The phytogenic cocktail (PC) is a unique combination of natural plant extracts consisting of coconut shell smoke, clove leaf extract, and mangosteen rind extract, predominantly containing phenol, eugenol, and α-mangostin. Chicken performance can be improved by its antibacterial properties. This study aimed to test PC as a replacement for antibiotic growth promoters (AGPs), assessing its impact on performance, intestinal microbes, and carcass traits in slow growth KUB chickens.  Two hundred and forty KUB chicks were distributed randomly to five dietary groups. Each group constituted six replicates, one replicate contained eight chicks. The treatments included the control diet (CD) with no additives, CD with 50 ppm Zinc bacitracin as an additive (AGPs), CD paired with 198 mL PC/ton feed provided for the initial 12 weeks (PC1), CD with 198 mL PC/ton feed given for the first 4 weeks (PC2), and CD supplied with 198 mL PC/ton feed for the first 8 weeks (PC3). Performance and mortality indicators were assessed during the feeding stage up to 12 weeks of age, while intestinal total microbial count and carcass characteristics were determined at 12 weeks. Duncan’s multiple-range test identified differences among the treatments in the randomized experiment.  The AGPs group weighed significantly more (p < 0.05) than PC2 but not significantly different (p > 0.05) from Control, PC1, and PC3 at 4 weeks. At 8 weeks, there was no significant difference (p > 0.05) in the body weight (BW) between the AGP, CD, and PC groups. The AGPs group had a significantly greater BW than PC1 and PC2 at 12 weeks (p < 0.05), but was comparable to CD and PC3 (p > 0.05). During the starter phase (0–4 weeks), dietary addition of AGPs or PCs significantly reduced feed intake (p < 0.05); however, no significant effect (p > 0.05) was observed during the later feeding periods (0–8 or 0–12 weeks). During the starter period, PC3 yielded the best feed conversion ratio, slightly surpassing AGPs and significantly (p < 0.05) outperforming CD. No significant variations (p > 0.05) were detected in the carcasses among the treatments. The reduction of abdominal fat relative weight was significant (p < 0.05) during the first 8 weeks of PC feeding. After the 12-week trial, no significant difference (p > 0.05) was observed in the proportionate weights of the crop, proventriculus, gizzard, pancreas, cecum, spleen, bursa of Fabricius, heart, and liver. The reduction in the intestinal microbe population due to AGPs or PC was not statistically significant (p > 0.05). About 100% viability was confirmed by the absence of mortality throughout the study.  PC supplementation in KUB chicken feed enhances their performance. The optimal feeding regimes were effective during the first 8 weeks of age. In the 0–4 week time frame, feeding the PC to the chicken worsened performance whereas no improvement was observed in the 0–12 week period. The application enhanced weight loss, feed efficiency, and reduced abdominal fat. Based on the research findings, the PC can replace AGPs as a feed additive due to comparable or superior improvement results. 

Dogs can detect specific cancer odors with their exceptional sense of smell. This study aimed to conduct a scientometric analysis of canine olfactory detection in oncology, identifying trends, visualizations, and patterns.  A retrospective observational study was conducted using a quantitative-scientometric approach. Scopus was comprehensively searched using terms related to canine olfactory detection in oncology. Documents indexed in Scival software (Elsevier) and published between 2013 and 2022 were included.  Claire M. Guest, Rob Harris, and Giuseppe Lippi authored significant academic work. Journals such as Journal of Breath Research and PLoS One rank highly in publications and citations due to significant citation ratios, according to CiteScore’s quartile-based impact analysis. According to Lotka’s and Bradford’s laws, a small group of authors and the Journal of Breath Research, respectively, dominate production in their fields.  This analysis forms a solid base for future research on canine olfactory detection in oncology. The collaborative essence of this multidisciplinary field is emphasized by the key contributors and identified patterns, with journals in the Q1 and Q2 quartiles of CiteScore holding significant importance. 

Small ruminants require vaccines to prevent and manage diseases. Unfortunately, no studies have been conducted in Bangladesh to assess the knowledge, attitudes, and practices (KAP) of small ruminant farmers (SRF) regarding vaccine use against infectious diseases, affecting the success of vaccination campaigns. The present study aims to assess SRF’s KAP regarding vaccines, revealing gaps and barriers to efficient vaccination.  Two hundred and twenty-eight SRF in northern Bangladesh were surveyed in a cross-sectional study. Data were collected from random participants through face-to-face interviews using a structured questionnaire. KAP levels were categorized as “good” or “poor” and “positive” or “negative” using a scoring method with a 60% cutoff. The analysis comprised the utilization of descriptive statistics as well as logistic regression models.  Results showed that most participants were female (60.5%), aged 31–40 (34.2%), with secondary education (28.1%), and vaccination training (22.8%). While 75% knew about vaccines, only 37.3% understood their role in preventing infectious diseases, and 63.6% in reducing antibiotic use 68.4% of farmers were aware of negative drawbacks, and 61.8% reported vaccinating their herds. About 42.1% of the farmers had good knowledge, 52.6% had a positive attitude, and 22.8% followed good practices. Female farmers with graduate degrees and 6–10 years of goat farming experience, but not those with vaccination training, demonstrated stronger knowledge. Female farmers with a graduate degree and 6–10 years of goat farming experience displayed positive attitudes. Female goat farmers from Thakurgaon had a higher likelihood of following good vaccination practices than those with vaccination training.  The study unearths disparities in KAP scores among farmers. To effectively address KAP gaps concerning vaccine usage and prevent potential infectious diseases, it is essential to design focused educational and training programs. About 52.6% of SRF hold a positive view toward vaccines. 

