useful as a subtyping tool for the epidemiological investigation of identical genotypes and low genetic diversity of B. anthracis. No other lineage of B. anthracis was circulating in East Indonesia. Other molecular methods are needed, such as extended MLVA, whole-genome sequencing, and canonical single-nucleotide polymorphism, for a more precise study of B. anthracis genetic diversity. Keywords: Bacillus anthracis, Indonesia, multilocus variable-number tandem-repeat analysis-8.

Cynomolgus monkeys (Macaca fascicularis) develop spontaneous infection of Papillomavirus (PV); thus, potentially beneficial for modeling human PV (HPV) infection study. Contrary to human origin, infection in cynomolgus monkeys does not always show evident clinical symptoms of cervical cancer. The absence of cervical cancer clinical symptoms leads us to investigate the molecular mechanism of the HPV infection in cynomolgus monkeys. This study aimed to investigate the messenger ribonucleic acid (mRNA) expression levels of KI67 and P53 genes, majorly known as biomarker oncogenesis of PV infection. The polymerase chain reaction (PCR) technique was used with MY11/MY09 primer to screen PV in cynomolgus monkey, further grouped as positive-PV and negative-PV infection groups. Real-time quantitative PCR was also applied to quantify the mRNA expression levels of KI67 and P53 genes in animals. Increased expression of mRNA level of KI67 genes was significantly higher in Positive- PV group than negative-PV group. In contrast, the P53 mRNA expression level increased markedly higher in the negative-PV group than in the positive-PV group. Our study describes the potential of cynomolgus monkeys as a spontaneous oncogenesis model of PV infection-type. However, we used a limited number of cancer genetic markers. So, further study of other genetic markers is required to prove that cervical cancer could be developed naturally in cynomolgus monkeys. Keywords: animal model, biomarker genes, cervical cancer, nonhuman primate.

In Iraq, stray dogs represent a critical population of free-roaming animals, which probably play a role in the transmission of different infections to other animals. Canine distemper is one of the most growing viral threats to carnivores in many countries worldwide, including Iraq. Therefore, this study was aimed to diagnose the disease using serological and molecular assay and the role of risk factors in the spreading infection. In all, 158 venous blood samples were collected randomly from stray dogs in rural and sub-urban areas of Iraq from May 2019 to December 2020. The samples were examined serologically using two enzyme-linked immunosorbent assay (ELISA) kits, immunoglobulin G (IgG) and immunoglobulin M (IgM), and molecularly by reverse transcription-polymerase chain reaction (RT-PCR) to detect and confirm chronic and acute infections. To determine the association between infection and various risk factors, the study animals were divided according to their locations, sexes, and ages. The age groups were ≤8 months (puppy), 1-3 years (young), and ≥3 years (old/mature). ELISA result shows that 6.96% and 19.62% of dogs were seropositive for acute and chronic distemper, respectively. The titer of chronic infections (0.421±0.027) was significantly higher (p≤0.025) than that of acute canine distemper (0.337±0.016). On RT-PCR, 8.86% of dogs were found positive for distemper. Using RT-PCR as the gold standard, the sensitivity and specificity of the IgM ELISA kit were 75% and 98.63%, respectively, whereas the positive and negative predictivity were 81.82% and 97.96%, respectively. A significant variation (p<0.05) was observed in the distribution of positive findings among the different epidemiological risk factors. Compared with rural areas, positivity was significantly higher (p<0.05) in sub-urban areas on IgM (26.92%) and IgG (64.15%) ELISA and RT-PCR (34.62%). On IgM ELISA and RT-PCR, no significant differences (p>0.05) were found among the three age groups; however, positivity was significantly higher (p≤0.048) in the ≥3 years group (22.73%) on IgG ELISA. Furthermore, only IgG ELISA showed a significantly higher (p≤0.032) positivity rate in female dogs (25.23%) than in male dogs (7.84%). To the best of our knowledge, this is the first Iraqi study to demonstrate acute and chronic canine distemper in stray dogs, suggesting that the prolonged shedding of virus from positive dogs is a critical point in the epidemiology of the disease. Additional studies in dogs or other carnivores are required to establish baseline data on the prevalence of the disease in Iraq. Keywords: epidemiology, Iraq, molecular, Morbillivirus, serology.

Bovine eimeriosis is a disease caused by apicomplexan parasites of the genus Eimeria. It is one of the most important and widespread bovine illnesses in the world. Some of the identified species of bovine eimeriosis have morphologically similar oocysts that are difficult to differentiate. For the identification of particular Eimeria spp., diagnostic laboratories are increasingly turning to DNA-based technology. This study aims to develop a multiplex polymerase chain reaction (mPCR) technique based on the internal transcribed spacer-1 (ITS-1) gene for the simultaneous identification of pathogenic Eimeria spp. in cattle from Sulawesi Island, Indonesia. Genomic DNA was extracted by the DNAzol reagent from the purified Eimeria oocysts. Species-specific primers targeting the ITS-1 region were used to amplify the distinct Eimeria spp. Using PCR ITS-1, this study showed that 36 of 120 fecal samples (30%) were infected by Eimeria spp. The multiplex PCR assay allowed for the simultaneous identification of six major Eimeria spp. in a single-tube reaction. The proportion of mixed Eimeria spp. infections was 100% (36/36). The maximum number of Eimeria spp. was five, and the minimum number was two. Identification of six pathogenic Eimeria spp. in cattle was successfully carried out by nested multiplex PCR using ITS-1 gene. In the future, a procedure to detect pathogenic Eimeria spp. in one tube reaction will offer economical and save diagnostic time. Keywords: bovine, diagnostic, eimeriosis, multiplex, nested, Sulawesi island.

