Canine monocytic ehrlichiosis (CME) is a tropical endemic tick-borne disease that causes fatality or chronic infection involving many organs in dogs. This study aimed to examine the prevalence, risk factors, and hematological and ultrasonographic changes in the liver, gallbladder, kidneys, and spleen following CME infection. This retrospective study used 30,269 samples collected from dogs at the hematology section of the pathology unit of a university veterinary hospital and 35 samples collected from dogs at the diagnostic imaging unit. CME was determined using the buffy coat smear method. Data were analyzed using descriptive statistics and odds ratios. CCl4 The data revealed that the average yearly prevalence of CME was 1.32%. Risk factors contributing to CME infection were a tick on the body during physical examination, lack of ectoparasite control, and outdoor living. All 148 dogs with CME infection had low platelet counts. The percentages of CME-infected dogs with elevated serum alanine aminotransferase, alkaline phosphatase, and both enzymes above the normal range were 33.6%, 65.9%, and 29.8%, respectively. The rates for elevated serum levels of blood urea nitrogen, creatinine, and both compounds were 33.1%, 19.1%, and 17.3%, respectively. The most common ultrasonographic changes were liver abnormalities (hyperechogenicity or hypoechogenicity, hepatomegaly, and hypoechoic nodules), hyperechogenicity of the kidneys, and an enlarged spleen. These ultrasonographic changes were consistent with the hematology results, which showed a greater elevation of serum liver enzyme levels than renal enzymes. Ultrasonographic changes during CME infection and after treatment with doxycycline can help to monitor and identify persistent pathological changes in the target organs resulting from immune response to CME. Keywords: dog, ehrlichiosis, hematology, monocyte, ultrasound.

Squamous cell carcinoma (SCC) is the most common form of carcinoma in cattle. Histopathological grading systems have been utilized over several decades for estimating the malignancy of cattle SCCs. This study aimed to detect p53 and Mdm2 expression in different SCC cases in cattle and correlate their expression with the SCC histopathological grading. Cattle SCC cases were collected at the Veterinary Teaching Hospital in Nineveh. The SCC grading system categorized the cases histologically based on their differentiation grade into three groups: Well, moderately, and poorly differentiated. The SCC cases were subsequently verified for p53 and Mdm2 immunoexpression. Fourteen of 16 examined cattle SCC samples tested positive for p53 expression. Moreover, 15 out of the 16 SCC samples tested positive for Mdm2 expression. The increased immunoreactivity of both p53 and Mdm2 was associated with a poor histological grading of the cattle SCC. There is a positive correlation between the nuclear expression of p53 and Mdm2, and the degree of differentiation and the number of mitotic figures in the examined cattle SCC samples. Our results demonstrate an increased p53 and Mdm2 expression in cattle SCC cases characterized by poor histopathological grading, thus suggesting an essential role of these molecules in the development of moderately and poorly differentiated SCC in cattle. Keywords: cattle, immunoexpression, Mdm2, p53, squamous cell carcinoma.

Swollen head syndrome (SHS) is a complex disease caused by various agents, including bacterial and viral pathogens, as well as environmental factors. Avian metapneumovirus (aMPV) is one of the most important causes of respiratory diseases and SHS in poultry and one of the most widespread viruses worldwide; however, it has not been recorded in Iraq. This study aimed at the molecular identification and subtyping of aMPV in poultry, with the objectives of investigating the prevalence of aMPV in infected broiler flocks with SHS and molecular typing using primers specific to the study of the prevalence of subtypes A, B, and C of aMPV. This study was performed on 67 broiler farms that reported typical SHS from September 2018 to August 2019. Swabs were collected from the trachea, infraorbital sinuses, and lung, then uploaded on FTA cards and subjected to an RNA extraction protocol. aMPV was detected in 16 (23.8%) samples. Molecular typing using primers specific to the attachment glycoprotein (G) gene showed that all positive samples belonged to subtype B, as assessed using the real-time polymerase chain reaction technique. aMPV may be the main etiological factor causing SHS in poultry. Moreover, this was the first report of the prevalence of subtype B aMPV strains in broiler farms in Iraq. Keywords: Avian metapneumovirus type (B), Iraq, middle Euphrates region, swollen head syndrome, upper respiratory tract infection, viral infection poultry.

Zebrafish have gained momentum as a leading experimental model in recent years. At present, the zebrafish vertebrate model is increasingly used due to its multifactorial similarities to humans that include genetic, organ, and cellular factors. With the emergence of novel research techniques that are very expensive, it is necessary to develop affordable and valid experimental models. This review aimed to highlight some of the most important similarities between zebrafish and humans by emphasizing the relevance of the first in simulating neurological disorders and craniofacial deformity.

Cysticercosis is a zoonotic disease with a global concern. Estimation of the prevalence and identification of potential risk factors are necessary for the prevention and control of the disease. This study aimed to estimate the seroprevalence of cysticercosis and the correlation of the increased prevalence with several potential risk factors. The seroprevalence of cysticercosis was conducted using an enzyme-linked immunosorbent assay (ELISA), developed by the Institute of Tropical Medicine Antwerp, to detect Cysticercus cellulosae. This study used serum samples from 62 pigs taken from two regencies on Timor Island. The data analysis was performed using SPSS software 20.0 (IBM Corp., NY, USA) to evaluate ELISA results and the strength of the relationship between risk factors and the prevalence of disease using the odds ratio (OR). Serum samples from 18 out of the 62 pigs were found to be positive; the seroprevalence of cysticercosis was 29%. The results showed that an extensive farming system led to a higher prevalence of cysticercosis compared to an intensive farming system, namely, 10 out of 18 (56.6%), and that the possibility of identifying cysticercosis in pigs in an extensive farming system was 5 times greater than that in pigs in an intensive farming system. In addition, the results showed that nine out of 18 households who did not have toilet facilities were found to be seropositive, indicating a significant relationship between the risk factor of toilet availability with cysticercosis in pigs, with an OR of 4.5. In addition, the results showed that there was no significant relationship between the risk factor of the feed source and the prevalence of cysticercosis in pigs. It can be concluded that the seroprevalence of cysticercosis was 29% in domestic pigs of Timor Island. The risk factors of an extensive pig farming system and toilet availability in community houses were significantly related to the possibility of cysticercosis on Timor Island. Keywords: cysticercosis, epidemiology, risk factor, seroprevalence.

