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A New Enzyme-linked Immunosorbent Assay for Serological Diagnosis of Seal Parapoxvirus Infection in Marine Mammals 一种新的用于海洋哺乳动物海豹副痘病毒感染血清学诊断的酶联免疫吸附试验
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0005
Y. Badr, M. M. Rahman, Y. Ohno, Keita Ishijima, Ken Maeda, K. Kohyama, Y. Kamatari, K. Shimizu, Ayaka Okada, Y. Inoshima
Abstract Introduction Seal parapoxvirus (SPPV) infection has been reported among pinnipeds in aquaria in Japan; however, its seroprevalence is unknown. Therefore, an enzyme-linked immunosorbent assay (ELISA) was developed for serological diagnosis of SPPV infection. Material and Methods The gene encoding the major envelope protein of SPPV was cloned into the eukaryotic expression vector pAcGFP1-N1, which encodes the green fluorescence protein (GFP), thereby producing a fusion protein (Env-GFP). Parental and cloned vector DNA was independently transfected into cultured seal cells for the expression of GFP and Env-GFP. The wells of an ELISA plate were coated with either GFP- or Env-GFP-transfected cell lysates. The light absorbance of each serum sample was adjusted by subtracting the absorbance of GFP-coated wells from that of Env-GFP-coated wells. Sera from two spotted seals (Phoca largha), six beluga whales (Delphinapterus leucas), three Pacific white-sided dolphins (Lagenorhynchus obliquidens), and ten bottlenose dolphins (Tursiops truncatus) from an aquarium in Japan were examined using the ELISA. Results Positive reactions were not observed, except in one preserved sample collected ten years ago from a naturally SPPV-infected spotted seal. Conclusion The established ELISA could be useful in screening marine mammal sera for anti-SPPV antibodies.
摘要日本水族馆报告了海豹副痘病毒感染病例;然而,其血清患病率尚不清楚。因此,建立了一种用于SPPV感染血清学诊断的酶联免疫吸附试验(ELISA)。材料与方法将SPPV主要包膜蛋白的编码基因克隆到编码绿色荧光蛋白(GFP)的真核表达载体pAcGFP1-N1中,产生融合蛋白(Env-GFP)。将亲本和克隆载体DNA分别转染到培养的海豹细胞中,分别表达GFP和Env-GFP。用转染GFP或env -GFP的细胞裂解物包被ELISA板孔。通过从env - gfp包被孔中减去gfp包被孔的吸光度来调整每个血清样品的吸光度。采用ELISA检测了日本某水族馆2只斑点海豹(Phoca largha)、6只白鲸(Delphinapterus leucas)、3只太平洋白边海豚(Lagenorhynchus obliquidens)和10只宽吻海豚(Tursiops truncatus)的血清。结果除10年前自然感染sppv的斑海豹标本外,未见阳性反应。结论建立的酶联免疫吸附试验可用于海洋哺乳动物血清抗sppv抗体的筛选。
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引用次数: 1
Estrogen-related Receptor α (ERRα) Functions in The Hypoxic Injury of Microglial Cells 雌激素相关受体α (ERRα)在小胶质细胞缺氧损伤中的作用
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0009
Chaowei Deng, Tingting Zhu, S. Lian, Jianfa Wang, Rui Wu, Jia-san Zheng
Abstract Introduction Hypoxia is a common pathological condition after spinal cord injury. Oestrogen-related receptor alpha (ERRα), as a key regulator of energy metabolism and mitochondrial functions, plays an important role in maintaining cell homeostasis. However, its role in hypoxic spinal microglia has not been fully elaborated. This study investigated the receptor’s activity when these cells are hypoxic and used as an in vitro model. Material and Methods In this study, microglia (BV2) were exposed to cobalt chloride as a hypoxic model, and the inverse agonist of ERRα, XCT790, and pyrido[1,2-α]-pyrimidin-4-one were used to regulate the expression of the receptor to explore the ERRα-related mechanisms involved in hypoxic spinal cord injury (SCI). Results ERRα promoted autophagy in BV2 cells and inhibited the activation of the p38 mitogen-activated protein kinase (MAPK) pathway and the expression of anti-inflammatory factors under hypoxic conditions. It also promoted the expression of fibronectin type III domain containing protein 5 (FNDC5). Conclusion When a hypoxic SCI occurs, ERRα may maintain the homeostasis of spinal cord nerve cells by regulating autophagy and the p38MAPK/nuclear factor-kappa B cell and FNDC5/brain-derived neurotrophic factor signalling pathways, which are beneficial to the recovery of these cells.
