Dietary fiber has distinctive effects on the environment and microbiota of the pig's intestinal tract. This study was conducted at the naturally ventilated facility of the experimental station, National Institute of Animal Sciences, Vietnam, to examine the effects of fiber sources in diets on the intestinal microbiota of two different pig breeds raised in Vietnam. A total of 18 native and 18 exotic pigs with average initial body weights of 9.5±0.4 and 16.5±0.4 kg, respectively, were each divided into three dietary treatments, including a low-fiber diet containing approximately 200 g NDF per kg dry matter (DM) and two high-fiber diets containing cassava by-products or brewer's grains containing approximately 300 g NDF per kg DM. At the end of the experiment (28 days), the bacterial diversity of digesta samples collected from the stomach, ileum, and colon segments was analyzed through DGGE analysis of the V3 variable regions of 16S-rDNA and by cloning and sequencing. Among the diets, significant differences were observed in the number of DNA bands in the stomach between the native and exotic pigs (p<0.05), but not in the ileum and colon. The dietary fiber affected the number of DNA bands in the ileum (p<0.05), but not in the stomach and colon. A significant interaction effect was found between diet and breed on the number of DNA bands in the ileum (p<0.05). Dietary fiber and breed had a greater effect on microbiota in the ileum and colon than that in the stomach. The fiber sources affected the number of DNA bands in the ileum, and breed affected the number of DNA bands in the stomach. The microbial compositions in the ileum and colon segments were significantly affected by the dietary fiber and breed. Keywords: bacterial diversity, brewer's grain, cassava by-products, exotic pig, fiber, native pig.

Umbilical cord blood (UCB) cells are an important source of mesenchymal stem cells (MSCs). It is known that the umbilical cord is rich in hematopoietic stem cells, which influenced research on ontogeny and transplantation (allogeneic transplantation). In recent years, stem cell research has emerged as an area of major interest due to its prospective applications in various aspects of both human and veterinary medicine. Moreover, it is known that the application of MSCs has several weaknesses. The use of these cells has limitations in terms of tumorigenesis effect, delivery, safety, and variability of therapeutic response, which led to the use of secretomes as an alternative to cell-free therapy. The main obstacle in its use is the availability of human UCB as an origin of MSCs and MSCs' secretomes, which are often difficult to obtain. Ethical issues regarding the use of stem cells based on human origin are another challenge, so an alternative is needed. Several studies have demonstrated that MSCs obtained from bovine umbilical cords have the same properties and express the same surface markers as MSCs obtained from human umbilical cords. Therefore, secretomes from MSCs derived from domestic animals (bovine) can possibly be used in human and veterinary medicine. This finding would contribute significantly to improve cell-free therapy. At present, the use of UCB MSCs derived from domestic animals, especially bovines, is very restricted, and only limited data about bovine UCB are available. Therefore, the aim of this review was to provide an updated overview of cell-free therapy and discuss the new possibilities introduced by the generation of this therapy derived from bovine umbilical MSCs as a promising tool in developing modern and efficient treatment strategies.

The pandemic (H1N1) 2009 influenza (H1N1pdm09) virus has affected both human and animal populations worldwide. The transmission of the H1N1pdm09 virus from humans to animals is increasingly more evident. Captive animals, particularly zoo animals, are at risk of H1N1pdm09 virus infection through close contact with humans. Evidence of exposure to the H1N1pdm09 virus has been reported in several species of animals in captivity. However, there is limited information on the H1N1pdm09 virus infection and circulation in captive animals. To extend the body of knowledge on exposure to the H1N1pdm09 virus among captive animals in Thailand, our study investigated the presence of antibodies against the H1N1pdm09 virus in two captive animals: Camelids and Eld's deer. We investigated H1N1pdm09 virus infection among four domestic camelid species and wild Eld's deer that were kept in different zoos in Thailand. In total, 72 archival serum samples from camelid species and Eld's deer collected between 2013 and 2014 in seven provinces in Thailand were analyzed for influenza antibodies using hemagglutination inhibition (HI), microneutralization, and western blotting (WB) assays. The presence of antibodies against the H1N1pdm09 virus was detected in 2.4% (1/42) of dromedary camel serum samples and 15.4% (2/13) of Eld's deer serum samples. No antibodies were detected in the rest of the serum samples derived from other investigated camelids, including Bactrian camels (0/3), alpacas (0/5), and llamas (0/9). The three positive serum samples showed HI antibody titers of 80, whereas the neutralization titers were in the range of 320-640. Antibodies specific to HA and NP proteins in the H1N1pdm09 virus were detected in positive camel serum samples using WB. Conversely, the presence of the specific antibodies in the positive Eld's deer serum samples could not be determined using WB due to the lack of commercially labeled secondary antibodies. The present study provided evidence of H1N1pdm09 virus infection in the captive dromedary camel and Eld's deer in Thailand. Our findings highlight the need for continuous surveillance for influenza A virus in the population of dromedary camels and Eld's deer. The susceptible animal populations in close contact with humans should be closely monitored. Further study is warranted to determine whether Eld's deer are indeed a competent reservoir for human influenza virus. Keywords: camel, Eld's deer, pandemic (H1N1) 2009 virus, serosurveillance, Thailand.

Fibrin forms in the coagulation process, enhancing local hemostatic properties and promoting wound healing. The study aimed to evaluate the efficacy of bubaline-derived fibrin glue in silk ligature-induced periodontitis rats. Bubaline blood–derived fibrin glue was prepared using cryoprecipitation and cryocentrifugation. Periodontitis was induced in rats by placing 5-0 silk ligatures around the mandibular first molars. The animals were divided into two groups: (1) Non-treatment and (2) bubaline fibrin glue–treated groups. Plaque, gingival inflammation, and mobility index were scored on days 1, 7, and 14 after intervention. Histological examinations were performed. The mRNA expression of inflammatory cytokines and growth factors was evaluated using a real-time polymerase chain reaction. Ligature-induced periodontitis was confirmed by the increase in inflammatory cell infiltration as well as histological bone and attachment loss. Compared to the non-treatment group, bubaline fibrin glue application reduced mononuclear cell infiltration into periodontal tissues corresponding to the reduction of collagen destruction. On days 7 and 14 after intervention, the inflammatory score and histological attachment loss were significantly lower in the bubaline fibrin glue–treated group than in the non-treatment group. A significant reduction in histological bone loss was observed in the treated group on day 7. Bubaline fibrin glue application led to a significant reduction of Tnfa and Il1b mRNA levels, while an increased expression of Pdgfa, Tgfb1, and Il10 was observed compared with the control. Bubaline fibrin glue could be beneficial in periodontitis treatment aiming to reduce inflammation and delay the progression of periodontal disease. Keywords: bubaline, fibrin, inflammation, ligature-induced, periodontitis.

