Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) are retroviruses associated with chronic and neoplastic diseases in domestic and non-domestic cats. There has been increasing interest in the clinical importance of feline retroviruses in Thailand and the identification of associated risk factors in domestic cats. To prevent the spread of retroviral diseases and improve the management of retrovirus-infected cats, risk factors and associated clinical laboratory data must be clearly understood. This study aimed to identify the influence of household, lifestyle, health status, sterilization, clinical presentations, and laboratory findings on FIV- and FeLV-infected cats in Bangkok, Thailand. A total of 480 cats were evaluated for FeLV p27 antigen and FIV antibodies using Witness FeLV-FIV Rapid Test and SNAP FIV/FeLV Combo Test at a veterinary hospital service. Of the 480 cats tested, 113 were positive for virus infection, including 60 for FeLV (12.5%), 40 for FIV (8.3%), and 13 for both FeLV and FIV (2.7%). The findings revealed that the risk factors for cats infected with FeLV, FIV, or both FeLV and FIV were significantly different compared with those for non-infected cats (p < 0.05). Multivariate analysis showed that multi-cat ownership is a risk factor for the high prevalence of feline retrovirus infection, as multi-cat households exhibited a higher prevalence of infection than single-cat households. Anemic and sick cats were also at a greater risk of testing positive for specific retrovirus infections. FeLV-infected cats had a higher risk of anemia and low erythrocyte and thrombocyte counts (p ≤ 0.0001), whereas FIV-infected cats were more likely to have anemia and leukocytopenia than controls. Knowledge of the risk factors for retroviral diseases and associated clinical and laboratory findings can be used to develop strategies to reduce FIV and FeLV infections in cats. Keywords: feline immunodeficiency virus, feline leukemia virus, hematology, risk factors, serum biochemistry.

This study examines methods to effectively control peste des petits ruminants (PPR), an emerging, highly contagious, transboundary disease that has been designated as a highly dangerous disease by the World Organization for Animal Health. Mathematical modeling was used as a predictive and preventive tool to assess the risk of PPR virus spread in the model area and the probability of its introduction into the territory of the Russian Federation. PPR risk assessment was performed by modeling the pathogen's ecological niche by performing linear regression analysis in the geographic information system ESRI ArcGIS Desktop and maximum entropy methods using MaxEnt software. The territories of Bangladesh, China, and Algeria were used as model countries because they have the highest number of confirmed PPR outbreaks, as reported by the Food and Agriculture Organization of the United Nations from 2009 to 2020. The prepared global model of the PPR pathogen's ecological niche was extrapolated onto the territory of the Far Eastern regions of the Russian Federation to assess the probability of virus introduction in that region. Global model analysis showed that two factors exerted the highest influence on the spread of the PPR pathogen on a global scale: The minimum temperature of the coldest month of the year and the density of roads per unit area, which reflect the overall economic activity within a region. The highest risk of PPR spread was observed in areas with a minimum annual temperature of 16°C and road density of 5000 m/km2. According to the model, areas with a dominant subtropical climate, where small livestock breeding is performed and where the average daily air temperature is >0°C throughout the year, are at the highest risk of PPR outbreaks. The risk of PPR spreading outside these areas is significantly reduced. Local extrapolation of the PPR ecological niche model demonstrates that the probability of epizootic development does not exceed 3–4% within the territories of the constituent entities of the Russian Federation adjacent to Mongolia and China. Keywords: ecological niche, environmental factors, goat, linear regression model, maximum entropy model, peste des petits ruminants, sheep, species distribution.

Clostridium perfringens type A is an anaerobic bacterium that produces four major toxins (alpha, beta, epsilon, and iota) that cause various diseases. Most of the important C. perfringens-associated diseases of farm animals are caused by alpha-toxin. This study aimed to produce a vaccine against alpha-toxin using C. perfringens type A (ATCC 13124) and investigate its potency, stability, and safety. The vaccine was formulated of its constituents for 1 h. Each milliliter of the final vaccine product contained alpha toxoid 15 lecithovitellinase activity (Lv) by adding (0.375 mL containing 40 Lv) and approximately 0.2 mL from 3% concentrated aluminum hydroxide gel, <0.001% W/V thiomersal, <0.05% W/V formaldehyde, and nearly 0.425 mL phosphate-buffered saline (pH 7.2). The vaccine efficacy was evaluated in rabbits and cattle by performing potency, stability, and safety tests. The vaccine produced approximately 8.8 and 4.9 IU/mL neutralizing antibodies in rabbits and cattle, respectively. These concentrations were higher than the lowest concentration recommended by various international protocols and the United States Department of Agriculture by 2.20-fold in rabbits and 1.23-fold in cattle. Interestingly, the formulated vaccine enhanced immune responses by 1.80-fold in rabbits compared with that in cattle; the difference was statistically significant (p < 0.0001). The vaccine was stable for 30 months. In vaccinated rabbits, the body temperature slightly increased temporarily during the first 10 h of vaccination; however, the temperature difference was not statistically significant (p > 0.05). This study describes a manufacturing process to obtain sufficient amounts of a vaccine against C. perfringens alpha-toxin. The formulated vaccine effectively elicited a higher level of neutralizing antibody response than the international standards. Furthermore, the vaccine was found to be stable, safe, and effective in preventing C. perfringens-related diseases in rabbits and cattle. Further studies are necessary to evaluate the efficacy of this vaccine in other farm animals. Keywords: alpha-toxin, Clostridium perfringens A, potency, safety, stability, toxoid.

Bartonella spp. are Gram-negative zoonotic bacteria that are transmitted to humans by several types of animal hosts, including rodents. Several studies have been conducted on the prevalence of Bartonella infections in rodents. However, the risk of rodent-associated Bartonella spp. infection in humans remains unclear. This study aimed to estimate the prevalence and genetic heterogeneity of Bartonella spp. in rodents and shrews from nine provinces of Thailand using culture and molecular techniques. A total of 860 blood samples from rodents and shrews across nine provinces of Thailand were collected from January 2013 to June 2016. Bartonella spp. were isolated from all samples using conventional culture techniques and polymerase chain reaction. Phylogenetic tree analysis was used to align the Bartonella sequences obtained from this study. The prevalence of Bartonella spp. in rodents and shrews was 11.5% (99/860, 95% confidence interval: 9.38–13.64%). The following nine species of Bartonella were detected: Bartonella tribocorum, Bartonella rattimassiliensis, Bartonella queenslandensis, Bartonella elizabethae, Bartonella chanthaburi spp. nov., Bartonella satun spp. nov., Bartonella coopersplainsensis, Bartonella ranong spp. nov., and Bartonella henselae. The prevalence of Bartonella-positive animals differed significantly among provinces. To the best of our knowledge, the three novel Bartonella spp. isolated from rodents and shrews across Thailand were detected for the first time in this study. Further studies on the epidemiology of Bartonella infection in rodents and its interaction with human health should be conducted in accordance with the Thai government's "One Health" approach to humans, animals, and the environment. Keywords: Bartonella spp., phylogenetic analysis, polymerase chain reaction, rodents.

The leptin (LEP) gene plays a role in the regulation of the activity required to obtain food, energy metabolism, and fat deposition and affects the body composition of animals. Lipogenesis is an ineffective process. A lot of energy from feed is expended on the synthesis of adipose tissue. This study aimed to determine the effect of LEP C528T and LEP C73T polymorphisms and pregnancy on adipose tissue formation and carcass grade in Aberdeen Angus heifers and first-calf cows. Heifers (n = 49) and first-calf cows (n = 30) were grouped according to their genotype for LEP C528T and LEP C73T polymorphisms at the age of 24 months. DNA samples were isolated from whole blood. Experimental animals were slaughtered at the age of 24 months, and a chemical analysis of samples of longissimus dorsi muscle and ground beef was performed. The maximum internal fat weight, back-fat thickness, and fat content of the longissimus dorsi muscle and ground beef were determined in heifers heterozygous for both LEP C528T and LEP C73T polymorphisms. The ranking of genotypes in terms of adipose tissue formation did not change in first-calf cows compared to heifers carrying the same LEP gene variants. Pregnancy had a more significant (p < 0.05–0.001) effect on slaughter parameters and lipogenesis in animals of different genotypes than studied polymorphisms. Heterozygosity of the LEP gene was much more strongly expressed in the carcass grade of heifers. "Prime" grades were assigned to 66.7% of carcasses heterozygous for LEP C528T and "Top Choice" to 58.8% of carcasses with LEP C73T nucleotide substitutions. LEP C528T and LEP C73T polymorphisms were shown to affect the extent of fat formation in Aberdeen Angus heifers and first-calf cows. Animals with heterozygous genotypes exhibited the maximum development of internal fat, back-fat, and intramuscular fat. Pregnancy had a more significant effect on slaughter parameters and adipose tissue formation than studied polymorphisms. First-calf cows had a significantly lower fat content in carcasses than heifers. These results can aid in the production of efficient mature herds of Aberdeen Angus cattle. Keywords: Aberdeen Angus, beef cattle, genotyping, leptin, polymorphism.