Calves have increased morbidity and mortality rates after dystocia. One cause is pain during birth, which reduces their colostrum intake. The administration of nonsteroidal anti-inflammatory drugs (NSAIDs) may break this causality. This study aimed to determine the consequences of a single administration of the NSAID meloxicam to dystocia calves after birth. Fifty Holstein–Friesian calves born with dystocia from four dairy cattle farms were included in this study. The animals were randomized into two groups. The animals in Group I (n=25, treatment group) received subcutaneous 0.5 mg meloxicam/kg body weight 2-8 h after birth. The animals in Group II (n=25, control group) received a control substance (Amynin®, bovine infusion solution, Merial) with the same volume. The newborn calves were clinically examined on the 1st and 10th days of life. The information regarding the days in between was gathered by questioning the farmer. There was a significant difference (p=0.04) only in calves with thin, mushy fecal consistency on the 10th day in the treatment group compared with the control group. Moreover, meloxicam had no effect on dystocia calves. Since NSAID administration did not produce a significant clinical effect, its necessity is questionable. Further studies should examine how modifying its application time would have an effect. The primary indicators of well-being, such as pain indicators in the blood, were not measured; however, these should be considered in subsequent studies. Keywords: calf, dystocia, nonsteroidal anti-inflammatory drugs.

Consumption of fatty acids (FA) can alter hepatic energy metabolism and mitochondrial function in the liver. Crocodile oil (CO) is rich in mono-and polyunsaturated FAs, which have natural anti-inflammatory and healing properties. In rat livers, we investigated the effect of CO on mitochondrial function for energy homeostasis. Twenty-one male Sprague-Dawley rats were divided into three groups at random. Group 1 rats were given sterile water (RO), Group 2 rats were given CO (3% v/w), and Group 3 rats were given palm oil (PO) (3% v/w). For 7 weeks, rats were given sterile water, CO, and PO orally. The researchers looked at body weight, food intake, liver weight, energy intake, blood lipid profiles, and mitochondria-targeted metabolites in the liver. The liver's histopathology, mitochondrial architecture, and hydrolase domain containing 3 (HDHD3) protein expression in liver mitochondria were studied. Body weight, liver weight, liver index, dietary energy intake, and serum lipid profiles were all unaffected by CO treatment. The CO group consumed significantly less food than the RO group. The CO group also had significantly higher levels of oxaloacetate and malate than the PO group. CO treatment significantly ameliorated hepatic steatosis, as evidenced by a greater decrease in the total surface area of lipid particles than PO treatment. CO administration preserved mitochondrial morphology in the liver by upregulating the energetic maintenance protein HDHD3. Furthermore, chemical-protein interactions revealed that HDHD3 was linked to the energy homeostatic pathway. CO may benefit liver function by preserving hepatic mitochondrial architecture and increasing energy metabolic activity. Keywords: crocodile oil, energy metabolism, liver, mitochondria, rat.

The use of antibiotics is prohibited in poultry feeding in many countries worldwide, which has resulted in the emergence of antibiotic-resistant bacteria. Therefore, probiotic supplements are a good alternative in these circumstances. Probiotics, including Bifidobacterium adolescentis and Lactobacillus acidophilus, can reduce the development of resistance and stimulate the growth of broiler ducklings. This study aimed to evaluate properties of the probiotic supplement containing these bacteria. We studied the response of broiler ducklings to the addition of a dietary probiotic supplement containing lactobacilli and bifidobacteria until they reached 7 weeks of age. All birds participating in the experiment received probiotic supplements on days 1-7, 15-21, 29-35. The state of the ducklings was assessed on day 21. At the age of 21 and 42 days, 4-5 ml of blood was drawn from the wing vein of 10 randomly selected birds (5 in each group). Blood samples were analyzed for total protein, concentration of glucose, hemoglobin, calcium and inorganic phosphorus, as well as the number of erythrocytes and leukocytes. Enzymatic calorimetric method, molybdate method, and haemocytometry according to Nutt method and haemoglobin cyanide method were used to analyze blood samples. The live weight of the experimental ducklings increased by 5.0%, showing a positive effect of probiotic supplementation, whereas their feed consumption per kilogram of weight gain decreased. Their hemoglobin content and red blood cell count increased. Although the number of lactobacilli and bifidobacteria increased, the number of Escherichia coli cells decreased 2.15-fold (p<0.05). The addition of the probiotic supplement contributed to improving the digestibility of protein and fat by 1.6%, fiber by 3.4%, and nitrogen-free extractive substances by 4.7%. The broiler ducklings had high meat quality indicators, including dressing percentage, which increased by 5.4%, whereas the fat content decreased by 1.3%. The introduction of the probiotic supplement in the diet of broiler ducklings improved their growth indicators and increased the number of lactobacilli and bifidobacteria while decreasing the number of E. coli cells in the intestine. It not only improved the meat quality but also increased the profit from $0.392 per bird in the control group to $0.472 per bird in the experimental group. Therefore, this probiotic supplement is a good alternative for raising ducklings in large enterprises and farms. The study limitations may be that the results are only applicable to broiler ducklings. The use of lactobacilli may yield different results in other bird breeds or broiler ducklings in different age groups. Keywords: metabolism, microbiocenosis, Pekin ducks, probiotic supplementation, production.