Understanding the regulations of rumen microbiota and their fibrolytic capabilities under different forages are essential to improve rumen fermentation and animal feed efficiency. This study aimed to evaluate the changes in the rumen fermentation and the structure and fibrolytic activities of rumen bacteria in camels fed barley straw and Egyptian clover hay. Three fistulated camels were fed a diet containing barley straw for 30 days; then transitioned to a diet containing Egyptian clover hay for 30 days. In addition, bacterial media enriched with xylan and different cellulose sources, namely, filter paper, wheat straw, and alfalfa hay, were used to evaluate the ability of camel rumen bacteria to produce xylanase and cellulase enzymes. The camel group fed Egyptian clover hay showed higher crude protein intake, rumen ammonia, total volatile fatty acids, and acetic acid. Moreover, the camel group fed barley straw showed higher neutral detergent fiber intake, rumen pH, and propionic and butyric acids. Principal component analysis showed that bacterial communities were separated based on the forage type. Forage type affected the composition of rumen bacteria and most of the bacterial community was assigned to phylum Bacteroidetes and Firmicutes. Egyptian clover hay diet increased the proportions of genus Prevotella and Ruminococcus; while fed barley straw diet increased the Butyrivibrio, RC9_gut_group, and Fibrobacteres. The bacterial culture of the Egyptian clover hay fed group produced the greatest xylanase and the bacterial culture of the barley straw fed group produced the maximum cellulase. Egyptian clover hay is recommended to feed camels in intensive production. Moreover, the bacterial community in the camel rumen is a promising source of lignocellulolytic enzymes. Keywords: Camelus dromedaries, cellulase and xylanase, forage type, hay and straw, rumen bacteria.

The crocodile is a model for studying relevant sources of environmental contamination. They were determined an appropriate biomonitoring species for various toxins. The cytosolic and microsomal fraction of crocodiles plays a role in detoxifying xenobiotics. Cytochrome P450 1A2 (CYP1A2) metabolizes aflatoxin B1 (AFB1) to aflatoxin M1, while glutathione-S-transferase (GST) catalyzes carcinogenic agents. This study aimed to investigate the GST activity in various organs of Crocodylus siamensis. Further, the fate of microsomal and cytosolic fractions from various crocodile organs against AFB1-induced apoptosis in human hepatocarcinoma (HepG2) cells was investigated. The liver, lungs, intestines, and kidneys tissues from a 3-year-old crocodile (C. siamensis) (n=5) were collected. The cytosolic and microsomal fraction of all tissues was extracted, and protein concentrations were measured with a spectrophotometer. Subsequently, a comparison of GST activity from various organs was carried out by spectrophotometry, and the protective effects of CYP450 and GST activity from various crocodile organs were studied. In vitro AFB1-induced apoptosis in HepG2 cells was detected by reverse transcription-quantitative polymerase chain reaction. Comparisons between the metabolisms of the detoxification enzyme in organs were tested using the Kruskal–Wallis one-way analysis of variance and Dunn's multiple comparison tests. All kinetic parameters were analyzed using GraphPad Prism software version 5.01 (GraphPad Software Inc., San Diego, USA). Total GST activity in the liver was significantly higher than in the kidneys, intestines, and lungs (p<0.05, respectively). The highest GST pi (GSTP) activity was found in the liver, while the highest GST alpha-isoform activity was in the crocodile lung. The kinetics of total GST and GST mu activity in the liver had the highest velocity compared to other organs. In contrast, the kinetics of GSTP enzyme activity was the highest in the intestine. The in vitro study of microsome and cytosol extract against apoptosis induced by AFB1 revealed that the level of messenger RNA expression of the Bax and Bad genes of HepG2 cells decreased in the treatment group in a combination of cytosolic and microsomal fractions of the crocodile liver but not for Bcl-2. Interestingly, the downregulated expression of Bax and Bad genes was also found in the microsome and cytosol of crocodile kidneys. The crocodile liver revealed very effective GST activity and expression of the highest kinetic velocity compared to other organs. The combination of liver microsomal and cytosolic fractions could be used to prevent cell apoptosis induced by AFB1. However, further study of the molecular approaches to enzyme activity and apoptosis prevention mechanisms should be carried out. Keywords: aflatoxin B1, apoptosis, Crocodylus siamensis, glutathione-S-transferase, human hepatocarcinoma cells.

Antibiotic resistance has been a progressively documented problem, resulting in treatment failure in humans and animals. This study aimed to investigate the antimicrobial susceptibility and virulence of extensively drug-resistant (XDR) Aeromonas spp. in wild Mugil cephalus and its surrounding seawater along the coastal road of Port Said, Egypt. Specimens were examined bacteriologically, confirmed biochemically, and tested for their sensitivity against 11 antimicrobial agents. Molecular confirmation of the obtained isolates by 16S rRNA was performed, followed by the detection of antimicrobial resistance and virulence genes. Aeromonas spp. was recovered from fish (44%) and water samples (36%). A. hydrophila was the most prevalent identified strain, followed by Aeromonas sobria, Aeromonas caviae, and Aeromonas schubertii. Moreover, 90% of the tested isolates were multidrug-resistant (MDR), while 26.67% were XDR. Tested isolates were resistant to β-lactams and sulfonamides (100%), oxytetracycline (90%), and streptomycin (62.22%) but completely susceptible to cefotaxime. XDR isolates successfully amplified resistance genes (blaTEM, sul1, and tetA(A)) but not the (aadA1) gene, although there was phenotypic resistance to streptomycin on plates. All XDR isolates carry the cytotoxic enterotoxin gene (act), but alt gene was detected in only one isolate (12.5%). Data in this study provide a recent update and highlight the role of wild mullet and seawater as reservoirs for MDR and XDR Aeromonas spp. that may pose a risk to humans as food-borne infection or following direct contact. Keywords: Aeromonas hydrophila complex, antimicrobial resistance, Mugil cephalus, resistance genes, Seawater, virulence genes.

