Veterinární medicína - In Press

A reproducible rat model for warfarin studies: Prothrombin time assessment with minimal blood using the CoaguChek XS and determination of an effective warfarin dosageOriginal Paper

ET Akyol, MT Hekim, M Erol

Warfarin research in rats is often challenged by methodological inconsistencies and large blood volume requirements. This study aimed to validate a refined, reproducible rat model for serial prothrombin time (PT) monitoring using the CoaguChek XS, in alignment with the 3R principles, and to determine an effective warfarin sodium dosage. Male Wistar albino rats received daily oral warfarin (0.125–0.750 mg/kg) for 5 days. PT was measured using the whole-blood CoaguChek XS and two laboratory plasma assays (Innovin, Thromborel S). Agreement was assessed using Bland–Altman and Passing–Bablok regression. The CoaguChek XS consistently yielded higher PT values than laboratory tests but showed strong agreement up to 25 seconds. A daily dose of 0.125 mg/kg resulted in a stable anticoagulant effect without mortality. Doses > 0.250 mg/kg were toxic, causing PT prolongation beyond the device’s range and death within 4–7 days after the initiation of treatment. In conclusion, the CoaguChek XS is a reliable tool for experiments requiring repeated measurements. Values remain reliable up to 25 seconds, and 0.125 mg/kg is a safe dose for short-term studies, effectively supporting animal welfare and the 3R principles.

Antimicrobial resistance in veterinary medicine: Mechanisms, regulation and control strategiesReview

P Matlakova, K Nedbalcova

Antimicrobial resistance (AMR) represents a major global threat to both human and animal health. Within the One Health framework, farm and wild animals serve as critical reservoirs of resistant bacteria and resistance genes, which can disseminate through food, water, and the environment. The excessive and inappropriate use of antibiotics in veterinary medicine exerts selection pressure that fosters the emergence and spread of antimicrobial resistance. Current mitigation strategies aim to reduce antibiotic consumption and promote prudent use through hygiene measures, vaccination, individualised therapy, pharmacokinetic-pharmacodynamic (PK/PD) dosing optimisation, and comprehensive AMR monitoring. European legislation (EU Regulations 2019/4 and 2019/6) provides stringent guidelines on antibiotic prescription, usage, and residue limits. In this context, antibiotic combination therapy represents a promising approach to enhance antimicrobial efficacy, broaden the antibacterial spectrum, and suppress the emergence of resistance by targeting multiple bacterial pathways simultaneously. In addition to developing new drugs, renewed focus on reviving older antimicrobial molecules – supported by updated PK/PD data and optimised dosing – could provide an effective alternative in combating multidrug-resistant pathogens.

Meloxicam blood levels are not affected by sex-related hormones in male and female goatsOriginal Paper

A Gokceoglu, Z Ozdemir Kutahya, GF Yarim, P Piner Benli, C Gokbulut

The present study aims to investigate the interaction between the non-steroidal anti-inflammatory (NSAID) drug meloxicam and sex hormones in seasonal polyoestrous male and female Saanen goats. It was hypothesised that sex hormones, both steroidal and non-steroidal, may influence the serum concentration of the lipophilic drug meloxicam. In the study, the relationship between plasma drug meloxicam concentration (μg/ml) and levels of Follicle-Stimulating Hormone (FSH), Anti-Müllerian Hormone (AMH), Testosterone, Estradiol (E2), and Progesterone (P4) was evaluated. For this purpose, six male and six female Saanen goats aged 1 year were intravenously administered a 0.5 mg/kg dose of meloxicam, and blood samples were collected at different time points (0, 5, 30 minutes, and 2, 8, 24, 48 hours) to obtain plasma and serum. Hormone levels were determined by ELISA method, following the manufacturer's instructions, and drug concentrations were measured by HPLC-UV technique. While the study found no direct correlation between meloxicam levels and sex hormones, significant differences in hormone levels by sex underscore the importance of considering sex-related physiological differences in veterinary pharmacology. These results will establish a scientific basis for future research on species- and sex-specific dosage adjustments.

