Although cats are not natural hosts for heartworm infections (Dirofilaria immitis), evidence suggests that feline heartworm disease can be detrimental because of a severe inflammatory response. Recent studies have found that infection with bacteria of the genus Wolbachia is the principal cause of acute inflammatory filaria disease; nonetheless, the prevalence of cats naturally infected with heartworms and Wolbachia remains unclear. This study aimed to estimate the prevalence and current distribution of feline heartworm disease and its association with Wolbachia infection in pet and semi-domesticated cats in Bangkok, Thailand. A total of 260 cats (130 pet cats and 130 semi-domesticated cats) were enrolled in this study. Blood samples were placed into ethylenediaminetetraacetic acid tubes for hematological analysis and DNA extraction. A polymerase chain reaction (PCR) was performed to analyze samples for the presence of D. immitis and Wolbachia infections. The prevalence (95% confidence interval [CI]) of D. immitis infection in pet, semi-domesticated, and all cats were 3.9% (1.3-8.8%), 27.7% (20.2-36.2%), and 19.6% (15.0-25.0%), respectively. The prevalence (95% CI) of Wolbachia infection in pet, semi-domesticated, and all cats were 18.5% (12.2-26.2%), 31.5% (23.7-40.3%), and 25.0% (19.9-30.7%), respectively. The prevalence of D. immitis and Wolbachia infections in semi-domesticated cats was significantly higher than in pet cats (p=0.002 and p=0.022, respectively). There was a significant association between D. immitis and Wolbachia infections (p<0.001). There was also a significant association between D. immitis infection and the presence of eosinophilia (p<0.045). From the PCR analysis, it can be concluded that semi-domesticated cats were at higher risk for D. immitis infection than pet cats. There was a significant association between positive D. immitis infection and positive Wolbachia infection. Combinations of anthelmintic and antimicrobial therapy should be considered in heartworm-positive cats. Keywords: cat, heartworm, prevalence, Wolbachia.

There is little understanding about antiparasitic drug prescription trends and implementation to reduce possible drug overuse or misuse worldwide. This study aimed to review sheep parasite control strategies and antiparasitic drug use habits in Latvia. To the best of the author's knowledge, this is the first study in the world that describes how antiparasitic drugs are used and what are the most common drug usage errors in a sheep farm. A semi-structured questionnaire was designed to collect relevant information from face-to-face interviews to assess 22 sheep farmers' knowledge and management procedures in farms. We collected information about animal feeding, herding, parasite diagnostics, and antiparasitic drug usage. The questionnaire summary included information on pasture use, parasite control management, and anthelmintic drug choice/use. Only 36% of farms regularly managed parasite control by analyzing fecal samples for parasites, but prophylactic deworming was employed in all farms. Ivermectin, albendazole, levamisole, and monepantel were used on the farms and most of the farms were multidrug users; 77.3% of the farms used albendazole and 72.7% used ivermectin. The results indicated a lack of parasitological examination and parasite control of the flock, mostly empiric drug selection, incorrect dosing, inaccurate drug administration, drug storage, and use errors. A proactive approach to herd health planning, regular parasitic control, and prophylactic measures may benefit farmers and veterinarians. Keywords: anthelmintics, drug resistance, drug usage errors, sheep.

Dairy ration formulations should consider the synchronization of the rumen degradable protein (RDP) to rumen undegradable protein (RUP) ratio (RDPR) with non-fiber carbohydrate (NFC) to achieve optimum microbial protein synthesis (MPS), reduce feed costs, and reduce N excretion to the environment. This study aimed to investigate the effect of RDPR and NFC synchronization on in vitro digestibility, fermentability, and MPS. The experiment used a 3×3 factorial randomized block design with four replications. The first factor was RDPR (RDPR1=50:50; RDPR2=55:45; RDPR3=60:40) and the second factor was NFC levels (NFC1=30%, NFC2=35%, NFC3=40%). The experimental diets were evaluated using a two-stage in vitro method. The examined parameters included rumen pH, NH3 concentration, total volatile fatty acid (VFA) concentration, the molar proportion of VFAs, rumen microbes (protozoa and total bacteria population), and MPS. Data were analyzed using ANOVA, followed by the Duncan test. The results show that neither RDPR nor NFC affected rumen pH, NH3, total VFA, and the rumen microbe population. The interaction between RDPR and NFC affected the molar proportion of acetate, iso-butyrate, and n-valerate. The combination of RDPR1 and NFC1 produced a lower molar proportion of acetate (49.73%) than the other treatment combinations (>54%). The acetate to propionate ratio was influenced by the NFC levels, in which NFC2 and NFC3 produced the highest ratio (p<0.05). MPS was affected by RDPR and NFC, but not by their interaction. Treatments NFC2 and RDPR3 produced the highest MPS. NFC affected the dry matter and organic matter digestibility (DMD and OMD), with treatment NFC3 resulting in the highest DMD and OMD. The combination of a 60:40 RDPR with 35% NFC resulted in the best synchronization of protein and energy available for MPS and digestion activity in the rumen. Keywords: dairy ration digestibility, microbial protein synthesis, rumen degradable protein, rumen undegradable protein, synchronization.

Brucellosis is a bacterial disease notorious for its ability to infect a wide range of domestic and wildlife animals, as well as humans. This study aimed to determine the seroprevalence of ovine and caprine brucellosis and the associated risk factors in the Province of Bam in Burkina Faso. The individual serological status of 300 unvaccinated sheep and 300 unvaccinated goats was determined by Rose Bengal and indirect enzyme-linked immunosorbent assay (iELISA) serological tests used in parallel. The frequency of behaviors conferring risk of developing this zoonotic disease was determined through two epidemiological questionnaires, which identified known risk factors for the transmission of brucellosis between animals and humans. Individual seroprevalence was estimated at 6.0% (18/300) in sheep and 4.3% (13/300) in goats. The "herd" prevalence of brucellosis was estimated at 60% in sheep while 40% in goats. Positivity in the iELISA serological test was significantly associated with age, sex, and husbandry system in sheep and goats. These results indicate that Brucella melitensis circulates in sheep and goat farms in the Province of Bam in Burkina Faso. As B. melitensis is highly pathogenic to humans, adequate measures must be taken to protect the population against this zoonotic disease. Keywords: brucellosis ovine and caprine, Burkina Faso, Province of Bam, seroprevalence, zoonosis.

Enrofloxacin and tylosin can be combined into an antibiotic formulation which is expected to have a broader range of antibacterial activity against various infections in broilers. Validation method analysis of the levels of these two active compounds needs to be done for future use in pharmacokinetic or residual studies. The present study aims to determine a suitable validation method of isocratic high-performance liquid chromatography (HPLC) to measure the concentration of antibiotic combinations in the broiler liver, kidney, and muscles. The combination of enrofloxacin and tylosin in the liver, kidney, and muscle was validated by HPLC method to find the procedures, processes, equipment, and systems used, consistently provides the appropriate results. The chromatography system consisted of an Octadecyl-silica column of 5 μm in diameter and 150 mm in length with a mobile phase of a mixture of 0.05 M monobasic sodium phosphate (pH 2.5) and acetonitrile (65:35 v/v). The solution was detected at a wavelength of 280 nm, 30°C, a flow rate of 1 mL/min, and an injection volume of 20 μL. The combination antibiotics powder was produced from PT Tekad Mandiri Citra, Bandung, Indonesia, and broiler tissues obtained from day-old chick broilers maintained for 30 days with free antibiotic feed. Validation of a combination solution of enrofloxacin and tylosin shows the linearity values of enrofloxacin and tylosin in the liver, kidney, and muscles as r2=0.9988, r2=0.9999, r2=0.9997, r2=0.9989, r2=0.9978, and r2=0.9962. The accuracy and precision values of enrofloxacin in the liver, kidney, and muscles were 5.53, 6.23, and 6.93, respectively. The values of accuracy and precision of tylosin in the liver, kidney, and muscles were 10.43, 4.63, and 7.16%, respectively. The retention times for enrofloxacin and tylosin were 1.945-2.000 min and 4.175-4.342 min. The limit of detection (LOD) and limit of quantity (LOQ) values for enrofloxacin were 3.03 and 10.1 μg/g, respectively. In contrast, the LOD and LOQ values for tylosin were 9.05 and 30.17 μg/g, respectively. The value of linearity, accuracy, precision, specificity, and sensitivity of the combined solution of enrofloxacin and tylosin showed promising results. Keywords: enrofloxacin-tylosin combination, high-performance liquid chromatography, validation method.

