Abstract Introduction Staphylococcus aureus is ubiquitous as a commensal bacterium and a frequent opportunistic pathogenic one in animals including livestock. The aim of this study was to analyse the expression levels of ovine cytokine genes and acute-phase proteins in response to the presence of S. aureus . Material and Methods Four different anatomical swabs, milk and blood samples came from 50 ewes from the native Uhruska and Świniarka breeds. The Uhruska breed exhibits high fertility and meat performance and the Świniarka breed is considered a primitive breed. Isolation and identification of S. aureus , haematological analysis, and analysis of cytokine and acute phase protein gene expression levels were performed. Results Staphylococcus aureus strains were isolated from swabs and milk from sheep of both breeds. A significant increase in IL6 and Hp (haptoglobin) gene expression was observed in leucocytes in samples from Uhruska sheep in which the presence of S. aureus was detected. Conclusion The expression level of selected cytokine and acute-phase protein genes have shown that even local carriage of S. aureus in small ruminants can lead to a systemic immune response, manifested by increased expression of IL6 and Hp genes in leucocytes.
Abstract Introduction Feline infectious peritonitis (FIP) is a fatal disease caused by feline coronavirus, most commonly associated with effusions in body cavities, while pericardial involvement remains rarely described. Material and Methods A three-year clinical study (2021–2024) included 256 cats with PCR-confirmed effusive FIP, of which 37 presented fluid accumulation exclusively in the pericardial sac. Diagnosis was based on real-time PCR analysis of pericardial fluid, echocardiographic examination and measurement of serum N-terminal pro-B-type natriuretic peptide (NT-proBNP) concentrations. Results Mutant feline coronavirus RNA was detected in pericardial fluid samples from all cats, with threshold cycle values ranging from 22 to 26 and highly specific PCR amplification confirmed by consistent melting temperatures (81.0–81.2°C). Ultrasonography revealed pericardial effusion in all 37 cats, with cardiac tamponade observed in 27% of cases and significantly higher NT-proBNP concentrations in affected individuals. Statistical analyses showed no significant associations between breed, age or sex and the occurrence of pericardial effusion, cardiac tamponade or NT-proBNP serum concentrations. Conclusion Pericardial effusion and cardiac tamponade may represent under-recognised manifestations of effusive FIP. Routine echocardiographic evaluation and molecular analysis of pericardial fluid should be considered in cats with suspected FIP and cardiopulmonary signs.
Abstract Introduction Disinfectants are essential for preventing infectious diseases in livestock, particularly in cattle footbath applications. Conventional culture-based methods for assessing bactericidal efficacy are reliable but time-consuming, leaving the need unmet for rapid laboratory-based screening approaches. Material and Methods Thirteen commercially available cattle footbath disinfectants were evaluated using the EN 1656 dilution–neutralisation method and flow cytometry (FCM) with propidium iodide (PI) staining. A binary logistic regression model was developed to relate the proportion of PI + cells to bactericidal efficacy. Results Disinfectants containing iodine or iodide, organic acids, hydrogen peroxide, quaternary ammonium compounds, glutaraldehyde or alcohol showed high bactericidal efficacy, whereas those with zinc sulphate were ineffective. The predictive model demonstrated good performance (AUC = 0.787) and identified an optimal threshold of approximately 80% PI + cells, yielding 85.47% sensitivity and 76.92% specificity. Agreement between FCM and the EN 1656 method was high, with 7.69% inconsistent classifications. Conclusion Flow cytometry represents a rapid and efficient laboratory-based screening method with the potential to complement culture-based approaches for evaluating bovine footbath disinfectant efficacy.