Anaplasma platys is a blood parasite that infects platelets, causing thrombocytopenia. Rhipicephalus sanguineus ticks are believed to transmit A. platys. To identify A. platys, nested polymerase chain reaction (PCR) has proven to be an effective diagnostic tool. In this study, the molecular prevalence of A. platys infection in dogs was investigated for the 1st time in the Khon Kaen region of Thailand. The association between risk factors and A. platys infection was also evaluated. A total of 130 blood samples were collected from dogs in Khon Kaen, Thailand. DNA from the samples was extracted and nested PCR was applied for molecular analysis. Platelet count and packed cell volume (PCV) levels were measured. Platelet counts were categorized into four grades: Non-thrombocytopenia (platelets >200,000 cells/μL), mild thrombocytopenia (platelets 150,000-200,000 cells/μL), moderate thrombocytopenia (platelets 100,000-150,000 cells/μL), and severe thrombocytopenia (platelets <100,000 cells/μL). Four categories for PCV levels of >37%, 30-37%, 20-29%, and <20% were defined as no anemia, mild anemia, moderate anemia, and severe anemia, respectively. DNA sequencing was analyzed using BTSeq™ (Barcode-Tagged Sequencing; CELEMICS, Seoul, South Korea) for similarity index. Among the 130 samples, 9 (6.9%) were positive for A. platys infection. There was an association between low platelet count and infection (p<0.05). PCV level was also associated with A. platys infection (p<0.05). DNA sequencing results of the nine positive samples showed similarity to known sequences of A. platys with 99.36-100% nucleotide identity. These results suggested low genetic diversity in A. platys infecting dogs in the Khon Kaen area. By amplifying 16S rRNA, A. platys infection was detected in the blood of Thai dogs. Further work should be performed to identify risk factors potentially associated with A. platys infection in dogs in Khon Kaen. Other related factors should also be considered, such as location and breeding, as well as the environmental characteristics of each locality. In addition, sampling a larger number of animals may reveal predictors for the positivity of A. platys in dogs in this region. Keywords: Anaplasma platys, molecular prevalence, nested polymerase chain reaction, thrombocytotropic anaplasmosis.

Actinomycetes are a group of Gram-positive bacteria with a fungus-like morphology. Their natural habitat encompasses terrestrial and water areas, including mangrove ecosystems. This study aimed to assess the PKS and NRPS genes as the producers of secondary metabolites and to determine the target bacterial species using molecular DNA tests. In this study, we isolated bacteria from sediment samples from mangrove forests located on Karimunjawa Islands and in Semarang city, purified bacteria, screened for antibacterial activity, extracted bacterial DNA, amplified the NRPS gene, detected and amplified the PKS-I and PKS-II genes, amplified and sequenced the 16S rRNA, processed molecular data, and simulated a map of secondary metabolite producing genes. Samples from the Karimunjawa Islands yielded 19 bacterial isolates, whereas samples from Semarang yielded 11 bacterial isolates after culture in different media. Further experiments identified three active isolates, which were termed PN.SB.6.2, S.SK.6.3, and S.SK.7.1, against pathogenic species of Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes. Isolate PN.SB.6.2 was determined to possess three biosynthetic gene clusters (BGCs), whereas the remaining two isolates, S.SK.6.3 and S.SK.7.1, only possessed two BGCs, namely, NRPS and PKS II. Products were estimated to be in the NRPS, thiopeptide, RiPP-like, siderophore, betalactone, terpene, Type III PKS, CDPS, and lassopeptide groups. DNA identification of the isolates found three species of actinomycetes with antibacterial potential, namely, Virgibacillus salaries, Bacillus licheniformis, and Priestia flexa. Keywords: actinomycetes, biosynthetic gene cluster, Karimunjawa Island, NRPS, PKS.

African swine fever (ASF) is a viral disease of pigs caused by ASF virus (ASFV). High mortality and the lack of available treatments have severely impacted the swine industry resulting in huge global economic losses. In response to the dire necessity for vaccines, this study aims to identify highly conserved cytotoxic T-cell epitopes in ASFV structural proteins pp220, pp62, p72, p30, and CD2v through immunoinformatics approach. The amino acid sequences of the structural proteins were retrieved from the National Center for Biotechnology Information protein database. The sequences were evaluated in CD-HIT Suite wherein resulting representative sequences were aligned in Clustal Omega. Highly conserved sequences were identified in the Protein Variability Server which were used as reference sequences for the cytotoxic T-cell epitope mapping. Epitopes were predicted using the tools in Immune Epitope Database. Peptides which bind to the swine major histocompatibility complex with IC50 binding scores >500 nM were filtered out. Epitopes which are classified to be potentially toxic and cross-reactive with the swine proteome sequences were all excluded from the study. The epitopes were docked with the swine leukocyte antigen-1*0401 (SLA-1*0401) wherein the binding affinity, the binding energy, and the root-mean-square deviation (RMSD) per residue of epitope-SLA complexes formed were determined and compared with the influenza epitope as positive control. A total of 112 highly conserved fragments with Shannon variability index ≤0.1 were identified. These include 66, 12, 26, 6, and 2 highly conserved fragments from ASFV proteins pp220, pp62, p72, p30, and CD2v, respectively. From these reference sequences, 35 nonameric peptides were selected for the list of candidate cytotoxic T-cell epitopes. These include 26 epitopes for pp220, 7 for pp62, 6 for p72, and one each for p30 and CD2v. Bioinformatics analysis classified the peptides as non-toxic. Further evaluations of epitopes showed that these are less likely to cross-react with the domestic swine proteome sequences. This study identified candidate epitopes from pp220 (IADAINQEF, FLNKSTQAY, QIYKTLLEY, and SLYPTQFDY), and pp62 (GTDLYQSAM, FINSTDFLY, and STDFLYTAI) which can bind to at least two widely distributed SLAs in pig populations. The immunogenicity of candidate peptides RSNPGSFYW, DFDPLVTFY, AIPSVSIPF, and VVFHAGSLY was validated by the acceptable binding affinities, binding energies, and RMSD of the peptide-SLA complexes formed. Results were also comparable with the crystal structure of an SLA-epitope complex in the database. This is the first study to identify highly conserved cytotoxic T-cell epitopes in the structural proteins of ASFV. Overall, the results of in silico evaluations showed that the identified highly conserved cytotoxic T-cell epitopes may be used as part of future vaccine formulations against ASFV infection in domesticated pigs. Nonetheless, these findings require in vitro and in vivo validation before application. Keywords: African swine fever virus, cytotoxic T-cell epitopes, immunoinformatics, in silico.

The aim of this study was to characterize Leucocytozoon caulleryi from backyard chickens in Khon Kaen Province, Thailand. Tissue samples were collected from backyard chickens suspected to have leucocytozoonosis and subjected to histopathology examination. The BLAST Tool at NCBI GenBank (Basic Local Alignment Research Tools) (http://www.ncbi.nlm.nih.gov/BLAST) was used to identify the nucleotide sequence of the partial cytochrome c oxidase subunit I (cox I) gene. A Phylogenetic tree for analysis of L. caulleryi was constructed by using MEGA 7.0 software (https:// www.megasoftware.net/). The necropsy results revealed the subcutaneous hemorrhages of pectoral muscles, multifocal hemorrhages of the thymus and pectoral muscles, hemorrhage of the proventriculus and peritoneal cavity, and megaloschizonts of the pancreas and intestine, including subcapsular hemorrhages of the liver. Microscopic examination revealed numerous megaloschizonts of Leucocytozoon spp. in the pectoral muscles, intestine, pancreas, and thymus. Molecular analysis of the partial cox I gene showed that the causal agent was closely related to L. caulleryi reported in Japan. From these results, L. caulleryi was determined to be the causal agent of leucocytozoonosis and was closely associated with L. caulleryi reported in Japan. Keywords: chickens, cytochrome c oxidase subunit I gene, leucocytozoonosis, megaloschizonts.