Pasteurella multocida B:2 is the causative agent of hemorrhagic septicemia (HS) in buffalo and cattle. Buffaloes are known to be more susceptible to HS than cattle, but the reason for this remains unknown. This study aimed to compare the in vitro efficiency with which buffalo and cattle macrophages can kill P. multocida B:2. Monocyte-derived macrophages of buffalo and cattle were used in this study. They were exposed to 1×106 colony-forming unit/mL of live P. multocida B:2 before the cells were harvested at 0, 30, 60, and 120 min post-exposure and viewed under a fluorescence microscope to count viable and non-viable macrophages and the macrophages with phagocytosing P. multocida B:2 cells. The phagocytosis, intracellular bacterial killing, and macrophage death rates were calculated and compared between the two species and sampling points. In general, the rates of phagocytosis, intracellular killing, and macrophage death increased with time of exposure for both animal species. No significant (p>0.05) differences were noted between the phagocytosis rates by the macrophages of buffalo and cattle throughout the experiment. However, the rates of intracellular killing were significantly (p<0.05) higher in cattle macrophages at 30 min and 120 min post-exposure than those of buffalo. The death rates of buffalo macrophages were significantly (p<0.05) higher than those of cattle at 60 min and 120 min post-exposure. With higher bacteria killing ability and lower macrophage death, cattle appeared to be more efficient at handling P. multocida B:2 infection than buffalo. Keywords: buffalo, cattle, in vitro efficiency, macrophages, Pasteurella multocida B:2.

The morbidity and mortality of Shigella infections remain a global challenge. Epitope-based vaccine development is an emerging strategy to prevent bacterial invasion. This study aimed to identify the ability of the 49.8 kDa pili subunit adhesin protein epitope of Shigella flexneri to induce an intestinal immune response in mice. Thirty adult male Balb/c mice were divided into a control group, cholera toxin B subunit (CTB) group, CTB+QSSTGTNSQSDLDS (pep_1) group, CTB+DTTITKAETKTVTKNQVVDTPVTTDAAK (pep_2) group, and CTB+ ATLGATLNRLDFNVNNK (pep_3). We performed immunization by orally administering 50 μg of antigen and 50 μl of adjuvant once a week over 4 weeks. We assessed the cellular immune response by quantifying T helper 2 (Th2) and Th17 using flow cytometry. In addition, we assessed the humoral immune response by quantifying interleukin (IL-4), IL-17, secretory immunoglobulin A (sIgA), and β-defensin using enzyme-linked immunoassay. Statistical analysis was performed using one-way analysis of variance and Kruskal–Wallis test. Peptide oral immunization increases the cellular immune response as reflected by the increase of Th2 (p=0.019) and Th17 (p=0.004) cell counts, particularly in the CTB_pep_1 group. Humoral immune response activation was demonstrated by increased IL-4 levels, especially in the CTB+pep_3 group (p=0.000). The IL-17 level was increased significantly in the CTB+pep_1 group (p=0.042). The mucosal immune response was demonstrated by the sIgA levels increase in the CTB+pep_3 group (p=0.042) and the β-defensin protein levels (p=0.000). All selected peptides activated the cellular and humoral immune responses in the intestine of mice. Further studies are necessary to optimize antigen delivery and evaluate whether the neutralizing properties of these peptides allow them to prevent bacterial infection. Keywords: epitope, immune response, oral, Shigella flexneri, shigellosis.

Gouty arthritis is a metabolic disorder involving monosodium urate (MSU) crystal deposition as a key initiator of acute inflammation. Dysregulation of the nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain-containing protein 3 (NLRP3) inflammasome is associated with the pathogenesis of gout through the maturation of interleukin-1β. Piperine (PIP) is a phytochemical with an anti-inflammatory activity that has the potential as an alternative treatment for gout. In this study, we examined the effect of PIP in immunosuppression of gout inflammation through the regulation of the NLRP3 inflammasome. An in silico study was done by pharmacodynamic modeling of PIP in suppressing MSU-induced inflammation through disruption of the NLRP3 inflammasome. In vivo tests, including inflammatory assessment, histopathology, cytology, estimation of lipid peroxidation index, and detection of systemic inflammatory reactants, were performed on two groups using preventive and curative protocols. In silico studies of molecular docking demonstrated the activity of PIP as a competitive inhibitor of the mitogen-activated protein kinases/nuclear factor-kappaB axis, upstream of the NLRP3 inflammasome. Analysis of gout models with curative and preventive protocols revealed the immunosuppression activity of PIP by reducing inflammatory symptoms, inhibiting tophus formation resulting from NETosis, reducing cartilage erosion, inhibiting leukocyte exudation, suppressing lipid peroxidation index, and inhibiting the production of C-reactive protein. The results demonstrate the activity of PIP as an immunosuppressant in gout flare. These findings indicate the potential of PIP as a candidate for prophylactic and therapeutic agent for the treatment of gouty arthritis. Keywords: gout, immunosuppression, monosodium urate, NLRP3 inflammasome, piperine.

Probiotics are beneficial microorganisms that play important roles by adhering to the gut and producing antimicrobial substances to inhibit pathogens. The objective of this study was to isolate and characterize the probiotic lactic acid bacteria (LAB) from Palmyra palm sugar, which can produce antimicrobial compounds against methicillin-resistant Staphylococcus aureus (MRSA), a new zoonotic and food-borne pathogens. Twenty-six LAB isolates were isolated from 30 Palmyra palm sugar samples. Three selected LAB were further characterized as probiotics. In addition, the antibacterial and anti-biofilm-forming activities of the probiotics' culture supernatants against MRSA and food-borne pathogens were investigated. Finally, the selected probiotics were identified by aligning 16S rRNA sequences. The three confirmed probiotics, WU 0904, WU 2302, and WU 2503, showed strong antibacterial activities against S. aureus, MRSA, Escherichia coli O157:H7, and Listeria monocytogenes, as measured by a broth microdilution assay. Among the LAB isolates, 82.22-86.58%, 91.83-96.06%, and 64.35-74.93% exhibited resistance to low pH, pancreatin treatment, and bile salts, respectively. It was found that 59.46% and 83.33% auto-aggregation was observed in 2 and 24 h, respectively. Moreover, 50.25-57.24% adhesion was detected after the incubation of the bacterial cells to Caco-2 cells. . Biofilm inhibition (82.81-87.24%) was detected after the treatment of MRSA with the culture supernatants, when compared with that to the control. By the alignment of 16S rRNA sequences, the isolate WU 2302 was identified as Lacticaseibacillus spp. with 98.82% homology when compared to the GenBank database. This study indicates that isolated probiotics can produce antimicrobial compounds against MRSA and food-borne pathogens. The obtained results strongly suggest that these probiotics are promising candidates for pharmaceutical products. Keywords: antibacterial activity, Lacticaseibacillus spp., methicillin-resistant Staphylococcus aureus, Palmyra palm sugar, probiotics.