Introduction:Mycoplasma gallisepticum (MG) imposes heavy losses on the goose industry in China, yet its goose-specific biology is poorly understood. In elucidation of the biology, the pathogenicity and immunogenicity of MG in geese were analysed. Material and Methods:Goslings were divided into three groups and either infected with the goose isolate MG-GD01/22, infected with chicken-derived MG R(low) or left untreated as controls. Clinical scores were recorded, and morbidity/mortality, air-sac lesions and tracheal and lung histopathology evaluated. Isolate membrane proteins were separated by SDS-PAGE and screened by Western blot with goose hyper-immune sera. Results:Both MG-GD01/22 and MG R(low) established infection, with morbidity rates of 90% and 80% and qPCR positivity of 100% and 80%, respectively. The MG-GD01/22 isolate caused 20% mortality and more severe respiratory signs, whereas MG R(low) induced milder disease without deaths. Both strains produced airsacculitis and interstitial pneumonia, but MG-GD01/22 caused more marked pulmonary haemorrhage. Gel electrophoresis showed similar bands at 50-130 kDa with differences at 25-50 kDa. Western blot revealed strong immune responses to conserved 75 kDa and 67 kDa proteins, particularly those of MG-GD01/22. Conclusion:Both strains are pathogenic in geese, with MG-GD01/22 exhibiting higher virulence. The conserved 75 kDa and 67 kDa proteins represent promising targets for the development of diagnostic assays and vaccines against Mycoplasma gallisepticum infection in geese.
Introduction:The tectum opticum, which is a section of the mesencephalon, is located between the cerebellum and the ventral hemispherium of birds. In birds, the tectum opticum has a well-developed, thick, layered structure. It is quite large compared to those of other vertebrates and is considered the primary pathway of visual information to the telencephalon. Heat-shock proteins (HSPs) are molecules that help fold newly synthesised polypeptides, assemble multiprotein complexes and transport proteins across cellular membranes. Material and Methods:In this study, 50 fertilised quail eggs were used to monitor embryonic development. The fixed tissues were blocked in paraffin, and Crossmonn's triple stain was applied to these sections for histological examination. The avidin-biotin-peroxidase complex technique was applied to determine the immunohistochemical localisation of HSP70. Results:Histologically, the tectum opticum was observed to consist of six main layers. They were formed starting from embryonic day 7 (E7). Ten sublayers of the stratum griseum et fibrosum superficiale layer became clear starting from embryonic day 12. Immunohistochemical staining showed HSP70 immunoreactivity in neuron bodies and nerve fibres starting from E7. Conclusion:The findings suggest that HSP70 contributes to the protection of cells and the continuity of developmental processes. The regulatory effect of HSP70 on cell cycle, apoptosis and differentiation processes during embryonic development is of vital importance for the healthy development of avian embryos. Therefore, it is clear that the mechanisms of HSP70 expression need to be known.
Introduction:Merkel cell carcinoma is an aggressive neuroendocrine skin malignancy rarely reported in non-domestic species. Material and Methods:A cutaneous nodule from an endangered Caucasian squirrel (Sciurus anomalus pallescens) was examined using histopathology, immunohistochemistry, imaging mass cytometry, spatial transcriptomics (10× Visium) and metagenomic sequencing. Results:Histology revealed a high-grade neuroendocrine carcinoma with frequent mitoses (52 per 2.37 mm2) and necrosis. Tumour cells were positive for cytokeratin 20 (paranuclear dot pattern), synaptophysin and chromogranin A, with a high Ki-67 index (68%). Spatial analyses delineated a distinct tumour core and combined invasive front and stromal compartments, revealing upregulation of neuroendocrine (atonal basic helix-loop-helix transcription factor 1 and neurogenic differentiation factor 1) and proliferative (marker of proliferation Ki-67) programmes, and activation of phosphoinositide 3-kinase-AKT serine/threonine kinase 1-mechanistic target of rapamycin and mitogen-activated protein kinase pathways. No evidence of Merkel cell polyomavirus was found. The tumour microenvironment was immune-excluded, with programmed-death ligand 1 expression on ~22% of tumour cells and CD8+ T cells restricted to the stroma. Conclusion:This study provides a comprehensive methodological framework for high-resolution tumour profiling in conservation pathology and highlights the emergence of neoplasia in threatened wildlife.