Although widely employed in traditional remedies globally, the safety and efficacy of Moringa oleifera remain inadequately documented through scientific research. This study evaluated the oral toxicity of M. oleifera leaf aqueous extract (MoAE) and its impact on gout-induced rats.  2000 mg/kg was given in a single dose during the acute oral toxicity test, while 100 mg/kg, 250 mg/kg, and 500 mg/kg were given daily for 28 days in the repeated dose toxicity test. 100 mg/kg, 250 mg/kg, and 500 mg/kg MoAE doses were administered during the assessment of its impact on gout caused by monosodium urate. In the hyperuricemia model induced by oxonic acid, serum uric acid levels were assessed and pain response was measured through acetic acid-induced writhing.  In acute oral and 28-day repeated dose tests, no indications of toxicity were detected, while MoAE alleviated ankle joint swelling and reduced serum uric acid concentrations in arthritic rats, causing a significant reduction in acetic acid-induced contortions.  No acute oral toxicity or toxicity in 28-day repeated doses was found for MoAE, while it exhibited antiarthritic, antihyperuricemic, and pain-relieving effects in the murine model. 

A piglet’s pre-weaning performance significantly influences both animal welfare and profitability in pig production. Understanding piglet pre-weaning performance influencing factors is key to enhancing animal welfare, reducing losses, and boosting profitability. The study aimed to evaluate the impact of parity, season of birth, and sex on within-litter variation and pre-weaning performance of F1 Large White × Landrace pigs.  Information regarding total litter size, number of born alive, number of stillbirths, piglet weight at birth, mortality, and count of weaned F1 Large White × Landrace piglets was acquired from the farm database (April 2022–February 2023). 2602 females and 2882 males, a total of 5484 piglets were utilized, with records from 360 sows. The coefficient of variation (CV) of birth weights among piglets within a litter was calculated. The general linear model analysis in MiniTab 17 was used to evaluate the data, with Fisher’s least significant difference test (p < 0.05) used for mean separation and Pearson’s moment correlation coefficient calculated to assess relationships between survival rates, mortality rates, litter size, birth weight, and birth weight CV.  Parity had a statistically significant impact on litter size, birth weight, and survival rate (p < 0.05). The sow’s parity did not significantly (p > 0.05) impact the number of piglets born alive or weaned. Multiparous sows had a significantly larger litter size (p < 0.05) than primiparous sows at birth. The litter weights for parities 2, 4, and 5 did not significantly differ (p > 0.05), with averages of 20.95, 20.74, and 20.03 kg, respectively. About 91.29% was the highest survival rate recorded in parity 2 (p < 0.05). The 1st week of life recorded an 8.02% mortality rate. The mortality rate in parity 3–5 group was significantly (p < 0.05) higher (11.90%) in week 1 than in the other groups (parity 1: 6.79%, parity 2: 5.74%, parity 3–5: 8.54 and 9.21%). The litter sizes in autumn (17.34) and spring (17.72) were significantly larger (p < 0.05) than those in summer (16.47) and winter (16.83). In autumn and spring, the survival rate (83.15 and 85.84%, respectively) was significantly lower (p < 0.05) compared to summer (88.40%) and winter (89.07%). In all seasons, the litter weights did not significantly differ (p > 0.05). The birth weight CV was significantly (p < 0.05) lower during summer (20.11%) than during spring (22.43%), autumn (23.71%), and winter (21.69%). The season of birth had no significant effect (p > 0.05) on the number of live piglets. Males (1.34 kg) were heavier (p < 0.05) than females (1.30 kg) at birth. Notably, the birth weight CV was similar between males (22.43%) and females (22.52%). Litter size was positively correlated with average litter weight (rp = 0.576, p < 0.001), birth weight CV (rp = 0.244, p < 0.001), and mortality rate (rp = 0.378, p < 0.001). An insignificant relationship was observed between average litter weight and birth weight CV (rp = –0.028, p > 0.05) and survival rate (rp = –0.032, p > 0.05).  In F1 Large White × Landrace pigs, birth uniformity among piglets declines as litter size grows larger. In parity 3–5, multiparous sows yield litters with reduced uniformity. With an increase in litter size, uniformity among piglets at birth worsens. A larger litter size and greater piglet birth weight variation are linked to a higher pre-weaning mortality rate. Producers need a balanced selection approach to boost litter size and must cull aging sows carefully to introduce younger, more productive females. 

The colostrum is essential for a kid’s survival and development. The metabolomic profiles of mammary secretion in goats are limited. This study investigated the metabolomic profiles of mammary secretion in purebred Black Bengal goats and their relationships with litter size and parity number.  500 MHz nuclear magnetic resonance was used to analyze the metabolomic profiles of 43 colostrum and milk samples collected on delivery day and day 7 after parturition, respectively.  Fifty-one metabolites were distinguished between colostrum and milk based on heatmap visualization and hierarchical cluster analysis. In colostrum, most compounds were present in significantly greater amounts than in milk. Milk of goats with multiple litter sizes had higher levels of lactose while fat, protein, total solids, solid not fat, and most of the metabolites were lower. The parity number of dams shows no difference in the composition of all components between primiparous and multiparous goats.  The components in colostrum were significantly more concentrated than those in milk. The multiple litter sizes in dams led to a significant impact on the composition of lactose and other milk metabolites.

The feeding and sanitary conditions significantly influence the productivity of farm animals. This study aimed to assess the impact of a lactic acid-producing microbial additive on broiler chicken productivity.  A 42-day experimental period utilized 120 1-day-old Cobb 500 chicks with an average weight of 46 g. In groups of 30 each, the chicks were randomly assigned to four experimental designs. The following treatments were assessed: T1 without intervention (control), T2 with bacitracin at a concentration of 0.5 g/L, T3 with a 5% probiotic mixture (PM), and T4 with a 7.5% PM. The birds were fed the commercial balanced feed without anticoccidials daily, while vaccines were administered according to the recommended biosecurity plan by the commercial house. Drinking water was treated with PM containing lactobacilli, yeasts, and short-chain organic acids.  In T4, a 7.5% PM resulted in a final weight of 2361.2 g (p < 0.05), a total weight gain of 1412.8 g (p < 0.05), and improved feed efficiency with a feed conversion of 2.00 (p < 0.05), during which feed intake was lower than in the other groups.  Microbial additives with lactic acid activity are a cost-effective and feasible solution for broiler chicken productivity. 

