Veterinary Research Forum

Abstract This study evaluated the protective effects of intraperitoneal tannic acid (TA) against ischemia-reperfusion (I/R) injury in a rat model of testicular torsion. Eighteen adult male Wistar rats were randomized into three groups (n=6 each): sham (surgery without ischemia), I/R (3 hr ischemia + 3 hr reperfusion), and I/R+TA (TA 50 mg kg-1; 100 µL i.p. 30 min before reperfusion). Testicular tissues were sampled immediately after reperfusion for biochemical assays to measure malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels. Epididymides were analyzed 60 days later for sperm count, motility, viability, morphology, and DNA integrity. I/R significantly elevated MDA and sperm DNA damage while reducing SOD, GPx, sperm parameters, testicular weight and spermatogenesis. TA administration reversed these changes and restored the parameters to levels close to those of the sham group. Overall, intraperitoneal TA mitigated I/R-induced oxidative stress and preserved reproductive function, indicating its potential therapeutic value in testicular torsion.

Abstract Urinary bladder cancer, the tenth most common cancer globally, exhibits significant variability in the reported expression of estrogen receptor alpha (ERα), a key prognostic marker. Accurate and reproducible assessment of ERα is critical for elucidating its role in bladder cancer biology and identifying therapeutic targets. Aim: This study compared manual scoring with automated analysis using ImmunoRatio software for quantifying ERα expression in a chemically induced bladder cancer model in rats, using N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN). Methods: Twenty male Wistar rats were randomly assigned to two groups: a control group (n=8) and a group treated with BBN (BBN; n=12), administered in drinking water at 0.05% for 20 weeks. Animals were sacrificed and urinary bladders were collected for macroscopic and microscopic evaluation. Samples underwent standard histological processing and immunohistochemical analysis to assess ERα expression, quantified both manually and automatically. Results: The immunoexpression of ERα, detected by ImmunoRatio was found to be significantly higher than that obtained through manual counting in both groups (p < 0.0001). ImmunoRatio analysis indicated a higher ERα expression in the control group than in the BBN group (p < 0.001), whereas manual counting showed the opposite tendency, without statistical significance (p > 0.05). Conclusion: Our findings demonstrated that ImmunoRatio resulted in significantly higher ERα immunoexpression values than manual counting in both control and BBN-treated groups, with inconsistent group-wise trends between methods. These findings suggest that, although digital pathology tools offer advantages in standardization and reproducibility, their application requires tissue-specific calibration and validation to ensure accurate and reliable biomarker quantification.

Abstract Hydronephrosis is an uncommon condition in cats and dogs, characterized by progressive dilation of the renal pelvis and subsequent atrophy of the renal parenchyma. This report describes an 8-year-old spayed female domestic longhair cat diagnosed with left-sided hydronephrosis and concurrent polycystic kidney disease (PKD) after comprehensive clinical, laboratory, and imaging evaluation. Because the contralateral kidney appeared functionally preserved, unilateral nephrectomy was selected as the most appropriate treatment option. During surgical manipulation, however, the affected kidney unexpectedly separated completely from its attachments before vascular ligation could be performed, without evidence of hemorrhage. The procedure was completed without complication, and no intraoperative or postoperative adverse events were recorded. The patient recovered uneventfully and remained stable throughout the perioperative period. To the authors’ knowledge, this represents the first documented case in veterinary medicine of spontaneous renal autoavulsion occurring during nephrectomy. This unusual finding underscores the severe structural fragility that can result from chronic obstructive renal disease and advanced parenchymal destruction. In addition, it suggests that renal autoavulsion may represent a previously unrecognized end-stage manifestation of longstanding hydronephrosis, particularly when associated with concurrent renal pathology such as PKD. This case emphasizes the importance of thorough preoperative assessment, careful surgical planning, and awareness of potentially extreme anatomic alterations in chronically diseased kidneys