Susceptibility of different rainbow trout (Oncorhynchus mykiss) genetic lines to infectious diseaseOriginal Paper

I Papezikova, V Vaibarova, L Pojezdal, K Matejickova, J Blahova, J Mendel, E Maresova, I Mikulikova, M Palikova, I Toulova

This study assessed the resistance of three rainbow trout (Oncorhynchus mykiss) genetic lines to three pathogens, the bacterium Flavobacterium psychrophilum and the parasites Ichthyophthirius multifiliis and Tetracapsuloides bryosalmonae, by monitoring mortality, immune parameters (leukocyte count and phagocytic activity) and pathogen load. Two of the fish lines tested carried quantitative trait loci associated with resistance to the selected pathogens. Unexpectedly, the line with declared genetic resistance exhibited the highest mortality rate following F. psychrophilum infection, suggesting the influence of additional factors such as differences in strain virulence or interactions between genetic background and environmental conditions. Following I. multifiliis infection, the resistant line initially exhibited a higher parasite load but later achieved more efficient parasite clearance. Overall, the findings suggest that disease resistance in trout is a complex trait influenced by interactions between genotype, pathogen and environment. Consequently, resistant lines must always be tested under specific conditions of the target aquaculture facility before implementation.

An experimental challenge model for infecting African catfish (Clarias gariepinus) with Edwardsiella tardaOriginal Paper

I Toulova, V Vaibarova, I Papezikova, K Matejickova, I Mikulikova, H Novotna, M Palikova

Edwardsiella tarda is an important fish pathogen that causes outbreaks leading to significant losses in aquacultural species such as African catfish (Clarias gariepinus). In order to study host-pathogen interactions and design preventive strategies, it is essential to prepare a reliable experimental infection model. The aim of this study was to develop a reproducible immersion challenge model for E. tarda infection in C. gariepinus. The present study consisted of preliminary experiments and a main experiment evaluating the different routes of infection and the bacterial concentrations and exposure durations in an immersion bath. Preliminary experiments demonstrated that infection could be induced by either intraperitoneal injection or immersion bath, and that infection was more successful at higher concentrations. The main experiment involved two concentrations (10⁸ and 2×10⁸ CFU/ml) of immersion bath and two immersion durations (1 and 2 hrs), each repeated twice. The fish were monitored for clinical signs and mortality throughout the experiment and samples were collected on day 10 post-infection for haematological, immunological, and pathological assessment. High-dose immersion reliably induced infection, with disease progression depending on bacterial concentration and exposure duration. This established model is a practical and reproducible tool for future studies on the pathogenesis and immunity of C. gariepinus, as well as the use of immunostimulants and therapeutics.

Efficacy of five peroral therapeutics against multiple phases of Ichthyophthirius multifiliis infestation in brook trout (Salvelinus fontinalis)Original Paper

K Matejickova, J Motlova, H Novotna, Z Mikulkova, P Ginterova, M Jerabek, L Pojezdal

Ichthyophthirius multifiliis is a widespread parasitic ciliate causing significant losses in intensive freshwater aquaculture. This study evaluated the efficacy of medicated feed containing amprolium hydrochloride (2 mg/kg bw/day), clopidol (2 mg/kg bw/day), praziquantel (20 mg/kg bw/day), toltrazuril (50 mg/kg bw/day), and doxycycline (40 mg/kg bw/day) administered at different stages of infection in experimentally infected brook trout fingerlings. Doxycycline and praziquantel demonstrated high protective efficacy (0‑5% mortality) when applied during parasite exposure or the preclinical stage of infection. In contrast, amprolium hydrochloride, toltrazuril, and clopidol showed limited or no protective effect. The timing of administration was critical, and prophylactic use of effective compounds via feed may offer a practical strategy for controlling I. multifiliis outbreaks. These findings support the potential of doxycycline and praziquantel as oral chemotherapeutics in aquaculture, warranting further investigation for therapeutic use in advanced infection stages.

The contribution of polymer additives to microplastic toxicity: A long-term study with Oncorhynchus mykiss exposed to polystyrene microparticles with different hexabromocyclododecane contentOriginal Paper

N Hodkovicova, J Blahova, K Stastny, M Charvatova, A Franc, F Tichy, L Sedlackova, J Mares, M Faldyna, Z Svobodova