Over recent years, antimicrobial-resistant Klebsiella species in humans, animals, food animals, food products, and agricultural environments have been the center of attention due to its role in the evolution of antimicrobial resistance. The emergence of resistance to fluoroquinolones and cephalosporins of third and higher generations in Klebsiella oxytoca has not received much attention in animal husbandry compared to that in Klebsiella pneumoniae. Reports on K. oxytoca are limited in the study area. Therefore, we investigated the antimicrobial susceptibility and resistance genes in K. oxytoca isolated from slaughtered pigs in Thailand. Microbiological examination was conducted on 384 Klebsiella spp. isolates recovered from slaughtered pigs in ten provinces of Thailand. Seventy-two K. oxytoca isolates (18.75%) were examined for antimicrobial-resistant genes (β-lactamase [blaTEM, blaCTX-M, and blaSHV]) and fluoroquinolone-resistant genes (qnrA, qnrB, qnrC, qnrD, qnrS, oqxAB, aac(6')-Ib-cr, and qepA). The most common genotype was blaCTX-M (58/72, 80.55%), followed by blaTEM with blaCTX-M (7/72, 9.72%) and blaTEM (6/72, 8.33%). The most common blaCTX-M group was blaCTX-M-1 (19/58, 32.76%), followed by blaCTX-M-9 (1/58, 1.72%). Plasmid-mediated quinolone resistance genes were identified in 13 (18.05%) isolates: qnrS (16.70%) and qnrB (1.4%). All 13 isolates had qnrS transferable to an Escherichia coli recipient, whereas qnrB was not detected in any transconjugants. Either blaCTX-M or blaTEM harbored by one K. oxytoca strain was transferable to an E. coli recipient. Analysis of antimicrobial susceptibility revealed that more than 90% of the blaCTX-M-carrying K. oxytoca isolates were susceptible to chloramphenicol, trimethoprim, ceftazidime, cefepime, cefotaxime, amoxicillin-clavulanic acid, piperacillin–tazobactam, and fosfomycin. All K. oxytoca isolates (13) harboring qnr were susceptible to carbapenem and ceftriaxone; however, 43 (74.13%) of the K. oxytoca isolates harboring blaCTX-M exhibited extended-spectrum β-lactamase activity. Most of the K. oxytoca isolates from pigs were highly resistant to ampicillin, azithromycin, and gentamicin. To prevent further transmission of Klebsiella spp. between food animals and humans, strict control of antibiotic use in clinical and livestock settings is necessary along with routine disinfection of the livestock environment and efforts to increase awareness of antimicrobial resistance transmission. Keywords: Klebsiella oxytoca, plasmid-mediated quinolone resistance, slaughtered pigs, Thailand, β-lactamase.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a persistent organic pollutant toxic to the human reproductive system. This study aimed to evaluate the effect of α-Tocopherol administration on the male fertility parameters of a rat model exposed to TCDD. Fifty healthy 12-week-old male rats were randomly divided into five groups. Rats in the control group were given corn oil twice daily in 4 h intervals. In the treatment groups, all rats were given TCDD at a dose of 700 ng/kg of body weight (BW)/day for 45 days. Four hours after receiving the TCDD, T0 rats were given corn oil, and T1, T2, and T3 rats were given α-Tocopherol at doses of 77, 140, and 259 mg/kg BW/day, respectively, for 45 days. On day 46, experimental animals were sacrificed to collect blood and testicular samples. TCDD exposure decreased superoxide dismutase activity, plasma membrane integrity, Leydig cell count, sperm cell count, sperm viability and motility, and increased malondialdehyde levels, serum testosterone levels, and sperm morphological abnormalities. The administration of α-Tocopherol mitigated the effects of TCDD exposure, and the 140 and 259 mg/kg BW/day treatments returned those male fertility parameters to normal levels. The administration of 140 mg/kg BW/day α-Tocopherol restored male semen quality in rats exposed to TCDD. We found dynamics serum testosterone levels in rats exposed to TCDD that need to be further studied. Keywords: α-Tocopherol, dioxin, fertility parameters, pollutant, reproductive system.

With the increased concerns about global protein supply, chicken meat, especially from male layer chicken, constitutes an alternative in terms of quality and carcass traits. Probiotics have been proposed for replacing antibiotic growth promoters (AGPs), which have been prohibited as poultry supplement feeds. The present study aimed to determine the efficacy of dietary supplementary probiotics during the starter period on growth performances, carcass traits, and immune organs of male layer chicken. In this study, one hundred and eighty 1-day-old male chicks from the strain ISA brown were used. They were divided into six groups according to the feed: 100% basal feed (T0), basal feed+2.5 g AGP/kg feed (T1), basal feed+probiotics 1 mL/kg feed (T2), basal feed+probiotics 3 mL/kg feed (T3), basal feed+probiotics 4 mL/kg feed (T4), and basal feed+probiotics 5 mL/kg feed (T5). Probiotics (Lactobacillus acidophilus, Lactobacillus plantarum, and Bifidobacterium spp.) were given at a concentration of 1.2×109 colony-forming unit/mL. Virginiamycin was used as AGP. ISA brown layer chicken was treated for 21 days. Growth performances (body weight, feed consumption, and feed conversion ratio [FCR]), carcass traits (weight at slaughter, weight of the carcass, breast muscles, liver, lungs, kidneys, and heart), immune organs (spleen, thymus, and bursa of Fabricius), and non-edible organs (head, legs, and wings) were analyzed. Probiotic supplementation at 4 and 5 mL/kg feed (T4 and T5) during the starter phase improved the body weight, FCR, and feed consumption. The weight at slaughter, weight of the carcass, breast muscles, and liver from the T4 and T5 groups were significantly greater than those in the other treatment groups. In addition, the weight of the heart, lungs, and kidneys was increased in the T1, T2, T3, T4, and T5 groups compared with that measured in the T0 group. Furthermore, there were significant differences regarding the immune organs between the T0 and the other treatment groups. The weight of the head, legs, and wings was also greater in the probiotic and AGP supplementation groups (T1, T2, T3, T4, and T5) than that in the basal feed group (T0). Probiotic (L. acidophilus, L. plantarum, and Bifidobacterium spp.) supplementation at 4 and 5 mL/kg feed during the starter period can be used to improve the growth, carcass traits, and weight of immune organs in male layer chicken. Keywords: feed supplementation, growth performance, ISA brown layer chicken, probiotic, starter period.

As a non-protein nitrogen source, urea is a popular, low cost, and easily obtained protein supplement. The objective of the present study was to perform a meta-analysis of the effects of urea supplementation on rumen fermentation and sheep performance. A total of 32 experiments from 21 articles were compiled into a dataset. The levels of dietary urea varied from 0 to 31 g/kg of dry matter (DM). Parameters observed were rumen fermentation product, nutrient intake, nutrient digestibility, and sheep performance. This dataset was analyzed using a mixed model methodology, with urea supplementation levels as fixed effects and the different experiments as random effects. Increasing levels of urea were associated with increases (p=0.008) in rumen pH, butyrate (C4) production, and ammonia (NH3–N) concentration. Urea supplementation had minor effects on total volatile fatty acids (p=0.242), total protozoa (p=0.429), and the microbial N supply (p=0.619), but tended to increase methane production (CH4; p<0.001). Supplementation of urea increased the intake of dry matter (DM; p=0.004) and crude protein (CP; p=0.001). Digestibility parameters, such as DM digestibility (DMD) and CP digestibility (CPD), also increased (p<0.01) as a result of urea supplementation. Retained N (p=0.042) and N intake (p<0.001) were higher with increasing levels of urea supplementation. In terms of animal performance, supplementation of urea increased average daily gain (ADG; p=0.024), but decreased the hot carcass weight percentage (p=0.017). This meta-analysis reports the positive effects of urea supplementation on rumen fermentation products (i.e., pH, C4, and NH3–N), intake (DM, CP, and N), digestibility (DMD and CPD), and ADG in sheep. Keywords: meta-analysis, sheep, supplementation, urea.