Toxoplasma gondii is a global zoonotic parasite infecting virtually all warm-blooded species, although a species-specific variability is evident referring to symptoms frame. Both the success of T. gondii and the outcome of infection depend on a delicate balance between host cellular pathways and the evasion or modulation strategies elicited by the parasite. The hormonal and molecular mechanisms involved in this delicate host-parasite balance are still unclear, especially when considering intermediate host species other than mouse. This study aimed to assess any correlation between T. gondii infection and selected molecular and hormonal factors involved in responses to infection in susceptible species such as swine. Moreover, blood counts and hematochemical assays (glucose, total cholesterol, and triglycerides dosage) were performed to evaluate the overall health condition of animals. Toxoplasmosis was diagnosed by enzyme-linked immunosorbent assay for antibodies determination and real-time polymerase chain reaction (RT-PCR) for T. gondii DNA detection. Target genes coding for key factors of cell responses to T. gondii infection were selected, and their transcription was assessed in various tissues by quantitative RT-PCR. 17-β estradiol concentrations were assessed by fluorimetric enzyme-linked immunoassay and the AIA- 360 automated immunoassay analyzer. Blood count and hematochemical analyses were performed by a blood cell counter and a spectrophotometer, respectively. The present research highlighted significant differences among infected and uninfected swine (control group) for both transcription profiles of some of the molecular factors considered and 17-β estradiol concentrations. Referring to the assessed hematological and biochemical parameters, no statistically significant differences were observed in infected swine compared to the control group. Our results contribute to the enrichment of data available about the subject and could be useful for a deeper knowledge of the interaction between this parasite and its hosts. However, more aspects are still unclear, such as the effective response of downstream molecules from the same pathways to the variation of factors observed in this study either assessing how the same factors respond to Toxoplasma gondii infection in other host speciesand further analyses should be performed on other host species. Keywords: biomarkers, host-parasite interaction, swine, Toxoplasma gondii.

Supplementation of AKBISprob (developed in a previous study) in feed can improve production efficiency and poultry health, especially laying hens. In addition, it can also increase cellulolytic lactic acid bacteria (LAB) in chicken intestines, but these bacteria are still unknown; thus, they need to be identified. This study aimed to identify cellulolytic LAB in the intestines of laying hens administered AKBISprob based on 16S ribosomal ribonucleic acid (16S rRNA) gene analysis. The samples used in this study were 13 LAB isolates from the intestines of laying hens that were given AKBISprob 4%. Cellulolytic LAB DNA was isolated and 16S rRNA gene was amplified by polymerase chain reaction, followed by sequencing, bioinformatics analysis, and phylogenetic tree construction. From 10 cellulolytic LAB isolates with a clear zone of >6 mm, four were selected and their DNA was amplified with BaCF and UniB primers ∼1500 bp DNA fragments. Of these, the P31H62 isolate was genetically close to Enterococcus hirae strain 1-1X-16 with 92.90% maximum identity, the P33S52 isolate had homology with Enterococcus mundtii strain ZU 26 with 96.76% maximum identity, and the P33S62 isolate was closely related to E. hirae strain SJ3 with 72.96% maximum identity. The phylogenetic tree revealed that the cellulolytic LAB isolates P31H62 and P33S52 were in one cluster closely related to the genus Enterococcus. This study suggests that the isolates P31H62, P33S62, and P33S52 from the intestines of laying hens administered 4% AKBISprob are cellulolytic LAB belonging to the genus Enterococcus. Keywords: 16S ribosomal ribonucleic acid, AKBISprob, Enterococcus, phylogenetic tree, polymerase chain reaction.

Heat stress (HS) can negatively impact farm animal productivity and adversely affect animal welfare worldwide, placing a major financial burden on global livestock producers. Dietary betaine (trimethylglycine) has been known to have several biological functions, which may aid in offering beneficial effects on livestock productivity during HS conditions. However, information on the role of dietary betaine in heat-stressed dairy heifer calves is yet to be documented. Therefore, this study aimed to assess the effects of supplementing dietary betaine on body temperature indices, blood metabolites, productive performance, and complete blood count (CBC) (hematological indices) in hyperthermic dairy heifer calves. In total, 14 Holstein heifer calves (4.0 ± 0.9 months old) were individually housed and randomly allocated to one of two dietary treatments: (1) a control diet (CON; n = 7) and (2) a control diet complemented with 21 g/d of natural betaine (BET; n = 7) top-dressed once daily. The experiment lasted for 28 d, during which all animals were subjected to natural cyclic HS conditions (26.1–39.2°C; 73.2–84.0 temperature–humidity index). Rectal temperature (RT) and respiration rate (RR) were measured twice daily (0700 and 1500 h), whereas dry matter intake (DMI) was measured once daily (0800 h). In addition, blood samples (collected from the jugular vein) were analyzed for metabolites and CBC on days 7, 14, 21, and 28. Relative to CON, BET supplementation was able to decrease RT on day 23 of the experiment (p = 0.04). Alternatively, RR was similar between the dietary treatments (p = 0.73). Feeding BET did not affect DMI compared with CON during HS conditions (p = 0.48). Furthermore, compared with CON, BET supplementation did not change leukocytes, neutrophils, lymphocytes, and hematocrit levels during HS conditions (p ≥ 0.17). However, a post hoc analysis indicated that hematocrit levels were decreased in BET-fed calves on day 7 of the study compared with CON calves during HS conditions (p = 0.05). Moreover, circulating glucose, albumin, and triglycerides were found to be similar between dietary treatments (p ≥ 0.55). BET supplementation slightly reduced RT and circulating hematocrit but did not affect other metrics in this HS experiment. More research into the effects of different doses of dietary BET on dairy heifer calves is needed. Keywords: betaine, dairy calves, heat stress.

The oviduct environment is of particular importance because it is the site of fertilization and early embryo development. The oviduct, as a component of the reproductive system, responds to ovarian hormone (estradiol [E2] and progesterone [P4]) stimuli depending on the estrous cycle phase. This study aimed to elucidate the effect of estrous cycle phases (follicular and early and late luteal phases) on gene expression patterns in bovine oviduct epithelial cells (BOECs). Oviducts were obtained from healthy slaughterhouse animals, corresponding to ipsilateral ovaries with dominant follicles or corpus luteum during early and late luteal phases. BOECs were recovered from the isthmus (IST) and ampulla (AMP), and the expression patterns of genes related to cytokinesis and mitosis mechanisms (rho-associated coiled-coil containing protein kinase and cellular communication network factor 2 [CCN2]), growth factors (insulin-like growth factor-binding protein 3, epidermal growth factor receptor [EGFR], vascular endothelial growth factor A, and EGFR), antioxidant mechanisms (glutathione peroxidase 4 [GPX4]), apoptosis (B-cell lymphoma 2), complement component (C3), energy metabolism (aldose reductase gene family 1-member b1 [AKRIB1] and solute carrier family 2), hormone receptors (estrogen receptor 1 and luteinizing hormone/choriogonadotropin receptor), and specific glycoproteins (oviductal glycoprotein 1) were analyzed. High P4 levels (late luteal phase) affected the expression of important genes related to antioxidant mechanisms (GPX4), energy metabolism (AKRIB1), growth factors (IGBP3 and EGFR), and cell growth regulation (CCN2) in the AMP. Low P4 levels (early luteal phase) affected the expression of AKR1B1, IGBP3, and CCN2. In addition, estrogen likely had an effect on OVPGP expression in the cattle oviduct. Differential gene expression patterns of BOECs in the AMP during the luteal phase (antioxidant mechanisms, energy metabolism, growth factors, and immunological regulators) and in the IST during the follicular phase (glycoproteins) may influence their renewal and population proportions, modulating the oviduct environment as well as gamete and embryo physiology. Keywords: bovine oviduct epithelial cells, estrus phase, follicular, gene expression, luteal, oviduct.

