New anticancer drugs are being developed to avoid the toxicity and chemoresistance of the currently available drugs. The Food and Drug Administration-approved anti-malarial drug atovaquone is known to act as a selective oxidative phosphorylation inhibitor in the mitochondria by competing with CO Q10 (mitochondrial complex II and III). This study aimed to investigate the effect of atovaquone by examining the Na+/K+-ATPase (NKA) activity in various canine cell lines. Canine cell lines were treated with various concentrations (2.5, 5, 10, 15, and 20 µM) of atovaquone for 24, 48, and 72 h. Human cell lines were used as a control to validate the canine cancer cell lines. The activities of the drugs against the cancer cell lines were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl- 2H-tetrazolium bromideassay. The cell metabolic activity was determined by measuring the activities of the nicotinamide adenine dinucleotide phosphate-dependent cellular oxidoreductase enzymes. The NKA activity was measured using the single-cell patch clamping assay. Atovaquone-induced apoptosis by elevating the concentration of reactive oxygen species (ROS) in the tumor cells, leading to cell death. Treatment of canine cancer cells with N-acetylcysteine (ROS inhibitor) reduced the activity of the drug. Furthermore, atovaquone inhibited more than 45% of the NKA ion current. This study demonstrated effects of atovaquone against canine cancer cell lines. The data may prove beneficial in repurposing the drug as a new anticancer agent in canine clinical trials, which might aid in fighting human cancer. Keywords: atovaquone, electrophysiology, Na+/K+-ATPase, oxidative phosphorylation, oxidative stress, plumbagin.

Coronaviruses (members of the Coronaviridae family) are prominent in veterinary medicine, with several known infectious agents commonly reported. In contrast, human medicine has disregarded coronaviruses for an extended period. Within the past two decades, coronaviruses have caused three major outbreaks. One such outbreak was the coronavirus disease 2019 (COVID-19) caused by the coronavirus severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). Over the 3-year COVID-19 outbreak, several instances of zooanthroponosis have been documented, which pose risks for virus modifications and possible re-emergence of the virus into the human population, causing a new epidemic and possible threats for vaccination or treatment failure. Therefore, widespread screening of animals is an essential technique for mitigating future risks and repercussions. However, mass detection of SARS-CoV-2 in wild animals might be challenging. In silico prediction modeling, experimental studies conducted on various animal species, and natural infection episodes recorded in various species might provide information on the potential threats to wildlife. They may be useful for diagnostic and mass screening purposes. In this review, the possible methods of wildlife screening, based on experimental data and environmental elements that might play a crucial role in its effective implementation, are reviewed.

Intranasal (IN) sedatives provide a non-invasive route for premedication drug administration. This study compared the cardiorespiratory and sparing effects of IN dexmedetomidine combined with morphine (DM) or tramadol (DT) on alfaxalone requirements for anesthesia induction in cats. Twenty-four cats were randomly assigned to three groups: Dexmedetomidine combined morphine (IN dexmedetomidine 20 µg/kg plus 0.2 mg/kg morphine), DT (IN dexmedetomidine 20 µg/kg plus 1 mg/kg tramadol), or control (no premedication). The intravenous dose of 1% alfaxalone for endotracheal intubation was recorded with sedation scores, cardiorespiratory parameters (heart rate and respiration rate), and side effects. Both DM and DT were associated with significantly higher sedation scores than baseline, and sedation scores were found to be highest 20 min after premedication. Sedation scores were comparable between DM and DT groups. Side effects, including hypersalivation, vomiting, and pupillary dilation, were observed in the DM and DT groups. The dosage of alfaxalone required in the DM group (1.5 ± 0.3 mg/kg) was comparable to that of the DT group (2.0 ± 0.6 mg/kg, p = 0.0861), and both groups required significantly less alfaxalone than the control group (3.0 ± 0.6 mg/kg; p < 0.01). Heart and respiratory rates were comparable between the DM and DT groups. Duration of anesthesia in the control group (11 ± 4 min) was significantly shorter than in the DM (29 ± 5 min, p = 0.0016) and DT (38 ± 14 min, p < 0.001) groups. Intranasal administration of DM or DT produces good sedation and offers an alternative, non-invasive route for cats undergoing general anesthesia. Keywords: alfaxalone, dexmedetomidine, intranasal, morphine, tramadol.

Canine monocytic ehrlichiosis is a vector-borne disease caused by the obligatory intracellular bacterium Ehrlichia canis, which is distributed across tropical and subtropical regions worldwide. Its prevalence within dog populations is high in municipalities located across the Pantanal biome, but it remains unknown in Barão de Melgaço, Mato Grosso, Brazil. This study aimed to determine the molecular prevalence and factors associated with E. canis infection in dogs domiciled in Barão de Melgaço. A cross-sectional study was carried out to investigate the prevalence of E. canis infection in 369 dogs from urban and rural areas in Barão de Melgaço, North Pantanal wetland, Brazil. Initially, the dogs were examined, and, through a questionnaire, the risk factors were investigated. Blood samples were subjected to DNA extraction and PCR was performed to estimate the prevalence of E. canis infection. The molecular prevalence of E. canis infection in dogs was 42.5% and none of the studied variables were significantly associated with polymerase chain reaction (PCR) positivity (p > 0.05). The high molecular prevalence demonstrates an increased transmission of the agent across the city. This also indicates that attention needs to be paid to E. canis infection and control measures should be introduced to prevent its transmission. The demographic and clinical risk factors commonly associated with E. canis infection in this study were not associated with PCR positivity. Keywords: bacterium, DNA, dog, ehrlichiosis, Pantanal.

