Lice infestation is a common ectoparasitic condition in sheep and may cause pruritus, wool loss, skin damage, and impaired productivity. This study developed a topical total alkaloid solution from Sophora alopecuroides and evaluated its formulation characteristics, stability, dermal tolerance, field efficacy, and target-animal safety in sheep naturally infested with lice. Total alkaloids were extracted from S. alopecuroides seeds using single-factor experiments and orthogonal optimization. Dermal safety was assessed using Buehler sensitization testing in guinea pigs and skin irritation testing in rabbits. Clinical efficacy was evaluated after medicated bath treatment in naturally infested sheep. The optimized extraction procedure used 70% ethanol, an 8-fold solvent-to-material ratio, 0.1% cellulase, and three extractions of 120 min each. The optimized formulation contained 6.5% total alkaloids, 10.0% sodium dodecyl sulfate, and 8.0% ethanol. The preparation met predefined quality criteria and remained stable under stress, accelerated, and long-term storage conditions. No treatment-related sensitization or irritation was observed in laboratory animals. In sheep, lice reduction rates at D4 were 95.27%, 100%, 100%, and 100% in the low-dose, medium-dose, high-dose, and diazinon groups, respectively, and all treated groups reached 100% lice reduction at D17. No obvious hematological, biochemical, or histopathological safety signals were observed. These findings support the potential of this plant-derived topical preparation for managing natural lice infestation in sheep.

Avian influenza virus (AIV) remains a major threat to poultry production in Bangladesh, where both highly pathogenic (HPAIV) and low-pathogenic (LPAIV) subtypes circulate. Although HPAIV is typically associated with high mortality, H9N2 LPAIV often causes mild clinical disease but has been associated with reduced egg production and economic losses in commercial poultry. This study describes the concurrent detection of H9N2 and Salmonella spp. in a 33-week-old Hy-Line Brown commercial layer flock in Moulvibazar, Bangladesh, in which a marked decline in laying performance was observed. A total of 50 birds were sampled for virological and bacteriological investigations. Oropharyngeal swabs were tested by real-time reverse transcription PCR (rRT-PCR) targeting the influenza A virus matrix (M) gene, followed by H9- and N2-specific assays. Cloacal swabs were cultured using selective enrichment and plating methods, and presumptive Salmonella spp. isolates were confirmed by PCR targeting the invA gene. Among the sampled birds, concurrent H9N2 and Salmonella spp. were detected in 11/50, while no birds were positive for H9N2 alone; Salmonella spp. alone was detected in 3/50 birds. The investigated flock exhibited markedly reduced egg production, remaining at 0% through 25 weeks of age and reaching a maximum of only 71.72% at 46 weeks, compared with the breed-standard value of 88%, while cumulative mortality reached 23.4% by 46 weeks, substantially exceeding the breed standard. These findings document the concurrent detection of H9N2 and Salmonella spp. and poor laying performance in the investigated flock. Given the single-flock design, absence of an H9N2-only comparison group and a contemporaneous healthy control flock, and incomplete assessment of other infectious and farm-level factors, the findings do not establish a causal relationship or determine whether Salmonella spp. contributed to the observed production losses. Further controlled and longitudinal studies involving multiple flocks and appropriate comparison groups are warranted to clarify the respective contributions of H9N2 and bacterial coinfections to production losses in commercial layer flocks.

A key aspect of controlling canine visceral leishmaniasis (CVL) is the availability of accurate diagnostic tools capable of reaching most of the population. In endemic areas with socially vulnerable populations, facilitating rapid CVL diagnosis by reducing sampling difficulties and diagnostic costs could provide substantial benefits. This study evaluated the performance of the Kalazar Detect™ Canine Rapid Test for CVL diagnosis using blood, serum, and saliva samples, and compared the results among sample types and with PCR detection in lymph nodes. Samples were collected from 100 dogs. Prevalence detected by the rapid diagnostic tests (RDT) was 21% using blood and serum and 17% using saliva, whereas PCR detected a prevalence of 46%. Agreement between serum and blood-based RDTs was 100%, while comparison between serum/blood and saliva showed a kappa index of 0.87, indicating almost perfect agreement. Using PCR as the reference standard, RDTs on serum and blood showed a sensitivity of 0.39 and specificity of 0.94, whereas saliva-based RDT showed a sensitivity of 0.33 and specificity of 0.96. The low sensitivity of the RDTs currently used by the National Leishmaniasis Program indicate that an additional diagnostic method should be incorporated into CVL surveillance in endemic areas. Nevertheless, our results suggest that saliva may be a suitable sample for the Kalazar Detect Canine Rapid Test, as it is easy to collect, less expensive, and less invasive. To our knowledge, this is the first study to evaluate the performance of this RDT for CVL diagnosis using saliva samples.