Organophosphate pesticides (OPs) used in agricultural production pose environmental and public health risks whenever non-target organisms are exposed to them. Oxon-type OPs, such as trichlorfon (TCF) and chlorpyrifos (CPF), are frequently used in Colombia and have been detected in water bodies in the vicinity of croplands; however, their effect on aquatic organisms, especially fish, is largely unknown. The neurotoxicity of OPs includes inhibition of esterase enzymes, neuronal damage, and increased glial reactivity. This study aimed to assess the astrocytic response in the brain tissue of juvenile red-bellied pacu (Piaractus brachypomus) exposed to TCF and CPF. A 25-day subchronic assay was conducted in which juvenile red-bellied pacu were exposed to CPF and TCF. After 25 days of exposure, the fish were killed and brain samples were collected and processed for immunohistochemistry to assess the morphology and reactivity of astrocytes; glial acidic fibrillary protein was used as a biomarker. The brain samples from animals under subchronic exposure to OPs for 25 days showed higher cellular density as well as changes in astrocyte phenotype characterized by shortening of cytoplasmic projections, hypertrophy, and ameboid morphology compared to those from nonexposed animals. Similarly, astrocyte hyperreactivity was detected in the optic tectum and medial longitudinal fasciculus of the exposed group. Immunoreactivity of brain glial cells under subchronic exposure to OPs measured through immunohistochemical tests as well as OPs-induced neuropathology may be useful as a biomarker for monitoring environmental pollution. The results also indicate that P. brachypomus is a suitable biomonitoring model for studying neurotoxicological and neurodegenerative diseases. Keywords: astrocytes, fish, neurotoxicity, organophosphates, toxicity biomarkers.

Studies on avian influenza virus (AIV) in Libya are few and limited. This study aimed to determine the presence of AIV in live bird markets (LBMs) in Tripoli and determine the risk factors associated with AIV spread. In total, 269 cloacal swabs were randomly collected from different bird species in 9 LBMs located in Tripoli and its surrounding regions. The target species were ducks, geese, local chickens, Australian chickens, Brahma chickens, turkeys, pigeons, quails, peacock broiler chicks, and pet birds. Total RNA was extracted from the swab samples and used for real-time polymerase chain reaction to detect AIV type A. Of the 269 samples, 28 (10.41% of total samples) were positive for AIV type A. The LBMs with positive samples were Souq Aljumaa, Souq Alkhamees, Souq Althulatha, and Souq Tajoura. The highest percentage (35.71%) of AIV was recorded in Souq Aljumaa. Positive results for AIV type A were obtained primarily in three species of birds: Ducks (14/65; highest percentage: 21.5%), local chickens (12/98; 12.24%), and geese (2/28; 7.14%). Furthermore, the following three risk factors associated with the spread of AIV type A were identified: Time spent by breeders/vendors at the market (odds ratio [OR] = 11.181; 95% confidence interval [CI] = 3.827–32.669), methods used for disposing dead birds (OR = 2.356; 95% CI = 1.005–5.521), and last visited LBM (OR = 0.740; 95% CI = 0.580–0.944). Restricting the movement of poultry vendors from one market to another may protect against AIV spread. The findings of this study indicate the high risk of AIV spread in LBMs and highlight the need for continuous surveillance of LBMs across the country. Keywords: avian influenza, live birds market, risk factors, Tripoli.

One of the most significant public health concerns is multidrug-resistant (MDR) microorganisms. Klebsiella spp. have been at the forefront of causing different types of infections such as bacteremia, urinary tract infections, pneumonia, enteritis, and sepsis in humans as well as animals. This study aimed to determine the genomic similarity between Klebsiella spp. isolated from wild animal samples and those described in the Institut Pasteur genomic database to verify the spread of resistant clones regionally in the state of Mato Grosso, and to compare the epidemiological data in different regions of Brazil and the world. Isolates from various sites of injury in wild animals were identified by sequencing the 16S rRNA gene. Antimicrobial susceptibility testing was performed using the disk diffusion method to verify the resistance profile, and then, multilocus sequence typing was performed to verify the population structure and compare the isolates from other regions of Brazil and the world. Twenty-three sequence types (STs) were observed; of these, 11 were new STs, as new alleles were detected. There was no predominant ST among the isolates. All isolates were MDR, with high rates of resistance to sulfonamides, ampicillin, amoxicillin, and nitrofurantoin and low resistance to meropenem, imipenem, and amikacin. Improving our understanding of the population structure of Klebsiella spp. in wild animals may help determine the source of infection during outbreaks in humans or animals, as the One Health concept emphasizes the interlinks between humans, animals, and environmental health. Keywords: antimicrobial, Klebsiella, multidrug resistance, multilocus sequence typing, susceptibility, wild animals.

The brown dog tick, Rhipicephalus sanguineus sensu lato, is the most common tick found on domestic dogs in Southeast Asia, including Thailand. Canine tick-borne pathogens are a public health concern worldwide. Tick-borne diseases are diagnosed by identifying pathogens based on the morphological or molecular analyses of dog blood samples. However, the collection of ticks, a non-invasive procedure, is easier than drawing blood. This study aimed to demonstrate the usefulness of collecting brown dog ticks for the diagnosis of tick-borne diseases and for estimating the prevalence of tick-borne pathogens among companion dogs in Khon Kaen, Northeast Thailand. Seventy brown dog ticks from 70 companion dogs in Khon Kaen Province, Thailand, were evaluated for molecular evidence of tick-borne pathogens, including Babesia spp., Ehrlichia canis, and Hepatozoon canis. Ticks were collected from dogs at a private animal hospital based on the presence of at least one of the three inclusion criteria: fever, anorexia, or lethargy. Molecular diagnosis was performed using conventional polymerase chain reaction for the detection of pathogens. Of the 70 ticks collected from 70 sick dogs, 55 (78.57%) were positive for tick-borne pathogens. The most common infection was a single infection with H. canis (65.71%) followed by Babesia spp. (31.43%) and E. canis (30.00%). Coinfection was observed in 14 ticks (20.00%), and coinfection with Babesia spp. and E. canis was the most prevalent double infection (n = 6). The prevalence of coinfection was identical for H. canis mixed with Babesia spp. and H. canis mixed with E. canis (n = 4). The present study showed that tick-borne pathogens are highly prevalent among companion dogs in Khon Kaen Province. Therefore, we encourage an increase in tick control or the reduction and prevention of tick-borne diseases in this region. Furthermore, this study revealed that ticks are valuable samples for the molecular detection of tick-borne pathogens. Keywords: brown dog tick, Rhipicephalus sanguineus, tick-borne diseases, tick-borne pathogens.

The efficacy of intranasal (IN) dexmedetomidine in cats as a premedication remains elusive. Thus, this study aimed to compare the perioperative and sparing effects of IN and intramuscular (IM) dexmedetomidine administration on propofol requirements for anesthetic induction in cats. This study randomly assigned 16 cats into two groups of IN or IM dexmedetomidine at 20 μg/kg. Sedation scores and side effects were recorded at time points of 0, 5, 10, 15, and 20 min after the dexmedetomidine administration. Anesthesia was induced with intravenous (IV) 1% propofol by titrating a bolus of 2 mg every 45 s and the total dose of the administered IV propofol to achieve endotracheal intubation was recorded. Cats receiving IM dexmedetomidine were significantly associated with higher sedation scores. All cats were sedated at 20 min after premedication; however, the average composite sedation scores in the IN group were significantly lower than those in the IM group during premedication. Pre-operative side effects, including vomiting, were more frequently observed in the IN group (5 cats, 62.5%) than in the IM group (3 cats, 37.5%; p < 0.05). Higher body temperature (>1°F compared to baseline) was more frequently observed in the IN group (6 cats, 75.0%) than in the IM group (1 cat, 12.5%; p < 0.05). The dosage of required propofol in the IN group was significantly higher (1.1 ± 0.3 mg/kg) than that in the IM group (0.7 ± 0.2 mg/kg; p < 0.05). The duration of general anesthesia was comparable between the groups. IN dexmedetomidine produces moderate sedation and cats may have side effects, including vomiting and higher body temperature. Higher sparing effects of propofol were identified in the IM group compared with the IN group. Nonetheless, IN administration of dexmedetomidine provides a noninvasive alternative to the IM route. Keywords: cats, dexmedetomidine, intranasal, propofol, sedative.