Avian pox is a contagious disease caused by the avian pox virus (APV). Mangostin and γ-mangostin in mangosteen rind (MR) and gingerol in red ginger (RG) exhibit antiviral activity. In this study, we evaluated the effect of MR and RG ethanolic extracts on APV based on pock lesions on the chorioallantoic membrane (CAM) of specific pathogen-free (SPF) embryonated chicken eggs (ECEs). Three APVs from chicken isolates (C1, C2, and C3), one APV from a pigeon isolate (P), 1.5% and 3% MR ethanolic extract, 5% and 10% RG ethanolic extract, and a combination of 1.5% MR and 5% RG at 0.1 mL/ egg were inoculated in ovo (7th day incubation, chorioallantoic route) in SPF ECEs. A control group inoculated in ovo with APV alone was also established. Each treatment consisted of three replicates. Parameters including embryo survival, CAM lesions, and average number of pock lesions were determined. In ovo inoculation of MR and RG ethanolic extracts was not harmful to the ECEs and did not induce CAM lesions. The average number of pock lesions in the control group (C1, C2, C3, and P) was 35, 14, 10, and 17, respectively, whereas in all treatment groups, the number was 0, except in the 5% RG group of C1, which had a value of 10. In ovo inoculation of 1.5% and 3% MR, 5% and 10% RG, and the combination of 1.5% MR plus 5% RG ethanolic extract s at 0.1 mL/egg inhibit APV by reducing the number of pock lesions on the CAM of the ECE. Keywords: avian pox virus, in ovo, mangosteen rind, pock lesions, red ginger.

CD 117 (c-KIT) internal tandem duplication (ITD), octamer-binding transcription factor 4 (Oct-4), and sex-determining region Y-box 2 (Sox-2) may govern the oncogenicity and aggressiveness of canine cutaneous mast cell tumor (MCT) in the crossbred dogs. Thus, a comprehension of this matter may help us establishing a novel platform to treat the disease in those dogs. However, evidence has lacked so far. Thus, this study aimed to survey CD 117 ITD, Oct-4, and Sox-2 expressions and their relations to the 2-tier grading in a group of Thai crossbreed dogs. The study was done using polymerase chain reaction (PCR), Reverse transcription PCR (RT-PCR), and immunohistochemistry. Thirty-three MCT specimens graded by the 2-tier histopathology grading were collected from the crossbred and purebred dogs. CD 117 ITD was detected by conventional PCR and immunohistochemistry. While, Oct-4 and Sox-2 expression levels were determined at the protein and mRNA levels by immunohistochemistry and RT-PCR, respectively. The expression magnitude of each parameter was then related to the grades and breeds. About 60.61% of specimens were low grade, while 39.39% were high grade. CD 117 ITD was not detected in all specimens. A significant increase of Oct-4 expression was found in the high-grade, crossbred dogs. Meanwhile, Sox-2 expressions were increased both in the purebred and crossbred dogs with high-grade MCT. The study finding has indicated that the level of Sox-2 expression may be a useful tumorigenic and prognostic biomarker because it correlates to the 2-tier grades but not dog breeds. Keywords: canine, CD 117, crossbred, mast cell tumor, octamer-binding transcription factor 4, sex-determining region Y-box 2.

Cercarial dermatitis or swimmer's itch is an allergic skin reaction caused by penetrating cercaria of animal blood flukes. It is considered as a zoonotic water-borne skin condition that is found globally. Among the schistosomatid trematodes, avian schistosomes are the most responsible for cercarial dermatitis. Very little is known regarding the occurrence of dermatitis-causing cercariae in Thailand. Therefore, the objective of this study was to preliminarily investigate the presence of larval blood fluke infection among local lymnaeidae snails in Phayao by the incorporation of morphological and molecular methods. Overall 500 Radix (Lymnaea) rubiginosa (Michelin, 1831) were collected from freshwater reservoirs near Phayao Lake in San Kwan village in Phayao, Thailand, from October to December 2020. The snails were examined for avian blood fluke infection by the cercarial shedding technique followed by morphological and molecular characterization. Only one type of furcocercous cercaria was observed to emerge from six infected snails (1.2%). Our molecular analyses demonstrated that the emerging cercariae showed most similarity to either the 28S ribosomal RNA gene (28S rDNA) or cytochrome oxidase C subunit 1 gene (cox1 or COI) sequences to those of Trichobilharzia species. In addition, phylogenetic tree analyses of both loci revealed similar results; the emerging cercariae were consistently clustered together with Trichobilharzia regenti. Our results clearly confirmed that the detected furcocercous cercariae belonged to the genus Trichobilharzia and displayed the highest homology to T. regenti. This study provides important data on the occurrence of dermatitis causing cercariae infection among local lymnaeidae snails, encouraging effective management, and control measures for this zoonotic infectious disease. Keywords: avian schistosome, cercarial dermatitis, furcocercous cercaria, swimmer's itch, Radix (Lymnaea) rubiginosa, Trichobilharzia regenti.

Antimicrobial resistance (AMR) and recently mobilized colistin resistance (mcr-1) associated colistin resistance among Escherichia coli isolates have been attributed to the overuse of antimicrobials in livestock production. E. coli remains an important pathogen, often associated with mortality and low carcass weight in poultry medicine; therefore, the need to use antimicrobials is common. The study aimed to determine the AMR profile and presence of mcr-1 and mcr-2 genes in avian pathogenic E. coli from poultry samples tested at a bacteriology laboratory for routine diagnosis. This is a first step in understanding the effectiveness of mitigation strategies. Fifty E. coli strains were assessed for resistance against ten antimicrobial drugs using broth microdilution. All isolates with a colistin minimum inhibitory concentration (MIC) of 2 μg/mL were analyzed for the presence of mcr-1 and mcr-2 genes by employing the polymerase chain reaction. For each isolate, the following farm information was obtained: farm location, type of farm, and on-farm use of colistin. Sixty-eight percent of the strains were resistant to at least one antimicrobial; 44% were multiple drug-resistant (MDR). Most E. coli isolates were resistant to doxycycline (44%), trimethoprim-sulfamethoxazole (38%), ampicillin (32%), and enrofloxacin (32%). None of the E. coli strains was resistant to colistin sulfate (MIC90 of 2 μg/mL). Only one E. coli isolate held the mcr-1 gene; none carried the mcr-2 gene. Resistance among E. coli isolates in this study was fairly high. Resistance to commonly used antimicrobials was observed, such as doxycycline, trimethoprim-sulfamethoxazole, and enrofloxacin. Only a single E. coli strain carried the mcr-1 gene, suggesting that mcr-1 and mcr-2 genes are common among isolates in this study. The prevalence of AMR, however, suggests that farmers must implement standard biosecurity measures to reduce E. coli burden, and antimicrobial use to prolong the efficacy life span of some of these drugs. Keywords: antimicrobial resistance, colistin, Escherichia coli, mobilized colistin resistance-1, poultry.

Ultrasound-guided fine-needle sample collection for cytology with manual restraint is frequently used for the primary assessment of diffuse liver disease in veterinary patients in Thailand. For better diagnosis, repeated collection of samples ensures the collection of adequate, representative samples, which increase diagnostic accuracy. However, in those that are unable to receive general anesthesia, it is difficult to collect the samples from several liver locations in manually restrained dogs and cats. The study aimed to compare the cytologic diagnosis of the ultrasound-guided fine-needle non-aspiration technique between the left and right liver lobes in dogs and cats with neoplastic and non-neoplastic diffuse liver disease. This prospective study included 25 client-owned dogs and cats with diffuse liver diseases. Two liver samples were randomly collected from the left and right liver lobes under ultrasound guidance for cytologic examination. All slides were subsequently examined blindly by experienced pathologists for cytologic analysis with cytologic agreement scores (CASs). Among all 50 samples obtained from ultrasound-guided fine-needle sample collection of the left and right liver, 78% were diagnostic and 22% were non-diagnostic. In the diagnostic group, 73.3% of fine-needle samples had concordant results between the left and right liver, which exhibited 100% cytologic agreement in lymphoma and 63.6% in non-neoplastic groups. Samples collected from the left liver had slightly higher CAS and higher cytologic quality than had those from the right liver lobe (p=0.053). The location and number of sample collections did not have a significant difference in the cytologic diagnosis of diffuse liver disease, especially in patients with lymphoma. For manually restrained patients, one time ultrasound-guided non-aspiration cytology procedure from the left liver lobe not only decreased restraint duration and minimized tissue trauma but also allowed for an adequate cytologic diagnosis in diffuse liver disease compared to multiple collections. Keywords: aspiration, cytology, diffuse, hepatic, liver, ultrasound.