Abstract Pasteurella multocida is a widespread zoonotic pathogen associated with severe hepatic inflammatory responses. Although traditional medicinal plants such as Tribulus terrestris L. (TL) have demonstrated anti-inflammatory properties, their protective effects against Pasteurella multocida-induced liver injury remain insufficiently studied. This study established a Pasteurella multocida-induced liver injury model to evaluate the protective efficacy of TL extract. Thirty-two male Kunming mice were randomly divided into four groups: control, model, penicillin-streptomycin, and TL extract. Therapeutic interventions were conducted by daily intraperitoneal injection of P. multocida (50 CFU/mL) followed by treatment with antibiotics or TL extract (1000 mg/kg/day) for 7 consecutive days. The therapeutic potential of T. terrestris extract was evaluated using histopathological analyses (hematoxylin and eosin (H&E) staining and Masan staining), RT-qPCR, Western blot, immunohistochemistry, and immunofluorescence to assess inflammatory cytokine expression and NF-κB pathway activation. The extract significantly attenuated hepatic fibrosis and inflammatory cell infiltration in infected mice. Following Pasteurella multocida infection, NF-κB pathway activation may indicate by upregulated NF-κB1 and downregulated IκBα expression in mouse liver, alongside increased pro-inflammatory cytokines TNF-α and IL-1β. Treatment with antibiotics or TL reversed these changes. TL likely suppressed NF-κB activation by downregulating receptor TNFRsf1a. Immunohistochemistry and immunofluorescence further demonstrated decreased phosphorylation of IκBα and reduced IL-1β and TNF-α after treatment, indicating effective inhibition of NF-κB signaling and inflammation. These effects were comparable to those observed with antibiotic treatment. Our research reveals that TL extract alleviates Pasteurella multocida-induced hepatic inflammation by inhibiting NF-κB activation, highlighting its potential as a plant-derived therapeutic agent for bacterial liver injury.

Abstract The thyroid gland is an integral part of the complex anatomy of the equine neck. Transcutaneous ultrasonography provides a non-invasive method for assessing the thyroid glands parenchyma and dimensions in horses. This study evaluated 30 clinically healthy Turkmen horses whose health status was confirmed through comprehensive physical examination, complete blood count, serum biochemical analysis, and thyroid hormone analysis. The horses were divided into two age groups: Group 1 under 36 months (three males and eight females), and Group 2: Thirty-six months and older (eight males and eleven females). Ultrasonographic assessment of the thyroid region was performed using a high-frequency linear transducer. Each thyroid lobe was systematically evaluated for its dimensions, volume, morphology, contour, parenchymal echogenicity and echotexture. Statistical analysis revealed no significant age-related differences in thyroid parameters; however, sex-based differences were observed: females had significantly greater thyroid lobe length and volume on both the left and right sides compared to males. Qualitative B-mode ultrasonography showed consistent morphological features; in the longitudinal view, all thyroid lobes exhibited a flattened, oval shape. On transverse images, most lobes (83.93%, n = 47) appeared elliptical, while the remaining lobes (16.07%, n = 9) were spherical. Irrespective of shape, all thyroid lobes demonstrated regular contour margins. Regarding echogenicity, most lobes were hyperechoic, with a few being isoechoic, relative to the sternocephalic muscle. Furthermore, all thyroid lobes exhibited homogeneous to mildly heterogeneous echotexture. This study establishes diagnostic baselines for thyroid evaluation in the Turkmen horse breed aiding in the detection and management of thyroid pathologies.

Abstract Foot-and-mouth disease virus (FMDV) continues to pose a major threat to the livestock industry worldwide due to its high transmissibility and substantial economic consequences. For diagnostic and vaccinal purposes, necessitate the exploration of novel immunogenic targets is pivotal. Here, antigenic determinant (an epitope) sequence comprising core of 12 amino acids from VP1 protein of foot-and-mouth disease virus (FMDV) was designed by immuno informatics. To improve the physio-chemical and stability characteristics of the final peptide, additional residues were incorporated until it resulted in the synthesis of a 23-amino acid peptide (VARTLPTSFNYGAIKATRVTDEC, 2.5 Kda and PI 8.17). After chemical synthetizing with 95 % purity, the peptide was conjugated to bovine serum albumin (BSA) via EDC/NHS cross linker (Peptide: EDC: Sulfo-NHS: BSA (carrier protein): 10:40:50:1 in MES buffer (50 Mm)) and formulated with an aluminum hydroxide adjuvant. Twelve male New Zealand White rabbits were randomly assigned into three groups and main group received the peptide-BSA conjugate. Sera were direct to Dot blot, double immunodiffusion (DID) and serum neutralization (SN) tests, and also indirect peptide ELISA. The synthetic peptide elicited strong antibody response by primary positive results in dot blot and DID test (positive in 1:4 serum dilution) as well as it showed cross-reactivity with whole antigen. Although only low SN titers (1:8) were observed. Moreover, the peptide ELISA confirmed specific antibody responses against O serotype (OD of 2.4 at day 55 in 1/10 dilutions). Altogether, rationally designed epitope are promising regimes for the future development of FMDV novel diagnostic tools and vaccine cocktails.

