Journal of Veterinary Research最新文献

Introduction: Since lagoviruses cannot be cultivated in vitro, using expression systems is an alternative and promising way of producing diagnostic viral antigens. It opens up their use as active immunogens for vaccine production.

Material and methods: Virus-like particles (VLPs) were produced in a baculovirus expression system in Spodoptera frugiperda 9 (Sf9) insect cells based on wild-type and mutated variants of the virus capsid VP60 protein from a Polish strain of European brown hare syndrome virus (EBHSV) and wild-type and mutated versions of this protein from a Polish strain of rabbit haemorrhagic disease virus 2 (RHDV2). The mutations were the substitution of an arginylglycylaspartic acid (Arg-Gly-Asp/RGD) motif in the P2 subdomain and, in the S or P2 domain, the substitution of three lysines. The VLPs were purified with sucrose gradient ultracentrifugation.

Results: Protein production was confirmed by Western blot analysis using rabbit or hare sera and ELISA tests with different types of monoclonal antibody. The haemagglutination properties of some VLPs were also evaluated. Electron microscopy of wild-type EBHSV, wild-type RHDV2 and the four VP60 variants produced in this experiment revealed the formation of characteristic VLP structures.

Conclusion: For the first time, mutated VLPs of RHDV2 with an RGD motif in the VP60 sequence were obtained, which could potentially be used to deliver cargo to eukaryotic cells. Virus-like particles based on the VP60 proteins of EBHSV and RHDV with a three-lysine substitution in the S or P2 domains were also obtained. Potential exists for VLPs of EBHSV and RHDV2 as vaccine candidates.

Introduction: The article presents a rapid and simple analytical procedure for determination of four sulfonamides (sulfadiazine, sulfamerazine, sulfamethazine and sulfamethoxazole), trimethoprim, tylosin and amoxicillin in animal medicated feed.

Material and methods: Eighteen medicated feed samples were analysed for active substances. The analytical protocol used a mixture of acetonitrile and 0.05 M phosphoric buffer, pH 4.5 for the extraction of seven antibacterial substances. After extraction, the samples were diluted in Milli-Q water and analysed by liquid chromatography with mass spectrometry. The developed procedure was subjected to validation in terms of linearity, selectivity, limits of quantification and determination, repeatability, reproducibility and uncertainty.

Results: The validation of the method was carried out in accordance with the criteria set out in Commission Implementing Regulation (EU) 2021/808 and ICH guidelines. This method provided average recoveries of 90.8 to 104.5% with coefficients of variation for repeatability and reproducibility in the ranges of 3.2-6.9% and 5.2-8.3%, respectively for all analysed antibacterial substances. The limit of detection and limit of quantification for all seven analytes ranged from 5.4 mg/kg to 48.3 mg/kg and from 10.4 mg/kg to 119.3 mg/kg, respectively. The uncertainty of the method depending on the compound varied from 14.0% to 24.0%. The validated method was successfully applied to the 18 medicated feeds.

Conclusion: The developed method can be successfully used to routinely control the content and homogeneity of seven antibacterial substances in medicated feed.

Introduction: Haemosporidian parasites are prevalent worldwide and can cause economic losses in poultry production. These parasites are arousing interest in Thailand and are found in many avian species. There is insufficient information on the genetic diversity of these alveolates from the largest families - Plasmodidae, Haemoprotidae and Leucocytozoidae - specifically parasitising ducks, turkeys, and geese.

Material and methods: Blood samples from 116 backyard poultry (60 ducks, 36 turkeys and 20 geese) in northeastern Thailand were investigated for Plasmodium spp., Haemoproteus spp. and Leucocytozoon spp. infections using microscopic examination and molecular approaches.

Results: A total of 37/116 birds (31.9%) had confirmed Plasmodium infections. The prevalence was 69.4% (25/36) in turkeys, 18.3% (11/60) in ducks, and 5.0% (1/20) in geese. Of these 37 positives, 86.5% were Plasmodium sp., 10.8% were P. gallinaceum and 2.7% were P. juxtanucleare. Sequence analysis based on the cytochrome b gene identified seven lineages, of which two were new lineages in backyard poultry.

Conclusion: This is the first report on the prevalence of haemosporidian parasites in backyard poultry in northeastern Thailand. The results provide important data for better understanding the molecular epidemiology of haemosporidian parasites infection in poultry in this region, which will be helpful in controlling these blood parasites.

Introduction: The broiler chicken digestive tract microbiome maintains the bird's immunity. Its composition has been shown to be important not only for the immune system but also for the gastrointestinal function and productivity of broiler chickens. If the microbiome is populated by supplementation with Lactobacillus, Pediococcus and Saccharomyces spp. - microorganisms with probiotic properties and alternatives to antibiotics - the immune system is stimulated. The use of probiotic supplements in the broiler production cycle can boost bird immunity and prevent adenovirus infection. The resilience of broiler chickens in different feeding schemes including supplementation with these microorganisms was assessed.