The transition period is extremely critical for pregnant producing animals. However, there is very limited research on the metabolic and immunological changes in Egyptian water buffalo cows during the transition period. Therefore, this study was conducted to investigate the immunometabolic changes occurring during the transition period in Egyptian water buffalo cows. A total of 50 multiparous pregnant Egyptian water buffalo cows were subjected to weekly blood sampling 3 weeks before calving and 3 weeks after calving and on the day of parturition to determine the complete blood count, including red blood cell count, total leukocyte count (TLC), differential leukocyte count, hemoglobin level, and packed cell volume (PCV). Some selected serum biochemical and immunological parameters were analyzed, including serum glucose, beta-hydroxybutyric acid (BHBA), non-esterified fatty acids, triglycerides, high-density lipoprotein, low-density lipoprotein (LDL), very LDL (VLDL), cholesterol, total protein, albumin, globulin, creatinine, blood urea nitrogen (BUN), aspartate aminotransferase, alkaline phosphatase, alanine transaminase, gamma-glutamyl transferase, Haptoglobin, and C-reactive protein and the pro-inflammatory cytokines interleukin β1, interleukin 6 (IL-6), and tumor necrosis factor-alpha. All data were statistically analyzed using the IBM Statistical Package for the Social Sciences statistics software. The neutrophil count showed a statistically significant increase at 2 weeks preparturition. There was also a significant increase in PCV, TLC, neutrophil count, and IL-6 and TNF-α level at the time of parturition and even at 2 weeks post parturition, except PCV that returned to normal levels in the 1st week post parturition. BHBA and BUN levels were increased significantly in the 2nd and 3rd weeks postcalving. Serum creatinine and VLDL levels were decreased significantly at the time of parturition, and VLDL levels showed a significant decrease even till the 3rd week postcalving, whereas creatinine levels gradually returned to the pre-calving levels in the 3rd week postcalving. Other parameters showed no significant changes. The most important immunometabolic changes occur in the first 2 weeks post parturition in Egyptian water buffalo cows, which exhibit a potent, remarkable physiological adaptation achieved by their functional liver, which can help the animal overcome the stressful conditions during the transition period. Keywords: Egyptian water buffalo, immunometabolic changes, transition period.

In tropical semiarid regions, supplementation with fungi could contribute to rumen modulation, promoting greater production of fibrolytic enzymes and degradation of forage. The objective of this study was to analyze the effect of supplementation with fungi, isolated from the bovine rumen, on the performance and microbiota of the digestive tract of Nellore calves. The experiment was conducted in randomized blocks evaluating eight Nellore calves that were daily supplemented with isolates of Aspergillus terreus and Trichoderma longibrachiatum, along with eight calves that were not supplemented. After 55 days, the animals were weighed, and samples of rumen fluid and feces were collected for analysis. The characteristics that showed normal distribution were subjected to analysis of variance and compared using Tukey's test. Whereas, the variables that did not show normal distribution were subjected to the Kruskal–Wallis test, and the frequencies of the bacterial and fungal genera were compared using the Chi-square test. Supplementation with fungi promoted the reduction in ruminal pH (p<0.05). However, the final live weight; average daily weight gain; total weight gain; rumen protozoa; and the count of Enterobacteriaceae, mycelial fungi, and yeasts of ruminal fluid and feces were not influenced by supplementation (p>0.05). Moreover, the protozoa Eodinium spp. was identified only in supplemented calves (p<0.05). Supplementation with the fungi presented the potential for use as possible additives because it did not alter the physiological parameters of the facultative anaerobic microbiota composition in the rumen and feces. In addition, it favored the presence of the ciliate genus Eodinium. However, further studies should be performed to better define suitable dosages for supplementation. Keywords: Enterobacteriaceae, ruminal microbiota, rumen protozoa, semiarid region, weaning.

The classification of diabetes mellitus (DM) in dogs has been controversial as currently canine insulin-dependent DM is classified together with absolute insulin deficiency, non-insulin-dependent DM, and relative insulin deficiency. Studies on human autoantibodies evaluated in canines with DM, such as anti-glutamic acid decarboxylase (GAD65), anti-islet antigen 2 (IA2), and anti-zinc transporter isoform 8 (ZnT8), have been inconclusive. Thus, this study was designed to establish the serological profile of anti-GAD65, anti-IA2, and anti-ZnT8 antibodies in a group of dogs with and without DM. Sixty-one dogs, including 31 patients with DM (with and without insulin treatment) and 30 patients without DM (normal weight and obese), were included for determining autoantibodies using a human enzyme-linked immunosorbent assay (ELISA) detection system for type 1 DM. This study found the presence of anti-IA2 antibodies in 58% of the sample (18/31 patients with DM); however, the presence of anti-GAD65 was not detected, and anti-ZnT8 was found in 3 (9.6%) patients with DM. This study showed a higher positive frequency of anti-IA2 antibodies in a sample of canine with DM, indicating that alterations in the signaling vesicle tyrosine phosphatase 2 lead to lower insulin release and thus to an increase in patients' glycemia. These preliminary results should be taken with caution and corroborated by a canine-specific assay when an ELISA is available for such determination. Keywords: autoantibodies, autoimmunity, diabetes, dogs.

Momordica charantia is mainly characterized by its antimicrobial and antioxidant properties. The current study aimed to evaluate the healing activity of gel and cream formulations based on M. charantia on induced wounds in mice. Acetonic extract of M. charantia was prepared and incorporated into gel and cream formulations. Mus musculus Balb/c (n=30) with induced injury were distributed into five groups: Group I (control – day 7), Group II (control – day 14), Group III (1% gel – day 7), and Group IV (1% gel – day 14) to which 1% M. charantia gel was dermally applied daily for 7 and 14 days, respectively, Group V (1% cream – day 7) and Group VI (1% cream – day 14) to which of M. charantia 1% cream were dermally applied daily for 7 and 14 days, respectively. Time of wound closure was determined during the experimentation; rats were euthanized with sodium pentobarbital 60 mg/kg/pc v.ip. for obtaining skin samples for histopathological analysis. Groups IV and VI showed a higher percentage of wound closure on day 14, and in histopathological analysis, effect was greater in Group VI with the presence of fibroblasts and abundant collagen and elastic fibers. M. charantia gel and cream showed wound healing activity on induced wounded mice; the most effective treatment was M. charantia 1% cream formulation. Keywords: histology, Momordica charantia, skin, topical administration, wound healing.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of coronavirus disease 2019 (COVID-19). This virus has become a global pandemic with unprecedented mortality and morbidity along with attendant financial and economic crises. Furthermore, COVID-19 can easily be transmitted regardless of religion, race, sex, or status. Globally, high hospitalization rates of COVID-19 patients have been reported, and billions of dollars have been spent to contain the pandemic. Angiotensin-converting enzyme (ACE) 2 is a receptor of SARS-CoV-2, which has a significant role in the entry of the virus into the host cell. ACE2 is highly expressed in the type II alveolar cells of the lungs, upper esophagus, stratified epithelial cells, and other tissues in the body. The diminished expressions of ACE2 have been associated with hypertension, arteriosclerosis, heart failure, chronic kidney disease, and immune system dysregulation. Overall, the potential drug candidates that could serve as ACE2 activators or enhance the expression of ACE2 in a disease state, such as COVID-19, hold considerable promise in mitigating the COVID-19 pandemic. This study reviews the therapeutic potential and pharmacological benefits of the novel ACE2 in the management of COVID-19 using search engines, such as Google, Scopus, PubMed, and PubMed Central.