Rattus spp. are the most common animals capable of adapting to their environment. They can be reservoirs or vectors of diseases that facilitate the transmission of zoonotic-borne parasites to humans. Hence, a study on the detection of parasites in rat populations in urban areas is crucial to prepare for emerging zoonosis. Therefore, this study aims to identify blood parasites, ectoparasites, and helminths in Rattus spp. from wet markets located in Klang Valley, an urban area with a high-density human population. A total of 32 rats were trapped in several wet markets in Klang Valley, Malaysia. They were anesthetized for morphometric examination followed by exsanguination. Various parasitological techniques such as perianal tape test, simple flotation, direct examination of the intestine, and fecal smear were performed for intestinal parasite detection; hair plucking, skin scraping, and full body combing for ectoparasite identification; and blood smear, microhematocrit centrifugation, and buffy coat techniques for blood parasite detection. The rats were identified as Rattus rattus (71.9%) and Rattus norvegicus (28.1%). The only blood protozoan found was Trypanosoma lewisi. The ectoparasites identified belonged to two broad groups, mites (Laelaps spp. and Ornithonyssus spp.) and fleas (Xenopsylla cheopis), known to be parasitic zoonotic disease vectors. The zoonotic intestinal parasites were cestodes (Hymenolepis nana), nematodes (Nippostrongylus brasiliensis, Strongyloides spp., Trichuris spp., Capillaria spp., and Syphacia spp.), and intestinal protozoa (coccidian oocysts and Giardia spp.). Microscopic images showing Giardia spp. are the first report of this organism in rats in Malaysia. Rats caught in this urban area of the Klang Valley harbor parasites can pose a potential zoonotic threat to humans, raising public health concerns because of their proximity to densely populated urban areas. Keywords: ectoparasites, endoparasites, Klang Valley, rats, zoonosis.

Bovine viral diarrhea (BVD) virus (BVDV) is an important viral pathogen of cattle that can infect diverse artiodactyl species. The clinical manifestations caused by BVDV in heterologous hosts, as they do in cattle, vary, although respiratory and reproductive failures are commonly reported. BVDV infections commonly result in reproductive failure in goats, with abortion being the primary clinical sign. In central Java, Indonesia, BVDV infection has been reported in two clinically healthy local goat species, and the testing indicated infection by BVDV Type 1. However, the genetic diversity of viruses has not been described in healthy or ill goats. The objectives of the present study were as follows: (1) To investigate the genetic variation of BVDV isolated from Sapera dairy goats with naturally occurring reproductive disorders in Yogyakarta, Indonesia, using the 5' untranslated region (5' UTR) and (2) to study the possible correlation between reproductive disorders and the presence of BVDV in the flock. Blood samples were collected in October 2021 from 39 goats that had been individually reported to have reproductive disorders. The serum samples were subjected to molecular detection and genetic characterization of BVDV based on the 5' UTR of the viral genome, followed by sequencing and phylogenetic analyses. Viral isolation was performed on BVDV-positive samples to analyze the viral biotypes. BVDV infection was detected in five out of 39 female goats. The clinical status of the BVDV-infected goats was abortion (n=2), metritis (n=1), and repeated breeding (n=2). All antigen-positive samples were confirmed as BVDV type 1a (BVDV-1a) and noncytopathic (NCP)-BVDV biotype. The BVDV-1a and NCP biotypes are the main subtypes and biotypes present in Sapera dairy goats exhibiting reproductive failure. This result is consistent with previous results in dairy cattle in Yogyakarta. The reported results can facilitate the design of methods for the prevention and control of BVD circulating in Indonesia. Keywords: 5'untranslated region, bovine viral diarrhea virus, reproductive disorder, Sapera dairy goats.