Abstract Background: Aging significantly impacts cardiovascular health, increasing the prevalence of heart diseases such as acute coronary syndrome (ACS) and coronary artery disease (CAD). This study explores the combined effects of melatonin and Nicotinamide Mononucleotide (N) on ischemia-reperfusion injury (IRI) in aged rats, focusing on inflammatory markers and hemodynamic parameters.
Methods: Five groups were created from forty-year-old male Wistar rats: Sham, IR, M (IR + Melatonin), N (IR + N), and N+M (IR + N + Melatonin). Rats underwent IR procedures, and hearts were analyzed for coronary flow (CF), heart rate (HR), left ventricular end-diastolic pressure (LVEDP), and left ventricular developed pressure (LVDP). Creatine kinase (CK) levels and inflammatory markers (IL-1β, IL-6, TNF-α, NF-κB) were also measured.
Results: IR led to reduced CF and HR, both of which improved with melatonin and N treatments. Both treatments lowered the increase in LVEDP and enhanced LVDP recovery, with the combined therapy showing the best results. CK, IL-1β, IL-6, TNF-α, and NF-κB levels were elevated in the IR group but significantly reduced by the treatments, especially the combined therapy, demonstrating stronger protective effects.
Conclusion: Melatonin and N, particularly in combination, mitigate the adverse effects of IR injury in aged rats, improving hemodynamic parameters and reducing myocardial damage and inflammation. These findings highlight their potential as therapeutic agents for age-related cardiovascular conditions.

Abstract An emerging global threat is the development of resistance to various antibiotic groups among pathogenic bacteria, including P. multocida, The excessive use of antibiotics and presence of antibiotic resistance carriers play a significant role in the development of multiple resistances to antimicrobial agents. Given the role and importance of plasmids in the development of antibiotic resistance, it is important to prevent the transfer, replication, and multiplication of plasmids by eliminating plasmids. In this study, the role of plasmids in the development and transmission of antibiotic resistance in P. multocida was investigated and evaluated. This study was performed on of P. multocida isolates were collected from apparently healthy and ailing animals (February to September 2014). Identification of P. multocida isolates using standard biochemical tests and kmt1 gene amplification. After initial tests and plasmid DNA extraction, five plasmid-containing isolates were selected for further work. To compare antibiotic resistance after plasmid removal, the disk diffusion method was used. Based on the results, it was determined that all plasmid-containing isolates that showed a high level of antibiotic resistance (especially with large plasmids) became sensitive to antimicrobial agents after plasmid removal. Therefore, according to the results obtained, the role and importance of plasmids in the development of antibiotic resistance in P. multocida is determined. It was also determined in this study that the existence and persistence of plasmids by P. multocida bacteria is of great importance in the role of plasmids in the development of antibiotic resistance.