Introduction:Parabens, organic substances used as preservatives in various branches of industry, penetrate the environment and living organisms. There is only scant knowledge of cats' exposure to parabens. Therefore, the aim of the present study was to assess the levels of common industrial parabens in samples of cat hair. Material and Methods:Concentration levels of four parabens (methylparaben - MeP, ethylparaben - EtP, propylparaben - PrP and butylparaben - BuP) were analysed in cat hair samples (n = 70) using a liquid chromatography-triple quadrupole mass spectrometer. The signalment, body condition score and possibility of outdoor access were recorded for each cat. Results:All four parabens were found in the cat hair. The concentrations of parabens in pg/mg were as follows: MeP - range 28.7-7,094.7, median 273.5 and mean 620.6 ± 1,193; EtP - range 10.8-6,458.2, median 43.1 and mean 188.2 ± 778.1; PrP - range 6.6-1,793, median 33.2 and mean 107.5 ± 246.9; and BuP - range 8.9-276.5, median 11.5 and mean 27.2 ± 49.5. Statistically significantly higher levels of PrP were noted in strictly indoor cats compared to cats with outdoor access. Statistically significant differences in paraben levels between animals of different sexes, ages and body conditions were not observed. Conclusion:The obtained results have shown that cats are exposed to parabens to a considerable extent, which suggests that the exposure to these compounds may negatively affect cats' health.
Introduction:The widespread distribution of the blood-sucking parasitic species Ashworthius sidemi among cervids highlights the need for increased understanding of how to prevent its transmission to livestock. Such knowledge should include detailed data on its life cycle, which may provide insights into the pattern of invasion and guide control measures. Material and Methods:Adult and subadult specimens of A. sidemi were obtained by necropsy examinations of 108 naturally infected cervids between 2011 and 2021. The month of host collection was used to divide the obtained data into two periods of the year: warm (May to October) and cold (November to March). The differences in the overall mean intensity of infection within a particular month and between periods were statistically analysed. Molecular analyses were conducted to confirm species identification. Results:A total of 16,574 specimens were found. The mean intensity of infection for adults and for subadults differed significantly, both within a particular month and between the two periods of the year (P-value < 0.01). Correspondence analysis revealed two distinct groups based on the month of collection from the host and the increased occurrence of adults and/or subadults in the abomasum. The occurrence of adult specimens was the greatest during the warm periods of the year, with their first appearance recorded in May. Conclusion:Although infection intensity remains relatively stable throughout the year, eggs shedding may be influenced by the occurrence of adults, what depends on the period of the year.
Introduction:Antimicrobial resistance (AMR) poses a global threat to public and animal health, and its progression is closely linked to the use of antimicrobials in veterinary medicine. In Europe, programmes such as the European Surveillance of Veterinary Antimicrobial Consumption (ESVAC) and subsequently European Sales and Use of Antimicrobials for Veterinary Medicine (ESUAvet) monitor and attempt to reduce the consumption of antimicrobials in food-producing animals. The study analyses the trends in the volume and sales structure of veterinary antimicrobials in Poland in 2011-2023 and compares them with data reported by other European countries. Material and Methods:The analysis was based on data from ESVAC reports (2011-2022) and the first ESUAvet report (2023), as well as data from the European Medicines Agency's Antimicrobial Sales and Use Platform. The data were evaluated by antimicrobial class, pharmaceutical form and Antimicrobial Advice Ad-hoc Expert Group (AMEG) categories, the latter being A (avoid), B (restrict), C (caution) and D (prudence). Results:While total sales of veterinary antimicrobials in Europe decreased by 48.0% between 2011 and 2023, in Poland they increased by 19.5% during the same period. In Poland, the predominance of AMEG category D antimicrobials persisted, although their share declined from 79% to 64% with a concurrent increase in sales of category C. The sales structure shifted significantly towards oral solutions, replacing oral powders. Penicillins became the dominant antimicrobial group in the later years, while tetracyclines sales declined. Conclusion:The Polish trend diverges from the European one of reducing veterinary antimicrobial consumption. To align with EU reduction targets, Poland should strengthen antimicrobial stewardship, tighten prescription control and promote preventive measures such as vaccination and higher biosecurity within the One Health framework.