Pomegranate is known to possess antibacterial properties, partly because of its punicalagin content. However, its effect on canine oral bacterial species has not yet been elucidated. In this study, we evaluated the effect of pomegranate extract present in pet dental products on the growth and survival of five canine oral bacterial species in biofilms. Five bacterial species, Neisseria shayeganii, Neisseria canis, Porphyromonas gulae, Porphyromonas macacae, and Porphyromonas crevioricanis, were individually cultured for biofilm formation and exposed to pomegranate extract (or control) for 15 min. Cell survival was analyzed using the 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide assay and was compared between different conditions using a student's t-test. In addition, the individual strains were grown in planktonic suspensions and exposed to serial dilutions of the extract to determine the minimum inhibitory concentration. At a concentration of 0.035% w/v, the extract significantly reduced the survival of P. gulae (–39%, p < 0.001) and N. canis (–28%, p = 0.08) in biofilms. At similar concentrations, the extract also completely or partially inhibited the growth of N. canis and Porphyromonas spp. in planktonic suspensions, respectively. The pomegranate extract found in some pet dental products can limit bacterial growth and survival in the biofilms formed by N. canis and P. gulae in vitro. As P. gulae is involved in periodontal disease progression, limiting its proliferation using products containing pomegranate extract could contribute to disease prevention. Further studies on dogs receiving such products are necessary to confirm these effects. Keywords: dental biofilm, dogs, Neisseria canis, oral hygiene, pomegranate, Porphyromonas gulae.

The Kalahari Red goat breed is the finest meat-producing species in South Africa, and its coat color ranges from light to dark red-brown. A practical approach to estimating their body weight (BW) using linear body measurements is still scarce. Therefore, this study aimed to determine the best data mining technique among classification and regression trees (CART), Chi-square automatic interaction detection (CHAID), and exhaustive CHAID (Ex-CHAID) for predicting the BW of Kalahari Red goats. This study included 50 Kalahari Red goats (does = 42 and bucks = 8) aged 3–5 years. Body length (BL), heart girth (HG), rump height (RH), height at withers (WH), sex, and age were the essential indicators to estimate BW. The best model was chosen based on the goodness of fit, such as adjusted coefficient of determination (Adj. R2), coefficient of determination (R2), root mean square error (RMSE), standard deviation ratio (SD ratio), mean absolute percentage error, Akaike information criteria, relative approximation error, and coefficient of variation. The SD values of the ratio ranged from 0.32 (CART) to 0.40 (Ex-CHAID). The greatest R2 (%) was established for CART (89.23), followed by CHAID (81.99), and the lowest was established for Ex-CHAID (81.70). CART was established as the preferred algorithm with BL, HG, and WH as critical predictors. The heaviest BW (73.50 kg) was established in four goats with BL higher than 92.5 cm. This study reveals that CART is the optimum model with BL, HG, and WH as the essential linear body measurements for estimating BW for Kalahari Red goats. The updated records will assist the rural farmers in making precise judgments for various objectives, such as marketing, breeding, feeding, and veterinary services in remote areas where weighing scales are unavailable. Keywords: body length, data mining algorithms, heart girth, rump height, withers height.

Several factors contribute to the unusual incidence of antibiotic resistance, which is now a primary public health concern. However, failure in managing preventive and therapeutic antibiotic use on farms is one of the most crucial factors. Therefore, this study aimed to evaluate the biosecurity of farms, farmers' competence, and practices related to antibiotics and their resistance in poultry and pig rearing in Togo. Through a cross-sectional survey, 121 commercial poultry farmers and 97 commercial pig farmers were questioned to evaluate the biosecurity of farms and farmers' competence and practices related to antibiotics and antibiotic resistance. Descriptive analyses, including the evaluation of proportions, were carried out. In addition, results from qualitative factors were evaluated in a defined grid and totaled up to assess cleanliness measures, awareness, and behavior regarding antibiotics and their resistance. The results demonstrated that most farmers working on poultry farms had a university education, while most working on pig farms had secondary education. Most poultry (69%) and pig (44%) farms were of small sizes (<1000 animals in poultry and <10 animals in pig farming). The footbaths were used in just 51% of poultry farms and 4% of pig farms, respectively, with 37% and 82% of poultry and pig farms having inadequate levels of hygiene. In poultry farms, respiratory issues and periodic decline in egg-laying were the main problems. Simultaneously, skin disorders (scabies) and plagues (African swine fever) were the primary health constraints in pig farming. Tetracycline is the most commonly used antibiotic by farmers. However, in poultry and pig farms, 21% and 67% of farmers were unaware of antibiotics. In addition, 39% and 57% were unaware of antibiotic resistance. Poultry and pig farmers' competence were substantially linked to their education level. Poultry farmers demonstrated better practices, including procuring antibiotics based on veterinary prescriptions (63%) and they knew where antibiotics should be bought (90%). Nevertheless, 43% of farmers asserted unpleasant activities – no application for laboratory testing (93%) and use of antibiotics for prevention (82%). In pig farming, most farmers (69%) reported inadequate incidents of the use of antibiotics. This study identified a crucial non-compliance with biosecurity measures and good practices toward antibiotic use on many farms. Therefore, training of farmers is mandatory for safe livestock products. Keywords: antibiotic resistance, antibiotics, biosecurity, knowledge, pig, poultry, practice.

The meat supply of local poultry for human consumption is greater than that of fast-growing poultry in Niger. However, meeting the protein needs of these local chickens is a major challenge due to the availability of protein sources and their cost. Nowadays, insect larvae such as houseflies are used and even recommended as animal feed; hence, the need to evaluate the effect of housefly (Musca domestica) larvae on the growth performance of local chickens. This study investigated the feeding effects of housefly larvae on the growth performance and carcass characteristics of local Nigerien chickens and determined the rate of fish meal substitution, in fresh or dry larvae form, whichever would be preferable. A total of 165 3-week-old local unsexed chickens of the salmon variety, weighing 120.3 ± 15.43 g, were used to evaluate the effect of housefly (M. domestica) larvae on their growth performance and carcass yield (CY). The experiment consisted of five treatments with three replicates, that is, 15 batches of 11 animals each. Five iso-protein-caloric diets were developed with 25%, and then 50% fish meal substitution with fresh and dried housefly larvae. The chicks were reared together during the first 3 weeks for their adaptation, during which they were fed an imported starter commercial feed, ad libitum. After that, they were weighed weekly for 12 weeks. Next, the body weights (BWs) were taken weekly for all chicks, feed daily intake and mortality were recorded daily, and average daily gain, feed conversion ratio (FCR), and viability rate were calculated. In the end, four chickens (two males and two females) per batch were slaughtered for the CY evaluation, breast meat, drumstick and tight (legs), and wings. Statistical analyses were performed using a linear mixed model for repeated data. The weight, FCR, and carcass traits were unaffected by either the rate or larvae state. Conversely, the growth rate was improved, and feed consumption was increased. Notably, the chickens consumed more feed but grew faster with fresh larvae and at a higher substitution rate. This study reported that 50% fresh or dried housefly larvae substituted into the fish meal in growing local chicken's diets had no effect on their BW, FCR, and carcass traits but increased the growth rate and feed consumption. Keywords: alternative feeds stuff, animals feeding, carcass, indigenous chicken, insect larvae, poultry diets, zootechny.

Brucellosis is a contagious livestock disease with a significant economic impact. This study aimed to compare the efficacy of antibiotics used alone or in combination with silver nanoparticles (AgNPs) against Brucella melitensis Rev 1 in vitro. AgNps conjugated with ciprofloxacin was synthesized and thoroughly characterized by ultraviolet visible spectrophotometry (UV-vis). The antimicrobial effect of ciprofloxacin alone and ciprofloxacin conjugated with AgNPs against B. melitensis Rev 1 was determined by minimum inhibitory concentration (MIC) and the erythrocyte hemolytic assay determined the capability of conjugation to cause hemolysis in human erythrocyte. The UV-vis spectra of both silver-drug nanoconjugates showed a characteristic surface plasmon resonance band at 420 nm. The MIC assays showed that AgNPs conjugation to antibiotics enhanced the antibacterial potential of the selected antibiotics against B. melitensis Rev 1 relative to non-conjugated antibiotics. The results show that low concentrations of AgNPs can kill B. melitensis Rev 1. The MICs of ciprofloxacin and ciprofloxacin–AgNPs were 0.75 and 0.05 μM, respectively. The conjugation of ciprofloxacin with AgNPs enhanced the antibacterial effects against B. melitensis Rev 1. In addition, this conjugation appears to inhibit the capability of this bacterium to adapt to the presence of antibiotics, thereby inhibiting bacterial resistance. Further studies are required to examine its potential as an in vivo treatment. Keywords: antimicrobial activity, Brucella melitensis Rev 1, minimum inhibitory concentrations, silver nanoparticles.