Acetate or lactate buffered, balanced isotonic rehydration fluids are commonly used for fluid therapy in dogs and may influence acid-base and electrolyte status. This study aimed to assess acid-base status, electrolyte levels, and lactate levels in dehydrated dogs after receiving acetate or lactate-containing intravenous rehydration fluids. In this prospective, randomized study, 90 dehydrated dogs were included and randomized to receive acetate [Sterofundin® ISO B. Braun Vet Care (STERO), Germany) or lactate (Ringer-Lactat-Lösung nach Hartmann B. Braun Vet Care (RL), Germany] containing intravenous fluids for rehydration. The exclusion criteria were as follows: Age <6 months, liver failure, congestive heart failure, and extreme electrolyte deviation. Physical examination, venous blood gas, and lactate levels were analyzed before and after rehydration. The two groups were compared using t-test and Chi-square test. The significance level was set at p≤0.05. Post-rehydration heart rate decreased in the STERO group (p<0.001) but not in the RL group (p=0.090). Lactate levels decreased in both groups STERO (p<0.001) and in group RL (p=0.014). Sodium and chloride levels increased during rehydration in group STERO (p<0.001; p<0.001) and group RL (p=0.002; p<0.001). There was a larger decrease in lactate levels in group STERO compared to group RL (p=0.047). Both solutions led to a mild increase in sodium and chloride levels and decreased lactate levels. The acetate-containing solution had an inferior effect on the decrease in lactate level. Keywords: acid-base disorders, crystalloid solutions, fluid therapy, hyperlactatemia, metabolic acidosis, rehydration.

Feed plays the most important role in supporting livestock productivity. There is a need for studies on the nutrient levels in feed absorbed by the body of livestock. The aim of this study was to investigate the effects of alfalfa (Medicago sativa L.) supplementation in feed on growth, small intestinal histomorphology, and digestibility in hybrid ducks. The study was conducted in vivo using 75 hybrid ducks, with three treatments and five replications. Each replication consisted of five ducks. The treatments were: T0=basal ration without any alfalfa supplementation; T1=basal ration+3% fresh alfalfa supplementation; and T2=basal ration+6% fresh alfalfa supplementation. Fresh alfalfa was quantified on the basis of dry matter content. Drinking water was provided ad libitum. The observed variables were growth performance, measured in terms of feed consumption, body weight gain, and feed conversion ratio; duodenum histomorphology, measured in terms of villus height, villus width, and crypt depth; digestibility, measured in terms of dry matter digestibility; and organic matter, crude protein, and crude fiber concentrations. The data were analyzed using variance analysis with a completely randomized design of one-way pattern with Statistical Product for Service Solution application of version 22. The data with significant differences were further analyzed using Duncan's new multiple range rest. The results of the study showed that 3% alfalfa supplementation increased feed consumption and body weight gain of the hybrid ducks at 35 days of age compared with 0% and 6% supplementation. Furthermore, 3% alfalfa supplementation presented the best result in terms of villus height and duodenal crypt depth. Meanwhile, 6% alfalfa supplementation decreased nutrient digestibility in the ducks. These findings show that supplementation of feed with fresh alfalfa could have a significant effect on hybrid ducks in terms of growth performance, small intestinal histomorphology, and digestibility. Keywords: alfalfa, digestibility, histomorphology, hybrid ducks, performance.

Escherichia coli is the cause of avian colibacillosis, a significant threat to the poultry industry and public health. Thus, this study investigated the prevalence of E. coli in diseased chicken broilers, pathological effects of these bacteria, and interleukin (IL) gene expression of different serotypes of E. coli (O78, O26, O44, and O55) on experimentally infected chickens. A total of 295 organ samples (liver, lungs, heart, and spleen) from 59 diseased broiler chickens were used for conventional identification of E. coli. Chickens were orally infected with one of the following E. coli serotypes (O78, O26, O44, or O55) and examined for clinical signs, mortality, macroscopic and microscopic lesions, and IL gene expression using real-time quantitative polymerase chain reaction. E. coli was isolated from 53.2% of broiler chicken organs with a high prevalence in lungs (26.1%). The most prevalent serotypes were O78, O26, O44, O55, O157, and O127 prevalence of 27.8, 22.2, 16.7, 16.7, 5.6, and 5.6%, respectively. In the experimental design, five groups (G1-G5) of birds were established. G1 served as the negative control group, while G2-G5 were challenged orally with E. coli O78, O26, O55, or O44, respectively. Chickens infected with E. coli O78 or O26 showed significant clinical signs in comparison to the other infected birds. Mortality (13.3%) was only observed in birds infected with E. coli O78. Necropsy of dead birds after E. coli O78 infection showed pericarditis, enteritis, airsacculitis, and liver and lung congestion. More severe histopathological changes were observed in intestines, spleen, liver, and lung from chickens infected with either E. coli O78 or O26 than for birds infected with other serotypes. On the 2nd day post-infection, E. coli challenge, particularly with E. coli O78, displayed significantly upregulated levels of ileal IL-6 and IL-8, but ileal IL-10 level tended to be downregulated in comparison to the control group. This study assessed the application of cytokines as therapeutic agents against infectious diseases, particularly colibacillosis. Keywords: broiler chicken, Escherichia coli, experimental infection, histopathology, interleukins.

Although wild boar hunting activities and the hunting dog trade in the Atlantic Forest and Cerrado biomes of Brazil overlap both with endemic and with non-endemic areas for visceral leishmaniasis, no study to date has focused on Leishmania spp. exposure among hunting dogs and hunters. The aim of the present study was to assess the presence of Leishmania spp. antibodies in hunting dogs and hunters in different anthropized areas of two Brazilian biomes. Blood samples were collected from 170 hunting dogs and 46 hunters between October 2016 and May 2018. The presence of antibodies against Leishmania spp. in hunting dogs was screened through a dual-path platform immunochromatographic test (DPP rapid test; Bio-Manguinhos/Fundação Oswaldo Cruz, Rio de Janeiro, Brazil) and in hunters through an rK39-based rapid immunochromatographic test. Both tests were used in accordance with Brazilian Ministry of Health recommendations. Overall, although antibodies were detected through the immunochromatographic test in 3/170 (0.02%) of these female asymptomatic hunting dogs, all living in anthropized areas of the Atlantic Forest biome in South Brazil, no sample was confirmed through the enzyme-linked immunosorbent assay. All the hunters were non-reactive in the rapid immunochromatographic test. Our study on three suspicious hunting dogs has suggested that Leishmania (Leishmania) infantum may circulate both in endemic and non-endemic areas in Brazil. In addition, a high rate of hunting dog replacement due to death and trade may have led to less chance of infection and transmission between animals and between animals and humans, which would corroborate the outcomes reported here. Further studies should be conducted to fully establish whether hunting dogs and hunters may be used as sentinels in other areas endemic for Leishmania spp. Keywords: Brazilian biomes, canine visceral leishmaniasis, hunting activities, Leishmania (Leishmania) infantum, rural dogs.