Peste des petits ruminants virus (PPRV) causes a highly contagious and economically devastating disease of small ruminants, particularly sheep and goats. Serological evaluation using recombinant protein-based Enzyme-linked immunosorbent assays (ELISAs) is essential for monitoring PPRV infection and assessing eradication programs. Although recombinant nucleoprotein (N) is widely used for serological evaluation, investigation of additional viral proteins may provide alternative antigenic platforms for PPRV antibody detection. The matrix (M) protein plays a central role in viral assembly by linking viral surface glycoproteins with ribonucleoprotein complexes. However, recombinant expression of full-length M protein in Escherichia coli (E. coli) is challenging because of low or truncated expression and inclusion body formation, often requiring complex refolding procedures. In this study, full-length M protein from the Nigeria 75/1 vaccine strain was successfully overexpressed as inclusion bodies (IBs) in E. coli and solubilized using N-lauroyl sarcosine (NLS) without refolding. The solubilized protein was purified by nickel-affinity chromatography, yielding 2.1 mg/L bacterial culture, and confirmed as a 38-kDa protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. An indirect ELISA was developed using purified M protein and optimized with commercial control sera. Optimal conditions were achieved with 1 µg/mL antigen and 1:20 serum dilution. Using pre-confirmed field sera, the assay demonstrated 94.59% sensitivity and 92.45% specificity at a cut-off of 0.25. Comparison with the commercial ID Screen[®] PPR Competition ELISA showed comparable performance and strong agreement. These findings demonstrate the recombinant PPRV M protein as a feasible diagnostic antigen for PPRV antibody detection which requires further validation for large scale application.

The present study aimed to evaluate the in vitro anthelmintic activity of Bambusa vulgaris leaves extract against Haemonchus contortus along with identification of phytochemical constituents and in silico evaluation. Ethanolic extract of Bambusa vulgaris leaves and their methanolic and chloroform fractions, obtained through column chromatography, were tested at concentrations ranging from 10 to 200 mg/ml. Anthelmintic activity was assessed in vitro by using the Adult Motility Test (AMT) and Egg Hatch Assay (EHA). The results of Adult Motility Test revealed that at 200 mg/ml, ethanolic extract and its methanolic, and chloroform fractions induced mortality at 17.3 ± 0.33, 18.3 ± 0.33, and 20.6 ± 0.33 min, respectively. The extracts also exhibited ovicidal activity, with nearly complete inhibition of egg hatching (96.42 ± 0.90, 94.94 ± 1.02%, and 95.98 ± 1.02%, respectively). Scanning Electron Microscopy (SEM) of treated worms revealed morphological alterations on cuticular surface with longitudinal and transverse wrinkles, compared to untreated controls. Gas Chromatography-Mass Spectrometry (GC-MS) analysis of ethanolic Extract identified several bioactive compounds, with Phytol, Neophytadiene, Undecane, 1,8-Nonanediol, 8-methyl, Decanoic acid, 2 methyl, Diethyl phthalate and β-Pinene are the phytoconstituents. Following this, in silico studies were performed to know the mechanism of action of the constituents against H. contortus. Molecular docking showed that all the compounds and especially Phytol, Neophytadiene and Diethyl phthalate showed good binding with Beta tubulin and Glutamate Dehydrogenase. Thus, these compounds have potential to disrupt energy metabolism and structural stability. Also, pharmacokinetic evaluation depicted that the compounds present in this plant are safe and Neophtyadiene and Phytol are poorly absorbed from GIT, that highlights their importance against gastrointestinal parasites. The findings suggest that Bambusa vulgaris leaves possesses, dose-dependent anthelmintic properties, supporting its potential as a natural source of anthelmintic agents.

