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Enrofloxacin decreases IL-6 and TNF-α production by lipopolysaccharide-stimulated porcine peripheral blood mononuclear cells
2016
Pomorska-Mól Małgorzata | Czyżewska-Dors Ewelina | Kwit Krzysztof | Pejsak Zygmunt
Introduction: The aim of the study was to explore the effect of enrofloxacin on production of selected cytokines by porcine peripheral blood mononuclear cells (PBMCs).
Show more [+] Less [-]Development and application of a TaqMan-MGB real-time RT-PCR assay for the detection of porcine epidemic diarrhoea virus strains in China
2016
Hou, Yi-Xuan | Xie, Chun | Wang, Kang | Zhao, Yu-Ting | Xie, Yang-Yang | Shi, Hong-Yan | Chen, Jian-Fei | Feng, Li | Tong, Guang-Zhi | Hua, Xiu-Guo | Yuan, Cong-Li | Zhou, Yan-Jun | Yang, Zhi-Biao
Introduction: A real-time RT-PCR method for identification and quantification of porcine epidemic diarrhoea virus (PEDV) strains in China was developed.Material and Methods: Based on the conserved sequence of the PEDV nucleocapsid (N) gene, a primer pair and probe were designed to establish a TaqMan-MGB real-time RT-PCR assay for quantitative detection of the virus. The sequence was cloned into the pMD18-T vector and a series of diluted recombinant plasmids were used to generate a standard curve with an R2 value of 0.999.Results: The developed quantitative PCR assay detected viral titres as low as 0.1 TCID₅₀ with high specificity and no cross-reaction with other porcine viruses (PoRV, TGEV, PRRSV, or CSFV). The intra-batch and inter-batch coefficients of variation were both less than 1%, which indicated good reproducibility. Thirty clinical diarrhoea samples obtained from pigs in Shanghai and Fujian were analysed using this quantitative PCR assay. Out of these samples, 93.3% were found to be PEDV positive.Conclusion: This approach is suitable for clinical sample identification and pathogenesis studies.
Show more [+] Less [-]Multiplex real-time PCRs for detection of Salmonella, Listeria monocytogenes, and verotoxigenic Escherichia coli in carcasses of slaughtered animals
2016
Denis, Edyta | Bielińska, Katarzyna | Wieczorek, Kinga | Osek, Jacek
Introduction: The study objective was to develop and evaluate a new TaqMan multiplex real-time PCR method for Salmonella, L. monocytogenes, and verotoxigenic Escherichia coli (VTEC) detection in slaughtered animal carcasses.Material and Methods: The procedure included an enrichment step, DNA extraction, and two multiplex real-time PCRs. The first PCR detected the invA and hly genes of Salmonella and L. monocytogenes respectively, the second the vtx1, vtx2, and eae genes of VTEC.Results: The validation of this method resulted in 100% relative sensitivity, specificity, and accuracy as compared to the reference ISO methods. The limit of detection per swab sample was established at 1 cfu for Salmonella and L. monocytogenes and 2 cfu for VTEC. The authors analysed 265 slaughterhouse-collected swabs from cattle, pig, and poultry carcasses. Among 125 from cattle, 51 were positive for VTEC, 29 for Salmonella, and 1 for L. monocytogenes. Among swabs from pig carcasses (n = 95), three, two, and one sample were positive for these pathogens respectively. None of the microorganisms tested for was identified in 45 samples of poultry origin.Conclusion: The obtained results showed that the method developed can rapidly identify the main bacterial pathogens that may contaminate carcasses of food-producing animals.
Show more [+] Less [-]Selected aspects related to epidemiology, pathogenesis, immunity, and control of African swine fever
2016
Woźniakowski, Grzegorz | Frączyk, Magdalena | Niemczuk, Krzysztof | Pejsak, Zygmunt
African swine fever (ASF) is currently one of the most severe viral infections of domestic pigs, wild boars, and other hosts belonging to Suidae family. ASF is also considered as the most complex and devastating infectious and haemorrhagic disease of swine due to its severe socio-economic impact and transboundary character. ASF it is a notifiable disease and due to the lack of specific treatment and vaccine, the disease can be only limited by the administrative measures comprising wild boar hunting and stamping out of affected pigs. ASF occurred for the first time in Kenya in 1921 while in Europe (Portugal) the virus was detected at the end of the 1950s. In spite of successful eradication of this threat in a number of affected regions, the virus remains endemic in both feral and domestic pigs in Africa and Sardinia. The ‘new era’ of ASF started in 2007 after its re-introduction to Georgia. Following its intensive expansion, the virus spread to other Caucasian countries, including the territory of the Russian Federation. In 2014 the virus reached Ukraine, Belarus, and, consequently, European Union countries: Lithuania, Latvia, Estonia, and Poland. The occurrence of ASF in wild boars and pigs had a severe impact on both epidemiology and economy because of the national and international transport and trade consequences. Up to date, starting from the February 2014, eighty ASF cases in wild boar and three outbreaks in domestic pigs have been diagnosed. Taking into account the diverse rate of spread in Poland, this review aims to present and discuss the current state of knowledge on ASF including its epidemiology, pathology, transmission, and perspectives of control.