Cryptococcal yeast cells are spread across different ecosystems through bird movement and are deposited in bird guano. These cells may be inhaled by humans and lead to cryptococcal pneumonia. In individuals with reduced immune T-cell populations, cells may disseminate to the brain and cause the often-deadly cryptococcal meningitis. In this study, we surveyed cryptococcal cells in bird droppings across the city of Bloemfontein, South Africa. We aseptically collected 120 bird dropping samples from 15 representative city sites. In the laboratory, samples were assessed with regards to location, weighed, and standardized to a mass of 1 g before suspension in 10 mL phosphate buffer saline. Samples were first screened using Calcofluor-white stain as it is a rapid technique for the detection of fungi via binding to cell wall components such as chitin. After this, positive Calcofluor samples were serologically assayed for the cryptococcal antigen (CrAg). To confirm assay data, CrAg positive samples were then cultured on bird seed agar and resulting colonies were assessed using Indian ink. We determined that 10/15 locations were positive for the CrAg. Pathogenic cells were identified on bird seed agar as brown colonies. When examined using microscopy, brown colony cells exhibited characteristic thick capsules representative of cryptococcal cells. This is the first proximate analysis showing the ecological distribution of cryptococcal cells in Bloemfontein. This is important as associated infections are acquired from the environment. Similarly, given the threat posed by cryptococcal cells to immunocompromised individuals, local authorities must initiate measures curbing the spread of these cells. Keywords: Bloemfontein, Calcofluor-white stain, cryptococcal antigens, cryptococcal cells, cryptococcal meningitis, cryptococcal pneumonia, South Africa.

Wild boars have recently been implicated as the maintainers and carriers of Amblyomma spp. ticks, which are essential for Rickettsia spp. transmission. Consequently, wild boar hunting may increase the risk of tick exposure and subsequent human tick-borne infection and disease. Therefore, this study was conducted to evaluate the risk factors for ticks and Rickettsia spp. exposure in wild boars, hunting dogs, and hunters in Brazilian biomes. The statistical relationship of Rickettsia spp. antibodies were evaluated using the Chi-square test in 80 wild boars, 170 hunting dogs, and 49 hunters. The only statistically significant difference in seropositivity found in this study was between male and female wild boars (p=0.034), probably associated with in-park exposure to Amblyomma brasiliense infected with Rickettsia spp. The absence of statistical differences in the associated risk factors for hunting dogs and hunters may indicate a random exposure to Rickettsia spp. Keywords: Brazilian spotted fever, hunting activities, wild boars.

Dermatophilosis is a bacterial infection of the skin of animals. It is prevalent worldwide and is caused by Dermatophilus congolensis. The study aimed to assess the therapeutic efficacy of different mixtures prepared with indigenous phytogenetic extracts from Benin in the management and treatment of Girolando cattle that showed high sensitivity to the disease compared to any other known cattle breed in Benin. Consequently, two types of extract mixtures (extract mixture 1=Elaeis guineensis kernel oil + essential oil of Ocimum gratissimum + sap of Jatropha multifidi; extract mixture 2= O. gratissimum + sap extract of J. multifida) of 20% concentration were prepared and tested on eight Girolando cattle. Two different parts of the affected skin with acute lesions of dermatophilosis were debrided, and 0.5 mL of each of the extract mixture was applied per square centimeter of a single zone of the affected skin. Both extracts mixtures were found to possess significant wound healing properties compared to the control (procaine G penicillin). However, the extract mixture 1, which was made up of Elaeis guineensis kernel oil, essential oil of O. gratissimum and sap of J. multifida (Linn), showed a better result. This was evident by increase in the rate of wound contraction and healing without recurrence 2 weeks after the end of the experiment and the subsequent immediate manifestation of hair or hair growth at the affected area. The preliminary findings of this study are very promising. Extract mixture 1 could serve as an alternative in the treatment or management of bovine dermatophilosis in Benin and other dermatophilosis endemic areas of the world. However, in vitro testing and sensitivity against isolated D. congolensis organism using extract mixture 1 as well as cost implications should be studied. Keywords: farmer resilience, indigenous plants, plant extract, sustainable cattle breeding.

Fasciolopsiasis is a parasitic infection caused by the flatworm Fasciolopsis buski. Since 1982, fasciolopsiasis has been reported in Indonesia's Hulu Sungai Utara (HSU) Regency, South Kalimantan Province. Fasciolopsiasis occurs when contaminated raw or undercooked aquatic plants are consumed. Cercariae of the parasite encyst in a variety of aquatic plants and grow into metacercariae that infect and reproduce in the human intestine. Until now, treatment for F. buski infection in the HSU Regency has been comparatively short, with patients receiving only a single dose of praziquantel, 30 mg/kg body weight, without further observation. A long-term effort through health promotion activities and intensive health education, particularly for elementary school children enrolled in the School Health Program, is ongoing to help prevent fasciolopsiasis from spreading and to improve environmental sanitation. Through 2018, intervention efforts successfully reduced the incidence of F. buski infection. Sustaining surveillance and investigation of fasciolopsiasis, including identification of new cases and community education, is critical for the elimination of the parasite from Indonesia. This review describes the spread of F. buski and its possible impact on public health to understand the critical nature of continuing F. buski surveillance and control efforts.

Salmonella causes most foodborne bacterial illnesses worldwide. It is found in various hosts, including pets, farm animals, and wild animals, as well as the environment. This study aimed to examine the epidemiological relationship between Salmonella isolates from aquatic environments and those from other avian hosts. The study examined 12 water samples, 210 aquatic animals, and 45 migratory aquatic bird samples collected from the protected area of Lake Qarun in El-Fayoum Governorate, Egypt, during migration seasons from different waterfowl migration areas (from October 2018 to January 2019). In addition, 45 fecal samples from domestic chickens were collected from the same geographic location from poultry farms. Bacteriological examination and polymerase chain reaction assay of two virulence genes (i.e., invA and stn) were performed to isolate and identify Salmonella. Salmonella was isolated from 58.3% (7/12) of Lake Qarun water samples, 13.3% (6/45) of migratory waterfowl, 6.6% of (3/45) of chickens (Gallus gallus domesticus), and 4.3% (3/70) of fish and pooled brine shrimp. In migratory aquatic bird species that were sampled, Salmonella were isolated from 23.1% (3/13) of Eurasian coot (Fulica atra), 12.5%, (1/8) of green-winged teal (Anas cardolinesis), 10% (2/20) of northern shoveler (Spatula clypeata), and 0% (0/4) of mallard duck (Anas platyrhynchos). In 35 Tilapia, Salmonella was isolated by (8.6%) 5.7% of external surfaces, 2.85% from the intestine, and 0% from the muscle. No Salmonella was isolated from the 175 brine shrimp samples. Phylogenetic analysis using the stn genes of Salmonella isolated from the aquatic environment, migratory aquatic birds, and chicken showed a strong association between these isolates. In addition, a higher nucleotide identity percentage was observed between the sequences recovered from migratory aquatic birds and Lake Qarun water samples. Salmonella distribution was confirmed through migratory aquatic birds, based on our phylogeny tree analysis, Salmonella considered a likely carrier of zoonotic bacterial pathogens. Furthermore, the close relationship between chicken and fish sequences highlights the scenarios of using chicken manure in fish farms and its public health implications. The presence of Salmonella in different environmental sources spotlights the urgent need to control and break down its epidemiological cycle. Keywords: migratory aquatic birds, fish, poultry, Salmonella, stn gene, Lake Qarun.