Humans and dogs with azotemia can develop coagulation disorders. Therefore, this study aimed to evaluate the coagulation profiles and thromboelastographic parameters in dogs with acute kidney injury (AKI) and chronic kidney disease (CKD). In this prospective study, 31 client-owned dogs with renal azotemia (creatinine >220 µmol/L) were enrolled. Clinical signs of hemostatic disorders, complete blood count, coagulation profile, D-dimers, thromboelastography, and 28-day survival data were obtained and analyzed using the t-test, Mann–Whitney U test, and Chi-square test. Statistical significance was set at p < 0.05. Seventeen dogs with AKI, 10 with CKD, and four with acute-on-chronic kidney injury (AoC) were enrolled. Ten dogs (AKI, 8/17; CKD, 2/10) had thrombocytopenia. Prothrombin time was prolonged in four dogs with AKI and longer in dogs with AKI than in dogs with CKD (p = 0.004). The activated partial thromboplastin time was prolonged in 23 dogs (AKI, 14/17; CKD, 7/10; AoC, 3/4) and was longer in azotemic dogs than in healthy control dogs (p = 0.003). Thromboelastographic tracings were hypocoagulable in three dogs with AKI and hypercoagulable in 16 dogs (AKI 4/17, CKD 9/10, AoC 3/4). The thromboelastographic values for maximum amplitude (p < 0.001) and global clot strength (p < 0.001) were lower in dogs with AKI than in those with CKD. Hypercoagulable thromboelastographic tracings were observed in dogs with CKD, whereas coagulation times were prolonged in dogs with AKI. However these findings should be validated by further studies. Keywords: acute kidney injury, canine, chronic kidney disease, coagulation, platelet function, viscoelastic test.

Cancer is one of the most important public health problems worldwide. Despite the great contribution of in-vitro studies for biomedical research, animals are essential to study diseases’ biopathology and diagnosis, and searching for new preventive and therapeutic strategies. Breast cancer is currently the most common cancer globally, accounting for 12.5% of all new annual cancer cases worldwide. Although the rat model of mammary cancer chemically-induced is widely used to study this disease, there is a lack of standardization in procedures for cancer induction, sample collection, and analysis. Therefore, it is important to provide a practical guide for researchers aiming to work with this model to make the analysis of results more uniform. Thus, in this review, we provide the researchers with a detailed step-by-step guide to implement a rat model of mammary cancer, based on our wide experience in this field, to obtain the best results, maximum throughput of each experiment, and easy comparison among researches.

The pathogenicity of Escherichia coli is determined by the presence of genes that mediate virulence factors such as adherence capacity and toxin production. This research aimed to identify the adhesion factors and antibiotic resistance capacity of E. coli strains associated with diarrhea in piglets in Colombia. Presumptive E. coli strains were isolated from the rectal swabs of piglets in swine farms between 4 and 40 days of age with evidence of diarrhea. Presumptive E. coli strains were tested for antibiotic resistance. The hemolytic capacity of presumptive E. coli strains was measured and molecularly identified. Strains confirmed as hemolytic E. coli was evaluated for the presence of five adhesion factors (F4, F5, F6, F18, and F41) and resistance to 11 antibiotics. Fifty-two putative E. coli strains were isolated, six of which showed a hemolytic capacity. The hemolytic strains were molecularly identified as E. coli. Adhesive fimbriae were found in five of six β-hemolytic E. coli isolates. Combinations of the adhesion factors F6–F18 and F6–F41 were linked to antibiotic resistance capacity. The phenomenon of E. coli strains resistant to multiple antibiotics on pig farms represents a constant risk factor for public health and pig production. Keywords: adhesive fimbriae, piglet diarrhea, enteroaggregative heat-stable toxin 1, enterotoxigenic Escherichia coli, hemolytic capacity.