Introduction:Inguinal hernias are a significant health and economic problem in pig farming, affecting animal welfare and production profitability. Suspected aetiological factors include disorders in the metabolism of extracellular matrix components, including collagen, which is responsible for the integrity and strength of connective tissue. The aim of the study was to compare the concentrations of type-I, -III and -IV collagen in the blood serum of pigs with inguinal hernias to those in healthy animals. Material and Methods:The study used 87 pigs, comprised of 59 hernia-affected individuals in the experimental group and 28 unaffected counterparts in the control group. The animals were of the same age and came from the same genetic line. The study used ELISA tests for the quantitative measurement of serum collagen concentrations. Results:The results showed significantly higher concentrations of type-I (P-value < 0.001) and type-III (P-value < 0.01) collagen in the experimental group, indicating an association between collagen metabolism and hernia status. For type-IV collagen, the differences were not statistically significant. Conclusion:Changes in type-I and type-III collagen levels may be associated with inguinal hernia status in pigs and warrant further investigation as potential risk biomarkers. Type-IV collagen showed no relationship with hernia status in the study population.
Introduction:Contagious ecthyma (CE) is a skin disease of goats, sheep and some other domesticated and wild ruminants worldwide, including Poland. The infection is caused by the orf virus (ORFV) of the Parapoxvirus genus. It is characterised by the appearance of skin lesions in the form of papules and pustules, most often in the area of the mouth and nostrils, hooves and genitals. The disease causes economic losses, especially in countries with high sheep and goat production. In Europe, the threat posed by this disease is rather underestimated, but following the re-emergence of foot-and-mouth disease (FMD) in the region in 2025, there is increased vigilance and a need to have differential diagnostic methods proven and ready. Material and Methods:DNA was extracted from lesions on mouth and hooves of sheep. The animals were on a farm located in southwestern Poland on which FMD was ruled out. A PCR was performed using two pairs of specific primers for detecting vIL10 and GIF gene fragments which were then sequenced. Results:Sequencing confirmed ORFV infection in all tested sheep samples. The sequences obtained for Polish viruses were also highly homologous to other parapoxviruses, indicating that the method used may be applicable for broad diagnosis of parapoxvirus infections not only in ruminants. Conclusion:This is the first molecular characterisation of ORFV in sheep in Poland. It signals a potential herd biosecurity problem in view of the neglected but re-emerging nature of CE. The marked genetic divergence between Polish field ORFV isolates and currently available vaccine strains may influence CE vaccine efficacy in Poland.
Introduction:This study compared the vaginal microbiota composition of dairy cows with follicular and luteal ovarian cysts using metagenomic analysis. Material and Methods:Ovarian cysts, which impair reproductive performance through endocrine disruption, were diagnosed by ultrasonography and serum hormone evaluation in Holstein cows 30-60 d postpartum. Forty-five cows were initially included and divided into follicular cyst, luteal cyst and control groups. Vaginal lavage samples were analysed using third-generation sequencing, and taxonomic classification was performed through 16S rRNA gene analysis. Results:A total of 258 operational taxonomic units (OTUs) were identified, with the highest diversity observed in the control group (mean of 56.8 OTUs) and the lowest in the luteal cyst group (mean of 49.0 OTUs). Proteobacteria was the dominant phylum across all groups (93.4%), followed by Tenericutes (5.9%). Firmicutes, Bacteroidetes and Fusobacteria accounted for less than 1%. At the family level, Burkholderiaceae (62.7%) and Pasteurellaceae (24.0%) were predominant, while of the genera, Ralstonia was the most abundant (62.2%). The luteal group had the highest relative abundance of Burkholderiaceae, whereas Pasteurellaceae was most abundant in the control group. Conclusion:These results indicate that cystic cows exhibit reduced microbial diversity and altered bacterial composition in comparison with healthy animals. The predominance of Proteobacteria and Ralstonia suggests a potential link between endocrine imbalance and changes in the vaginal microenvironment. Hormonal analyses supported the classification of cyst types, with follicular cyst cows showing low progesterone (0.31 ± 0.05 ng/mL) and high oestradiol-17β concentrations (55.57 ± 7.91 pg/mL), whereas luteal cyst cows exhibited higher progesterone (2.89 ± 0.74 ng/mL) and lower oestradiol-17β concentrations (6.19 ± 0.56 pg/mL) (P < 0.001). These results may support future studies evaluating vaginal microbial profiles as complementary indicators of ovarian status in dairy cows.