Acinetobacter baumannii presence in veterinary hospitals and companion animals is still poorly investigated despite its significance. The present study aimed to investigate the prevalence, antimicrobial resistance patterns, and resistance genes of A. baumannii in cats, veterinary hospital environments, and veterinary medical wastes in Bangladesh. Samples were collected from oral, rectal, and wound swabs of cats (n = 100) attending ten veterinary hospitals, as well as from environmental surfaces (n = 111) and wastes (n = 61) generated within these facilities. Isolates were identified by PCR targeting 16S rRNA (genus) and blaOXA-51-like (species) genes; antimicrobial susceptibility was assessed by disk diffusion and broth microdilution following CLSI standards, and resistance genes (blaOXA-23-like, blaNDM-1) were screened by PCR. A high prevalence of A. baumannii was observed in cats (37%), while 26.1% in environmental samples and 21.3% in waste samples. Apparently sick cats had significantly higher prevalence (p = 0.049) than apparently healthy cats. All isolates were imipenem resistant, consistent with carbapenem-resistant A. baumannii (CRAB). High resistance was also observed against cefotaxime and ceftriaxone. Multidrug resistance (MDR) was detected in 26.7%, 37.9%, and 30.8% of isolates from cats, environmental, and waste sources, respectively. The blaOXA-23-like gene was detected in only two cat isolates, whereas blaNDM was identified in one cat isolate and one environmental isolate. In conclusion, these findings identify cats, which live in close proximity to humans, as a possible CRAB reservoir, and veterinary hospital environments and waste as potential sources of persistence and dissemination, raising concern for zoonotic spread and underscoring occupational and public health risk.

While dopamine and dobutamine are widely used in anesthetized dogs, clinical evidence supporting their use remains limited. We aimed to compare the hemodynamic responses to clinical dosages of dopamine and dobutamine in dogs with hypotension undergoing elective orthopedic surgery under fentanyl-isoflurane anesthesia. The pressor effects at clinical dosages (2-8 µg/kg/min) of dopamine (n = 15) and dobutamine (n = 15) for hypotension were compared and confirmed using the ramped-rate infusion protocol in 30 dogs. The following hemodynamic parameters measured by invasive arterial blood pressure and transesophageal echocardiographic monitoring were analyzed: heart rate (HR), systolic (SAP), mean (MAP), diastolic arterial pressure (DAP), stroke volume index (SVI), cardiac index (CI), and systemic vascular resistance index (SVRI). The proportion of dogs that achieved the treatment goal (MAP: 70-89 mmHg) was 93% (dopamine responders, 14/15 dogs) in the dopamine group and 53% (dobutamine responders, 8/15 dogs) in the dobutamine group, with a significantly higher probability in the dopamine group (P = 0.035, a risk difference of 40% [95% confidence interval: 8.2-63.9%]). In dopamine responders, significant increases were observed in SAP, MAP, DAP, SVI and CI, whereas no significant changes were observed in HR and SVRI. In conclusion, the pressor effect was more likely to be observed at the clinical dosage of dopamine than that of dobutamine for treating hypotension in isoflurane anesthetized dogs. This study was approved by the Nihon University Animal Medical Center Research and Trial Ethics Committee (approval number: 1-007; approval date: September 29, 2022).