缺氧是脊髓损伤后常见的病理状态。雌激素相关受体α (estrogen-related receptor α, ERRα)作为能量代谢和线粒体功能的关键调节因子,在维持细胞稳态中起着重要作用。然而,其在缺氧脊髓小胶质细胞中的作用尚未得到充分阐述。本研究考察了这些细胞缺氧时受体的活性,并将其作为体外模型。材料与方法本研究将小胶质细胞(BV2)暴露于氯化钴作为缺氧模型,利用ERRα、XCT790和吡啶多[1,2-α]-嘧啶-4-one的逆激动剂调控受体的表达,探讨ERRα参与缺氧脊髓损伤(SCI)的相关机制。结果ERRα促进BV2细胞自噬,抑制缺氧条件下p38丝裂原活化蛋白激酶(MAPK)通路的激活和抗炎因子的表达。它还促进了纤维连接蛋白III型结构域蛋白5 (FNDC5)的表达。结论缺氧性脊髓损伤时,ERRα可能通过调节脊髓细胞自噬及p38MAPK/核因子- κ B细胞和FNDC5/脑源性神经营养因子信号通路维持脊髓细胞的稳态,有利于脊髓细胞的恢复。
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引用次数: 1
Dirofilaria Repens in Dogs and Humans in Slovenia 斯洛维尼亚的狗和人出现了Dirofilaria
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0008
T. Kotnik, A. V. Rataj, B. Šoba
Abstract Introduction The prevalence of Dirofilaria repens in dogs in countries bordering Slovenia ranges from 1.5% to 47.3%. The aim of this study was to estimate its prevalence in Slovenian dogs and to present the cases of dirofilariasis diagnosed in humans from 2010 to 2020. Material and Methods Epidemiological data were collected and blood samples were taken from 465 dogs older than one year and born in Slovenia. A real-time PCR was performed on all samples to detect filarioid DNA, and a D. repens-and D. immitis-specific real-time PCR was performed on positive samples. Blood samples from 446 dogs were tested for Dirofilaria spp. using a modified Knott’s test. Human cases were diagnosed from histological sections of excised subcutaneous nodules. Descriptive statistics were used to characterise the samples. The one-sample nonparametric chi-squared test was used to assess whether categories of a variable were equally distributed. Results Three dogs’ samples tested positive for D. repens using the species-specific real-time PCR, while D. immitis DNA was not detected. The modified Knott’s test was positive in two of the three PCR-positive dogs, two of which had never travelled outside Slovenia’s borders. Four human patients with D. repens dirofilariasis were diagnosed. Since their travel history was unknown, autochthonous transmission could not be confirmed. Conclusion Our study demonstrated a 0.64% prevalence of D. repens infection in dogs in Slovenia. Two cases could be autochthonous.