Human mycotic infections are one of the major health problems worldwide. Prolonged use of antimycotic drugs has contributed to the development of resistance in pathogenic fungi. This study was conducted to examine antioxidant and anticandidal activities of Moroccan propolis. Two ethanolic extracts of Moroccan propolis from the Fez-Meknes region were evaluated regarding the following physicochemical parameters: Yield, pH, total carbohydrates, total proteins, total lipids, minerals, total phenolic content, total flavonoid content, and antioxidant activity using ferric reducing antioxidant power (FRAP) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. In addition, we assessed the in vitro anticandidal activity against vulvovaginal candidiasis strains, that is, Candida albicans, Candida glabrata, Candida parapsilosis, and Candida krusei, using the broth micromethod according to the CLSI/M27-A3 reference guidelines. The propolis samples exhibited a mean yield of 16%, with an acidic pH ranging from 4.8 to 5.9; the sample from the Oued Amlil area (OAPEE) contained high levels of resin, balsam, moisture, total carbohydrates, and total lipids: 59.8%, 0.71%, 2%, 1.01 gGlcEq/g, and 120 mg/g, respectively. Moreover, the sample from the Sefrou area (SFPEE) was richer in total proteins and minerals, with values of 2.5 g/100 g and 1.84%, respectively. The total polyphenol and flavonoid content in the propolis extracts were 117.38 and 194.68 mg of gallic acid equivalent/g, and 17.45–27.79 mg of quercetin equivalent/g, respectively. Regarding the antioxidant activity, the most effective propolis extract was the sample from the Sefrou area, at 72.5 μg/mL and 118.78 μmoL Fe2+/g for ABTS-half-maximal inhibitory concentration and FRAP-half maximal effective concentration, respectively. The analysis of phenolic compounds using high-performance liquid chromatography with a diode-array detector revealed the presence of 13 polyphenols. The main compound in the OAPEE sample was epicatechin (310 mg/g), whereas in the SFPEE sample was apigenin (410 mg/g). Regarding the antifungal activity against Candida species, the minimum inhibitory concentration and minimum fungicidal concentration of the Moroccan propolis ethanolic extracts ranged between 31.2 and 62.5 μg/mL and 62.5 and 125 μg/mL, respectively, comparable with fluconazole (as a reference antimycotic). This study suggests that Moroccan propolis (31.2 and 125 μg/mL) may be an important source of bioactive molecules with anticandidal activity. Propolis may be a promising naturally-occurring candidate for the development of antimycotic drugs. Keywords: antioxidant activity, propolis, total flavonoids, total polyphenols, vulvovaginal candidiasis.

Antibiotic, improves the growth performance of laying hens when used as a feed additive; however, it has been banned in Europe. Furthermore, secondary metabolites used as a substitute for antibiotics are compounds produced by plants. Therefore, this aims to determine the effect of dates water extract (DWE) on the performance of laying hens. This study used dates containing secondary metabolites as a feed additive and substitute for antibiotics. A completely randomized design was used, dividing 400 Lohmann brown day old chick into five groups (each group has five replications and each replication consisted of 16 laying hens). Furthermore, there were two control groups such as mineral water control group and antibiotic growth promoters (basal feed+50 mg/kg of bacitracin), and three DWE groups such as 5% DWE (50 mg/mL), 10% DWE (100 mg/mL), and 20% DWE (200 mg/mL). Dates extract treatment was administered through drinking water for 54 days, whereby three laying hens from each replication were taken randomly and decapitated on the neck. Afterwards, a necropsy was performed for histological preparations of the small intestine, ovary, and lymphoid organs. The structure and morphology of the small intestine, and ovaries were observed through histological preparations, while lymphoid organs were observed through histological preparation and morphometry, and body morphometry, body weight, feed intake and weight gain were observed by measurements and weighing. Small intestine morphology, ovarian follicle, and growth performance of the DWE2 group increased significantly compared to the control group, but the lymphoid organs index was influenced by DWE1. The administration of 10% dates extract (100 mg/mL) in drinking water improves the morphology of the small intestine, ovarian follicles, lymphoid organs, and growth performance. Keywords: dates water extract, feed additive, ovary, performance, small intestine.

The reproductive performance of dairy cows is of a high economic value to maintain efficient and sustainable productivity. Postpartum anestrus is one of the major infertility problems that cause limitation of dairy cow reproductive performance and productivity. The previous studies have reported using a polyherbal formulation for different purposes, including reproductive performance. This study was conducted to examine the efficacy of polyherbal formulation administration on the reproductive performance of cows and to establish the oral administration of polyherbal formulations as a safe, effective, and economic treatment for dairy cows with postpartum anestrus due to nutritional disorders, negative energy balance, high milk production, and/or heat stress. A total of 14 dairy cows with postpartum anestrus were randomly divided into two groups, which were subjected to oral administration of distilled water that served as the control (Ctrl; n=5) or polyherbal treatment (polyherbal treated; n=9) for two shots of treatment (each for 3 successive days) with a 10-day interval. Blood sampling and ultrasonography were performed before treatment and after the first and second shots of treatment. Progesterone (P4) assay was also performed. The estrus induction rate was 66.7% after the first shot of treatment in the polyherbal-treated group, which increased significantly to 88.9% after the second shot of treatment, compared with the Ctrl group (20%). The estrus response was confirmed by ultrasonography and P4 hormone assay, wherein the polyherbal treatment significantly increased the P4 concentration in the polyherbal-treated group after the second shot of treatment compared to that before treatment and in the Ctrl group at all time points. On pregnancy diagnosis, the treated dairy cows showed conception rates of 66.7% and 20.0% in the polyherbal-treated and Ctrl groups, respectively. In contrast, the concentration of malondialdehyde, an oxidative stress marker, and the total antioxidant capacity remained unchanged between both groups before and after treatment. The polyherbal formulation containing tubers of C. rotundus, M. pterygosperma, rhizome of Z. officinale, and A. cepa has the potential to induce estrus response and luteal activity in dairy cows and is a possible treatment for ovarian inactivity in dairy farms. Keywords: estrous cycle, ovarian inactivity, phytotherapy, reproductive herbal medicine.

Bovine cysticercosis is defined as a foodborne parasitic zoonotic disease of cattle caused by the larval stage of the human tapeworm Taenia saginata. In Botswana, bovine cysticercosis has inflicted major economic consequences on the beef industry due to downgrading, condemnation, or treatment of infected carcasses. Thus, in this study, we aim to (1) estimate the prevalence of bovine cysticercosis in Botswana's southeastern districts, (2) describe the distribution of bovine cysticercosis through geospatial mapping, and (3) investigate the effect of seasonality on bovine cysticercosis occurrence. A retrospective study was conducted using abattoir records of cattle slaughtered from August 2015 to July 2016. In total, 13 licensed non-export abattoirs were selected for this study, wherein 26,827 cattle were slaughtered during this period. Detection of cysticerci from the carcass and offal was carried out by meat inspectors visually during meat inspection. Prevalence of bovine cysticercosis was calculated for the extension areas and veterinary districts for each month and form there used to establish its distribution and seasonality. Data were analyzed using SPSS version 25.0. The prevalence of bovine cysticercosis in the southeastern districts of Botswana during this study period was determined to be 6.2%. The prevalence in the veterinary districts differed significantly at p<0.05. Seasonality did not have a significant (p=0.651) effect on the prevalence of bovine cysticercosis. Geospatially, areas with greater than 8% prevalence were mainly located in the southernmost part of the study area. The prevalence of bovine cysticercosis was 6.2% during the study period. No previous studies on cysticerci prevalence in the study area was conducted; thus, it was not possible to determine whether there has been an increase or decrease in terms of prevalence rate. Therefore, the results of this study can be used as a baseline for the prevalence of cysticerci in the study area. Keywords: bovine cysticercosis, prevalence, geospatial distribution, meat industry, seasonal occurrence.

Humans are more frequently exposed to chemicals in daily life by inhalation of indoor and outdoor air. However, abusers and workers are the most exposed to those chemicals and their health risks particularly, liver diseases. The present study investigated the protective effects of pomegranate juice (PJ) (Punica granatum) and pomegranate peel aqueous extract (PAE) supplementation against toluene (Tol)-induced hepatotoxicity in Wistar rats. A phytochemical analysis and assessment of the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity were performed for the PJ and the PAE. The in vivo study was carried out using 70 Wistar rats that were divided into seven groups, each consisting of 10 rats. All groups were treated orally for 6 weeks as follows: Control (C), positive controls (CO: 1.25 mL/kg body weight [BW]; PJ: 4 mL/kg BW; and PAE: 400 mg/kg BW), Tol (550 mg/kg BW), and a mixture each of PJ-Tol and PAE-Tol. At the end of the 45th day of study, the hepatic biochemical markers (transaminases, alkaline phosphatase, total bilirubin, albumin, and total proteins) were auto-analyzed, as well as histology and oxidative stress markers of the liver were evaluated. The phytochemical analysis revealed that the DPPH scavenging activity and the total phenolic, flavonoid, and tannin contents were higher in the pomegranate peel extract versus the juice. The results also showed that Tol significantly increased liver enzyme activities and total bilirubin levels, whereas albumin and total proteins were significantly decreased. Similarly, Tol provoked a significant increase in hepatic malondialdehyde levels, with a decrease in glutathione content and glutathione peroxidase activity. The biochemical changes agreed with the hepatic histological alterations. A significant improvement in all parameters was observed in the PAE-Tol group compared with the PJ-Tol group. Exposure to Tol altered the hepatic antioxidant and biochemical parameters and histological profile of the rats, and PAE was more powerful than PJ in reducing Tol liver injuries through its antioxidant activity. Keywords: antioxidant activity, liver injuries, oxidative stress, Punica granatum, toluene.