Developing simple, cost-efficient sheep feed will improve farmers’ incomes. Including coffee skin in feed offers the most technical method of increasing sheep weight gain. This study aimed to evaluate varying proportions of ensiled coffee skin replacing dried water spinach and determine the optimal combination for the growth performance, physiological and hematological profiles, and rumen fluid of sheep. Eighty-four animals were randomly allocated to the treatments, arranged in a randomized block design using the initial weight as a block. Seven treatment diets were adjusted and a 12-animal replication was used for each treatment. The treatments were as follows: T0: 30% maize stover, 30% dried water spinach, 5% pollard, 20% coffee skin; T1: 30% maize stover, 25% dried water spinach, 5% pollard, 5% ensiled coffee skin; T2: 30% maize stover, 20% dried water spinach, 5% pollard, 10% ensiled coffee skin; T3: 30% maize stover, 15% dried water spinach, 5% pollard, 15% ensiled coffee skin; T4: 30% maize stover, 10% dried water spinach, 5% pollard, 20% ensiled coffee skin; T5: 30% maize stover, 5% dried water spinach, 5% pollard, 25% ensiled coffee skin; T6: 30% maize stover, 5% pollard, and 30% ensiled coffee skin. The sheep were reared for 70 days.The parameters observed during the early stage included growth performance (initial body weight, LW gain, final body weight, and feed intake). At the end of periods, a representative sample of ruminal fluid (approximately 150 mL) was collected from slaughtered sheep, duplicated, and then incubated for 18 h and blood samples were collected from the sheep (jugular vein) in ethylenediaminetetraacetic acid tubes. Then, used to analyze various blood biochemical parameters. The final body weights showed a linear curve increasing as the treatment increased (p < 0.05). The ensiled coffee skin tended to increase at 6 h incubation time, producing reduced methane gas (p < 0.05). However, in general, the use of ensiled coffee skin did not significantly alter the blood biochemistry of crossbreed sheep (p > 0.05). There was no significant effect on the protozoal population (p > 0.05). Increasing the level of ensiled coffee skin up to 30% replacing dried water spinach increased the final body weight of crossbreed sheep with no adverse effect. Keywords: biochemical blood, coffee skin, crossbreed sheep, ensiling, final body weight.

Avian metapneumovirus (aMPV) is a recently discovered respiratory virus in chickens. Avian metapneumovirus has been linked to respiratory syndromes, reproductive failure in affected chickens and turkeys, swollen head syndrome in chickens, and rhinotracheitis in turkeys. Wild birds are considered potential reservoirs of aMPV, particularly aMPV-C. However, little is known about the prevalence of aMPV in Saudi Arabia. Considering the relevance of backyard chickens in the transmission and sustainability of certain avian viral diseases, this study aimed to assess aMPV exposure in backyard chickens and wild birds circulating near selected locations. We collected 368 serum samples from unvaccinated backyard chickens in ten locations in Eastern Saudi Arabia. Furthermore, we collected 78 serum samples from species of free-ranging birds belonging to the Columbidae family, such as pigeons and doves, captured from the same areas. Using commercial enzyme-linked immunosorbent assay kits, we tested the sera of domestic backyard chickens and wild birds for antibodies against aMPV. Our results showed that 74/368 birds were positive for aMPV-related antibodies. Conversely, none of the tested wild birds seroconverted to aMPV. The antibody titers detected in the backyard chickens suggested recent exposure to aMPV. Considering these results, further large-scale serological and molecular studies are needed to evaluate the prevalence of aMPV in these birds and characterize the circulating strains of aMPV in this region. Keywords: antibodies, chickens, pigeons, Saudi Arabia.

Sulfadiazine, one of the sulfonamide group’s active compounds, is widely used for therapeutic production against several diseases. Veterinary drug residues can have a significant impact on human health conditions. This study aimed to develop a prototype of rapid test devices (RTDs) for detecting sulfadiazine residues on chicken carcasses based on the color indication. Seven samples of carcasses collected from traditional breeders in Surabaya-Indonesia were prepared and tested using RTDs. This sample represents the population considering that in the last report, the use of antibiotics was more than 40%, while the ability to monitor RTDs was estimated at 100. The standard color of purple by Hex code standard color or decimal code color was used to compare the positive samples. A light-emitting diode (LED) lamp was used to observe purple color. Analysis of sulfonamides resulting from RTDs was compared using a ultraviolet-visible spectrophotometer. Sulfonamides contamination levels of 50% and 100% were detected at concentrations of 0.472 µg/mL and 0.642 µg/mL, respectively. Sulfonamides contamination that was <0.395 µg/mL did not appear purple. The study’s findings showed that RTDs can be used to detect sulfonamides residues at a limit of detection 0.5 µg/mL after a 45 min exposure to an LED operating at a wavelength of 980 nm (p < 0.05). The limitation of RTDs was not being able to monitor the presence of residues bound in fat samples. Rapid test devices can be developed for commonly monitoring devices due to the limited technology available in the market. Keywords: diazotation, food safety, residues, sulfadiazine, veterinary drugs.

Colistin is used to treat avian pathogenic Escherichia coli (APEC), a microorganism that affects turkey meat production in the Gaza Strip and worldwide. However, the recent emergence of plasmid-borne mobile colistin resistance (mcr) genes in pathogenic E. coli strains is a serious antimicrobial resistance (AMR) challenge for both human and animal health. In December 2018, colistin was banned as a veterinary antimicrobial in the Gaza Strip. This study aimed to detect and track the prevalence of colistin-resistant APEC isolated from turkey flocks in the Gaza Strip. This study investigated 239 APEC isolates from turkey flocks in the Gaza Strip between October 2018 and December 2021 (at 6-month intervals). The colistin-resistant APEC strains were detected using the broth microdilution method. The mcr-1 gene was identified using a polymerase chain reaction. The overall colistin resistance among the isolated APECs was 32.2% during the study period. The average resistance in the first interval was 37.5%, which significantly decreased to 9.3% in the last interval. Among the 77 phenotypically resistant isolates, 32.4% were positive for mcr-1. The average abundance of mcr-1 in the first interval was 66.6%, which decreased to 25% in the last interval. To the best of our knowledge, this is the first study reporting the presence of the mcr-1 gene among the APEC isolates from turkeys in the Gaza Strip. Banned veterinary use of colistin significantly decreased the percentage of resistant APEC isolates from turkeys in Gaza Strip. Further studies are needed to investigate other colistin resistance genes and track the emergence of AMR. Keywords: avian pathogenic Escherichia coli, colistin, Gaza Strip, turkey.