Abstract The present study how riociguat supplementation modulates in a Tris–egg yolk extender on post-thaw ram semen quality. Semen samples were collected from eight healthy, sexually mature Tuj rams (n = 48 ejaculates) during the breeding season and cryopreserved after supplementation with riociguat at concentrations of 0 (control), 0.5, 5, 50, and 250 µM. Post-thaw sperm quality was evaluated using computer-assisted sperm analysis (CASA) for motility and kinematic parameters, alongside assessments of plasma and acrosomal membrane integrity, mitochondrial membrane potential, DNA integrity, and oxidative stress markers including malondialdehyde (MDA) and glutathione (GSH).
Riociguat supplementation (5–250 µM) significantly modulated mitochondrial membrane potential and intracellular antioxidant capacity, as evidenced by reduced MDA and increased GSH levels (P < 0.05). Moreover, all riociguat-treated groups exhibited significantly improved DNA integrity compared with the control (P < 0.05). However, acrosomal integrity declined at the highest concentrations (50 and 250 µM), indicating potential detrimental effects associated with excessive dosing.
In conclusion, riociguat modulates post-thaw ram sperm function in a dose-dependent manner, exerting both beneficial and detrimental effects depending on concentration. Although fertility outcomes were not assessed in the present study, the observed biochemical and functional changes provide preliminary evidence that riociguat may influence key pathways associated with sperm resilience to cryoinjury, thereby justifying further investigations incorporating in vivo fertility and reproductive performance assessments.

Abstract Swine colibacillosis, caused by colonization of the piglet small intestine with enterotoxigenic strains of Escherichia coli, imposes a significant economic burden on the swine industry. Due to high mortality rates and the emergence of antimicrobial resistance, it is considered one of the major welfare concerns in the livestock industry. This study elucidated the genetic basis and expression patterns of key antibiotic resistance genes in porcine E. coli isolates with colibacillosis. Intestinal contents from five affected piglets were subjected to high throughput sequencing, revealing 26 resistance genes across seven antibiotic classes. Four strains (428, 430, 432, 433) were further analyzed for mRNA expression levels of six resistance genes (blaTEM, tetA, tetB, tetM, qnrB, and qnrS) under exposure to ampicillin, tetracycline, and ciprofloxacin at three concentrations (0.01%, 0.001%, 0.0001% w/v). Our findings revealed five distinct expression patterns: (1) dose dependent upregulation/downregulation; (2) inverse response to concentration changes; (3) maximal downregulation at intermediate dose; (4) minimal downregulation at intermediate dose; and (5) sustained upregulation. Strain 428 uniformly followed Pattern 1 for all resistance genes; in strain 430, tetB followed Pattern 3 while blaTEM matched Pattern 1; strain 432 exhibited Patterns 2 for all tetracycline- and β-lactam-resistance genes, and Pattern 4 for qnrB; strain 433 showed Patterns 3 for both fluoroquinolone-resistance genes, Pattern 4 for tetB, and Pattern 5 for blaTEM. These findings highlight strain-specific and dose-dependent gene expression responses in swine colibacillosis. This work underscores the need for tailored antibiotic strategies, targeted interventions and well-validated monitoring frameworks for antibiotic resistance in veterinary.

Abstract The rise of antimicrobial resistance (AMR) poses a severe global health challenge, with multidrug-resistant (MDR) bacteria causing millions of deaths annually and diminishing the effectiveness of traditional antibiotics. Bacteriophage-derived endolysins, such as PVP-SE1gp146 from Salmonella phage PVP-SE1, emerge as promising alternatives due to their high specificity, low toxicity, and minimal resistance development. However, their activity against Gram-negative bacteria is limited by poor outer membrane (OM) penetration. This study investigates the structural and functional impacts of fusing the polycationic PCNP peptide (KRKKRKKRK) to PVP-SE1gp146 to create engineered endolysin that enhance OM permeability while preserving enzymatic function.
Using advanced computational tools including Phyre2, GalaxyWEB, and AlphaFold2, three-dimensional models of native and engineered endolysins were generated and validated with Verify3D. Sequence analysis via BLAST identified homologs, and structural superposition with TM-align confirmed high conservation (TM-score 0.921, RMSD 1.84 Å) despite the PCNP insertion. Active sites were predicted using PrankWeb, and molecular docking with AutoDock assessed interactions with peptidoglycan components (e.g., NAM-NAG dimer, tetramer). Results revealed superior binding affinities in the native endolysin (e.g., -10.35 kcal/mol for NAM-L-alanine) compared to the engineered variant (-9.65 kcal/mol for NAM-NAG-L-alanine trimer), with positive energies for larger ligands indicating steric hindrance.
This trade-off—reduced affinity for improved OM penetration—supports enhanced bactericidal activity against MDR pathogens like Pseudomonas aeruginosa. The findings highlight the potential of rational engineering for superior antimicrobials, emphasizing the need for molecular dynamics simulations and experimental validation to optimize Artilysin design in combating AMR.