To develop an environmentally friendly alternative to mosquito larvicides for vegetables, leaf extracts of Mangifera laurina, Mangifera casturi, Mangifera indica, Mangifera odorata, Mangifera caesia, and Mangifera foetida were prepared. This study aimed to determine the biological efficacy of several Mangifera leaf extracts on the mortality of Aedes aegypti mosquito and the inhibition of egg hatching. Extraction was performed in an organic solvent (methanol) using a Soxhlet extractor. The larvicidal potential of six leaves of Mangifera essential oil was evaluated against the third instar larvae of A. aegypti at concentrations of 1500, 2000, 3000, and 5000 ppm using the World Health Organization protocol. After Probit analysis, the 48 h LC50 and LC90 values of the essential oils were determined. The inhibitory effect on egg hatching was also tested at 160, 320, 480, and 640 ppm. The extraction of essential oils from several Mangifera species had excellent larvicidal activity and inhibitory activity against A. aegypti egg hatching. The LC50/LC90 values were: M. casturi, 241/1964 ppm; M. laurina, 2739/4035 ppm; and M. caesia, 1831/2618 ppm. The inhibitory effect on hatching was 78% for M. foetida, 70% for M. caesia, and 59% for M. casturi. The test results indicate the potential of some Mangifera species for use as larvicides and inhibitors of egg hatching; thus, they have the potential to control A. aegypti in the early stages of development. Keywords: Aedes aegypti, bio-efficacy, leaf extracts, Mangifera.

Motile Aeromonas septicemia is a crucial disease in freshwater fish. Aeromonas hydrophila is a disease agent associated with sporadic fish mortality, food safety, and public health. This study aimed to estimate the prevalence and the presence of the aerolysin gene and antimicrobial resistance profile of A. hydrophila isolated from milkfish in Gresik, Indonesia. A total of 153 milkfish gill samples were collected from 16 locations in Gresik and then cultured and identified using biochemical tests. The aerolysin gene was investigated using a polymerase chain reaction, and antimicrobial resistance profiles of the recovered isolates were investigated. Of the 153 examined samples, 35 (22.9%) were confirmed positive for A. hydrophila and 22 (62.9%) presented the aerolysin gene. The recovered isolates were resistant to the following antibiotics: Amoxicillin (62.9%), tetracycline (60%), streptomycin (54.3%), cefotaxime (51.4%), gentamycin (31.4%), kanamycin (28.6%), erythromycin (25.7%), chloramphenicol (20%), and trimethoprim (14.3%). Meanwhile, only ciprofloxacin, nalidixic acid, and imipenem were indicated as susceptible. The presence of the aerolysin gene is vital in determining the virulence of A. hydrophila. The study results indicated a high aerolysin gene prevalence. In addition, this study emphasized antibiotic use monitoring, food safety improvement, and negative impact reduction on human health and the environment. Keywords: aerolysin gene, Aeromonas hydrophila, antimicrobial resistance, milkfish, public health.

Staphylococcus aureus produces various superantigen exotoxins, including staphylococcal enterotoxin B (SEB). It causes fatal anaphylactic reactions and toxic shock. This study aimed to evaluate the reaction of leukocytes and histopathological changes in the respiratory organs of Balb/c mice after intranasal infection with enterotoxigenic S. aureus (SEB). The presence of the seb gene in S. aureus was established in this study using polymerase chain reaction-specific primer. Two groups of 8-week-old male Balb-c mice consist of six mice in each group. The treated group was infected with 50 μL and 100 μL of SEB intranasal on days 1 and 14, respectively. NaCl was administered in the second group and was considered as a control group. Blood samples were collected through the retro-orbital plexus on days 1, 4, 7, 14, and 22 after infections. Total cell counts were analyzed with an independent sample t-test and compared using the statistical package for the social sciences (SPSS) version 16.0 (IBM Corp., NY, USA). The infected tissues of the respiratory organ were observed descriptively and compared to the control group. The seb gene with a molecular size of 478 bp, indicating the SEB strain, is present in S. aureus used in this study. Intranasal administration of SEB showed increased leukocytes, lymphocytes, monocytes, and eosinophils on day 22 post-infection. Significant leukocytosis was seen on days 6 and 14; lymphocytosis on days 1, 4, 6, and 16; and eosinophilia on days 6, 14, and 22 compared with the control group (p > 0.05). In contrast, the neutrophil decreased after an increase of immature band cells compared to the control group, indicating a severe acute infection with SEB. The lungs and trachea of the test group had an inflammatory cell accumulation in the respiratory organ. Intranasal route infection of S. aureus containing seb gene significantly induced the cellular immune response and caused pathological changes in the respiratory tissues of the Balb/c mice model. The hematological changes were aligned with marked pathological changes in the respiratory tract. Balb/c mice could be an excellent experimental model to study toxic and anaphylactic shock against SEB to define the future therapeutic agents. Keywords: enterotoxin B, hematology, histopathology, intranasal, Staphylococcus aureus.

Surra is caused by Trypanosoma evansi. The detection method using conventional parasitological tests has not always shown positive results in blood parasite detection, although the livestock has presented with clinical signs. Therefore, a fast and accurate diagnosis is necessary to prevent the disease predominately in field isolates. This study aimed to investigate the sensitivity of molecular detection method using two different specific primers, namely, Internal Transcribed Spacer 1 (ITS-1) and Trypanosoma brucei repeat 1/2 (TBR-1/2) against T. evansi field isolates from Banten Province, Indonesia. The isolates of T. evansi used in this study were collected from Banten Province and cultured and preserved by the National Research Center for Veterinary Science, Indonesia. Eighteen experimental rats were divided into three equal groups, which were categorized as control, 1 × 101, and 1 × 104 infective doses. The isolates were injected into all experimental albino rats intraperitoneally. All samples were tested using conventional blood smear, card agglutination test (CATT), and polymerase chain reaction (PCR) method. The results of the CATT examination in all treatments showed negative results. However, PCR results showed that two different primers, namely, ITS-1 and TBR-1/2 had been successfully detected T. evansi from infected experimental rats, proven by positive PCR band appeared in 480 base pairs (bp) and 164 bp, respectively. Based on the molecular diagnostic test using PCR method, TBR-1/2 primer is more sensitive to detect T. evansi compared to ITS-1 primer. The present finding provides preliminary data for studying the efficiency of different primers if practically applied as a standard diagnostic test for trypanosomiasis, especially in Indonesian livestock. Keywords: infectious disease, ITS-1, surra, TBR-1/2, tropical disease.

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii. Cats are known to be the definitive hosts that can excrete these environmentally resistant oocysts. Other mammals, avians, and even humans can serve as the intermediate host. T. gondii infection is often asymptomatic in healthy individuals; however, it could result in serious health problems in immunocompromised and pregnant individuals. This study investigated the occurrence of T. gondii infection in cats in Khao Suan Kwang and Mueang Khon Kaen. In total, 100 serum samples from cats, that is, 62 owned cats (31 males and 31 females) and 38 adopted stray cats (21 males and 17 females), were examined for antibodies against T. gondii through rapid immunochromatographic tests (ICT). Owners were asked to sign a consent form and answer the questionnaires before sample collection. Demographic information about the cats and their owners was also recorded. The overall seroprevalence of cats positive for T. gondii antibodies was found to be 5%. Notably, the Toxoplasma antibody prevalence was significantly higher in the adopted stray cats (10.53% [4/38]) that roamed the zoo than in the owned cats (1.61% [1/62]) (p > 0.05). No significant difference was observed between male (8.33%) and female (1.92%) cats. The cat owners' questionnaire revealed that more than half had never heard of toxoplasmosis before (67.7%), whereas 30.6% knew nothing about the disease transmission routes. This study presented a low seroprevalence of antibodies to T. gondii in owned cats from the Mueang Khon Kaen District, whereas high seroprevalence was detected in the adopted stray cats from Khao Suan Kwang. Adopted stray cats can have a higher potential for T. gondii infection; thus, they could be a source of toxoplasmosis transmission to humans. Therefore, it is essential to control the number of stray cats, and a screening test for antitoxoplasmosis could be recommended before adoption. Although the total seroprevalence was noted to be low, the zoonotic disease was present. Therefore, raising the community's awareness and knowledge might reduce the disease transmission from animals to humans. Keywords: adopted stray cat, feline, Khon Kaen, toxoplasmosis, zoonoses.