Veterinarians are commonly exposed to occupational stressors, including excessive workload and financial constraints. These stressors can lead to psychological distress, which typically results in mental health disorders such as depression, anxiety, and burnout and can even culminate in suicide attempts or suicide deaths. Risk factors associated with poor mental health and high rates of suicide in veterinary practitioners include continuous exposure to challenging scenarios, such as interpersonal conflicts, performing euthanasia, and easy access to lethal means of suicide, such as opioids and anesthetics. The previous studies highlight the urgent need for a better understanding of predisposing factors, mental health-related improvements in the professional environment, and the subsequent establishment of primary mental health-related care policies. Effective ways to promote mental health and prevent suicide may include social support, resilience, developing coping skills, promoting a healthy work environment, and discouraging perfectionist behaviors. This review aimed to summarize findings in studies that have investigated mental health and suicide in veterinarians and veterinary students and highlight measures that could be implemented as options for mental health promotion and suicide prevention.

Paratuberculosis (PTB) or John’s disease is a chronic disease of ruminants impeding the reproduction and productivity of the livestock sector worldwide. Since there is a lack of pathological studies explaining the nature and development of the disease in camels, this study aimed to highlight the anatomopathological changes of PTB in camels, which may help in verifying and validating some diagnostic tests used to detect the etiology of the disease in camel tissues. In August 2017, at Alselaa border’s Veterinary Clinic of Al Dhafra Region, Western Abu Dhabi, UAE, one imported culled she-camel of 2 years old was subjected to clinical, microscopic, and anatomopathological investigations along with real-time quantitative polymerase chain reaction (q-PCR) to confirm the infection and correlate between clinical signs and pathological lesions of the PTB in dromedary camels. Clinically, typical clinical signs compliant with the pathognomonic gross and histologic lesions of PTB were seen in naturally infected dromedary camel. As presumptive diagnosis microscopically, acid-fast coccobacillus bacterium clumps were demonstrated in direct fecal smears as well as in scraped mucosal and crushed mesenteric lymph node films, and in histopathological sections prepared from a necropsied animal and stained by Ziehl-Neelsen stain. Free and intracellular acid-fast clump phagosomes were further confirmed as Mycobacterium avium subsp. paratuberculosis by q-PCR. Clinical signs and pathological lesions of paratuberculosis in a dromedary camel were found to be similar to those of the other susceptible hosts. Keywords: acid-fast bacteria, dromedary camel, Mycobacterium, paratuberculosis.

Human infections caused by Candida albicans are common and range in severity from relatively treatable skin and mucosal conditions to systemic, fatal invasive candidiasis. The treatment of fungal infections is challenged by major obstacles, including the scarcity of effective therapeutic options, the toxicity of available medications, and the escalating antifungal resistance. Hence, there exists an urgent need to develop new classes of antimicrobial agents. This study was conducted to investigate the effect of KW-23 peptide against standard and resistant strains of C. albicans alone and in combination with fluconazole. A conjugated ultrashort antimicrobial peptide (KW-23) was designed and synthesized. KW-23 was challenged against standard and multidrug-resistant C. albicans alone and in combination with fluconazole using standard antimicrobial and checkerboard assays. The toxicity of the peptide was examined using hemolytic assays. KW-23 positively affected the standard and resistant Candidal strains (at 5 and 15 µg/mL respectively), exhibiting potent synergistic antimicrobial activity against the standard strain when combined with fluconazole. The effect of the combination was additive against the resistant strain (0.6 µg/mL). Furthermore, the peptide exhibited negligible toxicity on human erythrocytes. KW-23 and its combination with fluconazole could be a promising candidate for developing anticandidal agents. Keywords: Candida albicans, drug combinations, fluconazole, synergism, ultrashort peptide.

Recent information on the occurrence of bovine hypodermosis in Kazakhstan is limited to the results of a few clinical studies in the northern and eastern regions. A first serological study aimed to obtain more data on its geographical distribution and to estimate the prevalence in this country. Serum samples collected from 891 dairy cows on 30 dairy farms in eight Kazakh provinces during the winter season 2015/2016 were examined for antibodies to Hypoderma spp. first-stage larval antigen using a commercially available enzyme-linked immunosorbent assay (IDEXX Bovine Hypodermosis Serum Antibody Test). Overall, 73.6% (95% confidence interval: 70.6%–76.5%) of the cows sampled were seropositive for Hypoderma, and antibody-positive cows were found in 28 of 30 farms and in seven of eight provinces. The results suggest a high prevalence of bovine hypodermosis in Kazakhstan, for which the socioeconomic changes in agriculture and village life following the country’s independence are considered to be responsible. Keywords: cattle, Hypoderma, Kazakhstan, myiasis, seroprevalence.