There is a limited amount of research conducted on quail breeding domestically and internationally, particularly at the molecular level. This study aimed to detect single-nucleotide polymorphisms in the growth hormone (GH) and insulin-like growth factor-1 (IGF-1) genes across two quail varieties and their hybrids correlate these genetic factors with body weight (BW) and growth rate at 0 and 6 weeks, and assess crossing effects.  White and Japanese quail were crossed. Simultaneously producing pure varieties and crosses (genotypes) was achieved through this breeding strategy. Fifty females from each genotype were randomly selected for blood sampling. Genomic DNA was extracted and amplified from the blood using the DNeasy blood kit (Qiagen, Germany). Nucleotide polymorphism between quail genotypes was determined through DNA sequencing.  Two types of alleles (A and B) for the GH gene in quails showed significant genotypic differences (AA, BB, and AB). The quail carried a mutated IGF-1 gene. For growth traits, substantial positive heterosis was detected.  The genotype AA had the highest BW and weight gain. The white variety can act as a sire, and both white and Japanese varieties can function as dams to improve growth traits. The growth characteristics of the hybrids surpassed those of the original varieties. 

Brain malaria, which results from Plasmodium falciparum infection, is responsible for substantial fatalities and health issues. These processes, including cytoadherence, rosetting, and sequestration, induce an immune response, hypoxia, brain microvascular obstruction, disruption of the blood-brain barrier, and cell death. Parasitemia level can reveal the presence of infection and its association with apoptosis-related genes. Neem (Azadirachta indica) leaves with antimalarial properties could replace ineffective Indonesian malaria medications. This study was designed to evaluate the impact of neem leaf extract on cerebral malaria-induced parasitemia and neuron cell apoptosis in mice through an in vivo approach.  13–16 weeks old C57BL mice received infection by Plasmodium berghei strain ANKA. Parasitemia was estimated daily from the mice’s tail blood. 8 mg, 12 mg, and 16 mg of a 96% ethanolic neem leaf extract were orally given for 6 days. Healthy, positive, and negative controls were included for treatment comparisons. On the 7th day, brain tissue was analyzed for (p > 0.05) gene expression. Through immunohistochemistry, both cell apoptosis in neurons expressing caspase-3 within a brain sample and the degree of parasitemia in a blood smear were assessed. The Pearson correlation test and one-way analysis of variance were employed to analyze the data.  Neem leaf extract reduces parasitemia and neuron cell apoptosis at multiple dosages (p < 0.000). Apoptosis in brain neurons and parasitemia show a strong positive correlation (r = +0.939). Neem leaf extract at doses of 12 and 16 mg was the most effective in reducing parasitemia levels and causing cell death.  Neem leaf therapy significantly reduced the degree of parasitemia and cell apoptosis in C57BL mice compared with the control group without treatment (p = 0.05). This shows that neem leaves have the potential to be a candidate drug for malaria. 

In tropical conditions, modeling the predictive parameters of live weight, including those at birth, pre-weaning, post-weaning, finishing, and maturing, and the average daily gain, is challenging. The heat load significantly influences the growth rate and final mature weights in the tropics. The study compared the growth rates of Kedah-Kelantan (KK), Brahman (BRAH), and Belgian Blue (BB) crossbred calves.  The study conducted growth analysis using the non-linear regression growth models as it approximates the sex, breed, and growth physiology changes in beef cattle. It is supported by the utility of the most common growth functions (Brody, Logistic, von Bertalanffy, and Richard’s model) in normal-muscled tropical breeds and double-muscled crossbred beef cattle in the tropics.  The BB crossbreds outperformed the KK and BRAH breeds by 50%–100% in live weight gains under tropical conditions. The crossbreds display the double-muscled effect and highlight the advantages of heterosis, making them suitable for upgrading local herds. The study’s findings on the growth characteristics of BB crossbred cattle were best described by the von Bertalanffy growth model, which had a high coefficient of determination (R2 > 0.8) and yielded estimated mature weights of 527.5 kg for males and 518.5 kg for females.  According to results, raising BB crossbreds in the tropics as a solution to ensure a sustainable beef supply could yield significant growth and economic benefits. 

In Central Vietnam, Anopheles dirus and Anopheles minimus are the primary malaria vector species. These Anopheles spp.’ distribution and prevalence are determined by environmental, climatic, and socioeconomic conditions. This study aimed to predict the potential distribution of these two Anopheles spp. in this region.  This study was conducted in 15 Central Vietnamese provinces. From 2014 to 2018, we utilized An. dirus and An. minimus presence records. Proxy data from the Google Earth Engine platform for the study area, encompassing environmental, climatic, and socioeconomic factors. MaxEnt software predicted the potential environmental, climatic, and socioeconomic suitability of these two Anopheles spp. in Central Vietnam.  The test area under the curve values for An. dirus and An. minimus MaxEnt models averaged 0.801 and 0.806, respectively, showing excellent performance. Minimum air temperature had the greatest impact on the distribution of both species. A negative correlation between precipitation and normalized difference water index influences the occurrence of An. dirus. In the temperature range of 13–19.5°C, An. minimus is most likely to be present, with nighttime light detrimentally influencing its distribution. The Central Highlands region is inhabited by both species, with some presence in North-Central and South-Central Coastal areas.  The importance of temperature in determining the presence of both species is emphasized by our findings, with subtle differences in the temperature-related factors shaping their distributions. The results highlight the need for focused malaria vector control and surveillance initiatives in the study area. 

