Journal of Veterinary Research最新文献

Introduction: Listeria monocytogenes (LM) is an important food-borne pathogen, and the risk of its ingestion is a serious public health issue. The better its environmental adaptation mechanisms and pathogenicity are understood, the better the risk it poses can be countered. The regulatory role of the small non-coding RNA (sRNA) rli106 in the environmental adaptation and pathogenicity of LM is still unclear and this study investigated that role through its biological function.

Material and methods: An LM-Δrli106 gene deletion strain and an LM-Δrli106/rli106 gene complementation strain were constructed using the homologous recombination technique. Then, the adaptation of these strains to temperature, alkalinity, acidity, salinity, ethanol and oxidative stressors, their biofilm-forming ability and their pathogenicity in mice were investigated to show the regulatory roles of sRNA rli106 in LM. The target gene of rli106 was also predicted, and the interaction between it and rli106 was verified by a two-plasmid co-expressing system based on E.coli and Western blot analysis.

Results: The adaptation of LM-Δrli106 to environmental stressors of pH 9, 5% NaCl and 8% NaCl, 3.8% ethanol and 5 mM H₂O₂ was significantly reduced when compared to the parental (LM EGD-e) and complementation strains. Also, the biofilm formation, cell adhesion, invasion, intracellular proliferation and pathogenicity of LM-Δrli106 in mice were significantly reduced. The results of two-plasmid co-expression and Western blot showed that rli106 can interact with the mRNA of the predicted DegU target gene.

Conclusion: The sRNA rli106 may positively regulate the expression of the DegU gene in LM. This study sheds light on its regulatory roles in environmental adaptation and pathogenicity, providing new insights into the molecular mechanism of sRNA mediation in LM .

Introduction: Many countries have reported severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) infections in mink, and transmission back to humans has raised the concern of novel variants emerging in these animals. The monitoring system on Polish mink farms detected SARS-CoV-2 infection first in January 2021 and has been kept in place since then.

Material and methods: Oral swab samples collected between February 2021 and March 2022 from 11,853 mink from 594 farms in different regions of Poland were screened molecularly for SARS-CoV-2. Isolates from those with the highest loads of viral genetic material from positive farms were sequenced and phylogenetically analysed. Serological studies were also carried out for one positive farm in order to follow the antibody response after infection.

Results: SARS-CoV-2 RNA was detected in mink on 11 farms in 8 out of 16 Polish administrative regions. Whole genome sequences were obtained for 19 SARS-CoV-2 strains from 10 out of 11 positive farms. These genomes belonged to four different variants of concern (VOC) - VOC-Gamma (20B), VOC-Delta (21J), VOC-Alpha (20I) and VOC-Omicron (21L) - and seven different Pango lineages - B.1.1.464, B.1.1.7, AY.43, AY.122, AY.126, B.1.617.2 and BA.2. One of the nucleotide and amino acid mutations specific for persistent strains found in the analysed samples was the Y453F host adaptation mutation. Serological testing of blood samples revealed a high rate of seroprevalence on the single mink farm studied.

Conclusion: Farmed mink are highly susceptible to infection with SARS-CoV-2 of different lineages, including Omicron BA.2 VOC. As these infections were asymptomatic, mink may become an unnoticeable virus reservoir generating new variants potentially threatening human health. Therefore, real-time monitoring of mink is extremely important in the context of the One Health approach.

Introduction: Bovine coronavirus (BCoV) is a causative agent of enteric and respiratory diseases in cattle. Despite its importance for animal health, no data is available on its prevalence in Poland. The aim of the study was to determine the virus' seroprevalence, identify risk factors of BCoV exposure in selected cattle farms and investigate the genetic variability of circulating strains.

Material and methods: Serum and nasal swab samples were collected from 296 individuals from 51 cattle herds. Serum samples were tested with ELISA for the presence of BCoV-, bovine herpesvirus-1 (BoHV-1)- and bovine viral diarrhoea virus (BVDV)-specific antibodies. The presence of those viruses in nasal swabs was tested by real-time PCR assays. Phylogenetic analysis was performed using fragments of the BCoV S gene.

Results: Antibodies specific to BCoV were found in 215 (72.6%) animals. Seropositivity for BCoV was more frequent (P>0.05) in calves under 6 months of age, animals with respiratory signs coinfected with BoHV-1 and BVDV and increased with herd size. In the final model, age and herd size were established as risk factors for BCoV-seropositivity. Genetic material of BCoV was found in 31 (10.5%) animals. The probability of BCoV detection was the highest in medium-sized herds. Polish BCoVs showed high genetic homology (98.3-100%) and close relatedness to European strains.