在与斯洛文尼亚接壤的国家,犬类中Dirofilaria repens的流行率为1.5%至47.3%。本研究的目的是估计其在斯洛文尼亚狗中的患病率,并介绍2010年至2020年在人类中诊断出的dirofil虫病病例。材料与方法收集了465只出生在斯洛文尼亚的一岁以上的狗的流行病学数据和血液样本。对所有样品进行实时PCR检测丝虫病样DNA,对阳性样品进行D. repens和D. immitis特异性实时PCR检测。对446只狗的血液样本进行了Dirofilaria sps检测,使用了一种改进的Knott检测方法。人类病例是通过切除的皮下结节的组织学切片诊断的。使用描述性统计来描述样本的特征。使用单样本非参数卡方检验来评估变量的类别是否均匀分布。结果3只犬标本经实时荧光定量PCR检测呈鼠棘球蚴阳性,未检出犬棘球蚴DNA。修改后的诺特氏试验在三只pcr阳性的狗中有两只呈阳性,其中两只从未到过斯洛文尼亚境外。诊断出4例人雷氏迪罗氏丝虫病。由于他们的旅行史未知,因此无法确认本地传播。结论:我们的研究表明斯洛文尼亚犬中repens感染率为0.64%。两种情况可能是先天性的。
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引用次数: 1
Prevalence of Infectious Diseases on Dairy Farms Classified on The Basis of Their Biosecurity Score 基于生物安全评分的奶牛场传染病流行情况分类
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0006
L. Perillo, G. Cascone, F. Antoci, G. Piccione, C. Giannetto, Rosario Salonia, F. Salina, E. Giudice, V. Monteverde, F. Licitra
Abstract Introduction The aim of the study was to investigate the relationship between biosecurity as scored on the Italian National Animal Welfare Reference Centre (Centro di Referenza Nazionale per il Benessere Animale – CReNBA) checklist and the prevalence of Mycobacterium avium subsp. paratuberculosis, Chlamydophila abortus and Neospora caninum on dairy farms located in Ragusa, Italy. Material and Methods The checklist was used to assign an animal welfare score to 31 dairy farms. Twenty-one farms with a moderate score (>33%, <66%) formed group 1, and 10 farms with a high score (>66%) were group 2. Blood samples were collected from all cows on each farm to investigate the titres of antibodies against the relevant pathogens. Two-way analysis of variance was applied to assess differences between the two experimental groups and the Mann–Whitney test was applied to evaluate prevalence differences in the tested parasites between the groups. Results All tested farms had a score that classified them as either good or excellent. A higher incidence of Neospora caninum was observed in group 1. The incidences of the other two parasites were no different between the two groups. Conclusion The CReNBA checklist represents an impartial, reproducible, functional and smart instrument based on risk analysis and assigns a farm a mathematical animal welfare score. Among the parasites tested for, only Neospora caninum had prevalence influenced by biosecurity. Our preliminary results highlighted the positive associations between good animal welfare, high levels of biosecurity, and the prevention of the infectious diseases caused by the parasites in our focus, which are common on dairy farms.
摘要简介本研究的目的是调查意大利国家动物福利参考中心(Centro di Referenza Nazionale per il Benessere Animale - CReNBA)检查表中生物安全性得分与鸟分枝杆菌亚种流行率之间的关系。在意大利拉古萨的奶牛场发现了副结核、流产衣原体和犬新孢子虫。材料与方法采用检查表对31家奶牛场进行动物福利评分。21个中等得分的农场(>33%,66%)为第2组。从每个农场的所有奶牛中采集血样,以调查针对相关病原体的抗体滴度。采用双向方差分析评价两实验组间的差异,采用Mann-Whitney检验评价两组间被测寄生虫患病率差异。结果所有受测农场的得分均为良好或优秀。第1组犬新孢子虫发病率较高。另外两种寄生虫的发病率在两组之间没有差异。结论CReNBA清单代表了一种基于风险分析的公正、可复制、功能和智能的工具,并为农场分配了一个数学动物福利评分。在检测的寄生虫中,只有犬新孢子虫受生物安全因素的影响。我们的初步结果强调了良好的动物福利、高水平的生物安全与预防由我们关注的寄生虫引起的传染病之间的正相关关系,这些疾病在奶牛场很常见。
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引用次数: 1
Antibiotic Susceptibility of Mycobacteria Isolated from Ornamental Fish 观赏鱼分枝杆菌的抗生素敏感性研究
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0011