Interstitial fibrosis is the final stage of chronic kidney injury, which begins with an inflammatory process. Crude Ganoderma applanatum polysaccharides are known to have anti-inflammatory properties. The potential role of crude G. applanatum polysaccharides in renal fibrosis through pro-inflammatory cytokines needs further investigation. This study aimed to determine the renoprotective effect of crude G. applanatum polysaccharide extract in mice with carbon tetrachloride (CCL4)-induced early kidney fibrosis. This study was conducted for 4 weeks using 24 male BALB/c mice selected for their metabolic stability. The mice were randomly divided into six groups, including control (CG), model (MG), silymarin group and crude G. applanatum polysaccharide extract groups comprising doses of 25, 50, and 100 mg/kg body weight. After sacrificing the mice, whole blood was analyzed for urea and creatine levels, and kidney tissue was prepared to assess tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), hyaluronic acid (HA), and laminin levels, both using enzyme-linked immunosorbent assay. Kidney histology was determined using hematoxylin and eosin staining, while the extracellular matrix (ECM) components were stained using Masson's trichome staining. The α-smooth muscle actin (α-SMA) concentration was determined using immunohistochemistry. These parameters were measured to determine the effectiveness of the crude G. applanatum polysaccharide extract in preventing interstitial fibrosis. Administration of crude G. applanatum polysaccharides effectively prevented increases in kidney weight and physiological enzymes, pro-inflammatory cytokines, and ECM production compared with those in the MG, as evidenced by the low levels of urea, creatinine, TNF-α, IL-6, HA, and laminin. Histopathological results also showed that crude G. applanatum polysaccharides prevented the occurrence of inflammatory infiltration, desquamated nuclei, cytoplasm debris, rupture at the brush border, dilatation of the glomeruli space and lumen of the proximal tubule, and necrotic cells compared with the MG. Masson's trichrome staining revealed lower collagen levels in the interstitial tubules of kidney tissue than those in the MG. Immunohistochemical analysis revealed low α-SMA expression in the crude G. applanatum polysaccharides treatment groups than that in the MG. The crude polysaccharide extract of G. applanatum has a protective effect that prevents the progression of kidney fibrosis in mice. Keywords: anti-inflammatory, carbon tetrachloride, fibrosis, Ganoderma applanatum, kidney.

Trypanosomiasis is a disease caused by unicellular protozoan parasites. Small ruminants succumb to trypanosomiasis in areas of high tsetse fly challenge, resulting in serious economic loss often to farmers in low-input smallholder systems. At present, trypanosomiasis is treated with trypanocidal drugs, but access to these can be limited, and increasing parasite resistance raises questions about their efficacy. The development of trypanotolerance in small ruminant flocks through targeted breeding strategies is considered a sustainable and economical option for controlling African trypanosomiasis. Recently, quantitative trait loci (QTLs) associated with trypanotolerance traits in sheep have been reported. The results of these studies form the basis for more studies to identify QTLs associated with trypanosomiasis resistance, particularly in African livestock species. For example, signatures of positive selection for trypanotolerance have been identified using genome-wide single-nucleotide polymorphism data. However, there are several challenges in performing genetic analyses using data from low-input smallholder systems, including a lack of recorded pedigree and production records and the need for large sample sizes when flock sizes are often fewer than 50 animals. Breeding strategies to improve trypanotolerance should also preserve existing genetic diversity as well as minimize excessive genetic introgression by trypanosusceptible breeds. This review discusses the possibilities of breeding for trypanosome tolerance/resistance in low-input/low-output small ruminant production systems. Potential challenges are outlined, and potential available genetic resources are described as a foundation for future work.

The Samosir goat has a high cultural value and is a source of germplasm in Indonesia. This study aimed to reveal the history and selection signatures of the Samosir goat. A total of 25 goats were divided into seven subpopulations of Indonesian goat breeds. Deoxyribonucleic acid (DNA) from blood samples was isolated with the use of the gSYNC™ DNA Mini Kit (Geneaid, Taipei, Taiwan). Cytb gene amplification was performed by the polymerase chain reaction (PCR) method, and the PCR products were sequenced. A phylogenetic tree was constructed by the neighbor-joining method using MEGA 11 software. A questionnaire was used to collect information related to the history and breeding practices of the Samosir goat on Samosir Island. Samosir goats are divided into four groups based on their coat color: Completely white, white with brown spots, white with black spots, and white with brown and black spots. The body form of the Samosir goat is similar to that of the Kacang goat. The space below a traditional Toba Batak house is used as a goat pen. The genetic difference between the Samosir goat and the Kacang goat based on the Cytb gene was approximately 0.1%. Phylogenetic analysis between Samosir goats and other indigenous Indonesian goats revealed that Samosir goats form a single clade, with a very close genetic distance from other local goats, such as the Kacang goat. The Toba Batak culture on Samosir Island has significantly influenced the selection and formation of the Samosir goat breed. Keywords: Batak tribe, ethnobiology, mitochondrial DNA, phylogenetic analysis, Samosir goat.

To improve overall milk quality in Thailand, dairy farmers and milk collection centers employ a payment program based on milk quality (PPBMQ) for milk trade. This study aimed to determine and compare the proportion of dairy farmers receiving benefits from the PPBMQ using data from selected dairy cooperatives located in northern and central regions in Thailand. Monthly data on milk components (n=37,077), including fat, solids not fat (SNF), and somatic cell counts (SCC) were collected from the two regions in 2018 and 2019. Based on the PPBMQ, farmers were classified into benefit-gain, benefit-loss, and no-benefit groups. A mixed-effects logistic regression model was used to compare the number of farmers in northern and central regions who received monthly benefits from the PPBMQ. More than 70% of dairy farmers benefited from the PPBMQ. The proportion of dairy farmers in the benefit-gain group was higher in the northern region (88.7%) than in the central region (57.1%). A high percentage of dairy farmers in the central region lost their benefits mainly due to SCC (40%) and SNF (44%). The PPBMQ benefited the vast majority of dairy producers in the northern region and approximately two-thirds of those in the central region. Thus, the efforts of authorities and stakeholders should be enhanced to support dairy farmers in the central region in improving milk quality. Keywords: benefits, bulk tank milk, milk compositions, milk quality, payment program, somatic cell counts.