Cryptosporidium spp. members of the phylum Apicomplexa are obligate protozoan parasites capable of infecting various vertebrate hosts, including rodents and chickens. Infection caused by these parasites may lead to zoonotic diseases in humans. The aim of this study was to estimate the prevalence of Cryptosporidium spp. in rodents and domestic chickens sampled in Franceville, Gabon.  Two hundred and eighty-five samples were collected, of which 185 samples were from rodents and 100 from domestic chickens. Microscopy after modified Ziehl-Neelsen staining and nested polymerase chain reaction targeting the small subunit (SSU) rRNA gene were used to examine Cryptosporidium spp.  The overall prevalence of Cryptosporidium oocysts was 55.8%, with a prevalence of 72.4% in rodents and 25.0% in domestic chickens. Molecular analysis showed that Cryptosporidium spp. were present in 4.0% of the samples. No significant correlation was observed between Cryptosporidium spp. carriage and sex or location in this study. These results indicate that Cryptosporidium spp. persist and circulate in the studied animal species in Franceville, Gabon.  Infection with Cryptosporidium is very common in rodents and chickens in Franceville. The potential risk of human contamination cannot be ruled out. More research should be conducted to characterize Cryptosporidium species circulating in rodents and chickens in Gabon. Such studies are essential to better understand the epidemiology of this protozoan and its potential impact on public health. 

No study has successfully isolated parvovirus in Vietnam. This study aimed to isolate and characterize parvovirus strains indigenous in Vietnam for vaccine development against porcine parvovirus (PPV).  We collected serum and stillbirth samples from six provinces in Vietnam, and PPV-positive samples were identified using a polymerase chain reaction. Parvovirus isolation was attempted using the PK-15 cells maintained in a minimum essential medium supplemented with 5% fetal bovine serum and 1% antibiotics (Penicillin-streptomycin). The cells were incubated at 37°C with 5% CO2. Virulence experiments were conducted on white primiparous sows to evaluate the virulence of the PPV strain through hemagglutination inhibition (HI) titers and fetus lesions.  We analyzed 360 serum and 32 stillbirth (liver and lungs) samples, revealing that 32/392 (8.2% ) of them were PPV-positive, all belonging to PPV1. Thirty-two PPV-positive samples were successfully isolated, with 100% identity as VP2 sequences. The phylogenetic tree revealed a close relationship with the Kresse strain (isolated from Canada in 1996) and the PPV1-0225-L-SD strain (isolated from China in 2022). Two PPV isolates (VC5 from Dongnai and TX7 from Thanhhoa) that exhibited high 50% tissue culture infectious dose titers were selected for the virulence experiment. On day 21, after injection, the HI antibody titers ranged from 10log2 to 12log2. On day 90, 71%–80% of fetuses were mummified.  This study showed that the PPV infection rate in Vietnam was 8.2%. Thirty-two isolates belonged to PPV1. Two PPV strains, VC5 and TX7, were determined to be highly virulent by the results of HI titers after injection into gilts. VC5 and TX7 were determined to be good candidates for further research on PPV vaccines. 

Mosquitoes carry numerous diseases of medical and veterinary significance. While citronella essential oil is safe as a mosquito repellent, extensive research does not document its ability to deter mosquitoes from animals. This study assessed the citronella essential oil bath bomb’s ability to repel Culex quinquefasciatus mosquitoes in dogs.  Citronella essential oil’s chemical composition was analyzed using gas chromatography-mass spectrometry (GC-MS). Through freeze-thaw testing, a bath bomb formulation containing 6% w/w citronella essential oil was assessed for its physical and chemical stability. Thirty-two healthy client-owned mixed-breed dogs were employed to test the mosquito-repellency effects of citronella essential oil (treatment group) and olive oil (control group) bath bomb formulations. Bath bombs were tested for irritation effects on animal skin for 15-day post-application.  Thirty-six compounds were identified through GC-MS, with citronellal (23.38%), δ-cadinene (12.25%), and geraniol (9.09%) being the most prevalent constituents. The bath bomb maintained its original physical properties after undergoing six freeze-thawing cycles and retained over 90% of its citronella essential oil. About 100%, 69.28%, and 65.58% mosquito repellency were displayed by the citronella essential oil bath bomb at 3 h, 6 h, and 8 h, respectively. None of the test animals exhibited skin irritation during the study.  The citronella bath bomb effectively repelled C. quinquefasciatus in dogs without irritating their skin. The formulation’s physical and chemical stability is demonstrated by the results of freeze-thaw stability testing. Further studies should be conducted to evaluate the repelling activity against other mosquito species. 

The intricate healing process involves distinct sequential and overlapping phases in thermal injury. To maintain the zone of stasis in Jackson’s burn wound model, proper wound intervention is essential. The extent of research on the histoarchitecture of thermal wound healing and the application of mesenchymal stem cell (MSC)-free-based therapy is limited. This study aimed to assess the efficacy of MSC-secretome-based hydrogel for treating partial-thickness cutaneous thermal burn wounds.  Eighteen male Wistar rats were divided into three groups, namely the hydrogel base (10 mg), hydrogel secretome (10 mg) and Bioplacenton™ (10 mg) treatment groups. All groups were treated twice a day (morning and evening) for 7 days. Skin tissue samples from the animals were processed for histological evaluation using the formalin-fixed paraffin-embedded method on days 3 and 7.  This study’s findings showed that secretome hydrogel expedited thermal burn wound healing, decreasing residual burn area, boosting collagen deposition and angiogenesis, guiding scar formation, and influencing the inflammation response facilitated by polymorphonuclear leukocytes and macrophages.  The secretome hydrogel significantly improves healing outcomes in partial-thickness cutaneous thermal burn wounds. The administration of secretome hydrogel accelerates the reduction of the residual burn area and promotes fibroblast proliferation and collagen density. The repairment of histo-architecture of the damaged tissue was also observed such as the reduction of burn depth, increased angiogenesis and epidermal scar index while the decreased dermal scar index. Furthermore, the secretome hydrogel can modulate the immunocompetent cells by decreasing the polymorphonuclear and increasing the mononuclear cells. Thus, it effectively and safely substitutes for thermal injury stem cell-free therapeutic approaches. The study focuses on the microscopical evaluation of secretome hydrogel; further research to investigate at the molecular level may be useful in predicting the beneficial effect of secretome hydrogel in accelerating wound healing. 