Bovine fasciolosis is a reemerging neglected disease with a worldwide distribution caused by the trematode Fasciola spp., which parasitize various hosts. Bovine fasciolosis is responsible for large economic losses in the bovine livestock sector. This study aimed to estimate the seroprevalence and risk factors of bovine fasciolosis in the municipalities of Colombia. This was a descriptive cross-sectional study with simple random sampling conducted on 1140 cattle from the municipalities of Chiquinquir´, San Miguel de Sema, and Ubaté for a duration of 3 months. Serum samples were processed using the commercial Fasciola hepatica Antibody Test Kit IDEXX® Fasciolosis Verification (IDEXX, United States), which identified immunoglobulin G antibodies for gf2 antigen purified from Fasciola extracts. The f2 antigen is extremely immunogenic and highly specific for F. hepatica. An epidemiological survey was performed to record variables related to the sampled animals and herd management practices. Data were processed using the statistical program Epi Info® (Centers for Disease Control and Prevention; Atlanta, Georgia). The prevalence ratio was estimated to evaluate the association between fasciolosis and the hypothesized causal factors and the significance of this association using Pearson’s Chi-square test. Finally, a logistic regression model was developed. The overall seroprevalence was 72.3%. The seroprevalence was 83.9% (323/385) in Chiquinquirá, 68.17% (257/377) in Ubaté, and 64.55% (244/378) in San Miguel de Sema. The seroprevalence was higher in male animals in Chiquinquirá and in female animals in San Miguel de Sema and Ubaté. Similarly, sex showed a statistically significant association with disease prevalence in Ubaté. The highest prevalence was found in cattle aged >2 years. The Holstein breed showed maximum seroprevalence in Chiquinquirá (p ≤ 0.05) and San Miguel de Sema, whereas crossbreed showed higher seroprevalence in Ubaté. Similarly, in Chiquinquirá, the association between the seroprevalence of fasciolosis and the presence of other species was statistically significant (95% confidence interval [CI]: 0.9601–3.4944; p = 0.0448). In Ubaté, the disease presentation was also associated with pasture rental (95% CI: 0.4047–1.0023; p = 0.003) and attendance to livestock expositions (95% CI: 0.2313–1.0636; p = 0.044). However, in San Miguel de Sema, water from the stream showed a statistically significant association with disease presentation (95% CI: 0.5209–1.0985; p = 0.00649785). Female sex and diarrhea occurrence were considered risk factors for fasciolosis. A high seroprevalence of antibodies to Fasciola spp. was detected in cattle in the study municipalities, indicating a high parasite distribution in these areas. Female sex and diarrhea were established as risk factors associated with fasciolosis in Ubaté and San Miguel de Sema, respectively. Further, research is necessary to establish prevention and control programs against parasitosis. Keywords: bovine, fasciolosis, risk factors, seroprevalence.

Antibiotic resistance, especially in Gram-negative bacteria, is a major public health risk affecting all industries requiring the use of antibiotics, including agriculture and animal breeding. This study aimed to use papaya extracts to synthesize silver nanoparticles (AgNPs) and evaluate their antimicrobial activity against various Gram-negative bacteria. Silver nanoparticles were synthesized from the aqueous extracts of papaya seed, root, and bark, with AgNO3 used as a reducing agent. The phytofabricated AgNPs were analyzed by ultraviolet–visible absorbance, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy, and photon cross-correlation spectroscopy (PCCS). The disc-diffusion method was used to perform antibacterial analysis, and the minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations were determined. We also investigated the antibiofilm activity of AgNPs and attempted to elucidate the potential mechanism of action on Escherichia coli ATCC 25922. Phytofabrication of AgNPs was successful with papaya root (PR-AgNPs) and papaya seed (PS-AgNPs), but not with papaya bark. Silver nanoparticles using papaya root and PS-AgNPs were both cubic and showed maximum absorbances of 2.6 and 0.3 AUs at 411.6 and 416.8 nm wavelengths and average hydrodynamic diameters X50 of 59.46 ± 7.03 and 66.57 ± 8.89 nm, respectively. The Ag in both AgNPs was confirmed by X-ray fluorescence by a distinctive peak in the spectrum at the silver Ka line of 22.105 keV. Both AgNPs exhibited broad-spectrum antimicrobial and antibiofilm activity against all Gram-negative bacteria, and PR-AgNPs were slightly better than AgNPs-PS. The MIC ranged from 16 µg/mL–28 µg/mL and 16 µg/mL–64 µg/mL, respectively, for PS-AgNPs and PR-AgNPs. The elucidation of the mechanism of action revealed interference with E. coli ATCC 25922 growth kinetics and inhibition of HM+-ATPase proton pumps. Papaya seed and root extracts were efficient reducing agents for the biogenic synthesis of AgNPs, with noteworthy antibacterial and antibiofilm activities. Future studies should be conducted to identify the phytochemicals and the mechanism involved in AgNPs synthesis. Keywords: antibiotic resistance, biogenic synthesis, Carica papaya, Gram-negative, silver nanoparticles.

