Journal of Veterinary Research最新文献

Introduction: In dairy cattle, oxidative stress is a predominant problem associated with diseases and reproductive health issues. This study aimed to detect the variation in the antioxidant biomarkers by adding different concentrations of β-hydroxybutyric acid (BHBA) and sought to elucidate its effects on the gene expression levels of growth hormone (GH) and antioxidant biomarkers in bovine hepatocytes.

Material and methods: Four antioxidant biomarkers, namely malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH Px) were evaluated using commercially available bovine ELISA kits. The expression levels of the bovine GH, its receptor (GHR), insulin-like growth factor (IGF), IGF-1, IGF-1 receptor, CAT, SOD, GSH-Px and β-actin (as a reference) genes in liver cell culture were determined by reverse transcriptase-PCR assay.

Results: With the increase of BHBA concentration and culture time, the activities of SOD, CAT, and GSH Px biomarkers in hepatocytes decreased. However, the content of MDA in hepatocytes increased gradually with the increase of hepatocyte culture time and BHBA concentration. The qPCR results revealed that after adding BHBA, gene expression levels of GSH-Px, SOD and IGF biomarkers in hepatocytes began to differ in the culture groups at 12 h, whereas the gene expression level of the CAT and GHR biomarkers in hepatocytes began to differ at 6 h.

Conclusion: Quantitative PCR results showed that the BHBA significantly downregulated the expression levels of the GHR gene and CAT, GSH Px and SOD antioxidant biomarker genes.

Introduction: The occurrence of pesticide residues in animal products deserves attention because of the contamination by environmental pollutants and pesticides that may be present in the food that animals are fed. The goal of this work was the validation of a method for detection of residues of multiple classes of pesticide and determination of their residues in chicken breast fillets.

Material and methods: Gas chromatography with mass spectrometry was used for analysis. A modified quick, easy, cheap, effective, rugged and safe (QuEChERS) method was put into practice for its validation and applied to real samples. The study optimised mass detection and investigated the effect of a freezing step during the preparation of samples. Pesticides were determined in samples from conventional and organic production.

Results: The impact of the matrix effect decreased, with the largest number of pesticides and satisfactory recovery determined by the application of mixed solvent acetonitrile and ethyl acetate for extraction. Detection of pesticide residues was achieved in a linear range between 5 and 50 µg/kg with satisfactory excellent correlation coefficients greater than 0.99. The recovery of all the pesticide residues ranged between 71.2 and 118.80%. The relative standard deviation was from 2.9% to 18.1% for all validated pesticide residues. The limits of quantification were in the range of 3.0-4.9 µg/kg. Out of 56 pesticide residues analysed in real samples, 5 were detected: α endosulfan, cypermethrin, endosulfan sulphate, permethrin and p,p´-dichlorodiphenyltrichloroethane (DDT) and their concentrations ranged from 4.9 to 15.2 µg/kg.

Conclusion: All tested samples were compliant with the evaluation criteria, and detected values of pesticide residues were lower than the maximum residual levels.

Introduction: Rabies is endemic in Europe and red foxes are the vector and reservoir of the rabies virus (RABV). Based on classification established in the early 1990s, four variants of the rabies virus have been distinguished in Europe. Rabies broke out in January 2021 in the Mazowieckie voivodeship in central north-eastern Poland. The virus spread rapidly, reaching the Świętokrzyskie voivodeship in the central southern part and the Lubelskie voivodeship in the eastern part in the next months. Nine rabies cases were reported in the Podkarpackie voivodeship in south-eastern Poland between 2021 and 2023, mainly in red foxes but also in dogs and wildcat. The aim of the study was the identification of RABV variants in wildlife and domestic animals in Poland between 2021 and 2023.

Material and methods: The study involved 157 animal brains tested positive for rabies using a fluorescent antibody test. From 10% w/v brain homogenates, RNA was isolated and full-length RABV genomes were high-throughput sequenced with an RABV-enriched approach. Complete genomes of RABV isolates were phylogenetically analysed and the variants were estimated.

Results: Molecular and phylogenetic studies revealed 147 (93.6%) of the RABV strains out of 157 which had rapidly spread in the wildlife of the Mazowieckie, Świętokrzyskie and Lubelskie voivodeships to be Central European strains. Nine RABVs (5.7%) detected in foxes, a wildcat and a dog in the Podkarpackie voivodeship were identified as North-Eastern European. A vaccine-induced rabies case was detected in a red fox in the Lubelskie voivodeship in May 2023.

Conclusion: Central European and North-Eastern European RABVs were circulating in Poland between 2021 and 2023.

Introduction: The main adaptive immune cells are T and B lymphocytes and they play key roles in the induction of immune responses against canine mammary tumours. Investigating these cell subpopulations may lead to more precise diagnosis of these malignancies.

Material and methods: The percentages of CD3⁺, CD4⁺ and CD8⁺ T cells and of CD21⁺ B cells in the peripheral blood of bitches with malignant mammary tumours were compared with those in the blood of healthy animals. The phenotypic features of peripheral blood leukocytes were evaluated by flow cytometry.