Abstract Interstitial cells of Cajal (ICC) play a crucial role in regulating gastrointestinal motility, functioning as pacemakers in the digestive system of equids. This study described and compared the ICC populations in different regions of the stomachs of equids (horses, donkeys and mules). A total of 30 healthy stomachs from horses, donkeys, and mules were evaluated using immunohistochemistry and high-resolution microscopy to assess various regions of the stomach in these species. The results showed the presence of ICC in all three equid species, with a predominance in the submucosa across the squamous mucosa, glandular mucosa and pyloric antrum. The density and morphology of ICC varied slightly between species, although no significant differences were found. Among the equids studied, donkeys showed the lowest ICC density, while horses exhibited the highest in the evaluated tissues. Four cellular morphologies were identified, three of which were included in the analysis, as toluidine staining revealed that one corresponded to mast cells. Further studies are needed to elucidate dynamics and possible alterations in response to gastric pathologies.

Abstract Pasteurella multocida and Mannheimia haemolytica are significant bacteria of respiratory tract infections in cattle. Identifying whether these bacteria are commensal or pathogenic is crucial for understanding their role in predisposing calves to respiratory tract infections. This study aimed to determine the prevalence and pathogenic characteristics of P. multocida and M. haemolytica in nasal swab samples of clinically healthy calves in Konya, Turkey, and to assess antibiotic resistance profiles. Using conventional culture methods, 5 samples (1.67%) from 4 different farms were positive for P. multocida and 3 samples (1%) were positive for M. haemolytica. PCR of nasal swab samples yielded P. multocida positivity in 17 (5.67%) samples and M. haemolytica positivity in 13 (4.33%), including all culture-positive samples. While all P. multocida isolates were atypical for the capsule, 18% of the PCR-positive samples were L2, 70% L3A, and 12% L8 serotypes. The ompA, omp87, hghA, and tbpA genes were the most prevalent, while the presence of other virulence genes varied. High rates of ampicillin/cloxacillin (100%) and streptomycin (80%) resistance were observed among the isolates. The rates of serotypes A1 and A6 in M. haemolytica PCR-positive samples were 53.85% and 46.15%, respectively. The ssa-1 gene was detected in 33.33% of the isolates. Moderate resistance to trimethoprim/sulfamethoxazole, streptomycin, and neomycin (66.67%) was recorded. The findings revealed the presence of P. multocida and M. haemolytica with various virulence characteristics and antimicrobial resistance profiles in the nasal microbiota of healthy calves, providing novel insights that could inform improved herd management, disease control, and treatment strategies.

Abstract Babesia caballi is a tick-borne protozoan parasite responsible for causing equine babesiosis, a disease with considerable variation in prevalence across different regions of the world. This report presented a confirmed case of B. caballi infection in a three-year-old mare originating from Gonbad-e Kavus, Iran, highlighting the clinical, laboratory and diagnostic aspects of the disease. The affected mare showed classical clinical signs including persistent fever, generalized weakness, tachycardia and pale mucous membranes, which are indicative of systemic infection and anemia. Laboratory evaluations revealed evidence of hepatic and renal compromise as well as hematological parameters consistent with active inflammatory processes. Microscopic examination of blood smears and molecular techniques such as PCR were employed to confirm the presence of B. caballi, while simultaneously excluding co-infections with other common equine pathogens including Theileria equi and Anaplasma phagocytophilum. This case underscored the critical need to combine conventional diagnostic approaches with molecular methods to achieve accurate detection, effective treatment and appropriate management strategies for equine babesiosis in endemic regions.