Introduction:Pasteurella multocida is an important pathogen in many species of domestic and wild mammals, as well as birds. It is also a commensal in the nasopharynx of clinically healthy animals. The aim of the study was to characterise and classify field strains of P. multocida isolated from rabbits in Poland. Material and Methods:A total of 115 field strains isolated from rabbits in 1999-2020 were used. The examined strains were evaluated morphologically and microscopically, and their ability to produce catalase and oxidase was determined. Identification of P. multocida strains was performed on basis of biochemical characteristics and serotype classification. Results:Based on the biochemical profile assessment using API 20E and the ID 32E (bioMérieux), the strains isolated from rabbits were identified as P. multocida. Among 17.4% were classified as P. multocida subsp. multocida, 80% as ornithine decarboxylase negative P. multocida subsp. multocida and 2.6% as P. multocida subsp. septica. The capsule of type A was found in 87.8% of strains, type D in 8.7%, and type F in 3.5%. The somatic antigen of serotype 12 was present in 64.3% of strains, and serotype 3 in 36.7% of strains. The capsular type A was predominant among strains from rabbits that were asymptomatic carriers, while the capsular type D and F were found mainly in strains isolated from sick animals. In turn, the somatic antigen of serotype 12 was present in strains from both healthy and diseased animals, while strains of serotype 3 were much more frequently isolated from diseased rabbits. Conclusion:The conducted research allowed to classify and obtain new in-depth data on the population structure of P. multocida strains isolated from rabbits in Poland.
Introduction Alternative methods of duck feeding can be good solutions for reducing production costs on small farms. Ensiled corn grain is a good source of energy that can be used in the rearing of broiler ducks. This research investigated productivity effects and meat quality parameters in experimental broiler ducks provided this material as feed portions.Material and Methods Broiler ducks were divided into a control group (CON) and two experimental groups (CSMIX and CD+SG). In the CSMIX group, birds were fed a commercial diet mixed with ensiled corn grain in a ratio of 80:20. In the CD+SG group, the commercial diet was limited, and ensiled corn grain was ad libitum.Results The highest body weight and weight gain were found in the CSMIX group. A lower feed intake was found in the control and CD+SG group than in the CSMIX group throughout the rearing. A higher final feed conversion ratio characterised ducks from the experimental groups and a higher pectoral muscle and total muscle weight than the CD+SG group was noted in carcasses of ducks from the CON group. In the pectoral and leg muscles of the CON and CSMIX groups, the content of polyunsaturated fatty acids was higher than in these muscles of the CD+SG group.Conclusion Duck feeding with ensiled corn grain (80:20) could be a feeding strategy for small farms because it yields favourable growth parameters, carcass composition and high polyunsaturated fatty acid content in meat, and reduces feed costs.
Introduction: Reproductive efficiency in swamp buffalo (Bubalus bubalis) remains constrained by the lack of molecular tools to identify fertile females before service. The aim of this study was to discover biologically grounded biomarkers of fertility. Material and Methods: Cervical mucus was profiled using high-resolution proteomic analysis, with quantitative and functional enrichment used to prioritise biologically relevant fertility markers. Results: The cervical-mucus proteome exhibited a conserved secreted core with marked quantitative shifts that separated favourable-quality from poor-quality mucus. Favourable mucus was characterised by a barrier- and adhesion-associated profile with antimicrobial and antioxidant features, its proteins including integrin/extracellular matrix interactors, redox enzymes, lipocalin 2 (LCN2), secretory leukocyte protease inhibitor (SLPI) and heat-shock protein B6 (HSPB6). Poor mucus showed a neutrophil-acute-phase axis with complement engagement and oxidative chemistry, typified by myeloperoxidase (MPO), pentraxin 3 (PTX3) and C-reactive protein (CRP). Functional enrichment emphasised extracellular and host-defence processes consistent with these opposing mucosal states. These six proteins were consistently the leading discriminants and had coherent biological roles. Conclusion: Swamp-buffalo cervical mucus is variously in a barrier-competent state and a neutrophil-dominant state. This dichotomy provides a mechanistic explanation for fertility differences at the cervix and yields actionable, secreted protein biomarkers - LCN2/SLPI/HSPB6 in favourable mucus and MPO/PTX3/CRP in poor mucus - for targeted assays and pre-service screening in tropical production systems.