Results: There was a significant difference in the mean percentages of CD3⁺ lymphocytes between healthy (66.7%) and metastatic dogs (46.1%), and between tumour-bearing non-metastatic (66.6%) and metastatic dogs. There was also a significant difference in CD4⁺ T helper cell percentages between healthy dogs (40.4%) and dogs with metastases (23.2%), and between the latter and dogs without them (35.5%). In the case of CD21⁺ lymphocyte subsets, a significant difference was noted between healthy animals (10.9%) and those with metastases (20.1%), and between the latter and patients without metastases (8.5%). There were also significant differences in CD3⁺/CD21⁺ ratios between the group with metastases (3.0), the healthy group (7.8), and the group without metastases (8.5). Similarly, a significant difference was noted in CD4⁺/CD8⁺ ratios between animals with metastases (1.4), bitches in the control group (2.2), and dogs without metastases (1.9).

Conclusion: Peripheral blood leukocyte phenotypic characteristics are putative novel biomarkers. These findings may be useful in future studies improving mammary tumour diagnostic procedures, especially in metastasis detection.

Introduction: The aim of the study was to monitor the occurrence of selected vector-borne diseases in anaemic dogs arriving in or returning to Poland from areas endemic for these diseases.

Material and methods: The study involved 497 dogs, of which 184 came to Poland from Ukraine with their owners fleeing the war. Other animals returned to the country from holidays spent in Croatia (n = 96), Turkey (n = 79), Italy (n = 48), Bulgaria (n = 42), Albania (n = 36) and Romania (n = 12). Molecular biology methods were used for detection of pathogens transmitted by the vectors.

Results: Molecular tests revealed the presence of vector-borne pathogens in 79 dogs. The most commonly diagnosed infection was caused by Babesia canis (27 dogs), followed by infections with Anaplasma phagocytophilum (in 20 dogs), Mycoplasma haemocanis (15 dogs), Bartonella henselae (7 dogs), Ehrlichia canis (4 dogs), Hepatozoon canis (3 dogs), Babesia gibsoni (2 dogs) and Leishmania infantum (1 dog). Most of the sick dogs (n = 39) came from Ukraine. In dogs spending holidays with their owners outside Poland, vector-borne diseases were most often detected after their return from Turkey (n = 16), and next in descending order from Croatia (n = 7), Italy (n = 6), Albania (n = 4), Bulgaria (n = 4) and Romania (n = 3).

Conclusion: The wider migration crisis and increasingly frequent trips of owners with their dogs to areas of endemic infectious and parasitic diseases observed in recent years are the main risk factors for the occurrence of these diseases in Poland. Therefore, constant monitoring of vector-borne diseases, especially in dogs returning from holidays and arriving in Poland from abroad, seems to be crucial for their early detection and introduction of appropriate therapy.

Introduction: This study focuses on perfluoroalkyl substance (PFAS) content in chickens' eggs and the livers of farm animals.

Material and methods: Chickens' eggs (n = 25) and the livers of cows (n = 10), chickens (n = 7) and horses (n = 3) were collected from various regions of Poland. Samples were analysed using the isotope dilution technique with liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS).

Results: The mean lower bound (LB) sum of four PFAS (∑4 PFAS) concentrations (perfluorooctanesulfonic acid (PFOS), perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA) and perfluorohexanesulfonic acid (PFHxS)) were the highest in cows' livers (0.52 μg/kg) and much lower in chickens' (0.17 μg/kg) and horses' livers (0.13 μg/kg) and chickens' eggs (0.096 μg/kg). The ratio of ∑4 PFASs to the limits set by Commission Regulation (EU) 2023/915 was <7% for liver and <6% for eggs. Linear PFOS was the compound with the highest detection frequency (8% in eggs and 48% in all livers). In cows' livers it was detected in 80% of samples. The estimated exposure to LB ∑4 PFASs via consumption of liver tissue from farm animals (assuming 50 g and 100 g portions) was <52% of the tolerable weekly intake (TWI) for children and <17% of the TWI for adults. Dietary intake via the average portion of three eggs led to low exposure of <15% for children and <5% for adults.

Conclusion: Neither eggs nor the livers of chickens or horses as analysed in this study are significant sources of PFASs, while cows' livers might contribute significantly to a child's overall dietary intake. Further investigation of PFOS in farm animal livers should be conducted.

Introduction: Chicken bones, a by-product of the poultry industry, can directly or indirectly enter the food chain. Bone meal and bone products could be sources of many contaminants. Considering the wide range of uses made of bones in the culinary and food industries, this material needs to be safe and antibiotic residue-free. To determine if such is the case, the concentration of doxycycline in chicken bones was investigated, this antimicrobial being one of the most commonly used in poultry production.

Material and methods: Ross 308 broilers were grouped into three experimental and one control group. Doxycycline was administered in drinking water at therapeutic and sub-therapeutic doses, as well as via spray treatment. The concentration of doxycycline in bones was determined post slaughter by ultra-high performance liquid chromatography-tandem mass spectrometry.