Conclusion: Infections with BCoV were more common than infections with BoHV-1 and BVDV. Bovine coronavirus exposure and shedding show age- and herd density-dependence.

Introduction: Carvacrol is an essential oil extracted from oregano which can be used as a natural additive in poultry litter and could have a positive impact not only on production rates but also on the quality of poultry meat. The aim of this study was to evaluate the effect of the addition of carvacrol to litter on weight gain and the occurrence of residues in chicken tissues.

Material and methods: One-day-old Ross 308 chicks were used for the study and were randomly divided into two experimental groups. For 42 days, one group was kept in a room with litter enriched with carvacrol and the second group was kept in a room with litter without carvacrol. After 42 days, the birds were sacrificed and necropsied. Carvacrol content was determined in homogenised organ tissue samples by liquid chromatography-mass spectrometry.

Results: Weekly weighing results showed that exposure to carvacrol in litter had no impact on chicken body weight. The analysis of plasma, muscle, liver and lung tissue after 42 days' exposure clearly indicated that there were residues of carvacrol in the analysed matrices.

Conclusion: Exposure of chickens to carvacrol left residues but did not affect body weight.

Introduction: Small ruminant lentivirus (SRLV) causes caprine arthritis-encephalitis in goats and maedi-visna disease in sheep. Transmission is via ingestion of colostrum and milk from infected dams or long-term direct contact between animals. Lifelong seroconversion can occur several weeks after infection via ingestion. However, sub-yearling lambs that ingest contaminated colostrum may be able to clear the infection and become seronegative. Whether a similar phenomenon occurs in goats remains unknown. Therefore, the serological status of goats was studied longitudinally from the moment of natural exposure to colostrum and milk of SRLV-positive dams through the age of 24 months.

Material and methods: Between February 2014 and March 2017 a dairy goat herd was studied which had been infected with SRLV for more than 20 years and carried maedi-visna virus-like genotype A subtype A17. Thirty-one kids born to dams seropositive for SRLV for at least a year beforehand were followed. They ingested colostrum immediately after birth and then remained with their dams for three weeks. The goats were tested serologically every month using two commercial ELISAs. The clinical condition of the goats was also regularly assessed.

Results: Out of 31 goats, 13 (42%) seroconverted at the age ranging from 3 to 22 months with a median of 5 months. Two goats seroconverted in the second year of life. The other eleven did so before the age of one year; two of these reverted to seronegative status. Only 9 out of 31 goats (29%) seroconverted in the first year of life and remained seropositive. They were early and stable seroreactors to which SRLV was transmitted lactogenically. The age at which they seroconverted ranged from 3 to 10 months with a median of 5 months. In 8 of the 18 persistently seronegative goats, a single isolated positive result occurred. No goats showed any clinical signs of arthritis. The level of maternal antibodies at the age of one week did not differ significantly between the stable seroreactors and the remainder.

Conclusion: Seroconversion appears to occur in less than 50% of goats exposed to heterologous SRLV genotype A via ingestion of colostrum and milk from infected dams and is delayed by 3-10 months. The natural lactogenic route of transmission of SRLV genotype A in goats appears to be less effective than this route of genotype B transmission reported in earlier studies.

Introduction: Previous gag and env sequence studies placed Polish small ruminant lentiviruses (SRLVs) isolated from sheep and goats in subtypes B1, B2, A1, A5, A12, A13, A16-A18, A23, A24 and A27. This study extended the genetic/phylogenetic analysis of previously identified Polish SRLV strains by contributing long terminal repeat (LTR) sequences.

Material and methods: A total of 112 samples were analysed. Phylogenetic analyses were carried out on the LTR fragment using the neighbour-joining, maximum likelihood, and unweighted pair group method with arithmetic mean methods.

Results: Polish caprine and ovine LTR sequences clustered within group A and grouped in at least 10 clusters (subtypes A1, A5, A12, A13, A16-A18, A23, A24 and A27). Most of the Polish strains (78%) belonged to the same subtype by the indication of the gag, env and LTR genomic regions. Discrepancies in affiliation depending on the particular sequence were observed in 24 (21%) strains, most of which came from mixed-species flocks where more than one SRLV genotype circulated. Sequences of the LTR reflected subtype-specific patterns. Several subtype-specific markers were identified, e.g. a unique substitution of T to A in the fifth position of the TATA box in A17, A27, A20 and B3.