Gold nanorods (AuNRs) have gained much attention in recent years due to their promising optical and chemical properties and are hence used in applied research and industrial nanotechnology. This study was designed to investigate the effect of gold nanoparticle shape (Gold nanorods vs. gold nanosphere) on immune response in rabbit. Thirty New Zealand white rabbits were divided into six groups (n=5 rabbits). The first group is the control negative received an intravenous (IV) injection of normal saline 0.9%; the second group (vaccinated) is the control positive, and the other four groups were vaccinated and received a single-dose or repeated five consecutive IV doses of 300 μg/kg body weight 50 nm AuNRs or 50 nm gold nanosphere (50 nm AuNSs) dissolved in ultrapure water. Blood and serum were collected for the hematological and biochemical analysis. White blood cells (WBCs) count, lymphocytes, monocytes, eosinophils, and basophils showed significantly (p<0.05) higher values with the repeated-dose AuNRs. γ-globulin levels showed a significant difference after 15 days in the single-dose AuNSs. Single-dose AuNSs significantly (p<0.05) increased the immunoglobulin G (IgG) and significantly (p<0.05) decreased the tumor necrosis factor-alpha. In addition, it elicited a significant (p<0.05) decrease in the malondialdehyde levels and a significant (p<0.05) increase of the superoxide dismutase, glutathione peroxidase, and catalase levels. Moreover, evoked red blood cells count, mean corpuscular volume, and mean corpuscular hemoglobin were significantly (p<0.05) lower than the control group. The platelet count, lysozymes, and nitric oxide were significantly (p<0.05) higher in repeated-dose AuNRs. The effect of AuNPs is shape and dose-dependent. The repeated 5 days IV 50 nm AuNRs doses over 15 days showed a significant antioxidant effect, with no considerable toxicity or vascular reactions. Keywords: antioxidant's activity, dose, gold nanorods, gold nanospheres, hematological, immunological parameters, rabbit.

Piroplasmosis is a serious disease that infects animals, inflicting significant economic losses in the livestock industry and animal trade worldwide. Anti-piroplasm drugs now on the market have demonstrated host toxicity and parasite resistance. As a result, developing more effective and safer anti-piroplasm drugs becomes an urgent issue. This study aimed to evaluate the inhibitory effect of Capsicum annuum methanolic extract (CA) against the growth of Babesia bovis, Babesia divergens, Babesia caballi, and Theileria equi in vitro and against B. microti in mice. Fluorescence-based SYBR Green I assay was used to evaluate CA's inhibitory effect in vitro and in vivo when used either as a monotherapy or combined with diminazene aceturate (DA). The hematological parameters (HCT, hemoglobin, and red blood cells counts) were determined in the blood of mice every 96 h using Celltac α MEK-6450 electronic hematology analyzer. The in vitro growth of B. bovis, B. divergens, T. equi, and B. caballi was inhibited by CA in a dose-dependent manner with IC50 values of 4.87±1.23, 44.11±8.03, 8.23±2.54, and 1.26±0.50 mg/mL, respectively. In B. microti-infected mice, a combination therapy consisting of CA and a low dose of DA showed a significant (p<0.05) inhibition of B. microti growth nearly similar to those obtained by treatment with the full dose of DA. The obtained results indicate that CA might be a promising medicinal plant for treating babesiosis, especially when used with a low dose of DA. Keywords: Babesia, Capsicum annuum, combination therapy, in vitro, in vivo, Theileria.

Promotions of goat farming by both public and private sectors encouraged considerable goat raising in central Thailand. Gastrointestinal nematodes (GIN) infection is a major health and economic problem; however, evidence of resistance to broad-spectrum anthelmintics is frequently reported. Investigation of anthelmintic resistance (AR) status and identification of factors related to the development of AR is important components for sustainable GIN control. However, no information is available on this topic in the study area. The present study aimed to gather information on GIN control practices and to evaluate the effectiveness of albendazole, ivermectin, and levamisole for treating GIN infestation in goat herds in Sing Buri Province. Twenty-nine herds were randomly selected. Information on management practices was collected by face-to-face interview using a structured questionnaire. Three field experiments for routinely used anthelmintics, including albendazole, ivermectin, and levamisole were conducted from June 2019 to November 2019. Fecal samples were collected pre- and post-treatment and examined for fecal egg count reduction to determine the status of anthelmintic resistance of goat GIN. Several improper practices were identified that lead to AR, especially chronic use of albendazole and ivermectin. All herds were considered resistant to albendazole and ivermectin, and levamisole resistant nematodes were detected in two herds. AR was strongly linked with the continuous use of anthelmintics. Levamisole, which was still effective in the province, should be used with caution to minimize the selection of resistant strains. Farmers should be provided with updated information for sustainable parasite control. Further, the efficacy of anthelmintics should be routinely monitored. Keywords: anthelmintic resistance, fecal egg count, gastrointestinal nematode, goat.

Aflatoxin M1 (AFM1) is a major fungal metabolite found in milk coming from aflatoxin B1 (AFB1) contaminated rations and is subsequently present in milk-based products demonstrating a serious public health hazard. This study aimed to investigate the levels of AFM1 and AFB1 in milk and some dairy products consumed widely by infants and children. This study investigated the incidence of AFM1 in 105 samples of processed cheese, Ras cheese, and raw milk (35 of each) retailed in the Egyptian markets. The degree of sensitivity and accuracy was evaluated using the enzyme-linked immunosorbent assay (ELISA) method followed by the estimation of the positive samples using the high-performance liquid chromatography (HPLC) with fluorescence detection. Mold count was determined in the examined samples by investigating AFB1 content using HPLC. AFM1 was found in all investigated Ras cheese, raw milk, and 82.86% of the processed cheese samples with mean values of 51.05±6.19, 40.27±3.996, and 10.77±1.39 ng/kg, respectively. Moreover, there was statistically no significant difference between AFM1 levels in the core and crust parts of the tested Ras cheese. AFM1 contaminated Ras cheese and raw milk samples were 48.57% and 25.71%, which exceeded the European and Egyptian tolerance levels. Results showed an acceptable correlation between ELISA and HPLC methods with no significant difference (p>0.05). Alternatively, none of the examined samples proved to be contaminated with AFB1 despite the presence of mold with mean counts of 3.79±3.29, 4.39±4.34, and 4.84±4.29 log CFU/g in the examined processed cheese, Ras cheese, and raw milk samples, respectively. Therefore, it is urgent to regularly inspect the contamination of animal feeds with AFB1 and apply special measures and novel techniques to protect the feed and food from public health hazards. Keywords: aflatoxin B1, aflatoxin M1, enzyme-linked immunosorbent assay, high-performance liquid chromatography, mold, sensitivity.