Muscle atrophy is common in Parkinson's disease (PD). Although myostatin has been implicated in muscle atrophy, its expression in PD skeletal muscle has not been investigated. Therefore, this study aimed to elucidate the influence of PD induction and exercise training on myostatin expression in the gastrocnemius skeletal muscle. Thirty albino mice were randomly selected and separated into three groups of 10 mice each: Sedentary control, sedentary PD (SPD), and exercised PD (EPD). 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine and probenecid were used to induce chronic parkinsonism in the PD groups. Immunohistochemistry was used to investigate the expression of myostatin and nuclear factor kappa B (NF-κB) in gastrocnemius muscles of all three groups. Myostatin expression and NF-κB nuclear localization, indicative of its activation, were significantly (p<0.01) higher in gastrocnemius skeletal muscle in the SPD group than in the control and EPD groups. Concomitantly, the average cross-sectional area of gastrocnemius muscle fibers in the SPD albino mice was significantly smaller (p<0.01) than in the control and EPD groups, indicating muscle atrophy. The present data are the first to indicate a correlation between PD induction and myostatin overexpression and NF-?B activation in the gastrocnemius muscle, potentially promoting the muscle atrophy commonly seen in PD. Additionally, the current data are the first to indicate the beneficial effects of exercise training on PD-associated myostatin overexpression, NF-κB activation, and muscle atrophy. Thus, our data are the first to suggest that myostatin and NF-κB might be regarded as potential therapeutic targets in an attempt to ameliorate skeletal muscle abnormalities commonly observed in PD. Keywords: gastrocnemius, immunohistochemistry, myostatin, nuclear factor kappa B, Parkinson's disease.

Tropaeolum tuberosum Ruiz and Pavón, a tuber native to South America, is characterized by its antioxidant, antimicrobial, and anti-inflammatory properties that contribute to wound healing. This study aimed to evaluate the healing effect of the topical T. tuberosum formulations (gel and cream) on induced wounds in mice. Here, an acidic ethanolic extract (1.5 N hydrochloric acid and 96% ethanol at the ratio of 15:85, v/v) was prepared with the tubers of T. tuberosum ecotype black and incorporated into topical cream and gel formulations at 1%. Thirty-twoBalb/c mice were divided into four experimental groups receiving daily topical treatments for 14 days: Group I (control; no treatment), Group II (a commercial ointment of neomycin, polymyxin B, and bacitracin), Group III (1% T. tuberosum gel), and Group IV (1% T. tuberosum cream). The wound closure in the mice was determined during the treatment; then, they were euthanized to obtain skin samples for histopathological analysis. Groups III and IV showed a higher percentage of wound closure from the 6th day of the treatment. From the 8th day, the effect was greater in Group III. The healing effect was confirmed by the histopathological changes. This study concludes that the topical formulations of T. tuberosum demonstrate wound-healing activity in mice, and the most effective treatment is the 1% gel formulation. Keywords: histology, histopathological changes, skin, topical formulations, Tropaeolum tuberosum, wound healing.

The emergence of drug-resistant strains of Eimeria spp. calls for the development of novel anticoccidial drugs. Plant extracts provide a possible natural source for such drugs. This study aimed to investigate the in vitro anticoccidial activity of encapsulated bromelain (EB) in chitosan nanocarriers on Eimeria spp. oocysts isolated from goats kept by farmers in Kenya. Bromelain was extracted from the peel of ripe pineapples using standard methods. Eimeria spp. oocysts were isolated from the feces of goats using a flotation method. The inhibition of sporulation was assayed after exposing the oocysts to solutions of EB, non-EB (NEB), and diclazuril (positive control) at concentrations between 4 mg/mL and 0.125 mg/mL for 48 h. The oocysts were examined under a microscope (40x) to determine the effects of the drugs on the sporulation process. The percentage of sporulation inhibition was calculated after 48 h and the inhibition concentration 50% (IC50) was determined by probit analysis. Bromelain manifested anticoccidial activity through the inhibition of the sporulation of coccidia oocysts. EB achieved inhibition with a lower dose compared with NEB. The IC50 values of diclazuril, EB, and NEB were 0.078 mg/mL, 0.225 mg/mL, and 0.575 mg/mL, respectively. There were significant differences (p<0.01) between the IC50 of EB and NEB compared with the standard treatment drug. This preliminary study showed that EB has anticoccidial activity supporting further evaluation at an in vivo level to develop a novel drug for the management of coccidiosis in goats. Keywords: anticoccidial activity, bromelain, chitosan, coccidia, goat, nanoencapsulation.

Pig production remains crucial to the livelihood of farmers in Zambia. However, low production continues to undermine efforts to reduce animal protein deficit; hence, the need emerges to improve production through biotechnology. To contribute to the prediction of their acceptance, this study assessed the attitudes and preferences of traditional farmers toward reproductive biotechnologies by exploring the socio-demographic characteristics of farmers. The study conducted a cross-sectional descriptive survey that employed a mixed-methods design with a qualitative-quantitative methodological triangulation. Quantitative data were obtained from 622 respondents using a questionnaire, whereas seven focus group discussions (FGDs) were conducted to obtain qualitative data. Descriptive statistics and thematic analysis were used to analyze quantitative and qualitative data, respectively. The majority (65.1%) of the respondents were low-income earners who mainly (64.8%) attained primary education. In addition, pig farming was dominated by middle-aged (43.7%) and elderly (40.7%) individuals. Moreover, most of the respondents owned (51.3% and 78.0%) more than 2 acres and six pigs, respectively. Furthermore, the respondents expressed a positive attitude (3.84±0.42) toward reproductive biotechnology application. However, despite supportive opinions (4.17±0.54) and favorable behavioral intentions (4.09±0.51), their feelings were generally neutral (3.10±0.89). In addition, the study observed various contrasting attitudes across socio-demographic factors. The respondents mainly preferred artificial insemination (AI; 66.2%). The results of the FGDs supported the survey findings. Nevertheless, the lack of information, knowledge and practical exposure, absence of peer influence, perceived beliefs and risks, poverty situations, and gender issues were pinpointed as the identified barriers to the biotechnology acceptance of the participants. The respondents generally supported reproductive biotechnology application and its contribution to improved production. However, further promotion of the favorable attitudes of the farmers will be required. In this case, interventions sensitive to their socio-demographic characteristics, perceived barriers, and identified contributing factors to favorable attitudes will be crucial. In addition, despite the overwhelming preference for AI, efforts to promote AI-supporting reproductive technologies are required because they contribute to AI success rate. Keywords: attitudes, indigenous pig production, reproductive biotechnology, survey, traditional farmers.