Aquaporin-2 (AQP2) and arginine vasopressin receptor-2 (AVPR2) are proteins that control water homeostasis in principal cells. Chronic kidney disease (CKD) is defined as the impairment and irreversible loss of kidney function and/or structure, which causes water imbalances and polyuria. The study aimed to know the expression of AQPs and AVPR2 in the kidneys of a canine with CKD. The kidneys were collected from two dog carcasses from Small Animal Teaching Hospital, Faculty of Veterinary Medicine, Chiang Mai University. The kidney tissue was prepared for immunohistochemistry and investigated the expression and localization of tissue's AQP2 and AVPR2. For statistical analysis, the Mann–Whitney U-test was applied to the data. By immunohistochemistry, AQP2 was expressed strongly in the basolateral and apical membranes of the principal cells, whereas AVPR2 was localized in the principal cell's basolateral membrane in both renal cortex and renal medulla. In the normal kidney, the semi-quantitative immunohistochemistry for the percentage of protein expression of AQP2 and AVPR2 was 5.062±0.4587 and 4.306±0.7695, respectively. In contrast, protein expression of AQP2 and AVPR2 in CKD was found to be 1.218±0.1719 and 0.8536±0.1396, respectively. The data shows that the percentage of AQP2 and AVPR2 expression was decreased, corresponding to a 4-fold and 5-fold in CKD (p<0.001). Our findings revealed that CKD was a marked decrease in AQP2 and AVPR2 expression. The central role of specific AQP2 and AVPR2 in regulating water homeostasis will provide correlations in case of CKD with polyuria. Keywords: aquaporin 2, canine, chronic kidney disease, immunohistochemistry, vasopressin receptor 2.

The coronavirus diseases-2019 (COVID-19) pandemic has caused a global lockdown, which has limited the mobility of the public, and thus, more time is spent with their pets. Unfortunately, many social media have blamed pet animals as a reservoir of severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2), the etiologic agent of COVID-19, triggering a panic abandonment of pets. However, no article has summarized the information regarding the role of pets as SARS-CoV-2 reservoirs. This study aimed to evaluate the role of pets as a reservoir of SARS-CoV-2 on the basis of research papers (i.e., animal model, surveillance, and case report) published in 2020. The review was conducted using articles from the PubMed database in 2020, using the keywords "COVID-19 in domesticated animals," which were screened and analyzed. Only the data from research articles were mimicked and transformed to conduct a meta-analysis. The meta-analysis was conducted regarding the effects of inhabitation and viral shedding in pets. In this study, we used 95% confidence intervals. A total of 132 papers in PubMed were related to the keywords, whereas only 12 papers were appropriate to answer the dynamics of the role of pets as the reservoir for SARS-CoV-2. Seven studies indicated the potential of cat-cat (4/7), human-cat (2/7), and human-dog (1/7) SARS-CoV-2 transmission. No study proved the presence of cat-human transmission. Another study showed that comingling did not affect SARS-CoV-2 viral shedding among a cat and dog. Furthermore, the viral shedding of cats and dogs caused asymptomatic manifestations and generated neutralizing antibodies within a short period of time. SARS-CoV-2 transmission is present in domesticated animals, especially in pet cats and dogs, and transmission occurs between animals of the same species (cat-cat). The reverse zoonosis (zooanthroponosis) was found from human to cat/dog (comingled) with asymptomatic clinical signs due to the representation of neutralizing antibodies. Keywords: asymptomatic, domesticated animals, pet, severe acute respiratory syndrome-coronavirus-2, transmission.

Although only a few studies have investigated about the development of animal prosthesis, currently, there is an increasing interest in canine limb prosthesis design and its clinical application since they offer an alternative to killing the animal in extreme situations where amputating the limb is the only option. Restoring normal function of amputated canine limbs with the use of a prosthesis is challenging. However, recent advances in surgical procedures and prosthesis design technology appear promising in developing devices that closely recreate normal canine limb function. Surgical advances such as evolution of osseointegration (bone-anchored) prostheses present great promise. Likewise, modern computer-aided design and manufacturing technology, as well as novel motion analysis systems are now providing improved prosthesis designs. Advances in patient-customized prostheses have the potential to reduce the risk of implant failure. The objective of this investigation is to present a general review of the existing literature on modern surgical approaches, design and manufacturing methods, as well as biomechanical analyses so that veterinarians can make more and better-informed decisions on the development and selection of proper canine limb prosthesis. Isolated research efforts have made possible an improvement in stability, comfort, and performance of canine limb prosthesis. However, continued multidisciplinary research collaboration and teamwork among veterinarians, engineers, designers, and industry, with supporting scientific evidence, is required to better understand the development of canine limb prosthesis designs that closely replicate the normal limb function.

The emerging concerns regarding the new Coronavirus's ability to cause infection in pets has led to animal testing and worrisome findings reported all over the world in domesticated and wild animals. This study aimed to investigate severe acute respiratory syndrome coronavirus (SARS-CoV)-2 by quantitative reverse transcription-polymerase chain reaction in dog and cat samples with the clinical presentation for respiratory or gastrointestinal disease in Brazil. One hundred and twenty-five samples were collected from 12 states of Brazil that originated from the gastrointestinal, upper respiratory tract, and other sites, including some pools of samples from before the onset of the pandemic including blood and/or urine samples. They were tested for RT-PCR detection of respiratory or gastrointestinal pathogens through Respiratory or Diarrhea RT-PCR Panels in the TECSA (Tecnologia em Saninade Animal - Animal Health Technology) Veterinary Medicine Laboratory. This work was conducted in compliance with ethical standards. Seven different microorganisms that can cause respiratory and/or gastrointestinal clinical signs were detected in cats (Feline Coronavirus [FCoV], Feline Parvovirus, Feline Leukemia Virus, Feline Calicivirus, Mycoplasma felis, Campylobacter spp., and Cryptosporidium spp.) and three in dogs (canine distemper virus, Cryptosporidium spp., and Babesia spp.). Although the samples corresponded to the beginning of coronavirus disease-19 spread in Brazil and clinically correlated with the expected viral replication sites, none of the animals tested positive for SARS-CoV-2; reassuringly, four cats tested positive or FCoV none of them were positive for SARS-CoV2. The epidemiological surveillance of SARS-CoV-2 in pets is considered a one health issue, important for monitoring the disease evolution, spread and minimizing the animal-human health impacts, and directing Public Health Policies. Keywords: companion animals, coronavirus disease-19, vigilance, virus.