Cardiac time intervals (CTIs) can provide important information on the electrical and mechanical properties of the heart. We hypothesized that cardiac function can be described using the combined power of electrocardiography (ECG) and phonocardiography (PCG) signals. This study aimed to (1) validate a novel custom device in measuring CTI parameters; (2) compare CTI parameters with a commercially available device and standard transthoracic echocardiography (STE); and (3) compare calculated systolic performance index (SPI) and myocardial performance index (MPI) with Tei index from the STE. This study determined CTIs based on simultaneous ECG and PCG recordings in 14 healthy Beagle dogs using the custom-built device. These CTI parameters were compared with a commercially available device (Eko DUO ECG + Digital Stethoscope; Eko DUO) and the STE. Agreement of CTI parameters between the custom device and the commercially available device or STE was evaluated. Calculated SPI and MPI based on Wigger's diagram were proposed, compared with SPI and Tei index, and correlated with STE parameters. We found that the ECG and PCG parameters measured from the custom-built device did not differ from the commercially available device and the STE. By combining ECG and PCG signals, we established CTI parameters in healthy dogs including indices for systolic function (SPI: QS1/S1S2) and global cardiac function {F1 ([QS1+S2]/S1S2), F2 ([RS1+S2]/S1S2), and F3 (RS1 + [QS2-QT]/S1S2)}. The SPI, F2, and F3 were comparable with echocardiographic parameters describing systolic (Pre-ejection period/left ventricular ejection time [LVET]) and Tei index ([MCOdur-LVET]/ LVET), respectively. Only SPI and F3 were correlated significantly with MCOdur and heart rate, respectively. We have validated the use of the custom-built device to describe CTIs that are comparable to the commercially available device and STE in healthy Beagles. The proposed SPI and MPI derived from CTI parameters can be useful in clinical practice to describe the cardiac function, especially in areas where access to STE is constrained. Keywords: cardiac time intervals, dogs, electrocardiography, healthy, phonocardiography.

Since the past decade, metagenomics has been used to evaluate sequenced deoxyribonucleic acid of all microorganisms in several types of research. Nitrite contamination originates from the natural environment in Swiftlet farmhouses (SFHs) and can influence nitrite levels in edible bird's nest (EBN). It is strongly speculated that the conversion process into nitrite is influenced by the bacteria present in SFHs. Nitrite can cause adverse effects on human health. The previous research has focused on the characteristics of bacteria that may influence the nitrite conversion process in SFHs. This study aimed to a metagenomics analysis of bacteria present in the dirt of SFHs and evaluated nitrite levels in EBN on Sumatera Island. In total, 18 SFHs on Sumatera Island were selected, and EBN and dirt samples were collected from each SFH, resulting in 18 EBN and 18 dirt SFH samples. Raw uncleaned white EBN and dirt from three areas of SFH were collected. The samples were analyzed for nitrite levels using a spectrophotometer, and the metagenomics sequencing of SFH dirt samples was performed using the MinIon nanopore method. The sequenced data were analyzed using the EPI2ME software. Of the 18 raw uncleaned white EBN samples, 9 (50%) had <30 ppm nitrite levels. The top five bacterial genera in SFH dirt samples in Group A (nitrite levels >30 ppm) were Aeromonas, Escherichia, Acinetobacter, Arcobacter, and Acetoanaerobium. Those in Group B (nitrite levels <30 ppm) were Aeromonas, Pseudomonas, Shewanella, Escherichia, and Acinetobacter. There were 12 genera of nitrifying bacteria in Group A and 8 in Group B. The total cumulative read of nitrifying bacteria in Groups A and B were 87 and 38 reads, respectively. This is the first study to show that characteristic bacteria present in the dirt of SFHs might significantly influence the conversion from nitrogen to nitrite. Approximately 50% of raw uncleaned EBN samples had <30 ppm nitrite levels. Aeromonas was the most dominant bacterial genus found in Groups A and B. The variations in genus and cumulative reads nitrifying bacteria in group A were greater than those in Group B. This study provides information on the characteristics of bacteria that may influence the nitrite conversion process in SFHs. Metagenomics data were obtained from the reading using the software EPI2ME. Further research is needed on the bacterial target species that can convert nitrite in SFHs. Keywords: analysis, edible bird's nest, metagenomic, nitrite, Swiftlet.

Canine pyometra, either the closed (closed pyometra [CP]) or open (open pyometra [OP]) cervix type, is a frequent uterine disease in intact old age bitches. Therefore, early diagnosis and appropriate medical and surgical treatments are crucial to avoid the life-threatening condition in these bitches. This study aimed to investigate clinical alterations, blood parameters, causative bacteria, antimicrobial susceptibility, and uterine histopathology obtained during aseptic surgical treatment on bitches with pyometra. Sixty bitches of various breeds and ages with presumptive pyometra diagnoses were included in the study. The diagnoses were based on history, clinical examination, blood parameters, radiography, and ultrasonography. All pyometra bitches were ovariohysterectomized as an emergency surgical treatment. In addition, uterine content and tissues were submitted for bacterial isolation, antimicrobial susceptibility, and uterine histopathological analysis. Except for abdominal CP distention, no specific clinical signs were linked to the pyometra type. The mean values of total white blood cell count (WBC) and plasma protein were predominantly raised in pyometra bitches regarding hematological parameters. Leukocytosis was found in both types; however, the WBC in CP was markedly higher than in OP. The mean value of blood urea nitrogen increased in the CP group. Klebsiella pneumoniae and Escherichia coli were the most frequent causative bacteria isolated in CP and OP, respectively. All isolated bacteria were 100% susceptible to imipenem, meropenem, and carbapenem. Marbofloxacin was the second most effective drug against isolated bacteria from both groups. Uncomplicated cystic endometrial hyperplasia (CEH) was not presented in the CP group. CEH and chronic endometritis (type IV), the most severe uterine histopathological changes, were discovered in the CP and OP. The CP and OP groups presented leukocytosis, increased plasma protein, and CEH and chronic endometritis. Depression, abdominal distention, and enlarged uterine size were the major characteristics of the CP group. Furthermore, abdominal distension is presented in other abnormalities in clinical practices, providing a differential diagnosis. Drugs in the carbapenem group were the most effective against isolated bacteria; however, they are not routinely used due to bacterial resistance concerns. Thus, marbofloxacin was recommended as an alternative medical treatment because it is convenient to manage by both oral and injection routes. Keywords: antimicrobial susceptibility, bacteria, canine, histopathology, pyometra.

African swine fever (ASF) is an infectious disease and a major viral pig disease that threatens pork production in several locations globally. The mortality rate of ASF in domestic pigs is very high, causing a decrease in pig populations and significant economic losses for farmers. Environmental or ecological risk factors are the most important associated with the spread of the ASF virus. Environmental (or ecological) niche models are commonly used to estimate the probability of an event using the maximum entropy (Maxent) method. This study aimed to estimate the probability risk of future ASF outbreaks in North Sumatra, Indonesia. Secondary data from the National Animal Health System Database (iSIKHNAS), including data on the ASF outbreaks of 2019–2020 in North Sumatra, Indonesia, were used in this study. The first analysis performed involved the identification of environmental risk factors using multiple regression analysis. The second analysis performed was the estimation of probability risk for future ASF outbreaks in North Sumatra, Indonesia, using the Maxent method. Data processing was performed using Microsoft Excel, ArcGIS version 10.5 software (ESRI, California, United States), Maxent version 3.4.4 software, and Rstudio (http://www.r-project.org/). The Maxent method was found to be highly accurate with a statistically significant area under the curve value of 0.860. The greatest contributing environmental factor identified by the model was the harbor, which contributed 57%. The range of high probability risk of future ASF outbreaks was found to be 0.723–0.84. The estimation of the highest probability risk of future ASF outbreaks in North Sumatra, Indonesia, was 0.723–0.84. The most contributing environmental factor identified using the Maxent method was harbors, at 57%. This methodology can be used to carry out subsequent ASF analyses and contribute to developing prevention and control strategies in this area. Keywords: African swine fever, environmental niche models, maximum entropy, probability risk.