Although microplastic toxicity in fish has been extensively studied, the role of polymer additives remains insufficiently understood. This study investigated the combined effects of polystyrene (PS) microplastics and the brominated flame retardant hexabromocyclododecane (HBCD) on the health of rainbow trout (Oncorhynchus mykiss) following six weeks of dietary exposure. Fish were exposed to PS particles with varying HBCD content (0.0 mg/g – PS-HBCDfree, 0.2 mg/g – PS-HBCDlow, 1.0 mg/g – PS-HBCDhigh) or to HBCD alone (HBCDonly). The combination of PS and HBCD induced the most pronounced biological responses at molecular, biochemical and histological levels. In the liver, oxidative stress and upregulation of pro-inflammatory cytokines (il8, il2) were observed, together with decreased glutathione reductase activity and increased catalase expression, indicating an imbalance in antioxidant defence. Histopathology confirmed liver dystrophy and renal lesions, while elevated vitellogenin expression suggested endocrine disruption. Although HBCD accumulation was confirmed only in the HBCDonly group, the polymer-bound form caused comparable physiological alterations, supporting its contribution to combined toxicity. Overall, the study demonstrates that PS microparticles can act as vectors enhancing the bioactivity of embedded additives such as HBCD, resulting in complex multi-organ effects and emphasising the importance of assessing additive–polymer interactions when evaluating the environmental risks of microplastics.

Heartworm removal using basket catheter technique in a 1.8-kg cat with pulmonary hypertension and right-sided heart failure: A case reportCase Report

S-Y Park, W-K Yoon, Y Oh

Feline heartworm infection poses significant diagnostic and therapeutic challenges owing to its distinct clinical features compared to canine cases. Even a single worm can cause life-threatening complications, including pulmonary hypertension and right-sided heart failure. There is no definitive treatment available at present. A 5-month-old intact male domestic shorthair cat (1.85 kg) exhibited respiratory distress, pleural effusion and ascites. Clinical examination revealed a grade 4/6 systolic murmur, jugular vein distension and 7% dehydration. Despite negative antigen and antibody tests for heartworm, echocardiography revealed hyperechoic structures in the right atrium consistent with Dirofilaria immitis, alongside pulmonary hypertension and right-sided heart failure. A 10-cm-long immature female heartworm was surgically removed through jugular venotomy using a basket device under fluoroscopic guidance. Postoperatively, the cat developed persistent pulmonary hypertension, tricuspid regurgitation and refractory hypotension, leading to death on day five. Necropsy confirmed no residual heartworms and showed eosinophilic arteritis, pulmonary oedema, alveolar histiocytosis and thrombus formation, all of which are consistent with heartworm-associated pulmonary vascular disease. Although basket device-assisted removal is feasible, existing vascular damage and postoperative complications significantly affect prognosis. This case report describes a patient managed for clinical diagnostic and therapeutic purposes. All procedures were performed as part of standard veterinary clinical care, and informed consent for publication was obtained from the rescuer. Since this report did not involve experimental interventions, approval from an institutional animal ethics committee was unnecessary.

Drugs and products in beekeeping: efficacy and perspectives of natural alternativesReview

I Szabo, L Sabova, R Sabo, M Sucik

Honeybee health is a key factor in sustainable apiculture, with the most significant colony losses attributed to the mite Varroa destructor and microsporidia of the genus Nosema spp. The aim of this review article is to summarise current knowledge on the use of pharmaceuticals and preparations in the prevention and treatment of these diseases and to highlight promising alternatives to synthetic acaricides. Commonly used active substances such as amitraz and pyrethroids show decreased efficacy in many countries due to the development of resistance. Therefore, organic acids (formic, oxalic, and lactic acids), essential oils, and plant extracts are increasingly applied as environmentally friendly, residue-free alternatives. Some plant preparations and extracts have been shown to be effective against Nosema ceranae. In addition to therapeutic interventions, the importance of preventive measures and immune support through supplements containing vitamins, amino acids, and probiotics is emphasised. The article highlights the need for integrated bee health management combining pharmacological, natural, and biotechnological approaches to limit resistance development, minimise residues in bee products, and promote overall colony vitality.

The efficacy of sap obtained from Commiphora gileadensis against Cadmium-induced liver toxicity in ratsOriginal Paper