The production of male calf beef cattle is an agricultural innovation needed to increase the farm's productivity as a provider of meat sources. This study aimed to determine the sex ratio of the offspring of cows inseminated with Y-bearing sperm enriched by Percoll density gradient centrifugation and swim up, combined with delayed fixed-time artificial insemination (FTAI). Ejaculates of Simmental bulls were divided into four equal portions and grouped as T0 (control, non-sexed semen), T1 and T2 were sexed semen using Percoll density gradient centrifugation three and five levels, respectively, and T3 was sexed semen using swim-up. After the sex was sorted, the semen was diluted in a tris egg yolk extender, packaged in French mini-straws containing 50 million live sperm cells, and frozen. Pre-sexed, post-sexed, and post-thawed spermatozoa were evaluated based on progressive motility, viability, intact plasma membrane, and abnormality. The post-thawed semen of T0 was artificially inseminated to recipient cows at 12 h after onset of estrus (not delayed FTAI). Meanwhile, the delayed FTAI was conducted 18 20 h after onset of estrus using the T0, the best of T1 and T2, and the T3 post-thawed semen. The Percoll density gradient centrifugation reduced motility, viability, and intact plasma membrane but increased sperm abnormalities. Meanwhile, the swim up process increased motility, viability, and intact plasma membrane of sperm cells but decreased sperm abnormalities. Post-thawed semen decreased motility, viability, and intact plasma membrane of sperm cells but increased sperm abnormalities. The sex ratio of the Simmental crossbred offspring was 96.08% and 100% in T1 and T3, respectively, compared to 48.25% and 67.39% in T0 not delayed and delayed FTAI, respectively. The Percoll density gradient centrifugation and swim-up methods are prospective for obtaining male offspring. Keywords: agricultural innovation, farm productivity, motility, pregnancy rate, sperm morphologic abnormality, viability.

Toxoplasma gondii is an intracellular protozoan that infects humans and animals. This study aimed to estimate the seroprevalence of T. gondii and the associated alterations in hematology and serum biochemistry of one-humped camels (Camelus dromedarius) in Mianwali district, Pakistan. A total of 350 blood samples were obtained from male and female camels of different ages (≤3 years old, 4-6 years old, and ≥7 years old). To validate T. gondii antibodies, the collected samples were subjected to indirect enzyme-linked immunosorbent assay using purified recombinant micronemal protein 3 as an antibody catching antigen. The prevalence of T. gondii was 50.2% higher in male camels than in female camels (16.5%) (p<0.001). Furthermore, the prevalence of T. gondii in camels was directly proportional to age (p<0.001). It was 63.33% (57/90) in camels of ≥7 years of age, 32.54% in 4-6 years old age group, and 23.08% in ≤3 years old age group. The hematological analysis of infected camels revealed a significant increase in the values of glucocorticoid-remediable aldosteronism, lymphocyte percentage, monocyte percentage (MONO%), corpuscular hemoglobin (MCH), and procalcitonin. Furthermore, substantially higher levels of liver enzymes alanine aminotransferase, aspartate aminotransferase, and the macro-mineral potassium were found in the serum of T. gondii-infected camels. The seropositivity of T. gondii is directly associated with the age and sex of camels, which may be considered as potential risk factors. Furthermore, T. gondii infection directly impacts the hemato-biochemistry of infected camels. Keywords: biochemistry, camel, hematology, public health, seroprevalence, Toxoplasma gondii.

Measuring blood progesterone (P4) concentration has become an essential diagnostic tool in small animal reproductive medicine. Methods enabling precise and rapid on-site measurements are in high demand, especially for the optimization of breeding management in bitches. This study aimed to compare two commercial on-site methods (Speed™ P4, Virbac [M1] and mini VIDAS®, bioMérieux [M2]) and a well-established radioimmunoassay (RIA), which was used as a reference method. Comparative measurements were performed on 52 blood serum samples collected from 45 clinically healthy bitches of different breeds. The dogs had been presented to determine the estrus cycle stage and predict the time of ovulation. Each sample was divided into three aliquots. In aliquot 1, P4 was measured immediately applying M2. Aliquots 2 and 3 were stored at –20°C until analysis was performed using RIA and M1. The consistency of the three methods was investigated by pairwise linear regression analyses. In RIA, the P4 concentrations ranged between 1.1 and 25.4 ng/mL. Regression analyses revealed highly significant (p<0.0001) positive correlations between the three methods applied (M1 vs. RIA: R=0.94; M2 vs. RIA: R=0.98; and M1 vs. M2: R=0.91). The results show that the two commercial on-site methods tested exhibit approximately equal, high consistency with the radioimmunological reference method and can, therefore, be used beneficially in a clinical setting. However, biological interpretation of data must be performed in a method-specific manner. Keywords: dog, enzyme-linked immunofluorescence, immunochromatography, progesterone, radioimmunoassay.