Coat color variations in cattle are known to be influenced by the melanocortin 1 receptor (MC1R) and receptor tyrosine kinase (KIT) genes. The presence of coat color abnormalities, such as white spots and albinism, in Bali cattle was the focus of this study. This study aimed to identify single nucleotide polymorphisms (SNPs) in the coding region of MC1R and exons 2 and 3 of KIT associated with coat color abnormalities in Bali cattle. The study included the analysis of 48 Bali cattle, including 20 individuals with standard coat color, 15 with white spots, and 13 with albinism. Total DNA was extracted using a DNA Extraction Kit, and MC1R (coding region) and KIT (exons 2 and 3) gene amplifications were analyzed using forward and reverse primers with polymerase chain reaction product lengths of 1071, 234, and 448 bp, respectively. The determination of MC1R and KIT gene diversity was analyzed through direct sequencing. Melanocortin 1 receptor and KIT gene sequence data were analyzed using BioEdit and MEGA6 to identify SNPs associated with standard and abnormal coat color phenotypes (white-spotted and albino) in Bali cattle. No SNPs associated with coat color abnormalities were found in the coding region of MC1R and exons 2 and 3 of KIT genes in Bali cattle. However, the intron two regions of KIT contained the SNP g.70208534A>G, which showed a high degree of diversity. The AA genotype frequency was highest in albino Bali cattle, whereas the G allele frequency was highest and the A allele frequency was lowest in white-spotted Bali cattle. The results indicated that standard, white-spotted, and albino coat colors in Bali cattle could not be distinguished by analyzing the MC1R and KIT genes. Keywords: Bali cattle, coat color, KIT gene, melanocortin 1 receptor gene, single-nucleotide polymorphism.

MicroRNAs (miRNAs) play an important role in various biological functions. According to many studies, miRNA expression is tissue-specific, strongly controlled throughout embryogenesis, and over- or under-expressed in numerous disorders, including cardiovascular pathologies. This study aimed to screen, characterize, and profile many induced biomarkers (miRNAs) in dog serum before and after experimentally inducing a regional myocardial infarction (MI) by occluding the coronary arteries under general anesthesia. A preclinical experimental animal study recruited 12 healthy canine dogs. The selected canine dogs were anesthetized with 1 mg/kg xylazine and 15 mg/kg ketamine before undergoing femoral arterial catheterization under fluoroscopic supervision. Commercial assay kits were used to purify total RNA and miRNA before the occlusion and 2 h after the occlusion according to the manufacturer’s guidelines, and the samples were stored in RNase/DNase-free water at –80°C. Data were analyzed by GraphPad Prism 5.0 software (GraphPad Prism, San Diego, CA) SPSS, and GenEx software (www.multid.se) or (REST V3). Among 325 transcribed genes, 20 were identified in 2 h. After MI, 14 biomarkers were negative, indicating downregulation, and 6 (3-F08, 3-B10, 4-A11, 1-A06, 2-E01, 3-F10) were positive, indicating upregulation. Polymerase chain reaction assay results showed a normalized fold-change in gene expression in the test sample. Fold values >1 represented a biologically significant change. Profiling of miRNAs before and after MI in a dog model revealed upregulation of six previously unidentified biomarkers (3-F08, 3-B10, 4-A11, 1-A06, 2-E01, and 3-F10), indicating various miRNA regulatory patterns. Keywords: dog model, heart infarction, microRNAs, veterinary.

Sperm maturation occurs in the epididymis through interactions with existing molecules inside the lumen. However, the mechanism of epididymis molecular transfer is currently unclear. This study was aimed to determine the necessity of the epididymal epithelial cells (EECs) in the process of sperm maturation in terms of sperm kinetics and tyrosine phosphorylation. A true experimental research design was used in this study. The medium tested was a primary culture of mice caput epididymal cells (cells and culture medium), conditioned medium (CM) (supernatant of EECs), and secretome (CM filtered at 0.22 µm). Sperm was cocultured in EEC culture, CM, and secretome for 1, 2, 3, or 4 h. The original culture medium was used as the control. Sperm kinetic analysis was performed after the indicated times using computer-assisted sperm analysis, and tyrosine phosphorylation was detected using the Western blot technique. A primary culture of caput EECs was successfully generated. The results showed increased sperm motility and progressive movement after 3 h of incubation (p < 0.05). There was a significant decrease in the average path velocity (VAP) values after 4 h of incubation (p < 0.05), but there was no significant change in the 1, 2, and 3 h incubation groups. The EEC culture-CM and secretome groups showed a significant increased progressivity and VAP percentage values compared with the control medium (p < 0.05). In terms of percentage motility, the culture and CM groups were significantly different from the control medium, but the secretome group was not. The sperm kinetics of sperm cultured in CM, secretome, and EEC were significantly increased after 3 h of incubation, suggesting that CM and secretome can be used to replace EECs, especially when analyzing molecules secreted by the epididymal epithelium during sperm maturation. The results of this study highlight the potential of CM and secretome as therapy mediums for sperm kinetic abnormalities. Keywords: conditioned medium, sperm kinetics, secretome, phosphorylation.