Newcastle disease (ND) has been recognized for a century as one of the most consequential viral diseases of poultry, and its scientific history provides a distinctive example of how sustained investigation of a single pathogen can drive advances across veterinary medicine, virology, immunology, epidemiology, molecular biology, vaccinology, and biotechnology. The severe outbreaks reported in Java, Indonesia, and Newcastle-upon-Tyne, England, during 1926 marked the beginning of the recognized scientific history of ND and initiated an international research effort that progressed from disease recognition and experimental characterization to diagnostics, vaccination, molecular pathogenesis, and genomic epidemiology. During the 1940s and 1950s, investigations of viral propagation, antigenicity, pathogenicity, and attenuation established the foundations of live and inactivated vaccination. From the 1960s onward, molecular investigations progressively elucidated the determinants of ND virus (NDV) replication, protein processing, and virulence, particularly the role of the fusion-protein cleavage site. The emergence of nucleotide sequencing and phylogenetic analysis during the 1980s and 1990s subsequently transformed strain characterization and provided the foundations for molecular epidemiology and genotype classification. Complete-genome sequencing and the establishment of reverse-genetics systems in the late 1990s further transformed NDV from an organism studied primarily through observation and isolation into a genetically tractable experimental system. Subsequent international efforts standardized NDV genotype nomenclature and enabled increasingly detailed analyses of viral evolution and geographic dissemination. In parallel, NDV research expanded beyond conventional disease control into recombinant vaccine development, systems-level host-virus studies, molecular diagnostics, and oncolytic virotherapy. This historical perspective traces these major scientific transitions from 1926 to 2026, emphasizing the investigators, experimental systems, conceptual advances, and technological innovations that collectively transformed NDV research from disease recognition into a multidisciplinary molecular and translational fields.

Cutaneous squamous cell carcinoma (cSCC) is a common canine skin tumour that occurs more frequently in high ultraviolet (UV) environments. Despite Australia's consistently high UV exposure, large-scale epidemiological studies on canine cSCC are limited. This study investigates the frequency of canine cSCC in Australia, its geographic patterns relative to UV index, and its association with animal-level risk factors. A matched case-control study was conducted using records from the VetCompass database from 1st January 2014 to 31st December 2023. In total, 295 histopathologically confirmed cSCC cases and 1,180 matched controls were included. Frequency statistics of cSCC and animal-level risk factors associated with cSCC were analysed and spatial patterns were mapped. The highest proportions of cases were recorded in Queensland (39.7%) and New South Wales (39.3%), with increased frequency along coastal areas of both states. cSCC cases occurred most frequently on body regions caudal to the thorax (54.9%), in senior dogs (58.6%), in desexed males (45.1%), and in dogs with white mixed coat colour (43.7%). In the final multivariable model, senior (seven to 11 years old) and geriatric (12 + years old) dogs had significantly higher odds of cSCC than mature (two to six years old) dogs (odds ratio = 7.4 and 4.8, respectively; p ≤ 0.001). Risk was also markedly elevated in Dalmatians, bulldogs, Staffordshire terriers, and schnauzers compared with mixed breeds. Dogs with white or white mixed coats had higher odds of cSCC than dark-coated dogs (p ≤ 0.001). cSCC demonstrated anatomical and geographic patterns consistent with chronic UV exposure, with age, breed, and coat pigmentation emerging as risk factors.

Rabies vaccination is conducted along the northern border of Peninsular Malaysia, but paired field serological data are limited. We analysed routine programme data from 154 owned dogs sampled immediately before vaccination and 30-83 days later (median, 43 days). Rabies virus-neutralising antibody levels were measured using the fluorescent antibody virus neutralisation test. Paired levels in IU/mL were compared using the Wilcoxon signed-rank test, and benchmark proportions were compared using the exact McNemar test. The proportion with levels ≥ 0.5 IU/mL increased from 72.7% (112/154; 95% confidence interval [CI], 65.2-79.1%) to 98.1% (151/154; 95% CI, 94.4-99.3%; p < 0.001). Three dogs remained below the benchmark. Median levels increased from 1.98 IU/mL (interquartile range [IQR], 0.38-5.92) to 13.5 IU/mL (IQR, 4.5-13.5); the median paired increase was 5.45 IU/mL (bootstrap 95% CI, 3.31-7.25; p < 0.001). Levels increased in 128 dogs, were unchanged in 19 and decreased in seven. Eighty follow-up values equalled the dataset maximum of 13.5 IU/mL, suggesting possible upper-end censoring. Vaccination history identified 99 previously vaccinated dogs and seven recorded as receiving their first vaccination, limiting comparison of these groups. Nearly all dogs retained at follow-up met the serological benchmark. Convenience sampling, incomplete follow-up and uncertain measurement limits restrict inference to this cohort and preclude estimates of population immunity or clinical effectiveness.