Some Culex mosquitoes are competent vectors for Japanese encephalitis virus (JEV) and Zika virus (ZIKV), which cause public health problems worldwide, especially in South-east Asia. Xenosurveillance of Culex mosquitoes remains limited compared with other common mosquito-borne diseases. This study aimed to identify JEV and ZIKV in field-caught Culex mosquitoes collected from Ubon Ratchathani province.  We investigated the presence of JEV and ZIKV in Culex mosquitoes from two districts in Ubon Ratchathani province, Thailand, and examined their role in viral interepidemic circulation. Female Culex mosquitoes (5,587) were collected using a mechanical aspirator from indoors and outdoors. The consensus sequences of the E and NS1 genes of JEV and the E gene of ZIKV were identified using real-time reverse transcription-polymerase chain reaction.  From 335 sample pools that contain a total of 5587 adult female Culex mosquitoes collected from Don Yung, Mueang district (4,406) and Phon Duan, Det Udom district (1,181), none of the collected mosquitoes tested positive for either JEV or ZIKV.  This study did not find JEV and ZIKV in Culex mosquitoes collected from the area of collection, which may be due to the low circulating amount of the virus in the vectors in the area, making it undetectable, or it may be because Culex mosquitoes are not suitable vector for the virus being tested. However, further xenosurveillance study of JEV and ZIKV in mosquito is suggested to prepare for the next outbreak. 

Bovine viral diarrhea (BVD), a highly pathogenic ribonucleic acid (RNA) virus, causes devastating financial losses and reproductive deaths among dairy cattle in Yogyakarta and globally. This study aimed to identify point mutations within the E2 structural protein of the acquired BVD virus (BVDV) isolates using genetic analysis.  The study period shows that we performed the research in 2023. We collected 118 serum samples from 2019 to 2023, among which only 10 BVDV positive were used and 108 were negative lacking the BVDV antigen. An anti-Erns monoclonal antibody-coated protein was used in indirect antigen capture enzyme-linked immunosorbent assay (I-ACE) to detect the BVD antigen present in positive BVDV serum specimens. In the initial step of the two-step reverse transcription polymerase chain reaction, the enzyme (superscript III reverse transcriptase) and the primer (random hexamer) were used to convert the RNA of the BVDV into complementary deoxyribonucleic acid (cDNA) during the process of reverse transcription. The final step involved the amplification of the E2 gene of the resultant BVDV cDNA through gene-specific primers (E2_fwd: 5′-TGGTGGCCTTATGAGAC-3′ and P7_rev: 5′-CCCATCATCACTATTTCACC-3′) and enzyme (platinum taq DNA polymerase high fidelity). For conducting Sanger sequencing, those 3 BVDV-1-positive isolates (about 2.6% of all isolates) were selected as a typical specimen for each site and year between 2019 and 2023 using a proportional computation. Therefore, only two BVDV isolates with complete genomes were chosen to perform their homological and genetic analysis based on the E2 gene by means of Blast and MEGA Version 11 in addition to the Bioedit 7.2.5 program.  By applying phylogenetic analysis relying on the E2 gene, a sum of 1011 nucleotides of the BVDV-1 isolates derived from each of the two BVDV-1 Indonesian isolates (n = 2) and its 23 reference BVDV strains were acquired from the National Center for Biotechnology Information (NCBI) database. The findings of the genetic analysis inside the phylogenetic tree revealed that the two BVDV Indonesian isolates were clustered into BVDV-1a subgenotype, while the reference BVDV strains were clustered into the five BVDV subgenotype, BVDV-1a (n = 6), BVDV-1b (n = 3), BVDV-1c (n = 11), BVDV-1m (n = 1), and BVDV-1n (n = 2). The branch exists in phylogenetic tree located before the division of our two BVDV isolates was divided into two branches with the same maximum bootstrap values of 99%, indicating a high degree of confidence, was seen. Next, we observed the branch near our study samples, which displayed the bootstrap value of 100, indicating that our 02 isolates were identical. In both isolates, V11 BVDV1/Indonesia/Yogyakarta/2023 and V16 BVDV1/Indonesia/ Yogyakarta/2023 with GenBank accession numbers PP836388 and PP836389, respectively, conserved D7E residues were mutated as well as cysteine changed/altered into serine (S) was identified at amino acid position 201.  We identified two isolates of BVDV belonging to the BVDV-1a subgenotype. Our findings indicate that the conserved D7E residues of isolates V11 BVDV1/Indonesia/Yogyakarta/2023 and V16 BVDV1/Indonesia/Yogyakarta/2023 were altered. The Indonesian BVDV isolates exhibited a cysteine to serine mutation at amino acid position 201, leads to vaccination failure, range of animal’s host will increase, and diagnostic kit will not be effective. 

Toxoplasmosis is caused by the parasite Toxoplasma gondii. Cats are the only known hosts that excrete resistant oocysts. Wild rats serve as crucial reservoirs and intermediate hosts for T. gondii’s survival and dissemination. Consuming soil and water containing oocysts can lead to illness. This study aimed to estimate the prevalence of toxoplasmosis in wild rats through molecular detection as an indicator of environmental contamination in Surabaya.  One hundred rats were collected from the three areas (housing, dense settlements, and traditional markets) and distributed into the five zones: West, East, Central, North, and South of Surabaya. Brain tissue samples were extracted using a Geneaid™ (New Taipei City, Taiwan) DNA isolation kit and analyzed through the loop-mediated isothermal amplification (LAMP) method.  The study analyzed brain tissue from 100 wild rats, consisting of 77 Rattus tanezumi and 33 Rattus norvegicus, displaying 30% LAMP positivity. The study revealed that 30% (30/100) of wild rats tested were infected with T. gondii. The molecular prevalence rate in male rats was 32.35% (22/68), compared to females with 25% (8/32). 41.9% of the housing population, 33.3% of traditional markets, and 22.6% of dense settlements had the highest molecular prevalence. The high positive molecular rate at the trapping site can be attributed to cats and dense populations.  Thirty percentage wild rats were tested positive for toxoplasmosis in Surabaya, East Java, Indonesia using LAMP method. Implementing strict control and monitoring is crucial in preventing the transmission of diseases from wild rats to humans. It is necessary to carry out further research related to genetic analysis of T. gondii to determine the type of T. gondii that infects animals and humans in Surabaya through bioassay and molecular test. 