The emergence of antimicrobial-resistant bacteria, such as Escherichia coli in milk, is a serious public health concern as milk is considered a complete food and an important part of daily human diet worldwide, including in Bangladesh. However, there have been no reports on the molecular characterization and antibiotic resistance profile of extended-spectrum beta-lactamase (ESBL)-producing E. coli from milk of healthy cows in Bangladesh. Therefore, this study aimed to detect and characterize ESBL-producing E. coli (ESBL-Ec) in milk samples from healthy cows in smallholder dairy farms in Mymensingh district, Bangladesh, and assess the potential risk of consuming this milk. A total of 100 milk samples were collected from apparently healthy cows on smallholder dairy farms. Escherichia coli was isolated from the collected samples using standard methods. The detection of ESBL-Ec was performed phenotypically using cultural methods and genotypically by ESBL genetic determinants using multiplex polymerase chain reaction. Antimicrobial susceptibility testing of the ESBL-Ec isolates was performed using the disk diffusion method with 15 common antimicrobials. In this study, out of the 100 samples tested, 70 (70%) were found to be positive for E. coli. Among these, 41 (58.6%) strains were identified as ESBL-producing, both phenotypically and genotypically, with the presence of blaCTX-M, blaTEM, and blaSHV individually or combined (blaCTX-M plus blaTEM plus blaSHV). The antibiogram of these ESBL-positive isolates revealed high resistance against commonly used antibiotics, such as ampicillin, cefotaxime, and gentamicin (100%), azithromycin (88%), oxytetracycline (27%), nalidixic acid, cotrimoxazole/trimethoprim (24%), and streptomycin (22%). In addition, one isolate showed resistance to 4th generation of cephalosporin (cefepime). Most importantly, extensive multidrug resistance was found in many ESBL-Ec isolates. However, the isolates were highly sensitive to drugs such as ceftriaxone (100%) and imipenem (100%). This is the first study to detect ESBL-Ec in raw milk from healthy cows on smallholder dairy farms in Bangladesh. More than 58% of the E. coli isolated from raw milk of healthy cows tested positive for ESBL production and showed resistance to most commonly used antimicrobials which may be alarming for human health. A limitation of our study is that we had a small size of sample collected from one district in Bangladesh. Therefore, a larger sample size covering a wider geographic area, and using multi-locus sequence typing and whole genome sequencing could provide a more comprehensive understanding of the prevalence and characteristics of ESBL-Ec in Bangladesh. Keywords: ampicillin, antibiogram, blaCTX-M, cefotaxime, multiplex PCR, multidrug resistance.

Anaplasmosis, an underestimated disease transmitted by ticks, is widespread in ruminants, such as the Arabian camel (dromedary camel). This study aimed to examine the presence of Anaplasma marginale in dromedary camels in the Al-Hiadyia region of the Al-Najaf desert, Iraq, using serological and molecular tests. Moreover, hematological and biochemical changes in infected animals were compared with those in healthy controls. The study was conducted on 30 healthy and 260 infected camels with severe anemia, pale mucus membranes, and progressive emaciation to investigate antibodies against A. marginale using indirect enzyme-linked immunosorbent assay, followed by polymerase chain reaction for selected positive samples targeting a specific region of A. marginale major surface protein 5 (MSP5). In addition, hematological and biochemical parameters were measured to indicate the effect of the disease on blood profile, mineral status, and liver and kidney functions. Enzyme-linked immunosorbent assay analysis and microscopic examination revealed that 115/260 (44.23%) and 87 (33.46%) camels were positive for Anaplasma spp., respectively. The MSP5 gene, which is unique to A. marginale, was amplified. The results of hematological analysis indicated a significant decrease in total red blood cells, hemoglobin, and packed cell volume and a significant increase in mean corpuscular volume in infected camels, but no difference in mean corpuscular hemoglobin concentration. Moreover, there was a significant increase in total white blood cells count, lymphocytes, erythrocyte sedimentation rate, and platelets. The results of biochemical analysis indicated a significant increase in the levels of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase, blood urea nitrogen, creatinine, and iron and a decrease in copper in infected camels. Cholesterol and triglyceride showed no significant variations between healthy and diseased camels. To the best of our knowledge, this is the first molecular study to demonstrate the presence of A. marginale in dromedary camels in Iraq. The MSP5 gene is a valuable and unique diagnostic target for identifying A. marginale. Keywords: Anaplasma marginale, biochemical parameters, dromedary Camel, hematological parameters, indirect enzyme-linked immunosorbent assay, polymerase chain reaction.

The emergence of antibiotic-resistant bacteria and hospital-acquired bacterial infection has become rampant due to antibiotic overuse. Virulence factors are secondary to bacterial growth and are important in their pathogenesis, and therefore, new antimicrobial therapies to inhibit bacterial virulence factors are becoming important strategies against antibiotic resistance. Here, we focus on anti-virulence factors that act through anti-quorum sensing and the subsequent clearance of bacteria by antimicrobial compounds, especially active herbal extracts. These quorum sensing systems are based on toxins, biofilms, and efflux pumps, and bioactive compounds isolated from medicinal plants can treat bacterial virulence pathologies. Ideally, bacterial virulence factors are secondary growth factors of bacteria. Hence, inhibition of bacterial virulence factors could reduce bacterial pathogenesis. Furthermore, anti-virulence factors from herbal compounds can be developed as novel treatments for bacterial infection. Therefore, this narrative review aims to discuss bacterial virulence factors acting through quorum sensing systems that are preserved as targets for treating bacterial infection by plant-derived compounds.