Human health problems due as a microbial resistance or tumors and cancers because consumption of the carcasses containing residues of tetracycline are main global problems in the context of fight against antimicrobial resistance phenomena. Explanation of the sustainable development goals, particularly point 3, is well recognized that all animal products for human consumption must be safe to live a healthy life. This study aimed to design a prototype of rapid test devices (RTD) based on principles of precipitate to obtain a specific color change after the process of reactions as an indicator to determine tetracycline residues in the carcass. Five samples of tetracycline-containing poultry carcasses using artificial add the tetracycline at pharmaceutics grade were examined using a prototype of a strong reaction solution for tetracycline fixation based on the concept bonded by ion Fe(III) at atom O in position atom C-1 at the ring of tetracycline and ion N+ as the functional branch of tetracycline. RTD detection was evaluated using a yellow color presentation and an absorbance spectrometric technique at a wavelength of 273 nm. The following chemicals were used to create the best-fixed tetracycline residue: HCl and H2SO4 dissolved in H2O, chromatographic grade of 0.1 N and 0.5 N of HNO3, and 1% Fe (III) Cl. The RTD had a higher limit of detection (LOD) than the ultraviolet-visible spectrophotometer. The results of this study revealed that RTD, as constructed in this study, can be used to detect residue at LOD 44.764 μg/mL during 120 min of exposure through a light-emitting diode at 980 nm wavelength (p<0.05). The necessity for using RTD was because of the apparent limitations of conventional devices. Keywords: antimicrobial resistance, color indicator, health lifestyle, light-emitting diode, precipitate principles.

Antimicrobial resistance (AMR) is a crucial and emerging multifactorial "One Health" problem involving human and animal health, agriculture, aquaculture, and environment; and posing a potential public health hazard globally. The containment of AMR justifies effective surveillance programs to explicate the magnitude of the problem across the contributing sectors. Laboratory-based AMR testing and characterization is the key component of an AMR surveillance program. An AMR surveillance program should have a "top management" for fund mobilization, planning, formulating, and multilateral coordinating of the surveillance activities. The top management should identify competent participating laboratories to form a network comprising a reference laboratory and an adequate number of sentinel laboratories. The responsibilities of the reference laboratory include the development of standardized test methods for ensuring quality and homogeneity of surveillance activities, providing training to the laboratory personnel, and in-depth AMR characterization. The sentinel laboratories will take the responsibilities of receiving samples, isolation and identification of microbes, and initial AMR characterization. The sentinel laboratories will use simple antimicrobial susceptibility test (AST) methods such as disk diffusion tests, whereas the reference laboratories should use automated quantitative AST methods as well as advanced molecular methods to explicit AMR emergence mechanisms. Standard guidelines set by Clinical Laboratory Standards Institute or the European Committee on Antimicrobial Susceptibility Testing, should be followed to bring about conformity and harmonization in the AST procedures. AMR surveillance program in animals is eventually similar to that in human health with the exception is that veterinary antibiotics and veterinary pathogens should be given preference here. Hence, the review study was envisaged to look deep into the structure of the AMR surveillance program with significance on laboratory-based AMR testing and characterization methods.

The use of antibiotics in animals for disease prevention and productivity has been banned in the European Union since 2006. Possible alternatives can be used prebiotics, probiotics, and synbiotics. These compounds can improve feed digestion and absorption in the gastrointestinal tract with identical nutrient uptake, while imparting the feeling of satiety, which reduces the activity of ghrelin-immunoreactive (IR) cells. The number of studies performed on the activity of ghrelin-IR cells in ruminants is insufficient. In particular, there are few such studies in calves during the transition period from being a relatively monogastric animal to a ruminant. The present study aimed to evaluate the effect of Jerusalem artichoke flour (containing ∼50% prebiotic inulin) and a new, commercially unavailable synbiotic (combination of Jerusalem artichoke flour and Saccharomyces cerevisiae strain 1026) on the amount of ghrelin-IR cells in the abomasum and intestines of 13-14-week-old calves. Fifteen crossbreed, Holstein Friesian and Red Holstein calves (Bos taurus) (32±4 days, 72.1±11.34 kg) were used. Calves were allocated into three groups: Control group (CoG, n=5) received the standard diet, prebiotic group (PreG, n=5) received 12 g of flour of Jerusalem artichoke (Helianthus tuberosus) per head containing 6 g of prebiotic inulin in addition to the standard diet, and synbiotic group (SynG, n=5) received a synbiotic in addition to the standard diet which consisted of two different products: 12 g of flour of Jerusalem artichoke per head containing 6 g of prebiotic inulin and probiotic 5 g of a yeast S. cerevisiae strain 1026. Feed additives were added to the concentrate once a day for 56 days. On days 1, 28, and 56, the live weight of the calves was determined. On day 56 of the experiment, three calves from each group were slaughtered. Histological samples were collected from the two parts of each calf abomasum: Pars pylorica and pars fundalis and the middle part of the duodenum and jejunum. Immunohistochemical tissue staining methods were used to detect ghrelin-IR cells. The live weight of the slaughtered calves on day 56 was 115.3±21.73 kg in CoG, 130.0±17.32 kg in PreG, and 119.0±7.94 kg in SynG. Ghrelin-IR cells were more abundantly localized in the cytoplasm of the abomasum muscle gland cells in pars fundalis and pars pylorica, and to a lesser extent in the duodenum and jejunum. The number of ghrelin-IR cells in the abomasal fundic gland area was significantly higher in the CoG, than in the PreG and SynG (p=0.0001), while the difference between the PreG and SynG was not significant (p=0.700). The addition of Jerusalem artichoke flour and its combination with the yeast S.cerevisiae stain 1026 in calves resulted in a lower number of ghrelin-IR cells in the abomasum, duodenum, and jejunum and, although insignificantly, increased live weight (p=0.491), suggesting that calves in these groups with the same feed intake as the CoG had a better breakdown of nutrients, thus having a longer feeling of satiety. Keywords: calves, ghrelin, inulin, Jerusalem artichoke, Saccharomyces cerevisiae.