The incorporation of herbs and species has been shown to enrich the food with antioxidants and bioactive antimicrobial compounds, thereby preserving the safety and productivity of broiler chicken production. This study aimed to determine the effects of three phytogenic feed additives (PHT) on certain zootechnical and hematobiochemical parameters in broiler chickens. Coriandrum sativum L. (coriander), Pimpinella anisum L. (green anise), and Trigonella foenum-graecum L. (fenugreek) were used to formulate the PHT. A total of 360 1-day-old Cobb broilers for 42 days were randomly assigned to four dietary treatment groups: A control group (CTLG) and three groups fed a basal diet supplemented with 3% of coriander (PHT1G), 3% of a combination 50% coriander-50% fenugreek (PHT2G), and finally, 3% of a combination 50% coriander-50% green anise (PHT3G), respectively, and each experimental group included three repetitions of 30 birds. Zootechnical parameters, carcass productivity, and hematobiochemical properties were measured. The birds in the PHT3G had the greatest body weight and organ weight (p < 0.05). However, the weight of abdominal fat remained unchanged. The same group of broilers had a significantly (p < 0.05) higher lymphocyte level of 120.103/μL, followed by the PHT2G, which had 80.103/μL. The levels of monocytes in the PHT2G and PHT3G were 66.103/μL and 60.103/μL, respectively. Regarding granulocytes, we observed 200.103/μL in the PHT2 group and 102.103/μL in the PHT3G. There was a statistically significant difference (p < 0.05) between the uric acid levels of the PHT1G, PHT2G, and PHT3G, with 50.4 mg/L, 59.84 mg/L, and 47.29 mg/L, respectively. All experimental groups had significantly lower uric acid concentrations than the control group (84.36 mg/L). The use of phytogenic feed additives may positively affect both weight gain and hematobiochemical parameters in broiler chicken, particularly the levels of various white blood cell subtypes and the uric acid rate. Keywords: broilers, blood parameters, natural feed additive, performances, spices seed.

Endogenous retroviruses (ERVs) found in all vertebrates, including non-human primates (NHPs), are known to be genetically inherited. Thus, recent studies have explored ERVs for human immunodeficiency virus vaccine development using human ERV (HERV) due to the hypervariability of exogenous retroviruses which cause conventional vaccines to be ineffective. HERV was also found to be able to induce an immune response in cancer patients. This study aimed to identify and molecularly characterize ERVs from Indonesian NHPs: Macaca fascicularis and Macaca nemestrina. Then, we described the phylogenetic relationship of these isolates with those of the simian ERVs (SERVs) characterized in other species and countries. First, 5 mL of whole blood samples was taken from 131 long-tailed macaques and 58 pig-tailed macaques in captive breeding facilities at Bogor, Indonesia, for DNA extraction. Next, the DNA samples were screened using the SYBR Green real-time polymerase chain reaction (PCR) technique with specific primers for env (simian retroviruses [SRV]1-5 7585U19 and SRV1-5 7695L21). Positive SERV results were those with cycle threshold (CT) values < 24 (CT < 24) and melting temperature (TM) ranges of 80°C–82°C. Then, whole-genome nucleotide sequences from two pig-tailed macaques samples detected as positive SERV were generated using a nucleic acid sequencing technique which utilized the walking primer method. Subsequently, the sequences were analyzed using bioinformatics programs, such as 4Peaks, Clustal Omega, and BLAST (NCBI). Subsequently, a phylogenetic tree was constructed using the neighbor-joining method in MEGA X. SYBR Green real-time PCR amplification results indicated that SERV (Mn B1 and Mn B140910)-positive samples had CT values of 22.37–22.54 and TM of 82°C. Moreover, whole-genome sequences resulted in 7991 nucleotide sequences, comprising long terminal repeat, gag, pro, pol, and env genes identical between the sequenced samples. Furthermore, the phylogenetic tree results indicated that both samples from M. nemestrina had 99%–100% nucleotide identities to the Mn 92227 sample identified at the National Primate Center University of Washington (NaPRC UW) which was imported from Indonesia in 1998, confirmed as a novel SERV strain. The phylogenetic tree results also indicated that although SERV whole-genome nucleotide and env amino acid sequences were clustered with SRV-2 (identity values of 82% and 79%, respectively), they had a 99%–100% nucleotide identity to Mn 92227. Meanwhile, the gag, pro, and pol amino acids were clustered with SRV-1, SRV-3, SRV-4, SRV-5, SRV-8, and SERV/1997, with 82% and 88% identity values. Based on the SYBR Green real-time PCR profiles generated, similarities with Mn 92227 were observed. Subsequent phylogenetic analysis confirmed that both samples (Mn B1 and Mn B140919) from pig-tailed macaques in the country of origin were novel SERV strains at NaPRC UW. Therefore, it could be used in biomedical research on ERVs. Keywords: Macaca fascicularis, Macaca nemestrina, retrovirus, simian endogenous retroviruses.

Ribonucleic acid viruses remain latent in different cell types, including mesenchymal stem cells; however, the distemper virus remains undetected in these cells. This study aimed to determine whether adipose stem cells (ASCs) from dogs with distemper disease are infected with the canine morbillivirus (CM). Twelve dogs with the neurological phase of the disease and who were positive for CM by reverse transcription polymerase chain reaction (RT-PCR), were studied. ASCs from adipose tissue of the lesser omentum of these infected dogs were isolated and characterized. Direct fluorescence was used to detect the viral antigen in cell cultures. Flow cytometry and RT-PCR identified detectable quantities of the virus in two cultures, while electron microscopy confirmed the CM particles within ASCs. This study revealed that ASCs of the omentum of dogs with distemper disease can be infected with CM, indicating their possible involvement in this virus latency and persistence. This suggests that its detection should be considered within the quality control process of stem cells intended for regenerative medicine. To the best of our knowledge, this is the first study that demonstrates that omentum ASCs from dogs with distemper disease can be infected with CM and may be involved in viral latency or persistence. Our study also suggests that the detection of CM should be considered within the quality control process of stem cells intended for regenerative medicine. Keywords: adipose stem cells, canine distemper disease, canine morbillivirus, cell therapy, viral latency.

The Indian and global poultry industries suffer significant economic losses due to Mycoplasma gallisepticum (MG) and Mycoplasma synoviae (MS) infections, which adversely affect egg production, hatchability, weight gain, and feed efficiency in farms, thus decreasing the overall production efficiency. This study aimed to determine the percent positivity and phylogenetic analysis of MG, MS, and co-infection of both mycoplasmas in commercial poultry farms across different states of India from 2017 to 2021. A total of 3620 tracheal or choacal swabs were collected from breeder and layer farms showing clinical signs of avian mycoplasma infections from commercial poultry farms across India, and the percent positivity for MG, MS, and co-infection of both mycoplasmas were determined by Polymerase chain reaction using the 16S rRNA and vlhA genes amplification, respectively. Phylogenetic analysis was carried out by sequencing the mgc2 and vlhA genes of 2 samples of MG and 24 samples of M. synoviae to gain insight into the genetic variability of Indian strains. The data were then compared with other Indian strains, vaccines strains, and strains from other countries. Our data shows the percent positivity of MG, MS, and co-infection of both MG and MS was 6.43%, 23.61%, and 15.49%, respectively. The phylogenetic relationship between MG and MS was determined using the vlhA and mgc2 genes, revealing two samples of MG and 24 samples of MS clustered with other Indian strains. M. synoviae MSM22 and previously studied M. synoviae MGS 482 clustered with vaccine strain M. synoviae MS-H. Mycoplasma synoviae infections in breeder, layer, and in both is predominant compared to MG across the states investigated in India. Sequenced samples of MG and MS showed evolutionary relationships with the previously studied Indian strains of MG and MS. These findings support our view that monitoring chickens for avian mycoplasma infections are of paramount significance. It further lends credence to the contention that such information will pave the way for the development of a home-grown vaccination control program and thus safeguard the poultry sector against mycoplasma infections. Keywords: epidemiology, India, Mycoplasma gallisepticum, Mycoplasma synoviae, phylogeny, polymerase chain reaction.

Cytology investigations are a frequent part of ophthalmological examination. We aimed to assess whether the cytological findings of healthy and conjunctivitis/keratoconjunctivitis samples differed based on the evaluator's experience. A study evaluated healthy eyes (n = 40) and eyes affected with keratoconjunctivitis and/ or conjunctivitis (n = 28) in dogs. An ophthalmological examination was performed before sampling the eyes using a sterile cotton swab. An evaluator with theoretical experience and one with undergone clinical pathology residency training performed cytology blinded to the clinical findings. In the healthy eyes group, the agreement between the evaluators for cellularity was nonexistent, while that for cell preservation and mucus content was fair. In the affected eyes group, the agreement for cellularity and mucus content was moderate, while that for cell preservation was fair. The inadequate sample rate differed significantly between the two evaluators in the healthy eyes group (p = 0.006) but not in the affected eyes group (p = 0.083). Bacterial presence was detected by both evaluators, and the findings did not differ statistically from the bacteriology results (p = 0.05). Significant variations were noted in the differential cell count; the mean count of the superficial epithelial cells and goblet cells of the healthy eyes group (p < 0.05) and that of the basal/intermediate cells and neutrophils of the affected eyes (p < 0.05) showed significant differences. The evaluator's experience significantly affected the differential cell count in both the healthy and affected eyes groups. Neutrophil degeneration was not observed by the less experienced evaluator, whereas bacteria were detected equally well by both the evaluators. Keywords: conjunctival cytology, microscopy experience, neutrophil morphology.