Cancer is one of the leading causes of death, the need for new anticancer herbal drugs is becoming more urgent considering the side effects of synthetic drugs. This study aimed to determine the anticancer activity of isolates derived from the methanol extract of Annona squamosa Linn. leaves and to identify the compounds that have an active effect against HeLa cells. The leaf metabolites of A. squamosa L. were extracted using methanol at room temperature (28°C) and were partitioned into n-hexane, chloroform, and n-butanol. The toxicity test of these extracts was conducted using a brine shrimp lethality assay. Furthermore, the most toxic extracts were separated and purified using silica gel column chromatography to yield four isolate fractions: FA, FB, FC, and FD. The most toxic isolates were tested for anticancer against HeLa cells, and their compounds were identified using liquid chromatography-mass spectrometry. The results showed that the most toxic isolate with an LC50 value of 100.00 ppm had a potency similar to that of an anticancer agent with an IC50 value of 70.9021 ppm. Furthermore, the five compounds identified in this isolate include (6S, 7aR)-6-hydroxy-4,4,7a-trimethyl-6,7-dihydro-5H-1-benzofuran-2-one or loliolide, cocamidopropyl betaine, N-[3- (dimethylamino)propyl]dodecanamide or lauramidopropyl dimethylamine, linolenic acid, and 1-dodecyl-2-azepanone or laurocapram. It can be concluded that the leaf isolates of A. squamosa Linn. had shown anticancer activities against cervical cancer. Keywords: Annona squamosa Linn, cervical anticancer activities, HeLa cells.

Collie eye anomaly (CEA) is a hereditary and congenital ocular disorder, which affects several dog breeds, including Collies, Collie-related breeds, and other purebreds. An intronic deletion of 7799-bp in the non-homologous end-joining factor 1 (NHEJ1) gene has been identified as the genetic defect causing CEA. This study aimed to investigate the prevalence of CEA based on NHEJ1 genotyping assay in Thailand. We clarified the prevalence of CEA in 224 dogs from five purebred dog breeds using a novel multiplex polymerase chain reaction (PCR)-based technique and confirmed the genotypic status with direct DNA sequencing. The highest frequency of the mutated NHEJ1 allele was 83.3% for Rough Collies, followed by 7.8% for Border Collies, 5.1% for Australian Shepherds, and 2.8% for Shetland Sheepdogs. The heterozygous mutated NHEJ1 genotype detected for Rough Collies, Border Collies, Australian Shepherds, and Shetland Sheepdogs was 33.3%, 15.6%, 10.3%, and 3.3%, respectively. The homozygous mutated NHEJ1 genotype was detected only in Rough Collies and Shetland Sheepdogs, accounting for 66.7% and 1.1%, respectively. Thai Ridgeback Dogs were not affected by this mutation. This study describes, for the 1st time, the genotypic survey of the NHEJ1 gene associated with CEA in dogs in Thailand. In addition, we successfully developed a new multiplex PCR assay with high accuracy, reproducibility, and cost-efficiency and validated its usefulness for determining NHEJ1 genotypes. Keywords: Collie eye anomaly, dogs, multiplex polymerase chain reaction assay, non-homologous end-joining factor 1 genotype.

The nasal cavity of a pig serves as an entry point and a habitat for the colonization of commensal microbes and pathogenic bacteria. Based on biochemical and serological tests, Streptococcus β-hemolytic Group C was identified as the Gram-positive bacteria, which resulted in the 1994 outbreak and death of thousands of pigs in Bali. Furthermore, this agent is zoonotic and frequently results in the development of meningitis lesions in the infected pig. Recently, a meningitis outbreak in humans was also reported after the consumption of pig-derived foods at Sibang Kaja, Badung-Bali. This study aimed to identify and characterize Gram-positive β-hemolytic organisms collected from nasal swab of pigs from the outbreak area, as well as to compare API Kit and 16S rRNA gene analysis methods. This study commenced with the cultivation of two isolates, Punggul Swab Nasal (PSN) 2 and PSN 19, which were characterized by β-hemolysis activity. These samples were then conventionally and molecularly identified using Kit API 20 Strep and 16S ribosomal RNA (rRNA) gene primers, respectively. Using the Kit API 20 Strep, both isolates were identified as Enterococcus faecium, which was previously classified as Group D Streptococci. Based on the 16S rRNA gene sequencing, PSN 2 and PSN 19 were molecularly confirmed to have 99 and 98.1% similarities with E. faecium (NR042054), respectively. Furthermore, both isolates share the same clade in the phylogenetic tree analysis. Using Kit API 20 Strep and 16S rRNA gene analysis, the PSN 2 and PSN 9 Gram-positive isolates with β-hemolysis activity from pig nasal swabs were identified as E. faecium. Keywords: 16S ribosomal RNA gene, Gram-positive bacteria, Kit API 20 Strep, nasal of pig, phylogenetic tree.

Trimmed asparagus by-products (TABP) is the resultant waste from asparagus possessing. TABP has fructans, such as inulins and fructooligosaccharide, which can be utilized as an alternative prebiotic. This study was conducted to examine the effect of TABP dietary supplementation on the productive performance, nutrient digestibility, gut microbiota, volatile fatty acid (VFA) content, small-intestine histology, and meat quality of broilers. A total of 320 1-day-old broiler chicks (Ross 308®) were raised under ambient temperature and assigned through a completely randomized design to one of four dietary treatments, with four replicates per treatment. The dietary treatments comprised corn-soybean basal diet supplemented with 0 (control), 10, 30, or 50 g/kg TABP. All birds were provided drinking water and feed ad libitum to meet the standard nutritional requirements of National Research Council for broiler chickens. TABP supplementation to the broilers significantly increased the apparent ether extract, crude fiber, and gross energy digestibility (p<0.05). TABP supplementation significantly increased lactic bacteria and Enterococcus spp. numbers as well as acetic, propionic, butyric, and total VFA levels (p<0.01); on the other hand, it also significantly decreased Salmonella spp. and Escherichia coli contents in the cecum compared with the control group (p<0.01). Moreover, TABP supplementation increased villus height in the duodenum and jejunum (p<0.01), cryptal depth in the jejunum and ileum (p<0.01), and villus surface areas in the duodenum, jejunum, and ileum (p<0.01). Overall, 0-35 day TABP supplementation significantly increased the feed intake (p<0.01) and average daily gain of broilers (p<0.05), but not significantly affected the viability, productive index, and economic benefit return (p>0.05). The carcass characteristics, pH, color, and water holding capacity of the chicken meat between groups were not significantly different (p>0.05). All levels of TABP supplementation appeared to be a feasible means of producing broilers with the lower serum low-density lipoprotein cholesterol and triglyceride levels as well as atherogenic indices of serum compared with the control (p<0.05). Cholesterol contents and palmitic acid, oleic acid, saturated fatty acids, and Monounsaturated fatty acids levels decreased with an increase of TABP supplementation (p<0.05). Furthermore, TABP supplementation decreased atherogenic index (AI) and thrombogenicity index (TI) of meat (p<0.05). Supplementation of 30 g/kg TABP in broiler diet could enhance broiler performance and provide chicken meat with beneficial properties, with decreased AI and TI resulted from altered cholesterol and fatty acid profiles. Keywords: asparagus by-products, broiler, functional feed, functional meat, prebiotic.