Show more [+] Less [-]Real-time quantitative PCR for detection and identification of Actinobacillus pleuropneumoniae serotype 2
2016
Dors, Arkadiusz | Kowalczyk, Andrzej | Pomorska-Mól, Małgorzata
Introduction: Porcine pleuropneumonia inflicts important economic losses on most commercial herds. Detection of subclinical or chronic infection in animals still remains a challenge, as isolation and identification of A. pleuropneumoniae serotypes is difficult and quantification of the bacteria on agar plates is often almost impossible. The aim of the study was to develop and evaluate a serotype-specific quantitative TaqMan probe-based PCR for detection of serotype 2 in pig lungs, tonsils, and nasal swabs.Material and Methods: The primers were designed from the capsular polysaccharide biosynthesis genes of A. pleuropneumoniae serotype 2. PCR specificity and sensitivity were evaluated using reference strains and several other bacterial species commonly isolated from pigs.Results: The real-time qPCR for detection of A. pleuropneumoniae serotype 2 was highly specific and gave no false positives with other serotypes or different bacterial species of pig origin. The detection limit for pure culture was 1.2 × 10⁴ CFU/mL, for lung tissue and nasal swabs it was 1.2 × 10⁵ CFU/mL, and for tonsils - 1.2 × 10⁵ CFU/mL.Conclusion: The method can be used to serotype A. pleuropneumoniae isolates obtained during cultivation and to detect and identify A. pleuropneumoniae serotype 2 directly in nasal swabs and tonsil scrapings obtained from live pigs or lung tissue and tonsils collected post-mortem.
Show more [+] Less [-]Enrofloxacin decreases IL-6 and TNF-α production by lipopolysaccharide-stimulated porcine peripheral blood mononuclear cells
2016
Pomorska-Mól, Małgorzata | Czyżewska-Dors, Ewelina | Kwit, Krzysztof | Pejsak, Zygmunt
Introduction: The aim of the study was to explore the effect of enrofloxacin on production of selected cytokines by porcine peripheral blood mononuclear cells (PBMCs).Material and Methods: Twenty pigs (10 control and 10 experimental) were used in this research. Pigs from experimental group received enrofloxacin at therapeutic doses. Blood samples were collected before, during, and after treatment with enrofloxacin. PBMCs were incubated with or without lipopolysaccharide (LPS). Production of IL-6, IL-10, INF-γ, and TNF-α were determined by ELISA.Results: Administration of enrofloxacin to healthy pigs for 5 d induced a transient reduction of the PBMCs response to LPS in terms of IL-6 and TNF-α secretion. The concentration of IL-6 returned to the day 0 level shortly after treatment, while TNF-α production remained reduced for 10 d after the treatment. The production of IL-10 was not affected by enrofloxacin. The level of IFN-γ was below the detection limit of the tests.Conclusion: The results indicate that enrofloxacin administered in vivo in therapeutic doses has an immunomodulatory effect through its capacity to inhibit secretion of IL-6 and TNF-α by porcine PBMC stimulated by LPS.
Show more [+] Less [-]Preliminary study on the detection of hepatitis E virus (HEV) antibodies in pigs and wild boars in Poland
2016
Weiner, Marcin | Tokarska-Rodak, Małgorzata | Plewik, Dorota | Pańczuk, Anna | Szepeluk, Adam | Krajewska, Monika
Introduction: Although HEV infection in pigs does not pose a major economic risk to pork production, the risk of zoonotic transmission to humans is an important aspect of public health. HEV genotype 3 infections were reported in developed countries in individuals who had consumed raw meat or meat products from deer, wild boars, or pigs. The aim of the study was the analysis of the occurrence of HEV-specific antibodies among wild boars and domestic pigs in Poland. Material and Methods: A total of 290 samples from wild boars and 143 samples from pigs were tested. The antibodies were tested by ELISA. Results: The presence of anti-HEV IgG was demonstrated in 44.1% of pigs and 31.0% of wild boars. Anti-HEV IgG antibodies were detected in 1.4% of samples from pigs and in 2.1% of samples from wild boars at borderline level. The statistical analysis shows significant differences in the positive results for anti-HEV IgG between the groups of pigs and wild boars (P = 0.0263). Conclusion: Regular surveillance of the occurrence of HEV in swine and wild boars should be performed in the future.