Pestivirus, a genus of the Flaviviridae family, comprises viruses that affect bovines, sheep, and pigs. Symptoms, including hemorrhagic syndromes, abortion, respiratory complications, and deadly mucosal diseases, are produced in infected animals, which cause huge economic losses to the farmers. Bovine viral diarrhea virus-1, bovine viral diarrhea virus-2, classical swine fever virus, border disease virus, Bungowannah, Hobi-like, and atypical porcine pestivirus belonging to the Pestivirus genus were selected for the study. This study aimed to estimate the codon usage bias and the rate of evolution using the glycoprotein E2 gene. Furthermore, codon usage bias analysis was performed using publicly available nucleotide sequences of the E2 gene of all seven Pestiviruses. These nucleotide sequences might elucidate the disease epidemiology and facilitate the development of designing better vaccines. Coding sequences of the E2 gene of Pestiviruses A (n = 89), B (n = 60), C (n = 75), D (n = 10), F (n = 07), H (n = 52), and K (n = 85) were included in this study. They were analyzed using different methods to estimate the codon usage bias and evolution. In addition, the maximum likelihood and Bayesian methodologies were employed to analyze a molecular dataset of seven Pestiviruses using a complete E2 gene region. The combined analysis of codon usage bias and evolutionary rate analysis revealed that the Pestiviruses A, B, C, D, F, H, and K have a codon usage bias in which mutation and natural selection have played vital roles. Furthermore, while the effective number of codons values revealed a moderate bias, neutrality plots indicated the natural selection in A, B, F, and H Pestiviruses and mutational pressure in C, D, and K Pestiviruses. The correspondence analysis revealed that axis-1 significantly contributes to the synonymous codon usage pattern. In this study, the evolutionary rate of Pestiviruses B, H, and K was very high. The most recent common ancestors of all Pestivirus lineages are 1997, 1975, 1946, 1990, 2004, 1990, and 1990 for Pestiviruses A, B, C, D, F, H, and K, respectively. This study confirms that both mutational pressure and natural selection have played a significant role in codon usage bias and evolutionary studies. This study provides insight into the codon usage bias and evolutionary lineages of pestiviruses. It is arguably the first report of such kind. The information provided by the study can be further used to elucidate the respective host adaptation strategies of the viruses. In turn, this information helps study the epidemiology and control methods of pestiviruses. Keywords: codon usage bias, evolutionary analysis, Flaviviridae, glycoprotein E2, India, Pestivirus.

Osteoarthritis (OA) is recognized as a degenerative joint disease that leads to chronic pain and low quality of life in animals. Captive elephants, the largest land mammals with a long lifespan, are more prone to develop OA due to restricted spaces and insufficient physical activity. This study aimed to investigate the effect of transforming growth factor-β1 (TGF-β1) and insulin-like growth factor 1 (IGF-1) on elephant chondrogenesis in a scaffold culture of articular chondrocytes. Elephant chondrocytes-seeded gelatin scaffolds were cultured in chondrogenic media with or without 10 ng/mL of TGF-β1 or IGF-1 alone or 5–10 ng/mL of their combination for up to 21 days. The mRNA expression of cartilage-specific anabolic genes, ACAN and COL2A1, was analyzed using a real-time reverse transcription-polymerase chain reaction. The amounts of sulfated glycosaminoglycans (sGAGs) in conditioned media and contents in cultured scaffolds were determined through dimethylmethylene blue assay. Cell morphology, accumulation of proteoglycans, and details of the cultured scaffolds were determined using hematoxylin-eosin staining, safranin O staining, and scanning electron microscopy (SEM), respectively. TGF-β1 alone significantly upregulated ACAN gene expression but not COL2A1, while IGF-1 alone did not enhance both ACAN and COL2A1 genes. The combination significantly upregulated both mRNA expression levels of ACAN and COL2A1 gene at day 14. The sGAGs accumulation and contents in the treatment groups, except IGF-1 tended to be higher than the controls, concomitantly with the production of the extracellular matrix, showed the formation of a cartilage-like tissue through histological and SEM analyses. Together, our results suggest that the single treatment of TGF-β1 has a selective effect on ACAN gene, while the combined growth factors seem to be an advantage on elephant chondrogenesis. This three-dimensional culture model is probably helpful for developing cartilage regeneration in vitro and is further applied in tissue engineering for OA treatment in vivo. Keywords: articular chondrocyte, chondrogenesis, elephant, insulin-like growth factor 1, scaffold, transforming growth factor beta 1.

Upper respiratory tract disease (URTD) is prevalent in cats, and diagnosis can be challenging. This study aimed to determine the most common causes of cat URTD in Latvia and describe computed tomography (CT) and laboratory diagnostic findings. The present retrospective study included a total of 94 cats who were diagnosed with URTD. All cats underwent CT, and 50 of them had additional diagnostic tests, such as histology and respiratory infection polymerase chain reaction (PCR) testing. The most common CT finding was rhinosinusitis (55.32%) followed by nasal neoplasia (26.6%) and nasopharyngeal polyp (14.89%), but in three cats, a cause of respiratory symptoms was larynx neoplasia, nasal dermoid cyst, and an oronasal fistula. PCR test showed that the most cause of rhinosinusitis was Mycoplasma felis. Nasopharyngeal polyp as the primary diagnosis was identified in 14 cats from 3 months to 6 years, with an average age of 1.85 ± 1.915 years, and 54% of cats were female. Nasal neoplasia as a primary CT diagnosis was determined in 25 cats at the age of 5–18 years, with an average age of 10.56 ± 3.416 years. Histology diagnosis included four types of neoplasia – squamous cell carcinoma, sarcoma, adenocarcinoma, and aplastic carcinoma. This study describes the most common CT and laboratory findings in cats with URTD. Included information will be helpful for general veterinary practitioners and researchers and will update their knowledge on feline URTD. Keywords: computed tomography, feline, nasal neoplasia, Mycoplasma felis, rhinitis.

Bacteria of the genera Vibrio and Aeromonas cause seafood-borne zoonoses, which may have a significant impact on food safety, economy, and public health worldwide. The presence of drug-resistant and biofilm-forming phenotypes in the food chain increases the risk for consumers. This study aimed to investigate the characteristics, virulence, biofilm production, and dissemination of antimicrobial-resistant pathogens isolated from seafood markets in Bangkok, Thailand. A total of 120 retail seafood samples were collected from 10 local markets in Bangkok and peripheral areas. All samples were cultured and the Vibrio and Aeromonas genera were isolated using selective agar and biochemical tests based on standard protocols (ISO 21872-1: 2017). The antibiotic susceptibility test was conducted using the disk diffusion method. The presence of hemolysis and protease production was also investigated. Polymerase chain reaction (PCR) was used to determine the presence of the hlyA gene. Furthermore, biofilm formation was characterized by microtiter plate assay and scanning electron microscopy. The bacterial identification test revealed that 35/57 (61.4%) belonged to the Vibrio genus and 22/57 (38.6%) to the Aeromonas genus. The Kirby–Bauer test demonstrated that 61.4% of the isolates were resistant to at least one antibiotic and 45.61% had a high multiple antibiotic resistance index (≥0.2). PCR analysis indicated that 75.44% of the bacteria harbored the hlyA gene. Among them, 63.16% exhibited the hemolysis phenotype and 8.77% showed protease activity. The biofilm formation assay demonstrated that approximately 56.14% of all the isolates had the potential to produce biofilms. The moderate biofilm production was the predominant phenotype. The results of this study provide evidence of the multiple drug resistance phenotype and biofilm formation capacity of Vibrio and Aeromonas species contaminating raw seafood. Effective control measures and active surveillance of foodborne zoonoses are crucial for food safety and to decrease the occurrence of diseases associated with seafood consumption. Keywords: Aeromonas spp., biofilm formation, drug resistance, foodborne, seafood, vibriosis.