Introduction: Contamination with herbicides is a frequent environmental issue. The aim of the study was to evaluate haematological and blood biochemical parameters, and the microstructure of selected organs in control and herbicide-exposed fish. Material and Methods: In this study, 108 common carp (Cyprinus carpio) were exposed to a commercial herbicide with nicosulfuron as its active substance. Three groups of fish were established: control, TAM1 (nicosulfuron at 1 mg/L), and TAM2 (nicosulfuron at 5 mg/L). Treatment lasted 1, 3 or 10 days. Functional (pathophysiological) and structural (histopathological) alterations in control and herbicide-exposed fish were evaluated. Results: Various haematological changes and some biochemical alterations were detected. The pathophysiological changes suggested erythrocyte damage, compensatory response (as indicated by an increase in red blood cell count and erythroblast percentage), stress response (as evidenced by an increase in glucose concentration) and possible lipid metabolism disorders (as indicated by a decrease in cholesterol concentration). More differences in pathophysiological parameters were detected between TAM2 fish and control fish than between TAM1 fish and control fish. Histopathological analysis revealed renal tubule necrosis, of which the severity depended on herbicide concentration and exposure duration. Conclusion: The exposure of common carp to the tested nicosulfuron-based herbicide formulation led to pathophysiological and histopathological alterations, indicating the need for systematic detection and quantification of nicosulfuron in aquatic environments.
Introduction:The purpose of this study was to determine how the duration of administration of β-glucans derived from the yeast Saccharomyces cerevisiae modifies cellular immunity in one-year-old leuciscids, specifically vimba bream (Vimba vimba). Material and Methods:The effects of two treatments were compared at three endpoints using a 2 × 3 factorial design, all groups in triplicate aquaria (n = 3). Kept in controlled recirculating aquaculture system conditions, three experimental groups were fed for two, four or eight weeks a commercial starter feed, and three other groups received the same feed supplemented with a commercial formulation of β-glucan polysaccharides from the cell wall of Saccharomyces cerevisiae yeast at 0.02%. Results for fish taking β-glucan were compared to the respective control groups of fish not given it. Results:Two-week administration of β-glucan in the diet resulted in significantly higher pinocytosis, respiratory burst activity and potential killing activity of head-kidney phagocytes in supplemented fish compared to phagocytic activities in fish not receiving β-glucan. However, in the groups receiving β-glucan for four and eight weeks, this activity was significantly lower than in the respective control groups. After eight weeks, T-lymphocyte activity was also lower. Conclusion:Under controlled recirculating aquaculture system conditions favourable for fish growth, the most beneficial effect of dietary β-glucan supplementation on vimba bream immune cells appears after two weeks. Extension of this period to four or eight weeks loses beneficial effects, and even significantly decreases some immune parameters. This finding is critical for animal nutrition, where glucans or their sources are increasingly used as permanent components of feed.