Results: Doxycycline was quantified at 135 μg/kg 22 days after the last day of antibiotic administration at therapeutic doses; 2,285 μg/kg after sub-therapeutic treatment for 27 days and 9.62 μg/kg 22 days after the end of spray application.

Conclusion: High concentrations and long persistence of doxycycline in bones were found in this study. Doxycycline can contaminate all bone-derived products in the food and fertiliser industries.

Introduction: No maximum residue limits in honey have been legislated in the EU for antimicrobial substances such as sulphonamides, and they are not permitted, therefore, for treating honey bees unless in a cascade system. Since sulphonamides are used illegally in apiculture to treat foulbrood, their residues can be found in honey and other apiculture products, including beeswax. The study aimed to assess the contamination of honey from beeswax containing residues of 10 sulphonamides (sulphadimethoxine (SDM), sulphadoxine (SDX), sulphamonomethoxine (SMM), sulphamethoxazole (SMX), sulphameter (SMT), sulphamethazine (SMZ), sulphamerazine (SMR), sulphadiazine (SDA), sulphathiazole (STZ) and sulphacetamide (SCA)).

Material and methods: Wax-based foundations fortified with 10 sulphonamides at 10,000 μg/kg were evaluated for sulphonamide concentrations and then placed in a beehive so that honey bees (Apis mellifera L.) could build honeycombs with them. Frames of capped honey were taken out of the hives one month later and honey was sampled from them. The honeycombs were subsequently incubated in a laboratory at 35°C for five months, and honey was sampled monthly. The honey sulphonamide concentrations were measured using liquid chromatography-tandem mass spectrometry and compared to the wax-based foundation concentrations.

Results: The maximum transfers to honey of the initial amount of SDM, SDX, SMM, SMX, SMT, SMZ, SMR, SDA, STZ and SCA in the wax-based foundations were 42.6, 34.3, 31.7, 30.1, 29.5, 25.2, 18.7, 16.1, 9.5 and 8.6%, respectively.

Conclusion: This study demonstrated that every tested sulphonamide could migrate from beeswax in antimicrobial-tainted honeycombs to honey, SDM having the highest migration potential and SCA the lowest.

Introduction: Exosomes are nanosized lipid bilayer membranous microvesicles, extracellularly released from a variety of mammalian cells. They mediate intercellular signalling by transporting several types of RNA, lipids and proteins and participate in the intercellular exchange of DNA, RNA, micro RNA, proteins and other components. These microvesicles are present in all body fluids in physiological and pathological conditions and reflect the state of the host organism. The aim of the study was the isolation and molecular determination of exosomes in blood and supernatant fluids of bovine dendritic cell cultures infected with bovine leukaemia virus (BLV).

Material and methods: Exosomes were isolated by ultracentrifugation from the blood sera, plasma and supernatant of bovine BLV-infected and uninfected control dendritic cell cultures and their presence was confirmed with scanning electron and transmission electron microscopy. Western blot analysis of the structural BLV glycoprotein 51 (Env) and protein 24 (Gag) and of the tetraspanin exosomal markers CD9, CD63 and flotillin-1 was undertaken in BLV+ and control BLV- cattle.

Results: In exosomes of leukaemic cattle both BLV proteins and exosomal markers were detected. In healthy control animals only exosomal markers were determined.

Conclusion: Proteins of BLV were released with exosomes and could be transferred into recipient cells as an alternative propagation route not requiring virus infection.

Introduction: Small ruminant lentiviruses (SRLV) cause multisystemic, degenerative and chronic disease in sheep and goats. There are five genotypes (A, B, C, D and E), of which A and B are the most widespread. The purpose of this study was to evaluate the serotyping efficiency of the Eradikit SRLV Genotyping ELISA and the molecular typing efficiency of a newly developed nested real-time PCR targeting the long terminal repeat-gag (LTR-gag) region using samples from animals infected with subtypes of SRLV known to circulate in Poland.

Material and methods: A total of 97 sera samples taken from 34 sheep and 63 goats were immunoassayed, and 86 DNA samples from 31 sheep and 55 goats were tested with the PCR. All ruminants were infected with known SRLV strains of the A1, A5, A12, A13, A16, A17, A18, A23, A24, A27, B1 and B2 subtypes.

Results: A total of 69 (80.2%, 95% confidence interval 71.6%-88.8%) out of 86 tested samples gave positive results in the PCR. In 17 out of the 86 (19.8%) samples, no proviral DNA of SRLV was detected. The differentiation between MVV (genotype A) and CAEV (genotype B) by PCR matched the predating phylogenetic analysis invariably. No cross-reactivity was observed. On the other hand, the proportion of samples genotyped the same by the older phylogenetic analysis and the Eradikit SRLV Genotyping ELISA was 42.3%. The test was unable to classify 40.2% of samples, and 17.5% of sera were incorrectly classified.

Conclusion: Our results showed that the Eradikit SRLV genotyping kit is not a reliable method for predicting SRLV genotype, while the nested real-time PCR based on the LTR-gag region did prove to be, at least for genotypes A and B.