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

One of the emerging viral diseases in freshwater fish is Tilapia lake virus (TiLV), which infects all stages of fish and results in mass mortalities. Previously, a TiLV case was detected in the wild environment in Malaysia that involved tilapia and tinfoil barb. Hence, this study aimed to determine the presence of TiLV in wild tilapia (Oreochromis niloticus) as well as tinfoil barbs (Barbonymus schwanenfeldii) at the similar lake after the initial outbreak in year 2017. Both fish species were sampled from this lake at a month interval for two years and subjected to TiLV detection using reverse transcriptase-polymerase chain reaction and cell culture isolation. Concurrently, bacterial isolation and water quality measurements were performed to deduce their correlation with TiLV occurrence. Other wild fish species and mollusk were also occasionally sampled during the fish inventory activity at this lake. The fish's weight, length, and associated clinical signs were noted throughout the entire study period. Mortality was not observed throughout the whole study period, and results indicated a moderate to high prevalence of TiLV infection in both tilapia and tinfoil barbs. There was no correlation between TiLV infection with the isolation rate of opportunistic bacteria such as Aeromonas spp., Plesiomonas spp., and Edwardsiella spp. in the study site. At the same time, the Pearson correlation test revealed a moderate negative correlation between the water pH with the presence of TiLV (R=–0.4472; p<0.05) and a moderate positive correlation between the water iron content with the monthly detection of Aeromonas spp. in wild tilapia. This is contrary to tinfoil barbs, where there was a moderate negative correlation between the water iron content with the monthly isolation of Aeromonas spp. (R=–0.5190; p<0.05). Furthermore, isolation of TiLV on cell culture-induced viral invasion was resulted in the cytopathic effects. Our results suggest that the wild fish may harbor TiLV for an extended period following a massive die-off event in 2017 without any obvious clinical signs and mortality. The persistency of viruses in the wild may need continuous and effective control as well as prevention strategies. Keywords: Malaysia, Tilapia lake virus, tinfoil barbs, wild tilapia.

Improper attachment of the grounding lead is one of the artifacts and causes difficulty in interpretation of ECG. This study aimed to examine the effects of the position of a ground electrode on electrocardiographic (ECG) parameters in horses. Sixteen Arabian horses without any cardiac problems were included in this study. The animals were divided into two groups, the base-apex lead method 1 (BA1 method) and the base-apex lead method 2 (BA2 method) with the reposition of the ground limb electrode to the xiphoid. ECG recordings (paper speed=25 mm/s and calibration=10 mm/ mV) were performed to obtain six limb leads (leads I, II, III, aVR, aVL, and aVF). The amplitude and duration of P waves and QRS complexes, PR interval, QT interval, and T duration were analyzed. T wave morphology was observed. Heart rate was evaluated by using R-R interval in each recording. Heart rate, P duration and amplitude, PR interval and T duration, and QRS duration and amplitude were not significantly different between the BA1 and the BA2 method, except that the BA2 method had a significantly higher QT interval than did the BA1 method (p<0.05). A higher significance of the percentage of coefficient of variation was seen on the P wave amplitude and the ORS amplitude in the BA1 method when compared to BA2 method (p<0.05). These data indicated that base-apex lead method with reposition of the ground limb electrode to the xiphoid can decrease variation of ECG configuration and might be useful for routine ECG evaluation and monitoring in horses. The limitation of this study was the evaluation of cardiac morphology and function by echocardiography to exclude cardiac problems. In further, the studies should examine the effect of breed, age, body weight, and sex on electrocardiography parameters in horses. Keywords: electrocardiogram, ground electrode, horses, position.