Show more [+] Less [-]Changes in circulating lymphocyte subpopulations in pigs receiving therapeutic doses of ceftiofur and tulathromycin
2016
Czyżewska-Dors, Ewelina | Kwit, Krzysztof | Pejsak, Zygmunt | Pomorska-Mól, Małgorzata
Introduction: The aim of the study was to evaluate the effect of administration of therapeutic doses of ceftiofur and tulathromycin on the circulating lymphocyte subpopulations in healthy pigs. Material and Methods: The study was conducted on thirty healthy 7- to 10-week-old pigs, assigned to three groups: the TUL group, injected with tulathromycin (n = 10); the CEF group, injected with ceftiofur (n = 10); and the C group, the control with no antibiotic administration (n = 10). Blood samples were collected before, during, and after treatment with antimicrobials. Lymphocyte subpopulations circulating in the blood were determined by immunostaining and flow cytometry analyses. Results: Following administration of a therapeutic dose of tulathromycin, there were no changes in the lymphocyte subpopulations circulating in blood. In contrast, administration of ceftiofur at the recommended dose decreased the absolute number of CD3+, CD21+, CD4+CD8-, CD4-CD8+, and double positive CD4CD8 cells. Conclusion: Results from the study indicate that ceftiofur possesses the ability to modulate the immune system in healthy pigs by decreasing lymphocyte subpopulations circulating in blood.
Show more [+] Less [-]Prevalence of intestinal parasitism of swine in a North Central State of Nigeria
2016
Julius Olaniyi Aiyedun | Oladapo Oyedeji Oludairo
Objective: Swine production is one of the most popular and promising livestock production industries in many parts of Nigeria. It prides itself in contributing to the supplementation of the overall national protein intake, gainfully engaging the citizenry in the industry and generating foreign exchange to the nation through export of produce and products. However, the swine production industry in Nigeria faced with religious and cultural constraints as well as several infectious diseases. The objective of this study was to investigate the prevalence of intestinal parasites in swine in the North Central State of Nigeria.Materials and methods: A total of 920 pigs comprising of 874 (95%) and 46 (5%) foreign and indigenous breed of pigs respectively were investigated in this study. Quantitative Mcmaster egg-counting technique was used for counting eggs of parasites. Semi-structured questionnaires was used to generate demographic data during October 2014 to March 2015. Results: Fifty five percent pigs were found to be positive for Hyostrogylus rubidus, while Ascaris suum, Trichuris suis and other unidentified eggs were present in 23, 17 and 5% pigs, respectively. and lsquo;Red Stomach Worm' infection in swine was the highest parasitic burden observed as compared to other parasites. Spreading of infection within sexes had a P value of >0.5.Conclusion: The prevalence of parasites in pigs obtained in this work is a consequence of improper husbandry measures and irregular veterinary medical intervention. [J Adv Vet Anim Res 2016; 3(3.000): 278-281]
Show more [+] Less [-]Identification and characterization of pig adipose-derived progenitor cells
2016
Zhang, Shuang | Bai, Chunyu | Zheng, Dong | Gao, Yuhua | Fan, Y. (Yanan) | Li, Lu | Guan, Weijun | Ma, Yuehui
Adipose-derived stem cells (ADSCs) are multipotent, and can be differentiated into many cell types in vitro. In this study, tissues from pigs were chosen to identify and characterize ADSCs. Primary ADSCs were sub-cultured to passage 28. The surface markers of ADSCs: CD29, CD71, CD73, CD90, and CD166 were detected by reverse-transcription polymerase chain reaction assays and the markers CD29, CD44, CD105, and vimentin were detected by immunofluorescence. Growth curves and the capacity of clone-forming were performed to test the proliferation of ADSCs. Karyotype analysis showed that ADSCs cultured in vitro were genetically stable. To assess the differentiation capacity of the ADSCs, cells were induced to differentiate into osteoblasts, adipocytes, epithelial cells, neural cells, and hepatocyte-like cells. The results suggest that ADSCs from pigs showed similar biological characteristics with those separated from other species, and their multi-lineage differentiation shows potential as an application for cellular therapy in an animal model.
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