Canine transmissible venereal tumor (CTVT), a unique transmissible cancer in dogs, affects the external genitalia and potentially spreads to other parts of the body. While somatic mutations in oncogenic and tumor-suppressing genes are linked to CTVT development, the impact of DNA methylation, which affects gene expression, remains unclear. This study explored whether DNA methylation in the promoter regions of the MYC oncogene and CDKN2B tumor suppressor genes in CTVTs is associated with their expression, both at the gene and protein levels.  To investigate promoter DNA methylation of MYC and CDKN2B in CTVTs, we analyzed frozen tissue samples from genital CTVT (GTVTs) and extragenital CTVT (ETVTs). Genomic DNA was extracted, bisulfite-treated, and analyzed using bisulfite polymerase chain reaction (PCR) and sequencing. The messenger RNA and protein of MYC and CDKN2B were also extracted and assessed by real-time PCR and Western blotting. Matching formalin-fixed, paraffin-embedded blocks were used for immunohistochemical staining to visualize protein distribution in GTVT and ETVT tissues.  Although both GTVT and ETVT samples showed MYC promoter methylation, the extent of methylation differed significantly. GTVTs displayed a much higher degree of methylation, potentially explaining the more pronounced downregulation of MYC gene expression and reduction in c-MYC protein levels observed in GTVTs compared with ETVTs. Our data revealed a prevalent hypermethylation pattern in the CDKN2B promoter across both sample types. However, DNA methylation, which was expected to have a suppressive effect, did not correlate with gene/protein expression. GTVTs displayed high protein levels despite significantly reduced CDKN2B expression. Conversely, ETVTs maintained regular CDKN2B expression but exhibited reduced protein production, suggesting a complex interplay between methylation and expression in these tumors.  MYC demonstrated a clear association between its promoter methylation status, gene expression, and protein levels; however, CDKN2B lacked this correlation, implying the involvement of methylation-independent regulatory mechanisms and highlighting the need for further investigation. 

Anemia, a clinical condition characterized by reduced erythrocytes, is often observed in cats. Regeneration indicates that the bone marrow can respond appropriately to anemia. The absolute reticulocyte count is the reference for differentiating regenerative and non-regenerative anemia, while red blood cell (RBC) indices and morphology provide supplementary information. This study aimed to identify anemia types and establish the most reliable RBC indices and morphology methods in agreement with the reference method.  One hundred forty-five cases of cat anemia were prospectively classified using two methods: RBC indices and RBC morphology, and subsequently compared with the absolute reticulocyte count.  Based on RBC indices assessment, 27 cases (19%) exhibited regenerative anemia. Based on RBC morphology, 29 (20%) cases were identified as having regenerative anemia. Using the reticulocyte absolute count as a reference method, 34 (23.4%) cases of regenerative anemia were identified. The findings indicated that RBC indices and RBC morphology did not align in evaluating medullary regeneration and that there is a good degree of agreement between RBC morphology assessment and the reticulocyte absolute count in identifying regenerative anemias.  Blood smear analysis of RBC morphology was more dependable for classifying regenerative anemia than RBC indices. Further studies should be conducted with a larger number of animals and that allow the identification of the cause of anemia and the monitoring of the animal. 

The excessive use of antimicrobials in livestock farming leads to the emergence and dissemination of antimicrobial-resistant organisms. This study aimed to detect extended-spectrum β-lactamase (ESBL)-producing Escherichia coli genes in integrated poultry-fish farms in Bogor, Indonesia.  A total of 256 samples were collected from six poultry-fish farms. One hundred and seventy-five chicken cloaca swabs, 60 fish skin swabs, six pond water samples, and 15 farmer’s hand swabs. ESBL-producing E. coli was confirmed through double-disk diffusion. The specific primers and probe genes for quantitative polymerase chain reaction detection of ESBL-producing E. coli targeted blaTEM, blaCTX-M, blaSHV, and blaOXA-48 genes.  Among the 256 samples tested, 145 (56.6%) were positive for E. coli, and 67.6% (98/145) were identified as ESBL-producing E. coli. The most ESBL-producing E. coli isolates were obtained from chicken cloaca (78.3%, 72/92), followed by pond water (66.7%, 4/6), fish skin (47.6%, 20/42), and farmer’s hand swabs (40%, 2/5). About 100% of the isolates carried the genes blaTEM and blaCTX-M, whereas 17.3% and 24.5% carried blaSHV and blaOXA-48, respectively.  ESBL-producing E. coli genes were investigated in chicken cloaca, fish, pond water, and farmers’ hands within an interconnected poultry-fish farming operation. The ESBL-producing E. coli in chickens can transfer resistant genes to aquatic environments. The transfer could harm other aquatic species and food chains, potentially threatening human health. 