Material and methods: Four groups of Ross 308 chickens vaccinated on the standard scheme were investigated over 42 days. Group P received probiotics, prebiotics and vitamins; group AO received antibiotics; group P&AO received probiotics, prebiotics, vitamins and antibiotics; and the control group C received none of these. The birds' immunocompetence against common viral poultry pathogens and their immune response to an experimental challenge with a field strain of infectious bronchitis was evaluated by ELISA and production parameters were recorded.

Results: Mortality was only observed in the control group and was 10%. All birds from the P, P&AO and AO groups responded to the challenge as would be expected of appropriately immunised chickens.

Conclusion: The obtained results indicated that supplementation with synbiotic products and vitamins can enhance broiler chicken immunity and result in better production parameters.

Introduction: Determination of morphological and biochemical blood indices facilitates assessment of the health and welfare of horses, their nutrient demand, the effects of training already undertaken, and the horses' suitability for exercise. Identification of the season-dependent components and the effects of sex and exercise on changes in frequently referenced haematological and biochemical parameters was the main goal of the current study.

Material and methods: The blood morphology of 21 healthy adult Shetland ponies (11 mares and 10 stallions) aged 6.5 ± 1.4 years from the central Pomeranian region in Poland was analysed. Blood samples were taken once per season for one year.

Results: No statistically significant season-dependent differences were found in the blood morphology parameters in either mares or stallions before or after exercise. Beta-coefficient results revealed the strength and type of the relationship of red blood cell distribution width (RDW) and granulocyte count (GRA) with the season, of red blood cell count (RBC), haematocrit, mean corpuscular volume and mean platelet volume with the sex, and of RDW, white blood cell count, GRA and RBC with the exercise factor. Biomarkers demonstrating the relationship between aerobic and anaerobic levels of energy metabolism in the blood did not show any sex dependency in regression analysis.

Conclusion: The sex-independence of energy metabolism biomarkers may indicate the universality of these parameters. Both seasonality itself and its combination with the exercise factor took part in the formation of effective adaptive reactions for maintenance of morphological blood indices in the ponies during exercise.

Introduction: Ixodes ricinus ticks are an important vector and reservoir of pathogenic microorganisms causing dangerous infectious diseases in humans and animals. The presence of ticks in urban greenery is a particularly important public health concern due to the potential for humans and companion animals to be exposed to tick-borne diseases there. The study assessed the prevalence of Borrelia burgdorferi and Anaplasma phagocytophilum infection in I. ricinus ticks feeding on dogs.

Material and methods: The study consisted in analyses of I. ricinus ticks collected in 2018-2020 from owned and stray dogs in the north-eastern part of Lubelskie province (eastern Poland). An AmpliSens PCR kit was used for qualitative detection and differentiation of tick-borne infections.

Results: Infections of B. burgdorferi and A. phagocytophilum were detected in 10.9% and 12.9% of the examined ticks, respectively. One tick (0.7%) was co-infected by both pathogens. Infection with B. burgdorferi was significantly more highly prevalent in ticks collected from the owned dogs than from the strays (18.7% and 2.8%, respectively), whereas the prevalence of A. phagocytophilum was similar in both groups (12.0% and 13.9%, respectively).

Conclusion: The co-infection observed in the study suggests the possibility of simultaneous infection by both pathogens from a single tick bite. The presence of pathogens in ticks collected from dogs is a factor in assessing infection risk not only to companion animals but also to their owners, who are in close contact with their dogs and visit the same green areas recreationally.

Introduction: Ergot alkaloids (EAs) are toxic substances naturally produced by Claviceps fungi. These fungi infest a wide range of cereals and grasses. When domestic animals are exposed to EAs through contaminated feeds, it is detrimental to them and leads to significant economic losses. For that reason, it is important to monitor feed for the presence of EAs, especially with methods enabling their determination in processed materials.

Material and methods: Ergot alkaloids were extracted with acetonitrile, and dispersive solid phase extraction (d-SPE) was used for clean-up of the extracts. After evaporation, the extracts were reconstituted in ammonium carbonate and acetonitrile and subjected to instrumental analysis using high-performance liquid chromatography with fluorescence detection. The developed method was validated in terms of linearity, selectivity, repeatability, reproducibility, robustness, matrix effect, limits of quantification and detection and uncertainty. The EA content of 40 compound feeds was determined.

Results: All the assessed validation parameters fulfilled the requirements of Regulation (EU) 2021/808. At least one of the monitored alkaloids was determined in 40% of the samples. The EAs with the highest incidence rate were ergocryptine, ergometrinine and ergocornine. The total concentrations of EAs ranged from under the limit of quantification to 62.3 μg kg^-1.