Parasitic invasions, especially gastrointestinal nematodes, are widespread and are one of the main problems in sheep farms. For this reason, sheep are dewormed more often than other livestock species, often several times a year. Concerns about antiparasitic resistance from the farmers and veterinarians have arisen because, on some farms, antiparasitic drugs are used routinely for very long periods. There are no data available on anthelmintic resistance in gastrointestinal nematodes in sheep in Latvia. Our work aimed to determine the most common endoparasites in sheep and the degree of anthelmintic resistance on sheep farms in Latvia. All sheep (577) underwent a coprological examination before the start of the study, and only sheep diagnosed with more than 200 McMaster eggs per gram of feces were included in the study. A fecal egg count reduction (FECR) test was performed on 20 sheep flocks in Latvia. In Latvia, sheep were most commonly infected with Eimeria spp. 97% (confidence interval [CI] 95% 96-98). The second most commonly diagnosed species were Trichostrongylidae 91% (CI 95% 89-93) and Strongyloides spp. 76% (CI 95%, 72-79). The ivermectin (IVM) FECR was 0.74% (0.73-0.74), showing resistance in all sheep farms included in this study. Albendazole (ABZ) FECR 0.89% (0.88-0.89) was effective. This study showed that the most popular deworming drugs (ivermectin, albendazole) in Latvia are ineffective in sheep. Additional studies on the use of IVM+ABZ combinations for deworming sheep should be performed. Keywords: anthelmintic resistance, gastrointestinal parasites, sheep.

Among several factors, the sperm quality of poultry is affected by the rooster's body size and the availability of antioxidants like vitamin E. This study aimed to determine the effect of dietary vitamin E supplementation on rooster sperm quality through a meta-analysis. After verification and evaluation, a total of 19 articles were included in this study. Data, including dietary vitamin E, semen volume, concentration, total sperm cells, pH, motility, viability, percentage of dead and abnormal sperm, vitamin E sperm content, malondialdehyde (MDA) content, and testosterone levels, were tabulated in a database; these were subsequently analyzed using mixed modeling with vitamin E dose as a fixed effect and study identity as a random effect. Dietary supplementation level of vitamin E significantly (p<0.001) affected sperm concentration, significantly affected motility (p<0.001), significantly affected sperm vitamin E (p<0.001), significantly affected viability (p<0.001), and significantly affected chicken sperm fertility (p=0.001). Vitamin E administration also significantly reduced the number of sperm cell deaths (p<0.001); however, increased dietary levels of vitamin E did not affect semen volume (p=0.853), pH (p=0.951), MDA (p=0.542), the percentage of abnormal sperm cells (p=0.343), nor testosterone levels (p=0.063). Dietary vitamin E supplementation is recommended for male chickens since it generally enhances the quality of their sperm. Keywords: meta-analysis, rooster, sperm, vitamin E.

Angiotensin-converting enzyme 2 (ACE2) is expressed and plays functional and physiological roles in different tissues of the body. This study aimed to distinguish the levels of expression of ACE2 in the lung tissue at different ages of rats. In this study, 18 male rats were used and divided into three groups according to age. Real-time quantitative polymerase chain reaction (RT-qPCR) was conducted to determine the levels of the quantification of eosinophil cationic protein mRNA transcript. In addition, tissue specimens of the lung were stained with routine hematoxylin and eosin stains. This study confirmed that RT-qPCR amplification plots of ACE2 gene exhibited clearly expression of the lung tissue of rats in the different groups and there are strong different threshold cycles numbers according to the age at 2 weeks, 2 months, and 6-8 months. Consequently, the expression of ACE2 was completely different between groups depending on the age of the rats. The RT-qPCR results showed that the older animal group (age of 6-8 months) had a significantly higher expression of ACE2 than the other animal groups (ages of 2 weeks and 2 months). In the same way, the second group (age of 2 months) had a significantly higher expression of ACE2 than the first group (age of 2 weeks). This study confirmed that the ACE2 expression is influenced by the age of rats. This study concluded that the expression of the ACE2 receptor of coronavirus disease 2019 would be different according to the age of rats, and this result suggested that expression of ACE2 in lung tissue could determine infection and pathogenesis of COVID-19 during different ages of rats or some individual differences. Keywords: angiotensin-converting enzyme 2, expression, rats, real-time polymerase chain reactions, severe acute respiratory syndrome coronavirus 2.

Milk produced from Awassi sheep is of high nutritive value; its production is relatively low in Awassi sheep, so the genetic improvement programs targeted milk production and its components are of high importance, especially when using genes that have an important signal to milk traits. This study was aimed at assessing the influence of alpha S1 (CSN1S1) and beta-casein (CSN2) genes genotypes interaction on Awassi ewes milk productivity. A total number of 391 milk yield and its composition records (taken through five consecutive years, 2007-2011) of 167 ewes were utilized for this study. DNA samples were extracted from the ewe's blood samples, then the polymerase chain reaction products of alpha S1 (CSN1S1) and beta-casein (CSN2) genes were sequenced. The obtained sequences were analyzed; thereafter, the detected variants were tested for their possible association with milk traits. The CSN1S1 and CSN2 variants allelic frequencies were 0.85 and 0.15, and 0.95 and 0.05, respectively. Lactose and solid not fat (SNF) % were associated with TC CSN1S1 genotypes. No association was found among CSN1S1 polymorphic genotypes with milk production, lactose, and SNF % were associated with TC CSN1S1 genotypes. Ewes of CSN2 AC genotype showed higher milk production traits, while no association was found between milk composition traits and CNS2 genotypes. Nevertheless, CSN1S1∗CSN2 interaction showed the highest SNF, fat percentages, and milk production. The substantial interaction effects between CSN1S1×CSN2 genes were significantly affected the amount of milk, fat, and SNF% produced. The detected variants should be included in the breeding programs of Awassi sheep that are designed for improving their milk quantity and quality. Keywords: Awassi sheep, fat, gene, interaction, milk yield, protein.

Neospora caninum is one of the most common infectious organisms worldwide that causes abortion in cattle. To the best of our knowledge, no previous studies have focused on N. caninum infection in the local Atlas brown cattle from Northeast Algeria. This study aimed to assess the importance of bovine neosporosis for causing abortion in Atlas brown cattle and to identify selected risk factors. A case-control study was performed on 60 control farms and 30 case farms. We collected 650 blood samples from 650 pregnant cows from 90 farms in five Algerian provinces; Jijel, Skikda, Annaba, El-Tarf, and Souk- Ahras. Sera samples were analyzed for the presence of antibodies against N. caninum using enzyme-linked immunosorbent assays. The seroprevalence of N. caninum infection in the cows was 36.2% (95% confidence interval [CI]: 32.7-39.8) and in the farms was 81.1% (95% CI: 73.0-89.2). Risk factors found by multivariable logistic regression included: Presence of dogs (odds ratio [OR] 4.7, 95 CI 2.9-7.3); age ≥84 months (OR 4.9, 95 CI 2.8-8.3); Jijel region (OR 2.2, 95 CI 1.1-4.5); white (OR 2.5, 95 CI 1.4-4.4) and gray (OR 2.5, 95 CI 1.4-4.5) coat; moderate (OR 2.30, 95 CI 1.4-3.8) and bad (OR 3.1, 95 CI 1.8-5.3) hygiene; and second (OR 2.5, 95 CI 1.4-4.4); and last (OR 2.3, 95 CI 1.3-4.2) stage of pregnancy. Our case-control study showed no significant association between seropositivity of N. caninum and abortion at the farms level (OR 0.9, 95 CI 0.3-2.7). Similarly, there was no significant association between seropositivity of N. caninum and abortion at the individual level (OR 0.8, 95 CI 0.6-1.2). This is the first study of N. caninum infection in pregnant local cows from Northeast Algeria. The prevalence rate of antibodies against N. caninum was high. Almost all risk factors studied for infection were significantly associated with seroprevalence. Our analysis showed no relation between N. caninum infection and abortion. Consequently, these local cows are resistant to abortion caused by N. caninum. Keywords: Algeria, Neospora caninum, pregnant cattle, seroepidemiology.