Citronella grass (Cymbopogon nardus) waste, produced by distilling citronella to produce essential oil, has a high potential for use as animal feed. However, the presence of high lignin content could limit its digestibility, prompting the need for treatment to improve its quality. This study aimed to improve the nutritional value and in vitro digestibility of ammoniated and fermented citronella waste (CW).  The treatments of CW included CW without treatment as a control (T0), ammoniation of CW with urea (T1), fermentation of CW with Trichoderma harzianum (T2), and a combination of ammoniation and fermentation (amofer) of CW (T3). This study employed a randomized block design with five replicates for each of the four treatments. If there was a significant effect (p < 0.05), a post hoc Duncan’s multiple range test was performed to analyze the variance of the data.  The process of ammoniation and fermentation led to a notable increase in crude protein (2%–6%) while decreasing crude fiber (2%–6%), neutral detergent fiber (NDF) (5%–14%), acid detergent fiber (ADF) (5%–9%), lignin (4%–9%), and cellulose (2%–10%). The treatments enhanced the digestibility of dry matter, organic matter (OM), NH3, and total volatile fatty acid by 4%–12%, 6%–19%, 0.9–10 mM, and 35–142 mM, respectively. The decrease in NDF, ADF, acid detergent lignin (ADL), and cellulose fractions was accompanied by an improvement in dry matter and OM digestibility in CW. Ammoniated-fermented (amofer) CW, followed by fermentation with T. harzianum and ammoniated urea treatment, significantly enhanced the nutritional content and in vitro digestibility. The decrease in NDF, ADF, ADL, and cellulose fractions led to an improvement in dry matter and OM digestibility in CW.  The application of amofer treatment with T. harzianum maximizes CW’s nutritional value and digestibility, making it the most efficient preservation method. Research is needed to explore the potential use of Aspergillus spp. and Pleurotus spp. for fermenting CW as ruminant fodder. 

Although reverse zoonotic transmission events from humans to domestic cats have been described, there is currently little evidence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) circulation in stray cats. Due to the evidence of natural and experimental infections in cats and the capacity to disseminate the virus among them, this study aimed to identify the SARS-CoV-2 antigen in stray cats from the Federal University of Sergipe in Brazil.  One hundred twenty six stray cats from the university were screened for SARS-CoV-2 antigens by random sampling. Throat swab samples were tested for the virus using rapid antigen detection tests.  Of the 126 animals tested, 30 (23.60%) were positive for SARS-CoV-2 antigens. To our knowledge, for the first time, this study detected the SARS-CoV-2 antigen in stray cats and confirmed the presence of SARS-CoV-2 infections in Brazil’s stray cat population.  The detection of SARS-CoV-2 in stray cats poses a risk for infected and healthy animals and possibly for humans who attend the university daily. As a limitation of the study, the small sample size necessitates caution when interpreting the results. This underscores the need for further research in this area to help control diseases in stray animals during potential pandemics. This highlights the need for monitoring and controlling the spread of the virus in stray animal populations. 

Ginger (Zingiber officinale) has great potential as a growth promoter and immunostimulant in ruminant nutrition. This study assessed the impact of ginger powder supplementation on Ossimi rams’ rumen fermentation, biochemical parameters, and antioxidant levels.  Fifteen Ossimi rams, aged 10 ± 1.3 months and weighing 30 ± 1.5 kg. Rams were randomly divided into three experimental groups: The control group (G1) received standard feed, while ginger powder (5 g and 7 g/kg body weight [BW] for G2 and G3, respectively) mixed in water was administered to groups G2 and G3 before their standard feed.  The control group recorded higher dry matter (DM) intake values (p < 0.05) than the ginger-treated groups. The ginger-treated groups showed superiority (p < 0.05) in weight gain and feed conversion compared to the control group. The digestion coefficients of DM, crude protein, and crude fiber were significantly (p < 0.05) increased by a high dose (7 g/Kg BW) of ginger supplementation, whereas organic matter, ether extract, and nitrogen-free extract digestibility remained unchanged. Compared to the control group, the rams given 5 g of ginger had significantly less (p < 0.05) total protein and globulin in their serum, but the rams given 7 g of ginger had significantly more (p < 0.05) of these proteins. In the ginger groups, these levels were significantly (p < 0.01) lower than those in the control group for serum creatinine, uric acid, urea, total lipids, triglycerides, total cholesterol, glucose, serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase. Rams given ginger had significant growth hormone, insulin-like growth factor-1, total superoxide dismutase, GSH-Px, TAC, immunoglobulin (Ig) A, and IgG enhancement (p < 0.01), and a decrease (p < 0.01) in malondialdehyde concentration compared to the control group. Significant increases in total short-chain volatile fatty acids, acetic, propionic, and isovaleric acids (p < 0.05), and significant decreases in NH3N and protozoa (p < 0.01).  Ginger powder (5 g and 7 g) can improve growth, immune responses, antioxidant status, and ruminal parameters in rams. Further study is needed to evaluate the effect of ginger on different types of animals (cow, buffalo, and goat) to develop new feed additives. 

Pseudomonas aeruginosa is an infectious agent of great importance for animals and humans. It causes serious infections that show high resistance to antibiotics. This study investigated the molecular detection of blaOXA-23 gene in antibiotic-resistant P. aeruginosa strains isolated from cows and humans.  In total, 120 samples, comprised 60 from cows (30 milk and 30 nasal discharge) and 60 from their owners (30 urine and 30 sputum), were individually collected, cultured, and tested for P. aeruginosa through molecular analysis targeting the blaOXA-23 gene. P. aeruginosa antibiotic-resistant isolates were identified by performing antibiotic susceptibility testing and detecting biofilm formation.  In total, 74.17% positive P. aeruginosa isolates, including 66.67% and 81.67% for cows and humans, respectively. Subsequently, positive cow isolates were detected in 60% of milk samples and 73.33% of nasal discharge samples; while positive human isolates were detected in 76.67% of urine samples and 86.66% of sputum samples. Targeting blaOXA-23 gene, 58.43% of cultured isolates were positive for P. aeruginosa by polymerase chain reaction. Respectively, positive isolates were detected in 66.67% and 45.46% of cow milk and nasal discharges as well as in 60.87% and 61.54% of human urine and sputum. The antibiotic susceptibility test revealed that all isolates were resistant to all applied antibiotics, particularly imipenem. Results of biofilm formation revealed 67.31% total positives, including 51.43% strong, 34.285% moderate, and 14.285% weak reactions. In addition, although values of the total positive cows and humans differed insignificantly, total positives showed insignificant variation between values of milk and nasal discharges of cows as well as between urine and sputum of humans; however, significant differences were identified in the distribution of strong, moderate, and weak positivity of these samples.  Antibiotic overuse contributes extensively to increasing the prevalence of resistant P. aeruginosa isolates carrying the blaOXA-23 gene in both cows and humans. Furthermore, studies in other Iraqi areas are necessary to support our findings. The main limitations include that the number of tested samples is relatively low, and there is a need to use a large number of samples from different sources. Also, the current methods for detection of resistant isolates are still culture-based approaches. 