Clostridioides difficile is a spore-forming pathogen that causes serious enteric disease in humans. Strains have been isolated from food animals and meat, including pork, which suggest a potential for foodborne transmission. Pork summer sausage is a popular fermented meat product, which is consumed cooked or cooked to a lower internal temperature due to acidification of the product. The effect of acidity and cooking on the viability of C. difficile spores in a fermented meat product has not been determined. Therefore, the aim was to study the survivability of C. difficile spores in fermented pork summer sausage. Fermented pork sausages were prepared according to a commercial recipe with or without starter culture and C. difficile spores followed by fermentation at 37°C for ∼12 h under 85% relative humidity until pH 5.0 was reached and further processed as cooked (>57°C) or uncooked (≤57°C) and stored at 4°C. C. difficile spores in sausages were enumerated at 1 h following inoculation and on days 0, 1, 7, 14, 21, 30, 60, and 90 of storage. It was observed that C. difficile spore viability in control unfermented treatment was significantly different on day 0 from the fermented, fermented cooked, and control unfermented cooked treatments (p<0.05); however, there was no significant difference among the latter three treatment groups throughout 90 days of storage (p>0.05). On day 90 of storage, the unfermented control sausages yielded ∼4.0 log colony-forming unit (CFU)/g of C. difficile spores compared to ∼3.5 log CFU/g recovered from fermented samples and the unfermented cooked control samples identifying spore viability in all treatment groups. C. difficile spores were found to survive the acidity and cooking of fermented pork summer sausage and storage at 4°C for 3 months, thereby highlighting the need for effective intervention strategies to reduce the risk of C. difficile contamination in pork products. Keywords: acidity, Clostridioides difficile, fermented pork sausage, spores.

Heat shock proteins (HSPs) are a group of proteins that play a significant role in protecting cells against cellular stress. HSP70 is a conserved, sensitive, and abundant gene associated with heat stress's physiological adaptability. The objective of this study was to reveal the polymorphisms of the partial sequences of the HSP70 gene (5' untranslated region [UTR]) in seven cattle populations in Indonesia. Polymerase chain reaction products (551 bp) of the HSP70 gene amplified from 102 animals representing seven cattle populations (Bali, Belgian Blue × Peranakan Ongole [PO] cross, Galekan, Jabres, Madura, PO, and Rambon) were sequenced by DNA sequencing method. Fourteen single-nucleotide polymorphisms (SNPs), generally found at a low frequency, were detected. Among these SNPs, only 1117G>A, 1125A>C, and 1204T>C were polymorphic in all the analyzed breeds. A Chi-square test showed that the majority of the loci were in Hardy–Weinberg equilibrium (p>0.05). Varying levels of observed (0.050-0.571) and expected heterozygosity (0.049-0.500) were noted. The polymorphism information content values (0.048-0.375) indicated that the SNPs in the HSP70 gene showed low-to-moderate polymorphism in the studied populations. Thirty-six haplotypes were defined according to the identified SNPs, of which haplotype Hap5 (CGACGAGAGTGTCC) and Hap4 (CGACGAGAGTGCCC) were generally dominant in the studied samples. The phylogenetic tree showed a close relationship between Bali and Rambon cattle and between Galekan and Jabres cattle, while the Belgian Blue × PO crossbred cattle were farther apart. The polymorphisms in the 5' UTR of the HSP70 gene identified in this study should be further investigated in a larger population to unravel the association between the SNPs and thermotolerance in Indonesian local cattle populations. Keywords: cattle, heat shock protein, heat stress, polymorphism, thermotolerance.

Kefir, a natural probiotic containing bacteria and yeast, is a fermented milk product, whereas glucomannan from porang tuber (Amorphophallus oncophyllus) is prebiotic in vivo. Simvastatin is a potent lipid-lowering statin that can be utilized for pharmacological therapy in obesity. This study aimed to determine the effect of goat milk kefir supplemented with porang glucomannan (synbiotic kefir) and goat milk kefir without glucomannan (probiotic kefir) on blood glucose, hemoglobin A1c (HbA1c), free fatty acids (FFAs), tumor necrosis factor-alpha (TNF-α), gene expression of peroxisome proliferator-activated receptor gamma (PPARγ), and insulin-producing cells in rats fed a high-fat and high-fructose (HFHF) diet. Male Sprague-Dawley rats were divided into five dietary groups: (1) Normal control, (2) rats fed HFHF, (3) rats fed HFHF+probiotic kefir, (4) rats fed HFHF+synbiotic kefir, and (5) rats fed HFHF+simvastatin. All of these treatments were administered for 4 weeks. There were no significant differences in plasma glucose levels in HFHF diet-fed rats before and after treatment. However, plasma HbA1c and TNF-α decreased, and FFAs were inhibited in rats after treatment with synbiotic kefir. Synbiotic kefir decreased the gene expression of PPARγ2 in HFHF diet-fed rats but did not affect the total number of islets of Langerhans and insulin-producing cells. Synbiotic kefir improved the health of rats fed an HFHF diet by decreasing HbA1c, TNF-α, and PPARγ2 gene expression and preventing an increase in FFAs. Keywords: health status, high fat-high-fructose diet, porang glucomannan, simvastatin, synbiotic kefir.