Bali Cattle (Bos j. javanicus) is a local breed originating in Indonesia, accounting for 32.3% of the total cattle population. To date, no studies of the genetic structure and demographic status of Bali cattle have been conducted, even though the breeding of Bali cattle has a long and unique history that is likely to have impacted its genetic diversity. Therefore, a study that used molecular breeding technologies to characterize the demography of Bali cattle would be timely. This study aimed to examine genome diversity in Bali cattle and estimate the linkage disequilibrium (LD) and effective population size (Ne) values in the cattle population. In this study, we explored the population structure and genetic diversity of Bali cattle using genomic-level analyses. Our study primarily studied cattle that had been bred in livestock breeding centers since these breeds had subsequently spread throughout Indonesia. We focused on characterizing the genetic structure, determining the level of LD present, and estimating the Ne of the Bali cattle population. The genomic data used for this study were obtained from DNA samples of 48 Bali cattle collected at the Breeding Center of Bali Cattle as well as 54 genomic samples from Bali cattle collected elsewhere in Indonesia that had been used in recent publications. This genomic dataset included exclusively 50K single nucleotide polymorphisms (SNP) array (Illumina Bovine 50SNP bead chip, Illumina, USA) data. We found that the LD values of Bali cattle from the breeding center and those raised elsewhere were 0.48±0.43 and 0.39±0.40, respectively. Subsequently, the Ne value of Bali cattle from the breeding center and farmers was 151 and 96, respectively. Our results suggest that the selection program of the breeding center is beneficial for maintaining the genetic diversity of Bali cattle. Keywords: Bali cattle, effective population size, genomic data, genetic structure, linkage disequilibrium.

Two endemic capripox infectious diseases, sheeppox (SP) and goatpox (GP) are common in Asia, Africa, and the Middle East. Sheep and goats, in general, are considered current assets of small and marginal farmers and have significant economic value in terms of meat, wool, and skin/hide production. Sheep and goat populations in India total 148.88 million and 74.26 million, respectively. Capripox caused US$ 2.3 million (Indian Rupee [INR] 105 million) in economic damages in Maharashtra (India) alone, and it took over 6 years for a flock to recover from the outbreak. The projected yearly loss at the national level is US$ 27.47 million (INR 1250 million). As a result, Capripox diseases put small and marginal farmers under much financial strain. The present study estimates the seroprevalence of SP and GP diseases in the Asian and African continents using systematic review and meta-analysis. The results of the study will help researchers and policymakers to understand the spatial and temporal distribution of the disease and its burden. In addition, the results are also helpful to design and implement location-specific prevention and eradication measures against these diseases. Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines of Cochran collaborations were used for systematic review and subsequently meta-analysis were used. The literature was collected from various databases. Initial search string resulted in more than nine thousand articles for the period 2000 to 2020 using the different combinations of keywords and Boolean operators (or not) asterisk∗ and quotation marks. Out of 9398 papers, 80 studies were chosen for complete test reviews and quality bias evaluation using the inclusion and exclusion criteria. Finally, 21 articles were used for the meta-analysis. The statistical study employed fixed effects and random effects models using R. Seroprevalence of SP and GP was calculated using studies with a cumulative sample size of 4352, out of which sheep and goats' samples together contribute 48%, followed by sheep (32%) and goat (21%). The result of the meta-regression revealed that detection techniques had a significant impact on the overall effect size at 5% level (Qm=14.12). Subgroup analysis of polymerase chain reaction (PCR) test with samples was further grouped into two categories based on the median, and it revealed that 62% of samples used PCR as a detecting test followed by group-II. From the study, it is concluded that SP and GP diseases are highly prevalent; hence, effective vaccines, proper education to farmers through extension activity, and transboundary disease movement restriction are necessary for the control and eradication of the disease. Keywords: Asia, Africa, capripoxvirus, meta-analysis, seroprevalence, subgroup analysis.

Fish diseases have a significant negative influence on the Malaysian aquaculture industry. Since the 1980s, the sector has grown in size, which has resulted in a rise in the prevalence of infectious outbreaks affecting both freshwater and marine cultured fish species. Demand for commercially available fish vaccinations is predicted to increase as infectious disease outbreaks continue to occur. In Malaysia, aquaculture vaccine research and development (R&D) are still in its infancy, with most efforts concentrating on producing vaccines against bacterial infections, most notably streptococcosis, vibriosis, and motile Aeromonas septicemia. Despite several attempts, no homegrown vaccine has been effectively introduced into the manufacturing pipeline to date. At the moment, only three imported aquatic vaccines have received full permission, a far cry from the 314 and 60 vaccines licensed in the poultry and porcine industries, respectively. This review will describe recent findings regarding the development of aquaculture vaccines for certain fish species and diseases in Malaysia. In our opinion, R&D on fish vaccines is critical to the aquaculture industry's viability.

Canine testicular tumors are among the most common reproductive tract tumors in male dogs and have been studied in many countries. However, to the best of our knowledge, studies with a large sample size have not been conducted in Russia. This study aimed to provide the latest information on the prevalence of canine testicular tumors in the Veterinary Oncology Scientific Center for Small Animals "Biocontrol" in Moscow, Russia, in 2010-2020 and the characteristics of the affected canine population. A retrospective review of patients and histological reports was collected and analyzed from 358 dogs with 447 testicular tumors within 11 years. The mean age of the affected dogs was 10.4 years, whereas that of dogs with Sertoli cell tumors was 9.4 years p=0.009. This study includes mixed-breed dogs (18.4%), Yorkshire Terriers (8.8%), Labrador Retrievers (7.9%), Golden Retrievers (5.0%), and Fox Terriers (3.4%). The most common tumors were interstitial cell tumors (n=227, 50.8%). In contrast, 107 (23.9%) seminomas, 80 (17.9%) Sertoli cell tumors, 19 (7.4%) mixed germ cell-sex cord-stromal tumors, and 26 (7.6%) testicular tumors developed from cryptorchid testes, which included 16 (61.5%) Sertoli cell tumors, 10 (38.5%) seminomas, and no interstitial cell tumors. This study provides baseline information on the prevalence of canine testicular tumors in the described population, including the median age of each tumor type and overrepresented dog breeds. We further found that the most common scrotal testicular tumor was interstitial cell tumor, whereas Sertoli cell tumor was the most common in cryptorchid testicles. Keywords: cryptorchid testes, interstitial cell tumors: mixed germ cell-sex cord-stromal tumors, seminomas, Sertoli cell tumors, testicular tumors.

Food of animal origin is considered a major source of foodborne diseases. In this context, multidrug-resistant (MDR) Escherichia coli pose a serious hazard to public health due to the consumption of food contaminated with antibiotics that are used for the treatment of various bacterial infections in farm animals. Therefore, this study aimed to determine the effect of the excessive use of antibiotics on the development of MDR E. coli strains in Egyptian poultry, dairy, and meat farms. A total of 1225 samples were randomly collected from poultry, dairy, and meat products intended for human consumption in different governorates. E. coli were isolated from the collected samples and subjected to biochemical identification and antibiotic sensitivity tests with antibiotics commonly used in human and veterinary medicine. Then, amoxicillin (AML)- and oxytetracycline (OT)-resistant E. coli isolates were subjected to a polymerase chain reaction test to detect the blaTEM and tetA genes, respectively. E. coli were isolated from 132 out of 350, 148 out of 350, 177 out of 350, and 35 out of 175 poultry, milk, meat, and human samples, respectively. Most of the isolates expressed multidrug resistance, and resistance genes (blaTEM and tetA) were detected in all the tested AML- and OT-resistant E. coli isolates. Foods of animal origin may represent a source of MDR E. coli, which can be a major threat to public health. Keywords: Escherichia coli, food of animal origin, misuse of antibiotics, multidrug resistance, poultry.

