Polish journal of veterinary sciences最新文献

Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be a major public health concern. Nucleocapsid (N) protein is the most abundant structural protein on SARS-CoV-2 virions and induces the production of antibodies at the early stage of infection. Large-scale preparation of N protein is essential for the development of immunoassays to detect antibodies to SARS-CoV-2 and the control of virus transmission. In this study, expression of water-soluble N protein was achieved through inducing protein expression at 25°C with 0.5 mM IPTG for 12 h. Western blot and ELISA showed that recombinant N protein could be recognized by sera collected from subjects immunized with Sinovac inactivated SARS-CoV-2 vaccine. Four monoclonal antibodies namely 2B1B1, 4D3A3, 5G1F8, and 7C6F5 were produced using hybridoma technology. Titers of all four monoclonal antibodies in ELISA reached more than 1.28×10 6.0. Moreover, all monoclonal antibodies could react specifically with N protein expressed by transfection of pcDNA3.1-N into BHK-21 cells in IPMA and IFA. These results indicated that water-soluble N protein retained high immunogenicity and possessed the same epitopes as that of native N protein on virions. In addition, the preparation of water-soluble N protein and its monoclonal antibodies laid the basis for the development of immunoassays for COVID-19 detection.

Neospora caninum ( N. caninum) is the etiologic agent of neosporosis, a potential cause of severe reproductive disorders in cattle, small ruminants, equines, wild animals and canids across the world. The current study is performed to estimate molecular prevalence of N. caninum in small ruminants and equines that had abortion in Kurdistan region of Iraq. A total of 64 tissue samples (brain, placenta, heart, lung and liver) were taken from aborted foetuses, with a total of 122 dam blood samples taken from 63 sheep, 39 goats, 12 mares and 8 jennies in local breed fields. Besides, a risk factor analysis for N. caninum positive animals was performed. The observed prevalence of N. caninum DNA in the blood of sheep, goats, horses and donkeys were 20.6%, 17.9%, 21.4% and 25.0%, respectively, and 19.3%, 17.6%, 18.1 and 20.0% in the aborted foetuses of the animals, respectively. Moreover, occurrence of N. caninum was 20.3% in the blood of aborted dams, while it was 18.7% in their aborted foetuses. Confirmatory analysis was also done through constructing a phylogenetic tree to compare the partial sequences of the Nc-5 gene in our isolates (OP771519, OP771520, OP771521 and OP771522) with the GenBank sequences. This showed 98-100% sequence identity with other N. caninum strains in the GenBank database. Older small ruminants and equines had a higher risk of being positive for N. caninum and exposure to dogs were considered as significant risk factors for N. caninum infection in the studied animals (p<0.05). Thus, the results of this study suggest that N. caninum is one of the microbial abortive agents in small ruminants and equines in Kurdistan region of Iraq. It is hoped that the results of this study will help to control animal abortion in livestock and reduce the economic losses.

Cryptosporidium spp., and Giardia duodenalis are intestinal protozoan parasites known to infect humans and various animals and cause diarrhea. This study aimed at determining the prevalence and genotype of Cryptosporidium spp. and Giardia duodenalis in sheep in different locations of Siirt province. The fecal material for this study was collected from 500 sheep in different locations of Siirt province, Turkey. Fecal samples obtained from sheep were examined for Cryptosporidium spp. by Kinyoun Acid Fast staining and the Nested PCR method. Microscopic and Nested PCR methods revealed a prevalence of 2.4% (12/500) and 3.6% (18/500), respectively. Sequence analysis revealed the presence of C. ryanae, C. andersoni, and zoonotic C. parvum. In terms of Giardia duodenalis, 8.4% (42/500) and 10.2% (51/500) prevalence was determined using Nativ-Lugol and Nested PCR methods, respectively. Using sequence analysis, zoonotic assemblages A and B as well as assemblages E and D were detected. As a result of this study, both the prevalence of Cryptosporidium spp. and Giardia duodenalis and the presence of species that appear to be host-specific, as well as those known to be zoonotic, were revealed. A large-scale study is needed to understand the impact of these agents on sheep farming and their consequences on human health.

The objectives of this research were to highlight the main factors, which have relevant significance for etiology of myopathies and to assess the incidence of myopathies in a representative population of broilers raised in Lithuania. Eighteen flocks were evaluated to assess the incidence of musculus pectoralis major myopathies (PMM) (total 54,000 broilers) and dorsal cranial myopathy (DCM) (total 124,200 broilers). Thirteen flocks (total 19,500 broilers) were evaluated to find out deep pectoral myopathy (DPM) occurrence in Lithuania. Investigated parameters of each flock were: average broiler live body weight (BW) at slaughter, average slaughter age, treatment and seasons. A correlation analysis was used to measure the strength of the linear relationship between the investigated traits and incidence of these myopathies. Overall, the incidence of PMM in Lithuania was 18.19%. DCM and DPM were 5.16% and 0.27%, respectively. The percentage of PMM in flocks was strongly associated with average broiler live BW at slaughter (r=0.898, p<0.001) and age at slaughter (r=0.693, p<0.001). The percentage of PMM in flocks was negatively related with treatment of broilers (rs=-0.535, p<0.05). The percentage of DCM was positively associated with average broiler live BW at slaughter (r=0.537, p<0.05) and with seasons (rs=0.658, p<0.01). However, our study results revealed, that the analyzed parameters are not so important in DPM etiology. Furthermore, predisposing factors of PMM, DCM and DPM are different. These findings suggest that not only broiler's heavy weight and age at slaughter could have influence for etiology of myopathies.