Introduction Leptin, a class I helical cytokine, regulates growth, appetite, energy homeostasis and reproductive functions in fish. The leptin gene (lep) is a candidate gene for growth trait influence in both farm animals and teleost fish.Material and Methods This study investigated variations in the lep gene and their associations with growth-related traits in European sea bass (Dicentrarchus labrax) using DNA sequencing.Results Twelve novel single-nucleotide polymorphisms (SNPs) were identified in the first and second introns and the third exon of the gene, including a non-synonymous third exon variant (g.11004767C>T) causing an arginine-to-tryptophan substitution. This locus was significantly associated with multiple growth traits, including total and fillet weights and standard, body and post-anal lengths (P-value < 0.05). Analysis revealed two haploblocks; block 1 having four haplotypes (GCG, GTG, ACA and GTA), with GTA associated with the highest total and fillet weights and body length measurements (P-value < 0.01). Block 2 comprised three haplotypes (AG, GG and AA), where AG carriers exhibited superior growth to GG and AA carriers (P-value < 0.01). In silico analysis of the non-synonymous third exon variant (g.11004767C>T) suggested its potential effects on local protein conformation and receptor binding.Conclusion These findings indicate that lep gene polymorphisms, particularly the g.11004714G>A locus and specific haplotypes, are associated with growth variability in European sea bass. The identified SNPs and haplotypes may facilitate molecular marker-assisted selection in growth performance breeding programmes in economically important aquaculture species.
Introduction:The pharmacokinetics (PK) and tissue distribution of enrofloxacin (EF) and its main metabolite, ciprofloxacin (CP), were determined in land snails (Cornu aspersum maxima) dosed singly and multiply orally or intrahaemolymphatically (IHL). Material and Methods:In the single-dose regimen, snails received EF via IHL injection (1 mg/kg), gavage (30 mg/kg) or medicated wafer (30 mg/kg). Haemolymph was sampled up to 72 h post administration. In the multiple-dose regimen, snails were treated with 10 mg/kg EF via gavage for five days. Tissues and body fluids were sampled up to 240 h. Ultra-high-performance liquid chromatography-tandem mass spectrometry simultaneously determined the concentrations of EF and CP. Results:Concentrations of EF in haemolymph after IHL administration remained above the limit of quantification up to 72 h post dose. Enrofloxacin had a 15-19 h elimination half-life. It was more bioavailable from medicated wafers (74.6%) than from gavage (27.7%); however, variability and small sample sizes prevented statistical analysis. Concentrations of CP were inconsistently detectable and often below quantification limits, suggesting limited EF biotransformation. After multiple dosing, EF accumulated predominantly in the hepatopancreas, and less in muscle, the albumen gland and mucus. Tissue contained detectable CP but haemolymph did not. Persistence of EF was noted in tissues beyond 240 h at concentrations often exceeding the 100-300 μg/kg maximum residue limits for other food-producing species. Conclusion:This preliminary study indicates that EF clears slowly and metabolises incompletely to CP in snails, resembling its PK patterns in aquatic and reptilian species. The persistence of EF in edible tissues suggests that withdrawal periods may need to be considerably longer than those for other terrestrial animals. Further depletion studies with extended sampling times, assessment of minimum inhibitory concentration values for snail pathogens and protein binding data are warranted to support safe and rational EF use in heliciculture.
Introduction African swine fever (ASF) is a devastating viral disease of domestic pigs and wild boar. As no safe vaccine is currently available, prevention relies primarily on strict biosecurity measures. Early diagnosis is essential to reduce the risk of ASF spread. Among the available methods, qPCR is considered the gold standard and is recommended by the World Organisation for Animal Health. The Flashtest system is designed to perform the complete workflow, from genetic material extraction to qPCR analysis, enabling rapid (under 1 hour) and accurate detection of various animal diseases in veterinary practice.Material and Methods In this study, the Flashtest system was evaluated in ASF detection using blood samples collected during ASF outbreaks in 2025 in Poland and samples from experimentally infected pigs. Its performance was compared with a validated commercial qPCR assay employed by the National Reference Laboratory for ASF in Poland.Results The system achieved up to 97% sensitivity, 100% specificity and reproducibility with a coefficient of variation below 3.5%. The limit of detection was estimated at 103 TCID50/mL, and a slightly reduced amplification efficiency was observed.Conclusion These results indicate that the Flashtest system reliably detects ASF in the majority of cases. Nevertheless, confirmatory testing is recommended for samples with low viral loads.