The current stable housing of Colombian Creole Horses severely restricts the animals' locomotion and natural behaviors. In addition, their feed consists of a combination of high-energy concentrates with considerably little forage which potentially leads to locomotor or oral stereotypies. This study aimed to report the frequency of locomotor and oral stereotypies in Colombian Creole Horses in Girardota (Antioquia, Colombia) and associated risk factors. A prospective cross-sectional study was conducted from 2019 to 2020, in which 102 stabled horses aged 28 months and older participated. A questionnaire was developed to collect information on the horses' daily barn routines. The horses were observed twice a day for 2 h for 3 consecutive days to record information related to stereotypy behaviors. The Fisher's exact test and the Mann–Whitney U test were utilized for data analyses. Associations were considered statistically significant at p<0.05. Among the horses evaluated, 32.35% presented at least one stereotyped behavior. The most common was crib-biting (i.e., cribbing), with 17.65% exhibiting this behavior. Age, weight, gender, type of feeding, visual contact between horses, and natural lighting were all associated with oral stereotypies. Crib-biting was most common in young horses (U=1.36, p≤0.05), wind-sucking was more common in lighter weight animals (U=1.45, p=0.01), and lip-smacking was more common in stallions (χ2=9.10, p≤0.01). It is noteworthy that their feeding diet included bran, molasses, and gopher. Horses that did not have visual contact with other horses and those that did not have natural lighting were associated with pica (χ2=9.52, p≤0.02; χ2=3.72, p≤0.05; and χ2=3.72, p≤0.05, respectively). Of locomotor stereotypies, kicking the wall was significant in young animals (U=1.54, p=0.03) and walking in circles in lactating mares (χ2=13.20, p≤0.02). Housing conditions in this study were found to have several risk factors affecting horses that exhibit stereotypic behaviors, and all these factors resulted in a higher frequency of stereotypies. Establishing risk factors for the presentation of abnormal behaviors allows for the implementation of better management practices in the production systems of the Creole Colombian Horse and will help improve their overall welfare. Keywords: behaviours disorders, equines, stereotypies, welfare.

Canine tick-borne pathogens (CTBPs) are an important cause of morbidity in dogs in Thailand. This study aimed to evaluate the occurrence of three CTBPs in clinically normal, owned dogs to understand the risk for the general canine population. We also examined sex, age, tick infestation, and packed cell volume (PCV) of the animals in association with active infection of the CTBPs. A total of 139 dogs were included in the study. Blood samples were collected for thin blood smear, PCV and nested polymerase chain reaction (PCR) assay. Statistical analyses were performed to examine the association between individual factors and CTBP infection status determined by PCR. In addition, sensitivity, specificity, and Cohen's kappa were calculated to assess the utility of routine blood smear. The PCR results showed that 31 dogs (22.3%) were infected with at least one of the three pathogens. The occurrence rate for Ehrlichia canis, Anaplasma platys, and Hepatozoon canis was 2.2% (3/139), 18.7% (24/139), and 2.8% (4/139), respectively. There were two cases of coinfection with A. platys and E. canis. The univariate analyses did not yield any associations between recorded variables and the active infection. Microscopic examination showed good sensitivity and agreement only for H. canis (Sn: 75%, 95% confidence interval: 24.9-98.7, κ=0.85). Our findings confirmed the endemicity of the CTBPs in owned canine population in the study site. In-depth epidemiological investigation would be warranted to elucidate environmental risk factors for CTBP infection. Keywords: Anaplasma platys, canine tick-borne disease, Ehrlichia canis, Hepatozoon canis, polymerase chain reaction, thin blood smear.

Caprine arthritis encephalitis virus (CAEV) is a virus that affects goats all over the world and causes enormous economic losses; as a result, screening for the disease is a priority, especially in Iraq. The present study aimed to estimate the prevalence of CAEV in infected goats using the précised PCR method in Babylon, Iraq. A total of 85 blood samples from goats aged 1 month to ≥6 years were analyzed for CAEV infections using molecular methods. The polymerase chain reaction primer was designed to amplify a 573 bp region of the proviral pol gene. The CAEV tests revealed that five out of 85 goats were positive for CAEV. There were no significant differences in CAEV infection according to goat sex and significant differences according to age. Based on these results, the present study is the first molecular survey to confirm the current CAEV genome in an Iraqi goat flock. Keywords: caprine arthritis encephalitis virus, goats, Iraq, molecular detection, polymerase chain reaction.

Candida albicans is the most prevalent human fungal pathogen. In biofilms, C. albicans becomes more resistant to antifungal agents because of the production of an extracellular matrix (ECM) that protects the yeast cells. This study aimed to determine the effects of hydrolase enzymes and the Bgl2 ligand on monomicrobial and polymicrobial biofilms. Biofilm induction in rats was carried out using streptomycin (25 mg/kg) and gentamicin (7.5 mg/kg) administered orally once per day for 5 days. Rats were injected subcutaneously with cortisone acetate (225 mg/kg) as an immunosuppressant on day 5. In addition, rats were orally administered C. albicans for the single microbial model and a combination of C. albicans with Escherichia coli for the polymicrobial model. Following the biofilm production, the groups were treated with glucosamine (8.57 mg/kg body weight) and Achatina fulica hydrolases (1.5 mL) orally for 2 weeks. The reduction of the biofilm was measured using confocal laser scanning microscopy (CLSM). Data were analyzed using a t-test, with a significance value of 95%. CLSM images revealed a strong association between C. albicans and E. coli in the polymicrobial biofilm. On the contrary, the combination treatment using glucosamine and A. fulica hydrolases reduced the ECM of the single microbial biofilm (53.58%). However, treatment effectiveness against the matrix (19.17%) was reduced in the polymicrobial model. There is a strong association between C. albicans and E. coli in the formation of polymicrobial biofilms. The combination of glucosamine and the A. fulica enzyme can reduce the single microbial biofilm ECM; however, it is ineffective in the polymicrobial model. Keywords: Achatina fulica hydrolases, Bgl2 ligand, Candida albicans, Escherichia coli, intestinal polymicrobial biofilm.