Water plays a pivotal role in the body. Alteration of the fluid balance promotes metabolic disorder, thus leading to the development of various diseases, such as diabetes mellitus (DM). Hydrogen-rich water (HW) is recognized as a novel antioxidant. This study aimed to investigate the role of HW on insulin, insulin receptor (IRs), and superoxide dismutase (SOD) levels in streptozotocin (STZ)-induced diabetic rats. A total of 30 male Wistar rats were randomly divided into five groups: Normal (N), DM rats, DM+metformin (DM+Met, 45 mg/kg body weight [BW]), DM+Met+HW, and DM+HW. DM rats were induced by feeding them a high-fat diet for 30 days and then injecting with repeated low doses of STZ (35 mg/kg BW) intraperitoneally. Fresh HW was administered orally and ad libitum for 14 days. Insulin, IRs, and SOD were observed in each group. HW therapy increased the level and expression of insulin and IRs. In addition, treatment with HW also elevated the SOD levels in the serum and liver. The study results indicated no significant differences between the administration of HW and metformin. HW has antioxidant activity in STZ-induced DM rats, increasing insulin, IRs, and SOD. Keywords: diabetes mellitus, hydrogen-rich water, insulin receptor, insulin, superoxide dismutase.

Dermatitis is a soft-tissue infection caused by Staphylococcus aureus. The recurrence of inflammatory skin is linked to clinical manifestations. Anti-inflammatory cytokines, which are essential for tissue damage, are released by bacteria through skin tissues. Oxidative stress causes inflammatory cells to necrotize and reduces their antioxidant profile, resulting in toxic damage to surrounding tissues. Although studies on the antibacterial effects of Thunbergia laurifolia Lindl., Curcuma longa L., Garcinia mangostana L., and Andrographis paniculata (Burm.). Bacterial infection of S. aureus have been conducted, most of these studies have been in vitro and were not related to the rabbit model. In addition, anti-inflammatory and antioxidant studies need to be evaluated. Thus, this study aims to compare the antibacterial, anti-inflammatory, and antioxidant properties of four local herbs with a standard antibiotic in S. aureus-induced rabbit dermatitis model. The skin of New Zealand white rabbits were artificially wounded using a sterile blade and then infected with S. aureus. The rabbits were divided into seven groups, each with three rabbits (Total 21 rabbits): The first group was the no infection group (no infection and no treatment with scarification), the second group was the no treatment group (S. aureus infection of the wound but no treatment), and the other five treated groups were T. laurifolia, C. longa, G. mangostana, A. paniculata, and bacitracin cream, all of which involved wound infection and treatments. The treatment lasted for 7 days. The antibacterial, anti-inflammatory, and antioxidant properties after treatment were measured. The efficacy of T. laurifolia, C. longa, G. mangostana, and A. paniculata was similar to that of an antioxidant and free radical scavenging property. The bacterial infection process gradually reduced the activities of antioxidant systems (i.e., enzymatic levels and gene expressions) and total glutathione. However, the activities of the antioxidant system were steadily increased when treated with herbal extracts. During bacterial invasion of the skin, the concentration of thiobarbituric acid reactive molecules, the level of lipid peroxidation, and the expression of anti-inflammatory cytokine genes were increased. All these were decreased when herbal extracts were used to treat the lesion. It can be concluded that T. laurifolia, C. longa, G. mangostana, and A. paniculata extract have antibacterial, anti-inflammatory, and antioxidant properties and are effective antibacterial agents. G. mangostana is the most effective herbal extract for antidermatitis and has the potential to be used as an alternative topical treatment. Keywords: antibacterial, antidermatitis, anti-inflammatory, antioxidant, Garcinia mangostana, lipid peroxidation.

Toxoplasma gondii tachyzoite is the infective stage that causes acute infection, leading to severe toxoplasmosis. The tachyzoite stage has been extensively used for several inoculation purposes, including antigen production, immunological studies, nutrition mechanisms, and in vitro drug trials. The use of fresh tachyzoites is required for inoculation in either in vitro or in vivo studies. However, there is a lack of information on preserving live tachyzoites during transportation from laboratories to inoculation sites. Therefore, this study aimed to validate suitable preservative conditions for maintaining live parasites by determining the survival and viability of T. gondii tachyzoites on the basis of different media, temperatures, and incubation times. The free live T. gondii tachyzoites were evaluated on their viability when maintained in different media without 5% Carbon dioxide (CO2). The purified tachyzoites of the RH and PLK strains were individually suspended in normal saline (NS), phosphate-buffered saline (PBS), minimum essential medium (MEM), and MEM with 10% fetal bovine serum (MEM-FBS) and incubated for 6 h at ice-cold (IC; 3-9°C) and room temperature (RT; 25°C). Parasite survival was measured at the 0, 1st, 2nd, 3rd, 4th, 5th, and 6th h post-incubation using the trypan blue exclusion test. The viability was in the range of 85.0%–91.0% for IC using NS and 81.0%–85.1% (IC) and 75.3%–77.5% (RT) using PBS. The viability was approximately 75.0%–83.0% (IC) and 70.0%–79.0% (RT) using MEM and MEM-FBS. There was a significant difference in the viability between the seven periods on the basis of one-way repeated Analysis of variance and Friedman analyses. Parasite survival slightly reduced (20.0%–30.0%) in NS and MEM-FBS at both temperatures during incubation. Notably, PBS could not support tachyzoite viability after 3 h post-incubation. NS was a suitable preservative for maintaining purified T. gondii tachyzoites during transportation at IC and RT without 5% CO2 supplementation. This could be a valuable medium for parasite transportation, especially when there is a large distance between the laboratory and inoculation site. Keywords: preservation time, T. gondii tachyzoites, temperature, transportation, viability.