Conclusion: This study provides valuable insights into the genetic diversity of SRLV field strains in Poland, their phylogenetic relationships and their position in the recently established SRLV classification. Our results confirmed the existence of the ten subtypes listed and the readier emergence of new SRLV variants in mixed-species flocks.

Introduction: Haemorrhagic enteritis virus (HEV) is a common turkey pathogen which suppresses the immune function. The immunosuppressive potential of both field and vaccine strains of HEV makes it necessary to seek substances which can limit or prevent this phenomenon. The aim of the presented work was to investigate the effect of two immunomodulators in the immune response of HEV-infected turkeys. The immunomodulators were synthetic methisoprinol and a natural preparation containing 34.2% β-glucans (β-1,3/1,6) and 12% mannan oligosaccharides (MOS).

Material and methods: The synthetic immunomodulator was administered to female Big 6 turkey chicks at a dose of 200 mg/kg b.w. in drinking water i) for 3 days before, ii) for 5 days after, or iii) for 3 days before, on the day of infection, and for 5 days after experimental HEV infection in turkeys. The natural counterpart was also given to female Big 6 turkey chicks at a dose of 500 g/tonne of feed i) for 14 days before, ii) for 5 days after, or iii) for 14 days before, on the day of infection, and for 5 days after infection. Their effect was evaluated on the synthesis of interferon gamma (IFN-γ) by splenic CD4+ and CD8α+ T cells in response to mitogen stimulation in vitro. Samples were taken 3, 5 and 7 days after infection and analysed by intracellular cytokine staining assay.

Results: Methisoprinol was shown to increase the CD4⁺IFN-γ⁺ and CD8α⁺IFN-γ⁺ T cell count in these birds over the same cell count in control turkeys. A similar effect was obtained in turkeys that received the natural immunomodulator.

Conclusion: The evaluated immunomodulators may be used to attenuate the effects of immunosuppression in HEV-infected turkeys.

Introduction: Escherichia coli is a widespread environmental pathogen frequently causing dairy cow mastitis. This bacterium is particularly capable of acquiring antimicrobial resistance, which can have severe impacts on animal food safety and human health. The objective of the study was to investigate antimicrobial resistance and genetic correlations of E. coli from dairy cow mastitis cases in northern China.

Material and methods: Forty strains of E. coli from 196 mastitis milk samples were collected, susceptibility to 13 common antibiotics and the prevalence of resistance genes were tested in these strains, and the genetic characteristics were identified by multilocus sequence typing.

Results: The results showed that most isolates were multidrug resistant (MDR) (75%), and the resistance rates to cefazolin, trimethoprim-sulfamethoxazole and ampicillin were 77.5%, 55.0%, and 52.5%, respectively. The representative genes of the isolates were aadA (62.5%) and tet(B) (60.0%). Multilocus sequence typing showed 19 different sequence types (STs) and 5 clonal complexes (CCs) in the 40 isolates, mainly represented by ST10 and CC10. The strains of the same ST or CC showed a high level of genetic relatedness, but the characteristics of their antimicrobial resistance were markedly different.

Conclusion: Most E. coli isolates in the study were MDR strains. Some strains of the same ST or CC showed diverse resistance characteristics to common antimicrobials. Therefore, E. coli from dairy cow mastitis in northern China should be investigated to elucidate its antimicrobial resistance and genotypes.

Introduction: The intracellular bacterium Coxiella burnetii is the aetiological agent of Q fever, a zoonosis affecting many animal species worldwide. Cattle and small ruminants are considered the major reservoirs of the bacteria and they shed it through multiple routes.

Material and methods: A total of 2,180 sera samples from 801 cattle herds in all Polish voivodeships were tested by ELISA for the presence of specific antibodies. Milk samples were obtained from seropositive cows in 133 herds as part of a separate study. The milk samples were examined by ELISA and real-time PCR tests.

Results: Seroprevalence at the animal level was 7.06% and true positive seroprevalence was 6.0% (95% confidence interval (CI) 1.1-9.4). Seroprevalence at the herd level was estimated at 11.1% and true positive seroprevalence was 10.5% (95% CI 3.2-15.8). Shedding of the pathogen in milk was detected by real-time PCR in 33 out of 133 tested herds (24.81%, 95% CI 17.74-33.04%) and the presence of C. burnetii antibodies was confirmed in 85 of them (63.9%, 95% CI 55.13-72.05%). The highest level of conformity between ELISA and real-time PCR results was obtained for bulk tank milk samples.

Conclusion: Coxiella burnetii infections are quite common in cattle herds across the country, which emphasises the crucial roles of surveillance and adequate biosecurity measures in the prevention and limitation of Q fever spread in Poland.