Abstract The excessive production of reactive oxygen species during the freezing and thawing process triggers lipid peroxidation in the sperm membrane, resulting in oxidative harm and a decline in semen quality. The aim of this study was to examine the impact of different concentrations of canthaxanthin on the quality of goat spermatozoa throughout the process of cryopreservation. Semen was collected from each goat using an artificial vagina for a continuous duration of 3 weeks. The semen was divided into five equal portions and mixed with an extender having different concentrations of canthaxanthin: 0.00 (control), 5.00, 15.00, 25.00, and 35.00 µM. The computer-assisted semen analyzer was used to determine the sperm motility and kinetic parameters. The findings indicated that the inclusion of canthaxanthin at a concentration of 5.00 µM resulted in a minimal enhancement in sperm kinematic factors such as overall motility, progressive motility, fast progressive motility, curve-line velocity, distance curve line, amplitude of lateral head displacement, beat-cross frequency, wobble, linearity, and straightness. Nevertheless, these enhancements did not demonstrate statistical significance compared to the control, 15.00, 25.00, and 35.00 µM. The introduction of 5.00 µM of canthaxanthin resulted in a significant rise in straight-line velocity, average path velocity and head activity compared to the control group. Hence, the addition of 5.00 µM of canthaxanthin could be employed to improve the cryopreservation of goat spermatozoa.

Abstract Bovine tuberculosis is a chronic bacterial disease primarily caused by Mycobacterium bovis, a member of the Mycobacterium tuberculosis complex (MTBC), with significant zoonotic implications. This study aimed to detect MTBC in wildlife species, specifically nilgai (Boselaphus tragocamelus) and sambar deer (Rusa unicolor), using gross pathology, histopathology, acid-fast staining, and molecular confirmation. Necropsied tissue samples were collected during post-mortem examination of a nilgai and a sambar deer from the Assam State Zoo, Guwahati, India. Macroscopically, multiple granulomatous tubercles of varying sizes were observed in the lungs and liver, with creamy white caseous material marked upon sectioning. Ziehl-Neelsen staining of the tissue smears from granulomatous lesions confirmed the presence of acid-fast bacilli. Microscopic examination of tuberculosis granulomas revealed a central necrotic mass surrounded by inflammatory cell infiltration, including Langerhans-type giant cells. Molecular confirmation of MTBC infection was achieved by amplifying hsp65 and IS1081 in tissue samples, further validated by Basic Local Alignment Search Tool for nucleotide analysis following Sanger dideoxy sequencing. In conclusion, this study confirmed the presence of tuberculosis in these wildlife species through an integrated approach combining pathology, microbiology, and molecular diagnostics, highlighting the need to understand pathogen entry into the herd and prevent potential spillover.

Abstract Wild birds play a significant role in the dissemination of micro-organisms, including fungal agents, through feathers, skin debris, and feces, both locally and across regions via migration. Identifying fungal carriage in wild birds is crucial for understanding the prevalence of pathogenic fungi and developing effective strategies to prevent and control mycotic infections. This study investigated fungal carriage in wild pigeons, doves, water birds, birds of prey, and small passerines in Balıkesir province of Türkiye, an ecologically important region characterized by key wetlands, two national parks, and major migratory bird routes. A total of 46 wild bird samples were analyzed by culture method, as the “gold standard”, and several clinically important fungal isolates were obtained and subsequently further analyzed by PCR. Fungi were detected across all bird categories, with 27 birds (58.70%) testing positive for at least one fungal species. In total, 10 distinct fungal taxa were isolated from the wild birds. The most frequently identified organisms were Mucor spp. (28.26%) and Aspergillus niger (6.52%), followed by Sporothrix schenckii (4.35%), Candida glabrata (4.30%), Arthroderma terrestre (2.17%), Trichophyton mentagrophytes (2.17%), Nannizzia gypsea (2.17%), Aspergillus flavus (2.17%), Aspergillus fumigatus (2.17%), Lichtheimia spp. (2.17%), and Beauveria spp. (2.17%). Prevalence was the highest in birds of prey and small passerines (77.78%), moderate in pigeons and doves (58.82%), and the lowest in water birds (27.27%). These results emphasize the role of wild birds as potential reservoirs and vectors of fungal agents, highlighting the importance of continued surveillance and biosecurity measures to mitigate the risk of environmental and zoonotic transmissions.