Sporothrix schenckii is the causative agent of sporotrichosis, which most commonly causes lymphocutaneous infections in immunocompromised hosts. This pathogen infects dogs, cats, cattle, and buffaloes and can potentially infect humans. Diagnosis by fungal culture is lengthy, and although there are several clinical diagnoses and molecular methods, these are complicated and time-consuming for veterinarians. This study aimed to develop a visual diagnostic assay that is less time-consuming and can be used by veterinarians to screen for sporotrichosis. To develop a loop-mediated isothermal amplification (LAMP) assay for sporotrichosis, primers specific for fragments of the 18S rRNA gene of S. schenckii were designed. Then, the time and temperature were optimized to successfully achieve LAMP. Ten-fold serial dilutions of DNA were used to determine the detection limit using both LAMP and nested polymerase chain reaction (nPCR) assays. The optimal LAMP conditions were incubation at 73°C for 30 min. Agarose gel electrophoresis revealed a ladder-like pattern of the LAMP product, and a sky-blue color indicated a positive result. A comparison of the LAMP assay with nPCR revealed that it was 10 times more sensitive than nPCR, with a detection limit of 10 pg. The use of a heat box compared with a thermocycler gave the same results. Loop-mediated isothermal amplification gives good results and may represent a future alternative diagnostic tool for screening fungal pathogens before the results of conventional fungal cultures are received. However, this method should be further studied to clarify its use with clinical samples. Keywords: loop-mediated isothermal amplification, molecular detection, rapid detection, Sporothrix schenckii, sporotrichosis, visual detection.

Mastitis in dairy cattle is associated with a high rate of morbidity and death, which has major implications for milk production and quality. This study aimed to investigate the protein component and the activity of matrix metalloproteinase-2 (MMP-2) and -9 (MMP-9) in raw milk samples with different testing scores determined using the California mastitis test (CMT). Thirty cows were employed in the study, and milk from each quarter was tested for subclinical mastitis (SCM). According to the results of CMT, raw milk samples were classified into five categories: Healthy (score 0), trace (score T), weakly positive (score 1), distinctly positive (score 2), and strongly positive (score 3) for somatic cell count (SCC). The total milk protein was analyzed using the Bio-Rad protein assay, and the milk protein composition was determined using the sodium dodecyl sulfate-polyacrylamide gel electrophoresis technique. In addition, gelatin zymography was used to evaluate changes in proteolytic abilities. Milk samples with CMT scores of 1 and 3 had the highest total milk protein levels (32.25 ± 12.60 g/L and 32.50 ± 7.67 g/L, respectively), while the samples from healthy cows (CMT score 0) were only 6.75 ± 1.64 g/L. Globulin and lactoferrin were significantly increased in samples with a CMT score of 3 compared with those with other CMT scores. The bovine serum albumin level in samples with a CMT score of 2 was significantly (p < 0.05) higher than those with other CMT scores. No significant differences in casein abundance were found among samples with different CMT scores. Results from analysis of proteolytic activities demonstrated that the level of MMP-9 in samples with a CMT score of 3 was significantly (p < 0.05) higher than those with other CMT scores. The protein content and gelatinolytic activity of milk were drastically altered by the number of SCC, mainly due to SCM. Keywords: milk protein, protease activity, subclinical mastitis, Thai-crossbred dairy cows.

Hematology and serum biochemical analyses are integral parts of the clinical evaluation of sick animals. This is especially true regarding the clinical care of wildlife species, where clinical signs and historical data relating to the particular illness are often not available. Therefore, this study was designed to report various hematology and serum biochemistry parameters in Arabian oryx (Oryx leucoryx). Various hematology and serum biochemistry parameters were determined in 49 Arabian oryx of various ages and sexes. Hematology parameters included total red blood cells (RBC), packed cell volume (PCV), hemoglobin concentration, mean corpuscular volume, mean corpuscular hemoglobin (MCH), MCH concentration, and total leukocyte count (white blood cell) using an automated hematology analyzer. Serum biochemistry variables included glucose (Trinder method), total protein (biuret method), albumin using the Bromcresol Green (BCG) method, and blood urea nitrogen (colorimetric method). In addition, serum electrolyte concentrations of calcium, magnesium, and phosphorus were determined using colorimetric methods. There was a significant difference in RBC count, PCV, and serum glucose concentration between adult and young Arabian oryx. The RBC count was significantly higher in males than in females, whereas the serum glucose concentration was significantly higher in females. Results of this study showed significant differences in RBC, PCV, and serum glucose concentration between apparently normal young and Adult Arabian oryx. Similar differences were also detected between normal males and females. Knowledge of these data could prove vital in the clinical evaluation of the health status of this wildlife species. Keywords: clinical examination, general health status, laboratory analysis, wildlife.