Methicillin-resistant Staphylococcus aureus (MRSA), an important opportunistic pathogen, is a Gram-positive coccus known to be resistant to β-lactam antibiotics. Its virulence depends on a large range of factors, mainly extracellular proteins, such as enzymes and exotoxins, that contribute to causing a wide range of diseases in human and animal species. The major reasons for the success of this pathogen are its great variability, which enables it to occur and thrive at different periods and places with diverse clonal types and antibiotic resistance patterns within regions and countries. Infections caused by antibiotic-resistant S. aureus bring about serious problems in the general population (humans and animals). Infections with these pathogens can be devastating, particularly for the very young, adults and immunocompromised patients in both humans and animals. This study aimed to determine the presence of MRSA in both apparently healthy and sick sheep brought to the veterinary hospital as well as veterinary staff and students on clinical attachment in the hospital. A total of 200 nasal swab samples were collected aseptically from sheep and humans (100 each) for the isolation of MRSA. The samples were processed by appropriately transporting them to the laboratory, then propagated in nutrient broth at 37°C for 24 h followed by subculturing on mannitol salt agar at 37°C for 24 h, to identify S. aureus. This was followed by biochemical tests (catalase and coagulase tests) and Gram staining. MRSA was isolated using Clinical Laboratory Standard Institute (CLSI) guideline and confirmed by plating onto Oxacillin (OX) Resistance Screening Agar Base agar. The antimicrobial susceptibility pattern of the MRSA isolates was determined using the disk diffusion method against 12 commonly used antimicrobial agents. The total rate of nasal carriage of S. aureus and MRSA was found to be 51% and 43% in sheep and humans, respectively. The MRSA prevalence in male and female sheep was 18% and 8%, while 9% and 8% were for male and female human samples, respectively. The antimicrobial susceptibility test showed 100% resistance to OX, cefoxitin, oxytetracycline, cephazolin, and penicillin-G (Pen) by MRSA isolates from humans. Conversely, there was 100% susceptibility to ciprofloxacin, imipenem, and gentamicin; for linezolid (LZD), it was 87.5%, norfloxacin (NOR) (71%), and erythromycin (ERY) (50%) susceptibility was recorded. The MRSA isolates from sheep recorded 100% resistance to the same set of drugs used for human MRSA isolates and were equally 100% susceptible to gentamicin, imipenem, LZD, ciprofloxacin, NOR (92%), and ERY (50%). This study determined the presence of MRSA in sheep and humans from the Veterinary Hospital, Maiduguri. It appears that certain drugs such as ciprofloxacin, imipenem, and gentamicin will continue to remain effective against MRSA associated with humans and sheep. Reasons for the observed patterns of resistance must be explored to reduce the burdens of MRSA resistance. Furthermore, the present study did not confirm the MRSA resistance genes such as mecA and spa typing to ascertain the polymorphism in the X-region using appropriate molecular techniques. Hence more studies need to be conducted to elucidate these findings using robust techniques. Keywords: antimicrobial resistance, humans, methicillin-resistant Staphylococcus aureus, sheep.

To effectively control dengue hemorrhagic fever (DHF), it is necessary to assess the risk of vertical virus transmission in Aedes aegypti mosquitoes. This study aimed to detect dengue virus (DENV) transovarial transmission in A. aegypti collected from DHF patients' residences in Denpasar, Bali. A. aegypti samples were acquired by rearing A. aegypti eggs collected from ovitraps placed in the homes of DHF patients. Ovitraps were installed for 7 days and viewed using a loupe to determine whether there were Aedes spp. eggs present. An immunocytochemical method was utilized with 200 samples, and virus detection was performed using a reverse transcriptase-polymerase chain reaction (RT-PCR). Of the 10 DHF patient houses fitted with ovitraps, four produced positive ovitraps from which larvae developed (house index=40%). Of the 50 ovitraps mounted in the 10 homes, 14 ovitraps were positive and contained A. aegypti eggs (ovitrap index=28%). Of these 14 positive ovitraps containing A. aegypti eggs, 10 ovitraps produced larvae. Immunocytochemical tests were conducted on A. aegypti eggs from the four houses under study. It was found that from the 200 samples collected, 197 samples could be observed, and 11 samples (5.6%) were positive for DENV antigen. RT-PCR examination conducted on mosquitoes reared from the four houses studied obtained a negative virus content result. This study found the presence of DENV antigen to be as high as 5.6%. This means that potential for transovarial transmission exists within DHF patients' homes in Denpasar, Bali. Aedes control strategy in Denpasar should address this finding, in addition to the current approaches which have focused primarily on the elimination of larval breeding habitats and control of adults using insecticidal fogging during outbreaks. Keywords: Aedes aegypti, dengue virus, immunocytochemical, transovarial transmission.