Abstract Equine glandular gastric disease (EGGD) has limited prevalence studies and scarce understanding of the predisposing factors and pathophysiological mechanisms involved. The objectives of this study were to determine and compare the post-mortem prevalence of EGGD and other gastric lesions in Colombian equids, specifically horses, donkeys, and mules. The study utilized a sample of 300 equids stomachs, with 100 specimens from each group, collected immediately after slaughter. The post-mortem examination included a thorough inspection of the ventral fundus, adjacent area of the margo plicatus, and pyloric antrum of the glandular mucosa, carefully identifying any lesions or abnormalities. The results showed that the overall macroscopic presence of EGGD in the equid population was 68.30% on average, with specific rates of 45.00% in horses, 77.00% in donkeys, and 83.00% in mules, with lesions predominantly affecting the fundus and pyloric antrum regions. Inflammatory processes were observed to be similar between donkeys and horses, while mules exhibited distinct inflammatory patterns in the glandular mucosa. Additional gastric findings, such as gastritis (85.30%), gastric impaction (10.00%), and parasitic infections (13.30%), were also documented during the study. The findings suggested that EGGD was prevalent across all three groups of equids. Although the inflammatory conditions and their distribution were similar in horses and donkeys, differences in the location, frequency, and severity of mucosal lesions were observed among the three species.

Abstract This study investigated the effects of zinc oxide nanoparticles (ZnO-NPs) on the growth, blood parameters, skin mucus, and serum biochemistry of juvenile common carp (Cyprinus carpio). A total of 300 fish with an average weight of 12.00 ± 0.33 g were randomly distributed in 12 fiberglass tanks with a capacity of 300 L and fed with diets supplemented with ZnO-NPs at concentrations of 0 (control), 10.00, 50.00, and 100 mg kg^-1 for 60 days. The results indicated that 50.00 mg kg^-1 ZnO-NPs supplementation significantly improved fish weight, length, specific growth rate, and feed conversion ratio. There was no significant difference in survival rates between the treatments and control group; however, the highest survival rate was observed in the treatment with 50.00 mg kg^-1 ZnO-NPs. This treatment also led to the highest red blood cell counts, hemoglobin levels, mean corpuscular hemoglobin concentration, and lymphocyte percentages. In contrast, the highest neutrophil counts were observed at 100 mg kg^-1 ZnO-NPs. The highest serum biochemical parameters, including total immunoglobulin, lysozyme, and Zn levels were observed in the treatment with 50.00 mg kg^-1 ZnO-NPs. While, in the skin mucus, total immunoglobulin, lysozyme, and alkaline phosphatase significantly increased at groups fed with 10.00 mg kg^-1 ZnO-NPs. Overall, the findings suggest that dietary inclusion of 50.00 mg kg^-1 ZnO-NPs can effectively promote growth and enhance immune responses of C. carpio juveniles, indicating their potential application in aquaculture practices. However, caution is advised at higher concentrations due to the potential adverse effects on fish health.

Abstract This study is the first to apply the behaviour, immunity, nutrition and environment (BIENE) protocol to assess dairy ewe welfare in intensive farms in Algeria. A total of 1,964 ewes from 50 farms in the Algiers region were evaluated between December 2022 and March 2023 using 13 animal-based indicators related to nutrition, housing, health, and behavior. Moderate tooth loss affected 54.67 ± 3.32% of ewes, and 20.38 ± 2.78% had a low body condition score (< 2). Housing problems included fleece moisture (11.28 ± 2.55%) and over 40.00% soiling of flanks and hindquarters. Health issues, such as mild anemia (23.55%), lameness (26.45%), nasal discharge (35.47 ± 4.43%), and labored breathing (8.79 ± 1.47%), were observed. In terms of behavior, most ewes appeared attentive and active, although 40.55% showed fear and 19.90% aggression. Flight time increased from 2.50 ± 0.07 sec to 3.80 ± 0.07 sec, and animals maintained greater distance from the corridor (2.79 ± 0.08 m) than from the observer (1.85 ± 0.06 m). Principal component analysis of qualitative behavior assessment identified three clusters, including well-being, discomfort, and vitality. Spearman correlation analysis showed that relaxed ewes had better body condition score (r = 0.29) and were less likely to be lame (r = – 0.32) or anemic (r = – 0.37). Vigorous ewes showed fewer signs of respiratory distress (r = – 0.32), while apathetic ones were more prone to poor conditioning (r = 0.31) and injuries (r = 0.30).The BIENE protocol proved effective in detecting major welfare issues in intensive Algerian sheep farming.