Odontogenic tumours are rarely reported in teleost fishes, and detailed histopathological descriptions of these lesions remain limited. This study describes a rare case of compound odontoma in the striped poison-fang blenny mimic, Petroscirtes breviceps (Valenciennes, 1836). Among 30 examined individuals, one male specimen measuring 13 cm in total length and weighing 12 g presented with a distinct tumour-like growth in the oral cavity. Gross examination revealed a localized, pale pink, fleshy odontogenic lesion attached to the left side of the lower jaw. The mass contained numerous toothlet-like structures of varying sizes, suggesting progressive tumour-associated odontogenic differentiation. Histopathological analysis showed a well-circumscribed, non-invasive mass composed of multiple irregular denticle-like structures embedded within a moderately cellular fibrovascular stroma. These denticles exhibited features consistent with dental tissue organisation, including eosinophilic dentine-like mineralised matrices surrounding central pulp-like cores, with enamel-like caps observed in some areas. The lesion also contained proliferating fibroblasts, mesenchymal cells resembling dental papilla, and scattered nests of odontogenic epithelium. No evidence of cytological atypia, necrosis, vascular invasion, inflammatory infiltration, or significant mitotic activity was observed, confirming the benign nature of the tumour. The presence of well-differentiated tooth-like structures composed of dentine-, cementum-, and enamel-like materials supported a definitive diagnosis of compound odontoma. To the best of our knowledge, this represents the first documented report of odontoma in P. breviceps from Rameswaram Island, India.

The bovine uterus undergoes marked structural remodeling during the early luteal phase, yet quantitative information on post-ovulatory changes in uterine horn echotexture remains limited. This study characterized uterine horn echotexture during the first follicular wave following synchronized ovulation in Holstein cows and evaluated its association with uterine horn classification and antral follicle count (AFC). Thirty-six Holstein cows (Bos taurus; parity 2-6; 60-65 days in milk) synchronized using an Ovsynch protocol were examined on Days 0, 1, 3, 6, and 9 after ultrasonographically confirmed ovulation. Mean gray level (MGL), gradient (GR), homogeneity (HOM), and contrast (CON) were quantified in both uterine horns (360 observations) and analyzed using linear mixed-effects models. AFC was defined as the mean number of follicles ≥ 3 mm across five examinations. All echotexture parameters changed significantly over time (p ≤ 0.007); MGL, GR, and CON increased, whereas HOM decreased. No significant differences occurred between ipsilateral and contralateral horns (p ≥ 0.094), and horn-level variance within cows remained below 5.7%. Anatomically, MGL was 3.05 units higher in the right horn (p = 0.043), although 27 of 36 ovulations occurred in the right ovary, limiting separation of anatomical from functional laterality. AFC was associated only with MGL; each 10-follicle increase corresponded to a 4.84-unit increase (95% CI: 1.51-8.17; p = 0.006), with no day × AFC interaction (p = 0.184). Quantitative echotexture analysis provides an objective approach for monitoring physiological uterine remodeling, whereas ovarian follicle reserve shows a modest, parameter-specific association with uterine echogenicity.