The use of frozen goat semen for artificial insemination frequently results in a decline in sperm quality following thawing, which can be attributed to cold shock from cryopreservation, reduced motility, and possible DNA damage. Freezing may compromise mRNA stability due to the presence of free radicals. Despite strong post-thaw motility and no visible DNA fragmentation, sperm can still exhibit altered gene expression patterns. To reduce the damaging impact of free radicals during cryopreservation, antioxidants are typically added to the freezing medium. This study assessed the impact of adding coenzyme Q10 (CoQ10) to frozen sperm diluent on the ATP5F1A and CPT2 gene expression, sperm motility, and viability post-thawing.  CoQ10 was added to sperm at six different concentrations: 0 mg/dL (P0), 6.25 mg/dL (P1), 12.5 mg/dL (P2), 25 mg/dL (P3), 50 mg/dL (P4), and 100 mg/dL (P5). The Statistical Package for the Social Sciences (SPSS) software version 22 was used to conduct comparative tests using one-way analysis of variance followed by Duncan’s test for motility and viability and Kruskal–Wallis test followed by pairwise comparison test for membrane integrity and gene expression.  The addition of CoQ10 to semen diluent has a notable impact on the post-thawed quality of sperm. The most significant outcomes were observed with a 25 mg/dL dosage (P3) for cell viability, membrane integrity, and ATP5F1A gene expression, and with a 50 mg/dL dosage (P4) for sperm motility, membrane integrity, and CPT2 gene expression.  Incorporating CoQ10 into frozen semen diluent improves gene expression and prevents deterioration of the cell quality of thawed goat spermatozoa. While the study demonstrates the benefits of CoQ10, the precise molecular mechanisms through which CoQ10 enhances gene expression and cell quality were not fully elucidated. Further investigation is needed to understand these mechanisms in detail. Comparative studies with other antioxidants and cryoprotectants can help establish the relative efficacy of CoQ10 and potentially develop more effective combinations. 

Wolves (Canis lupus) play a role in nature, including the regulation of the number of ungulates and the use of dead animals. In addition, wolves are a natural link and carrier for the spread of many parasitic invasions. Hence, the main task in preventing the spread of parasitic invasions is to regulate the wolf population. This study aimed to monitor the endoparasitological fauna of wild wolves inhabiting Northern and Central Kazakhstan.  Overall, 81 wolves were investigated for parasitic worms using the K. I. Scriabin method. Wolf intestinal materials were collected from the following six regions: North Kazakhstan, Pavlodar, Kostanay, Akmola, Ulytau, and Karaganda. The genetic diversity of the parasites was identified using a polymerase chain reaction with specific primers. After data collection, a comprehensive statistical analysis was performed.  Several helminth types were identified in wolves, including Echinococcus granulosus, Taenia hydatigena, Mesocestoides spp., Toxascaris leonina, Trichinella nativa, Alaria alata, and Dirofilaria repens. Based on the results of this study, young male wolves aged 1–4 years were the most vulnerable to helminthiasis. Wolves living in steppe and semi-desert regions are often exposed to helminth infections. The prevalence of T. nativa in the wolves was 20.4%. This study also revealed the presence of echinococcosis among wolf populations in Karaganda and Kostanay, with prevalence rates of 4.1% and 4.7%, respectively. The overall prevalence of tapeworms in wolves was 54.3%.  This study highlights the significance of understanding the potential risks associated with helminth infections in wild carnivores because helminths can act as disease reservoirs and pose a threat to humans, livestock, and other wild carnivores. These results can contribute to the development of effective control and management strategies for helminth infections in wolves, which can infect humans and livestock. 

Aeromonas hydrophila infections in fish result in significant financial losses within aquaculture. Previous research indicates black soldier fly (BSF) prepupae provide immunomodulatory benefits through their fatty acids, chitin, and proteins. The study evaluated the impact of hexane extract from black soldier fly prepupae (HEBP) on interleukin (IL)-4 and IL-10 cytokine expression in zebrafish, both infected and uninfected with A. hydrophila.  Adult zebrafish (aged 4–5 months) was assigned to a negative control group (fed commercial feed), a positive control group (commercial feed + A. hydrophila infection at 107 colony-forming unit/mL), and three treatment groups (T1, T2, T3) that received HEBP at doses of 1000; 2000 and 4000 mg/kg feed for 30 days, respectively. A. hydrophila infection was introduced on day 31 through immersion. Analysis of IL-4 and IL-10 expression in the head kidney trunk region (body without head and tail) through quantitative polymerase chain reaction was conducted on day 33.  The HEBP modulated the immune response to A. hydrophila infection at a concentration of 1000 mg/kg feed, as evidenced by an increase in IL-4 and IL-10 expression in the groups not infected with the bacteria. However, these cytokines were decreased in the infected groups.  A feed concentration of 1000 mg/kg HEBP was identified as optimal for cytokine modulation. This discovery marks a significant advancement in the development and benefit of a natural extract-based immunomodulator in a zebrafish model, which is potentially immunotherapeutic against bacterial infections in fish for the aquaculture industry.