Mastitis is considered a significant disease of lactating animals. There are new attitudes for recognizing genes responsible for causing this disease to overcome and change the manipulation of this problem. This study aimed to isolate and identify Staphylococcus aureus strains from mastitic bovine animals and detect some specific biofilm-forming genes (icaA, icaD, and biofilm-associated protein [bap] genes clfA, fnbA, agrI, agrII, agrIII, agrIV, and cna). A total of 121 mastitic milk samples were analyzed using biochemical tests (catalase test, oxidative-fermentative test, and coagulase test) and Gram stain. Multiplex polymerase chain reaction was applied to characterize biofilm genes (icaA, icaD, bap, clfA, and fnbA) in addition to (agrI, agrII, agrIII, agrIV, and cna). Among the 121 milk samples, 35 staphylococci isolates were derived with an incidence of 28.92% (35/121); among them, 19 are coagulase positive. Ninety percent of the isolates had ica genes (icaA and icaD) while bap gene was not recognized in any isolate. In addition, the incidence of fnbA, can, and clfA was 89.5% each. The prevalence of agr specific groups (agrI, agrII, agrIII, and agrIV) was 78.9%, 52.6%, 10.5%, and 15.8%, respectively. This study concluded that S. aureus has variant mechanisms of pathogenicity to form biofilm devoid of carrying a specific gene. Keywords: biofilm genes, mastitis, molecular identification, Staphylococcus aureus.

At present, tick-borne borreliosis is the most common infectious disease transmitted by ticks in Europe, Asia, and North America. This study aimed to examine the epizootiological aspects of the natural nidality of tick-borne borreliosis in Moscow region (the Russian Federation). A total of 2,537 ticks representing two species were collected, namely, Ixodes ricinus and Dermacentor reticulatus. The activity, number of ticks, and Borrelia infestation rates were investigated during a high season, that is, from early spring to mid-autumn. In May, amount of I. ricinus spp. was found 2.5 times more than those representing D. reticulatus spp. (p≤0.01). In June, August, and September, the amount of I. ricinus was 9.0 (p≤0.0001), 2.0 (p≤0.05), and 5.0 times higher, respectively, compared to D. reticulatus. In the first 10 days of April, the amount of D. reticulatus was 3 times higher than that of I. ricinus (p≤0.02); in the next 10 days, their amounts were equal (p≥0.05) and in the last 10 days the amount of I. ricinus exceeded that of D. reticulatus (p≤0.05) by 1.5 times. In general, Borrelia afzelii, and Borrelia garinii, were detected. In addition, the naturally occurring tick-borne borreliosis pesthole was revealed in the Moscow region. Borrelia infection rates for ticks comprise 30%. An increase in Borrelia tick infestation was detected within the vicinity of populated areas. The amount of ticks directly depends on the temperature (20°C-25°C) and moisture (from 50%) values. Keywords: borreliosis, Dermacentor reticulatus, infection, Ixodes ricinus, Ixodes ticks.

The search and development of disinfectants is promising worldwide. However, there are currently no international regulations governing the testing and registration of germicidal agents. Moreover, the number of safety requirements for disinfectants for human, animal, and environmental health has increased. This research aimed to evaluate the prospects of using a collection of bacteriophages for disinfectant purposes. The objects of research were bacteriophages isolated from a total of 129 environmental samples obtained from seven sources in and around livestock buildings: (1) Feed residues from feeders and automatic drinkers; (2) washouts from floors, walls, and posts; (3) soil from underneath floors; (4) bedding; (5) sewage; (6) ponds; and (7) soil from paddocks. The corresponding strains were used as indicator test cultures for bacteriophages. The authors employed the following methods to work with bacteriophages: (a) Bacteriophage isolation methods, (b) the Appelman method (i.e., serial dilutions), (c) the Grazia method (i.e., agar layers), (d) phage titration on solid media, and (e) the bacterial phagotyping method. The results of the analysis on the bacteria of the Enterobacteriaceae family isolated 11 bacteriophages; one bacteriophage is specific to Pseudomonas aeruginosa, and another one is specific to Brucella abortus. The results also indicate that all bacteriophage strains of the Enterobacteriaceae family demonstrate lysis at a pH of 7.0. In addition, this polyphage lyses all strains of sensitive bacterial cultures. The optimum temperature for the cultivation of bacteriophages is 35°C. While using electron microscopy to study the consortium of bacteriophages, clearly distinguishable virions of bacteriophages were found in the microscope field of view. The main parameters for the production of polyphages include the ratio of the bacteriophage and its corresponding bacteriophage-sensitive culture, the pH of the cultivation medium, and the cultivation time of the bacteriophage system as well as the sensitive bacterium. With regard to the aforementioned parameters, the results indicate that the average value for all bacteriophages is 1:2, and the average cultivation medium pH is 7.0 for all bacteriophages. The average cultivation time for all bacteriophages is 18-24 h. Keywords: bacteriophages, biological preparation, disinfection, lytic activity, strain, titer.

Ehrlichia canis is a well-known cause of both anemia and thrombocytopenia in dogs. There are insufficient epidemiological data on this blood parasite in Thailand and the association of infections with hematological abnormalities. This study aimed to analyze the molecular characteristics and to identify E. canis as well as the risk factors associated with E. canis infection in dogs in Khon Kaen, Thailand. Blood samples from 126 dogs that visited animal clinics were subjected to molecular detection using nested polymerase chain reaction for E. canis 16S rRNA gene. The risk factors and hematological profiles associated with the infection were analyzed using the logistic regression test in program SPSS version 19. Forty-one dogs were infected, indicating a 32.5% molecular infection rate of E. canis. The factors significantly associated with E. canis infection include animal housing status, low packed cell volume, low red blood cell count, and low platelets (p<0.05). Ten positive samples were amplified, sequenced, and phylogenetically analyzed. Sequence and phylogenetic analysis confirmed the current ten samples as E. canis compared with reference sequences in GenBank, using the BLAST program hosted by NCBI, which showed 99.74-100% similarity. This study provided the first data of infection rate of E. canis using nested PCR and molecular characteristics of E. canis in randomly selected domestic dogs in Khon Kaen, Thailand. Keywords: Ehrlichia canis, molecular characteristics, nested polymerase chain reaction, phylogenetic analysis, Thailand.