Escherichia coli (E. coli) is a significant livestock and foodborne pathogen. This study was designed to isolate E. coli from raw cow milk, evaluate its antimicrobial susceptibility, and investigate the antimicrobial activity of silver nanoparticles (nAg), zinc oxide nanoparticles (nZnO), and Moringa nano-emulsion (MNE) against multidrug resistant (MDR) strains. A total of 146 individual milk samples were obtained from dairy cows in Egypt. E. coli were isolated by standard culture methods and pathotyped using molecular techniques. The antibiotic resistance of the isolates was determined, and the antimicrobial activity of the selected nanomaterials was evaluated using the broth microdilution method on the identified MDR strains. E. coli was detected in 54.8% (80/146) of the samples; 42 out of 80 (52.5%) were virulent strains, including STEC (15/42; 35.7%), ETEC (12/42; 28.57%), EPEC (12/42; 28.57%), and AE-STEC (3/42; 7.1%). The isolated strains showed the highest resistance rate to sulfa-trimethoprim (52.5%), tetracycline (45%), and ampicillin (42.5%); and the lowest against marbofloxacin (8.75%) and ceftiofur (27.5%). Overall, 42.5% of the E. coli strains exhibited MDR, including virulent and non-virulent strains at 45.2% (19/42) and 39.47% (15/38), respectively. The minimum inhibitory concentration (MIC) values against MDR E. coli isolates were lowest for nAg (1.56-12.5 µg/mL), followed by nZnO (12.5-50 µg/mL). In contrast, Moringa nano-emulsion (MNE) requires much higher concentrations (125-250 µg/mL) for growth inhibition. Raw cow milk is a potential source of pathogenic and MDR E. coli, emphasizing continuous surveillance and prudent antimicrobial use. Although selected nanomaterials showed consistent in vitro efficacy, further studies are required before considering practical veterinary applications.

Newcastle disease (ND) poses a serious threat to poultry, with vaccination remaining the most effective control strategy. However, the widely used ND vaccine strain exhibits significant antigenic mismatch with prevalent genotype VII Newcastle disease virus (NDV), necessitating increased vaccination dosages and frequencies. This study evaluated optimization of immunization schedules for the ND marker vaccine based on genotype VII MG7 strain. Three groups were tested: Group A (4 vaccinations with inactivated ND marker vaccine), Group B (5 vaccinations with combined inactivated and live ND marker vaccines), and Group C (7 vaccinations with LaSota genotype II vaccine). Results showed that Groups A and B maintained hemagglutination inhibition antibody titers above 9 log2 before laying and ≥ 8 log2 during the four-month laying period, providing superior immune persistence compared to Group C. The study has confirmed that the specific ELISA test successfully distinguished between chickens that received the marked vaccine and those that did not. Furthermore, the marker vaccine strategy did not interfere with immune response to H5 and H7 subtype avian influenza vaccines. Notably, Group B achieved the highest production efficiency, with 3.01 CNY higher benefit per chicken versus Group C, while reducing vaccination stress and improving flock welfare. These findings demonstrate that the genotype VII marker vaccine is effective and provides a key tool for future ND eradication.

During cryopreservation, donkey semen is highly vulnerable to oxidative damage, impacting fertility. Although removing seminal plasma (SP) is standard practice for freezing, it eliminates the main antioxidants protecting against reactive oxygen species (ROS). This study aimed to evaluate the oxidative/antioxidant status and cryotolerance of donkey sperm cryopreserved with different permeable and non-permeable cryoprotectants. Thirty semen samples from ten donkeys were evaluated across eight treatments combining 5% DMF, 200 mM sucrose, 1% BSA, and 10% homologous SP, as follows: T1: DMF; T2: DMF/SP; T3: SUC/BSA; T4: SUC/BSA/SP; T5: DMF/SUC/BSA; T6: DMF/SUC/BSA/SP; T7: DMF/BSA; and T8: DMF/BSA/SP. Semen was frozen using a conventional freezing protocol. Post-thaw oxidative/antioxidant markers were evaluated, including total antioxidant capacity (TAC), ROS, the superoxide anion (O2[-]) and the hydroxyl radical (OH[-]). Sperm cryotolerance was assessed by motility, mitochondrial membrane potential (ΔΨM), and viability of frozen-thawed semen. For statistical analysis, mixed models were fitted and means were compared by the Tukey's test. The results showed that T6 and T8 showed higher total motility. Treatments containing SUC/BSA/SP exhibited higher TAC. Treatments T4, T7 and T8 showed higher ROS production, while T7 produced the highest O2[-]. Treatments T7 and T8 had the highest proportion of cells with high-ΔΨM; however, they exhibited the lowest sperm viability (p < 0.05). It is concluded that the use of different combinations of permeable and non-permeable cryoprotectants during semen freezing, including DMF, BSA, SUC, and SP, influences the oxidative/antioxidant status, as well as the cryotolerance of donkey sperm.