AS Aljohani, S Al Dakhil, AS Soliman, M El-Alreshoodi, IM El-Ashmawy

Cadmium (Cd), a toxic heavy metal, ranks among the top ten environmental threats to human health, adversely affecting vital organs such as the central nervous system, kidneys, liver, pancreas, lungs, and testes. The antioxidant potentials of Commiphora gileadensis (CG) sap in vitro were estimated by total phenolic and flavonoid content, DPPH (2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity) and ABTS (2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid) free radical scavenging. The reducing activities were also assessed using copper (cupric reducing antioxidant capacity, CUPRAC) and ferric (ferric reducing antioxidant power, FRAP). The protective effects of CG sap against Cd-induced liver and kidney damage in male rats were investigated in vivo. Forty-two healthy adult male albino rats were randomly divided into seven groups and treated for 60 days as follows: G1 (negative control), G2 (50 mg/l cadmium chloride in drinking water), G3 and G4 (100 mg/kg and 400 mg/kg CG sap, respectively), G5 (50 mg/l cadmium chloride + 100 mg/kg CG sap), G6 (50 mg/l cadmium chloride + 400 mg/kg CG sap), and G7 (1 % Tween 80, 1 ml/rat as a vehicle). The in vitro investigation revealed a potential antioxidant (DPPH, APTS) and reducing (CUPRAC, FRAP) activities of sap obtained from Commiphora gileadensis. Additionally, the in vivo experiment showed cadmium exposure resulted in a significantly increased malondialdehyde, reduced catalase and antioxidant capacity, and impaired liver and kidney functions. However, co-administration of CG sap markedly ameliorated these adverse effects in a dose-dependent manner, improving antioxidant capacity and restoring hepato-renal function by improving biochemical parameters and histological structure. It could be concluded that CG sap has potential as a natural therapeutic agent to mitigate cadmium toxicity and its associated environmental hazards.

Microplastic-induced changes in the rainbow trout gut: Insights from transcriptomic, proteomic, and microbial approachesOriginal Paper

N Hodkovicova, A Hollerova, J Gebauer, M Crhanova, K Stastny, Z Svobodova, M Faldyna

This study examined the intestinal effects of polyethylene (PE; size 46.6 ± 11.3 µm) and polystyrene (PS; size 52.5 ± 11.5 µm) microparticles at three different concentrations (0.5%, 2%, and 5%) on juvenile rainbow trout (Oncorhynchus mykiss) using an integrative approach combining transcriptomics, proteomics, and microbiome analysis. After a six-week experiment, molecular analyses identified concentration-dependent transcriptional responses, including downregulation of genes involved in ion exchange (slc9a1b) and appetite regulation (ghrl), alongside upregulation of immune- and stress-related genes (il10, tfa), particularly at higher concentrations. Proteomic profiling showed a more pronounced effect of PS compared to PE, including suppression of digestive enzymes, disruption of lipid and energy metabolism, and activation of proteins associated with oxidative stress and immune responses. Microbiome analysis confirmed plastic-induced dysbiosis, characterized by reduced microbial diversity, depletion of beneficial taxa (e.g., Bacteroidetes, Actinobacteriota), and shifts in short-chain fatty acid–producing bacteria. By integrating multiple levels of biological organization, this study provides new insights into how microplastics interfere with intestinal physiology, beyond acute toxicity. The findings emphasize the relevance of polymer type and exposure concentration in shaping biological responses and highlight the vulnerability of freshwater species to environmental microplastics.

Protective effect of Loranthus tanakae Franch. & Sav. on allergic asthma induced by ovalbuminOriginal Paper

S-H Park, S-W Pak, W-I Kim, B-R Jin, Y-K Cho, T-W Kim, J-W Ko, J-S Kim, J-C Kim, A Yeong Lee, I-S Shin

Allergic asthma is a widespread disease with elevated eosinophil levels. Although corticosteroids are widely prescribed for allergic asthma, numerous patients experience limited sensitivity and side effects. Loranthus tanakae Franch. & Sav., a traditional herbal plant, has anti-inflammatory and antioxidant properties in pulmonary inflammation caused by Asian sand dust and cigarette smoke condensate. To assess protective effects of L. tanakae, we examined the influence of L. tanakae ethanol extract (LTE) in ovalbumin (OVA)-induced asthma. Mice received intraperitoneal sensitization with OVA, and challenged using OVA inhalation. LTE was consecutively orally gavaged for 6 days. Following sacrifice, bronchoalveolar lavage fluid (BALF) and lung tissue was analyzed. LTE treatment considerably dropped inflammatory cell counts, proinflammatory cytokines in BALF, and immunoglobulin E as opposed to the OVA group, along with a reduction in airway hyperresponsiveness. LTE also improved airway inflammation and suppressed mucus hypersecretion within lung tissues. Additionally, expression of MMP-9 and activation of Erk, JNK, and p-38 were notably diminished in the LTE groups. This study revealed that reduced airway inflammation in OVA-induced asthma via suppressing MMP-9 and mitogen-activated protein kinase associated factors. Consequently, our findings demonstrated that LTE is suggested as a potential remedy for allergic asthma.