The aim of this study was to evaluate the plasma levels of chemokines CCL2 and CXCL12 in female dogs with malignant mammary gland tumours without and with metastases. The concentrations of CCL2 and CXCL12 were determined in 25 female dogs with malignant mammary gland tumours (15 without metastases and 10 with metastases) and 10 healthy control animals using a specific canine ELISA assay. The mean plasma concentrations of CCL2 and CXCL12 were significantly higher (p<0.05) in the metastatic group compared to the control group. Moreover, the concentrations of these chemokines were markedly higher in the dogs with metastases than in those without metastases; however, a statistically significant difference was not found. The concentrations of both tested chemokines were numerically increased in the dogs with grade 2 and grade 3 carcinomas compared to the dogs with grade 1 carcinomas but the differences did not reach statistical significance. In conclusion, the results of our study demonstrate that plasma concentrations of chemokines CCL2 and CXCL12 are significantly increased in the dogs with metastatic malignant mammary gland tumours compared to the healthy dogs and show an upward trend compared to those without metastases. However, clarifying whether the increase of these chemokines is a cause or an effect of metastasis in female dogs with malignant mammary gland tumours as well as their potential role in metastatic process requires further research.

The aim of the study was to compare the serum protein profile of Bernese Mountain Dogs (BMDs) reacting positive for Bb in snap testing with the serum protein profile of dogs of other breeds (healthy and with clinical borreliosis) using the MALDI time-of-flight (MALDI-TOF) technique. The observations included five groups of dogs. BMDs reacting positively to Bb in snap serological testing and showing symptoms of borreliosis (group 1), BMDs for which no borreliosis symptoms were determined but with seropositivity for Bb determined with snap serological tests (group 2), clinically healthy BMDs with no antibodies for Bb found in the serum (group 3), five dogs of different breeds, reacting positively in serological testing, in which borreliosis symptoms were observed (group 4), clinically healthy dogs of different breeds with negative reaction in tests towards Bb (group 5). A proteomic analysis demonstrated the presence of five identical protein fractions among all five groups. An additional two protein fractions of approximately 7.630 and 15.260 kDa were found in all the serum samples obtained from the dogs positive for borrelia in a snap test, both in those exhibiting symptoms of borreliosis, and seropositive BMDs not presenting symptoms of the disease. These two additional protein fractions may be used to differentiate between seropositive and seronegative B. burgdorferi dogs and may be considered a seropositivity marker, however, it cannot be used to differentiate between animals with the clinical form of the disease and those that are only seropositive.

Montmorillonite (MMT), a natural absorbent agent, has widely been accepted for its antidiarrhea function in human and farm animals; however, its specific physicochemical property limits its biological function in practical use. In the current study, raw MMT was loaded by andrographolide, namely andrographolide loaded montmorillonite (AGP-MMT). The microstructure of AGP-MMT was observed by scanning electron microscope (SEM) and X-ray diffraction (XRD). The effect of AGP-MMT on the growth performance, intestinal barrier and inflammation was investigated in an enterotoxigenic Escherichia coli (ETEC) challenged mice model. The results show that the microstructure of MMT was obviously changed after andrographolide modification: AGP-MMT exhibited a large number of spheroid particles, and floccule aggregates, but lower interplanar spacing compared with MMT. ETEC infection induced body weight losses and intestinal barrier function injury, as indicated by a lower villus height and ratio of villus height/crypt depth, whereas the serum levels of diamine oxidase (DAO), D-xylose and ETEC shedding were higher in the ETEC group compared with the CON group. Mice pretreated with AGP-MMT showed alleviated body weight losses and the intestinal barrier function injury induced by ETEC challenge. The villus height and the ratio of villus height/crypt depth, were higher in mice pretreated with AGP-MMT than those pretreated with equal levels of MMT. Pretreatment with AGP-MMT also alleviated the increased concentration of serum tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), and the corresponding genes in the jejunum induced by ETEC infection in mice. The protein and mRNA levels of IL-1β were lower in mice pretreated with AGP-MMT than those with equal levels of MMT. The results indicate that AGP-MMT was more effective in alleviating intestinal barrier injury and inflammation in mice with ETEC challenge than MMT.

Mycoplasma bovis is a highly contagious pathogen that causes clinical or subclinical mastitis. The present study was aimed for the isolation, molecular characterization and antibiogram determination of M. bovis from raw milk samples. Milk samples were collected randomly from lactating cows and buffaloes from different tehsils of district Faisalabad, Pakistan. Samples were inoculated on modified Hayflick medium and biochemical tests were performed for further confirmation of isolated M. bovis. Out of total 400 milk samples, 184 (46%) samples were found positive for culture method. The 16S-rRNA gene polymerase chain reaction was performed for molecular characterization of isolated M. bovis strains. Out of total 400 milk samples, 240 (60%) positive for M. bovis through PCR method were examined. The 16S-rRNA gene PCR positive isolated M. bovis strains were sequenced and results were compared using Maximum-likelihood method and sequenced strains of M. bovis were aligned and analyzed by Clustal W software. Antibiogram of isolated M. bovis strains was analyzed by disc diffusion assay against eight commonly used antibiotics. Tylosin (30μg) and Tilmicosin (15ug) showed inhibition zones of 32.34 ± 1.10 mm and 17.12 ± 0.93 mm respectively against isolated M. bovis which were found sensitive. Isolated M. bovis was found resistant to other commonly used antibiotics. Statistical analysis revealed that p-value was < 0.05 and the odds ratio was >1.0 at 95% CI. This study complemented the lack of epidemiological knowledge of molecular characterization, comparative effectiveness and resistance trends of isolated M. bovis strains against commonly used antibiotics.