Necrotic enteritis (NE) is one of the most prevalent diseases in broiler poultry caused by Clostridium perfringens connected with significant economic losses. A cross-sectional survey was conducted in Mymensingh district of Bangladesh to assess the prevalence of C. perfringens through toxinotyping molecular assay and confirm the risk factors for NE, including antimicrobial-resistant (AMR) status of the isolates. We included 40 small-scale commercial broiler farms randomly selected from two subdistricts of Mymensingh district of Bangladesh. As an individual sample, 240 cloacal swabs, and as a pooled sample, 40 drinking water, 40 workers' hand washing, 40 litter swab, and 40 feed samples were collected and evaluated by culture, biochemical, and molecular assays. A pretested semi-structured interview questionnaire was employed to capture flock-level data on risk factors from the farm owners. The flock-level data on risk factors were assessed through univariable and multivariable logistic regression analyses with p<0.05 was considered statistically significant. Overall flock-level prevalence of C. perfringens was estimated to be 10.3% (95% confidence interval [CI] 7.5- 13.6%). Litter swab (pooled) was found to be highly contaminated with C. perfringens (25.0%, 95% CI: 12.7-41.2%) followed by the cloacal swab (10.4%, 95% CI: 6.9-15.0%) and feed sample (5.0%, 95% CI: 0.6-16.9%). History of coccidia infection (Adjusted odds ratio =33.01, 95% CI: 2.14-507.59, p=0.01) was significantly associated with flock-level C. perfringens infection status. In this study, 78.1% isolates were found as multidrug-resistant as they demonstrated resistance to 3-5 antimicrobial agents. Evidence-based control options need to be taken through the uses of prebiotics and probiotics, biosecurity, and hygienic measurement, including control of coccidia infection, is needed to lessen the NE infection and AMR related to this pathogen in small-scale commercial broiler poultry. Keywords: antimicrobial-resistant, Bangladesh, broiler, Clostridium perfringens, necrotic enteritis, prevalence.

The recent coronavirus disease (COVID-19) outbreak is one of its kind in the history of public health that has created a major global threat. The causative agent, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has a zoonotic source and hence, reverse zoonosis (disease transmission from humans to animals) increases the risk and rate of SARS-CoV-2 infection. Serological and molecular analyses and experimental infection studies have identified SARS-CoV-2 infection in several animal species in various countries. Different domestic and wild animals, including cats, dogs, tigers, lions, puma, snow leopard, minks, and pet ferrets, are infected naturally with SARS-CoV-2, mostly through suspected human to animal transmission. In addition, in vivo experimental inoculation studies have reported the susceptibility of cats, ferrets, hamsters, Egyptian fruit bats, and non-human primates to the virus. These experimentally infected species are found to be capable of virus transmission to co-housed animals of the same species. However, SARS-CoV-2 showed poor replication in livestock species such as pigs, chickens, and ducks with no detection of viral RNA after the animals were deliberately inoculated with the virus or exposed to the infected animals. As the pets/companion animals are more susceptible to COVID-19, the infection in animals needs an in-depth and careful study to avoid any future transmissions. The one health approach is the best inter-disciplinary method to understand the consequences of viral spread and prevention in novel host populations for the betterment of public health. Further in this review, we will explain in detail the different natural and experimentally induced cases of human to animal SARS-CoV-2 infection.

Enterohemorrhagic Escherichia coli (EHEC) is an important foodborne pathogen with worldwide distribution. Data regarding its presence, distribution, virulence, and antimicrobial susceptibility among various animal species and humans in Jordan are lacking. Therefore, the objectives of this study were to isolate and characterize EHEC from human and animal diarrhea fecal samples and ground beef samples. A total of 100 and 270 diarrhea fecal samples from humans and animals, respectively, were collected. In addition, 40 ground beef meat samples were collected from retail markets. EHEC was positively identified by detecting Shiga toxins (stx1 and stx2) genes using multiplex polymerase chain reaction (PCR). Antimicrobial susceptibility patterns were determined using the disk diffusion test. Beta-lactamase production was detected using the double disk diffusion test and the extended-spectrum beta-lactamases (ESBLs) were identified by detection of blaTEM, blaSHV, and OXA-1 genes using multiplex PCR. Pulsed-field gel electrophoresis (PFGE) was used to investigate the relatedness of EHEC isolates from different sources. Out of 410 samples, 194 E. coli isolates were positively identified, of which 57 isolates (29%) were classified as EHEC. Thirty-five (61%) of EHEC isolates were serotyped as O157 (19: O157:H7 and 16: O157:NM). The stx1 gene was detected only among the sheep and goats isolates at a rate of 7.6% and 5.2%, respectively, while the stx2 gene was detected in only one ground beef meat sample. EHEC isolates showed high resistance patterns against amoxicillin, gentamycin, cephalexin, and doxycycline. Twenty-four out of 32 EHEC isolates were determined as ESBL producers, among which 14 isolates expressed the blaSHV gene and 19 isolates expressed the blaTEM while four expressed both genes. PFGE analysis revealed two clusters with high similarity (92%) originated from ground beef meat and cattle fecal samples. No similarities were found between human and animal E. coli isolates. Results of this study indicate widespread ESBL EHEC among humans, animals, and ground beef meat samples. These results represent an important alarm that requires the implementation of appropriate preventative measures by both human and animal health sectors to prevent the transmission of this important foodborne pathogen. Keywords: animal health, Enterobacteriaceae, foodborne pathogens, public health.

The mutation in the wild-type tumor suppressor gene p53 is the most common genetic change in human tumors. In addition, the normal function of p21, which is both antiproliferative and an inhibitor of the cell cycle, is disrupted in some types of cancer. Meanwhile, cyclin D1 is a member of the cyclin protein family that is involved in regulating cell cycle progression. This study aimed to assess the expressions of the cell cycle inhibitory proteins p21, cyclin D1, and tumor suppressor gene p53, as well as their influence on the expressed histopathological changes in breast cancer tissues. Overall, 40 breast tissue specimens were investigated in this study, 30 of which were cancerous, while 10 were healthy tissues. p53, p21, and cyclin D1 expression patterns were detected using an immunohistochemistry (IHC) system. The IHC reactions for p53 were positively observed in 27/30 (90%) cancerous tissues, compared with 2/10 (20%) normal breast tissues. For p21, reactions were observed in 28/30 (93.33%) cancerous tissues and 3/10 (30%) control tissues. For cyclin D1, reactions were observed in 25/30 (83.33%) cancerous tissues and 1/10 (10%) control tissues. The differences between the breast cancer tissues and the control tissues were statistically highly significant (p<0.01). The high expression rates of p21, cyclin D1, and p53 in malignant breast cancer cells with little or no regulatory role might imply mutational events in these proteins operating in concert with a variety of other genetic mutations in these tissues, which may play a molecular role in the development and/or progression of breast carcinogenesis. Keywords: breast cancer, D1 protein, Iraq, P21, P53.

Despite worldwide case reports, including Brazilian cases, no frequency study on infection of pets by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been conducted to date in Brazil. Accordingly, the present study was aimed to assess dogs and cats belonging to positive owners in Recife, Northeastern Brazil. This was a longitudinal prospective study on dogs and cats in the city of Recife whose owners were in isolation at home due to a confirmed laboratory diagnosis of SARS-CoV-2 through reverse-transcriptase polymerase chain reaction (RT-qPCR). Oral and rectal swabs from the pets were tested for the presence of SARS-CoV-2-specific RNA by means of RT-qPCR. Among the pets tested, 0/16 dogs and 2/15 cats were positive for SARS-CoV-2. Interestingly, the two positive cats were owned by two unrelated asymptomatic veterinary students, which, therefore, post a warning to veterinarians worldwide. The findings herein indicate that cats may act as sentinels for human cases, particularly sharing households with asymptomatic human cases. Although with small sampling and convenient recruiting, the presence of infected cats by SARS-CoV-2 was most likely due to close cat-human contact with positive owners, posting a human-animal health threat when pets share the same bed and interact with owners without protection, particularly during owner self-isolation. Thus, infected owners should follow the same human preventive guidelines with their pets to avoid spreading infection. Keywords: Brazil, coronavirus, domestic animals, pandemic.