From grain to forage: A decade of mycotoxin contamination in ruminant feed in the Czech Republic (2013-2023)Original Paper

J Cahova, L Kvasnickova, J Illek, T Palenik, A Staffa, V Vlasakova, Z Svobodova, L Ravonjison, M Svoboda

This study is based on analytical data focusing on mycotoxin contamination in ruminant feeds in the Czech Republic over a ten-year period (2013­—2023), specifically deoxynivalenol (DON), zearalenone (ZEN), T-2 toxin (T-2/HT-2), and ochratoxin A (OTA). Analyses were performed in accredited State Veterinary Institutes using validated immunochemical and chromatographic methods. Samples included forage and concentrate feeds for dairy cows, calf feed and fattening cattle feed. Across the monitoring period, DON was detected within higher concentration ranges, reflecting the susceptibility of field crops to Fusarium infection under local environmental conditions. Forage feeds showed significantly higher concentrations of DON (472.6 ± 13.2 µg/kg), ZEN (61.7 ± 4.4 µg/kg), and T-2/HT-2 toxin (36.2 ± 2.7 µg/kg) than concentrates for dairy cows (P < 0.05). Additionally, ZEN levels differed significantly between concentrates for dairy cows (17.1 ± 2.3 µg/kg) and calf feed (16.2 ± 3.3 µg/kg) (P < 0.05). Although mycotoxin concentrations generally remained below recommended limits, a small proportion of samples exceeded guideline values for DON (0.13%) and ZEN (0.56%). These findings highlight the importance of continued monitoring and preventive strategies to manage mycotoxin risks.

Comparison of the effect of Mospilan SP and its active substance on Artemia franciscanaOriginal Paper

M Falis, K Benova, M Spalkova, L Leskova

Contaminants formed after the use of pesticides raise concerns for human health and aquatic ecosystems, i. a. due to their high persistence in surface waters and their further migration to other components of the environment. In our study, we compared the effect of the preparation Mospilan SP and its active substance acetamiprid on the aquatic crustacean Artemia franciscana. For Mospilan SP LC50 after 120 hours was 367.44 mg/l, and after 144 hours 102.82 mg/l. For the active substance acetamiprid LC50 after 144 hours was 148.04 mg/l and after 168 hours 28.84 mg/l. The commercial preparation Mospilan SP showed a significant suppressing effect on the measured morphological parameters (body length, body width, intestinal diameter, and intestinal wall thickness) which were at a statistically significant level after 10 days of exposure. When comparing the changes after the application of the preparation and the active substance itself, it is impossible to confirm the same effect on the morphological parameters in given time intervals.

Impact of histological and molecular subtype on the potential therapeutic effect of Buparlisib in canine mammary gland tumoursOriginal Paper

A Baykal Ugur, G Guney Eskiler, O Turna

This study aims to evaluate the response of primary cells to Buparlisib, a PI3K inhibitor, at varying concentrations and exposure durations across different histological and molecular subtypes of CMGTs, and to assess PI3K/Akt/mTOR pathway activity by measuring Akt and mTOR expression. Three carcinomas (C), three sarcomas (S), and two carcinosarcomas (CS) tumours were collected from the dogs. The primary cells were produced from tissues and treated with Buparlisib at different doses. Subsequently, the WST-1 assay, Annexin V, and AO/PI staining were performed sequentially to evaluate the effects of Buparlisib. PI3K/Akt/mTOR signalling pathway inhibition was revealed at the gene level in C, S, and CS cells following 5 µM Buparlisib treatment by RT-PCR analysis. Our results demonstrated that C1 and C2 (basal-like) cells were more sensitive than C3, CS1, and CS2 cells (Luminal A) upon Buparlisib treatment. Liposarcoma S2 cells responded more to Buparlisib than undifferentiated S cells (S1 and S3). Buparlisib also induced apoptosis and inhibited Akt and mTOR mRNA levels in CMGT cells (P < 0.05). A higher rate of apoptotic cell death was observed in the C histological subtype and basal-like cells, with 62.9 ± 0.8% apoptosis in C1 and 79.1 ± 0.3% in C2. The efficacy of Buparlisib was more pronounced in C2 basal-like CMGT cells and liposarcoma S2 cells with the downregulation of Akt and mTOR mRNA levels (P < 0.001).  Therefore, PI3K inhibitors could be used to treat CMGT, particularly the basal-like molecular subtype.