Conclusion: The results demonstrated that the developed method was suitable for simultaneously determining twelve EAs in compound feed and could be used for routine analysis.

Introduction: Strains of Leptospira interrogans belonging to two very closely related serovars, Icterohaemorrhagiae and Copenhageni, have been associated with disease in mammalian species and are the most frequently reported agents of human leptospirosis. They are considered the most pathogenic serovars and represent more than half of the leptospires encountered in severe human infections.

Material and methods: Nineteen such isolates from the United Kingdom - human, domestic and wildlife species - were typed using three monoclonal antibodies (F12 C3, F70 C14 and F70 C24) in an attempt to elucidate their epidemiology. They were further examined by restriction endonuclease analysis (REA), multiple-locus variable-number tandem repeat analysis (MLVA) and lic12008 gene sequence analysis.

Results: Monoclonal antibody F12 C3, which is highly specific for Icterohaemorrhagiae and Copenhageni, confirmed that all the strains belonged to these two serovars. Sixteen strains were identified as Copenhageni and three as Icterohaemorrhagiae serovar. Only one restriction pattern type was identified, thus confirming that REA is not able to discriminate between the Icterohaemorrhagiae and Copenhageni serovars. Variable-number tandem-repeat analysis found three loci with differences in the repeat number, indicating genetic diversity between British isolates. Sequences of the lic12008 gene showed that all isolates identified as the Icterohaemorrhagiae serotype have a single base insertion, in contrast to the same sequences of the Copenhageni serotype.

Conclusion: Copenhageni is the predominant serovar in the Icterohaemorrhagiae serogroup isolated in British Isles. There is a genetic diversity of MLVA patterns of the isolates but no genetic tool used in the study was able to determine serovars.

Introduction: Maedi-visna virus and caprine arthritis encephalitis virus are two closely related lentiviruses which cause multisystemic, progressive and persistent infection in goats and sheep. Because these viruses frequently cross the species barrier, they are considered to be one genetic group called small-ruminant lentiviruses (SRLV). They have in vivo tropism mainly for monocytes and macrophages and organ tropism with unknown mechanisms. Typical clinical signs are pneumonia in sheep, arthritis in goats, and mastitis in both species. Infection with SRLV cannot currently be treated or prevented, and control programmes are the only approaches to avoiding its spread. These programmes rely mainly on annual serological testing and elimination of positive animals. However, the high genetic and antigenic variability of SRLV complicate their early and definitive diagnosis. The objective of this review is to summarise the current knowledge of SRLV genetic variation and its implications for tropism, the development of diagnostic tests and vaccines and the effectiveness of control and eradication programmes.

Material and methods: Subject literature was selected from the PubMed and the Google Scholar databases.

Results: The high genetic diversity of SRLV affects the performance of diagnostic tools and therefore control programmes. For the early and definitive diagnosis of SRLV infection, a combination of serological and molecular tests is suggested. Testing by PCR can also be considered for sub-yearling animals. There are still significant gaps in our knowledge of the epidemiology, immunology and biology of SRLV and their impact on animal production and welfare.

Conclusion: This information may aid selection of the most effective SRLV spread reduction measures.

Introduction: The molecular contamination of an animal facility was investigated during and after an infection with highly pathogenic African swine fever virus (ASFV) among domestic pigs. The investigation evaluated the risk of indirect transmission of the disease and indicated points that may facilitate cleaning and disinfection processes.

Material and methods: Six domestic pigs were infected oronasally with the highly pathogenic Georgia 2007 strain. Environmental samples from the floors, walls, rubber floor mats, feeders, drinkers, high-efficiency particulate-absorbing filter covers and doors were collected 7 days post infection (dpi), 7 days later and 24 h after disinfection of the facility. The samples were investigated by real-time PCR and in vitro assays to find genetic traces of ASFV and infectious virus.

Results: Typical clinical outcomes for ASF (i.e. fever, apathy, recumbency and bloody diarrhoea) were observed, and all animals died or required euthanasia before or at 9 dpi. No infectious virus was found in environmental samples at the sampling time points. Genetic traces of ASFV were found in all locations except the doors. The initial virus load was calculated using real-time PCR threshold cycle values and was the highest at the drain. A statistically significant decrease of virus load over time was found on non-porous surfaces mechanically cleaned by water (the floor and drain).

Conclusion: The gathered data confirmed different routes of virus excretion (oral and nasal, faeces and urine, and aerosol) and showed virus locations and different initial concentrations in the animal facility. Maintaining the facility with mechanical cleaning and using personal protection (gloves) and hand disinfection may efficiently minimise the risk of further virus spread. Together with the results of previously published studies, the present investigations' failure to isolate infectious virus may suggest that if stable environmental conditions are assured, the time needed before the introduction of new herds into previously ASF-affected farm facilities could be shortened and in this way the economic losses caused by the disease outbreak mitigated.