We previously developed the gene-editing by electroporation (EP) of Cas9 protein method, in which the CRISPR/Cas9 system was introduced into porcine in vitro fertilized (IVF) zygotes through EP to disrupt a target gene. This method should be further developed, and a combination of EP and MI methods should be evaluated in pigs. This study aimed to determine that a combination of microinjection (MI) and EP of CRISPR/Cas9 system could increase the rates of biallelic mutation for triple-gene knockout in porcine blastocysts. We targeted the pancreatic and duodenal homeobox1 (PDX1) gene using cytoplasmic MI 1 h before or after EP, which was used to edit alpha-1,3-galactosyltransferase (GGTA1) and cytidine 32 monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) genes in porcine zygotes. We introduced guide RNAs targeting PDX1, GGTA1, and CMAH with the Cas9 protein into IVF zygotes (one-cell stage) through EP 10 h after the start of IVF (IVF; EP group) or in combination with MI (1 h before, MI-EP group, or after EP treatment EP-MI group) and evaluated the blastocyst formation rate and efficiency of target mutations in the resulting blastocysts. Our results revealed a significant reduction in the rate of blastocyst formation in the two groups that underwent MI before and after EP (MI-EP and EP-MI group), compared with that in the groups treated with EP alone (EP group) (p=0.0224 and p<0.0001, respectively) and control (p=0.0029 and p<0.0001, respectively). There was no significant difference in the total mutation rates among the treatment groups in the resulting blastocysts. As an only positive effect of additional MI treatment, the rate of blastocysts carrying biallelic mutations in at least one target gene was higher in the MI-EP group than in the EP group. However, there was no difference in the rates of embryos carrying biallelic mutations in more than 2 target genes. These results indicate that although a combination of MI and EP does not improve the mutation efficiency or biallelic mutation for triple-gene knockout, MI treatment before EP is better to reduce mortality in porcine zygotic gene editing through a combination of MI and EP. Keywords: clustered regularly interspaced short palindromic repeats/Cas9, electroporation, gene editing, microinjection, porcine zygotes.

Hypertrophic cardiomyopathy (HCM) is a common heart problem that affects many cats. Although cats with HCM are symptomatic, some die suddenly or develop congestive heart failure. Therefore, this study aimed to estimate the prevalence of myosin-binding protein C3 (MYBPC3), A31P, and A74T polymorphisms in Maine Coon cats to assess risk factors for diagnosing HCM in cats. Forty-nine Maine Coon cats of at least 10 months of age were enrolled in this study. First, clinical parameters, such as heart rate, systolic blood pressure, and echocardiography, were evaluated. Then, polymerase chain reaction, followed by DNA sequencing, was conducted using specific primers for amino acid substitutions caused by genetic variants of MYBPC3-A31P and -A74T polymorphisms. Investigations showed that the prevalence of MYBPC3-A31P and -A74T mutations in this study was 16.33% and 24.45%, respectively. Moreover, HCM in cats with MYBPC3-A31P and A74T mutations increased with age, body weight, high heart rate, and prolonged isovolumic relaxation time. Therefore, we propose that Maine Coon cats develop HCM due to multiple genetic factors and underlying clinical characteristics in individual cats. Furthermore, relaxation time assessments can be a sensitive technique for HCM screening during its preclinical phase and can help identify the risk of developing HCM. However, further studies are warranted to evaluate the effect of MYBPC3 mutations on the phenotypic expression of HCM. Keywords: feline, hypertrophic cardiomyopathy, mutation, myosin-binding protein.

In 2020, Indonesia, which has the highest global production of edible bird's nest (EBNs), exported up to 1312.5 tons of this product at a value of USD 540.4 million. Recently, food safety aspects related to EBNs, including contamination with heavy metals, have become a serious concern. However, data on the presence and concentration of heavy metals in EBNs in Indonesia are not yet available. This study aimed to determine and compare the presence and concentrations of arsenic (As), mercury (Hg), lead (Pb), cadmium (Cd), and tin (Sn) in EBNs originating from several primary Indonesian islands. The study also analyzed the effect of washing on the heavy metal content in EBNs. A study on 44 swiftlet farmhouses (SFHs) was conducted to determine the concentrations of heavy metals in EBNs. The number of samples from the SFHs was allocated proportionally to the main EBN-producing islands in Indonesia, that is, Kalimantan, Sumatera, Sulawesi, and Java (22, 13, 7, and 2, respectively). The concentrations of the above five elements in the samples before washing (raw–unclean EBNs) and after washing (raw–clean EBNs) were determined by inductively coupled plasma mass spectrometry. Washing was conducted according to the general procedures at an EBN processing plant. The raw–unclean EBNs from the four islands contained As, Pb, Cd, and Sn at varying concentrations. However, Hg was not detected in the raw–unclean EBN samples from Sulawesi. The raw–unclean EBNs from Kalimantan had lower concentrations of Pb and Cd compared with the other islands. The concentrations of As, Pb, Cd, and Sn in the EBNs decreased significantly after washing with clean water. Heavy metals (As, Hg, Pb, Cd, and Sn) were detected at a low level in most of the raw–unclean EBNs originating from the main Indonesian island where they were produced. The concentrations of all the heavy metals reviewed in the raw–unclean EBNs samples decreased significantly after washing. Keywords: edible bird nests, heavy metal, swiftlet farmhouse, washing.

Water pollution caused by industrial waste and human activities has disrupted the reproductive health of aquatic organisms. This study aimed to analyze the effects of water pollution caused by polystyrene particles (PP) on the steroid (estradiol and testosterone) levels and histopathology of male tilapia gonads. In addition, we also analyzed the potential of supplementary feeding to remove and neutralize oxidants. Thirty-six tilapia fishes were taken for the study and were divided into 12 groups (n=3), including a control group (fed with commercial pellets only) and groups fed with a mixture of commercial-probiotic pellets (200 mL/kg, 1×108 colony-forming unit [CFU]/mL) and commercial vitamin C pellets (100 mg/kg), respectively. The PP concentrations used for this study were 0, 0.1, 1, and 10 mg/L, and the treatment time was 2 weeks. The testosterone and estradiol concentrations were analyzed by enzyme-linked immunosorbent assay and histopathological analysis of the gonads. Laboratory analysis performed using tilapia fishes showed that exposure to a PP concentration of <74 μm, mixed with feed for 14 days, could decrease estradiol and testosterone levels. Exposure to plastic particles could change the structure, shape, and size of male gonads. It can also affect the spermatogenic cell number and alter the diameter inside the cysts. Originally, plastic particles were believed to reduce the permeability of the cyst membrane, and this damages the membrane or ruptures the cyst. Supplementary feed containing probiotics (200 mL/kg, 1×108 CFU/mL) and vitamin C (100 mg/kg) can ameliorate the impact of PP exposure on steroid levels. The steroid levels increase with a concurrent improvement in cysts and seminiferous tubule structures. Overall, this study indicates that PP concentrations in the aquatic environment negatively affect tilapia reproduction, and this may pose a potential threat to the fish population in freshwater. Provision of supplementary feed containing probiotics and vitamin C may serve as an alternative way to counter the negative impact caused by plastic particles. Keywords: fish, fisheries, freshwater, polystyrene, probiotics, testicular.

Serological assays are widely used to monitor the performance of foot-and-mouth disease (FMD) vaccines to estimate vaccination coverage and to ensure that vaccinated animals generate adequate immune responses. This study aimed to measure the FMD virus (FMDV)-specific responses in cattle and sheep after a single dose of a trivalent FMD vaccine containing serotypes A, O, and Asia-1, and to use these sera to calibrate virus neutralization tests (VNTs) and serotype-specific serological enzyme-linked immunoassays (ELISAs) that can measure post-vaccination responses. Sera were collected from cattle (n=10) and sheep (n=10) on 0, 21, and 56 days after immunization with an imported aqueous formulated FMD vaccine. These samples were tested by VNT using field FMDV isolates that are representative of the epidemiological risks in Central Asia (A/ASIA/Iran-05, A/ASIA/GVII, O/ME-SA/Ind-2001, O/SEA/ Mya-98, O/ME-SA/PanAsia, and Asia-1 Shamir). Heterologous VNT antibody responses were compared to those measured using commercial FMDV-specific ELISAs for serotypes O, A, and Asia 1. Administration of the FMD vaccine increased FMDV-specific antibody titers for both species in sera collected on day 21, but these elevated titers were short-lived and were decreased by day 56. These results highlight the short duration of immunity with a single dose of this aqueous vaccine and motivate further studies to assess immune responses in cattle and small ruminants after a two-dose course vaccination schedule. Further comparative data for VNT and serotype-specific ELISAs are needed to define cutoffs that can be used to monitor post-vaccination immune responses in low-containment laboratories where it is not possible to handle live FMDVs. Keywords: cattle, enzyme-linked immunoassay, foot-and-mouth disease, immunogenicity, post-vaccination monitoring, sheep, virus neutralization test.