L. Guz, K. Puk
Abstract Introduction Nontuberculous mycobacteria (NTM) are increasingly recognised as causative agents of opportunistic infections in humans for which effective treatment is challenging. There is very little information on the prevalence of NTM drug resistance in Poland. This study was aimed to evaluate the susceptibility to antibiotics of NTM, originally isolated from diseased ornamental fish. Material and Methods A total of 99 isolates were studied, 50 of them rapidly growing mycobacteria (RGM) (among which three-quarters were Mycobacterium chelonae, M. peregrinum, and M. fortuitum and the rest M. neoaurum, M. septicum, M. abscessus, M. mucogenicum, M. salmoniphilum, M saopaulense, and M. senegalense). The other 49 were slowly growing mycobacteria (SGM) isolates (among which only one was M. szulgai and the bulk M. marinum and M. gordonae). Minimum inhibitory concentrations for amikacin (AMK), kanamycin (KAN), tobramycin (TOB), doxycycline (DOX), ciprofloxacin (CIP), clarithromycin (CLR), sulfamethoxazole (SMX), isoniazid (INH) and rifampicin (RMP) were determined. Results The majority of the isolates were susceptible to KAN (95.95%: RGM 46.46% and SGM 49.49%), AMK (94.94%: RGM 45.45% and SGM 49.49%), CLR (83.83%: RGM 36.36% and SGM 47.47%), SMX (79.79%: RGM 30.30% and SMG 49.49%), CIP (65.65%: RGM 24.24% and SGM 41.41%), and DOX (55.55%: RGM 9.06% and SGM 46.46%). The majority were resistant to INH (98.98%: RGM 50.50% and SGM 48.48%) and RMP (96.96%: RGM 50.50% and SGM 46.46%). Conclusion The drug sensitivity of NTM varies from species to species. KAN, AMK, CLR and SMX were the most active against RGM isolates, and these same four plus DOX and CIP were the best drugs against SGM isolates.
摘要介绍非结核分枝杆菌(NTM)越来越被认为是人类机会性感染的病原体,有效的治疗是具有挑战性的。关于波兰NTM耐药流行情况的信息很少。本研究旨在评价NTM对抗生素的敏感性,NTM最初是从患病观赏鱼中分离出来的。材料与方法共分离99株,其中快速生长分枝杆菌50株(其中四分之三为chelonae分枝杆菌、peregrinum分枝杆菌和fortuitum分枝杆菌,其余为neoaurum分枝杆菌、septicum分枝杆菌、脓肿分枝杆菌、mucogenicum分枝杆菌、沙门氏菌分枝杆菌、saopulense分枝杆菌和塞内加尔分枝杆菌)。其余49株为生长缓慢的分枝杆菌(SGM)分离株(其中只有1株为苏氏分枝杆菌,其余为海洋分枝杆菌和戈登分枝杆菌)。测定了阿米卡星(AMK)、卡那霉素(KAN)、妥布霉素(TOB)、多西环素(DOX)、环丙沙星(CIP)、克拉霉素(CLR)、磺胺甲恶唑(SMX)、异烟肼(INH)和利福平(RMP)的最低抑菌浓度。结果大部分菌株对KAN (95.95%: RGM 46.46%和SGM 49.49%)、AMK (94.94%: RGM 45.45%和SGM 49.49%)、CLR (83.83%: RGM 36.36%和SGM 47.47%)、SMX (79.79%: RGM 30.30%和SMG 49.49%)、CIP (65.65%: RGM 24.24%和SGM 41.41%)、DOX (55.55%: RGM 9.06%和SGM 46.46%)敏感。对INH (98.98%: RGM 50.50%, SGM 48.48%)和RMP (96.96%: RGM 50.50%, SGM 46.46%)耐药较多。结论NTM的药敏因种而异。KAN、AMK、CLR和SMX对RGM最有效,而这4种药物加DOX和CIP对SGM最有效。
{"title":"Antibiotic Susceptibility of Mycobacteria Isolated from Ornamental Fish","authors":"L. Guz, K. Puk","doi":"10.2478/jvetres-2022-0011","DOIUrl":"https://doi.org/10.2478/jvetres-2022-0011","url":null,"abstract":"Abstract Introduction Nontuberculous mycobacteria (NTM) are increasingly recognised as causative agents of opportunistic infections in humans for which effective treatment is challenging. There is very little information on the prevalence of NTM drug resistance in Poland. This study was aimed to evaluate the susceptibility to antibiotics of NTM, originally isolated from diseased ornamental fish. Material and Methods A total of 99 isolates were studied, 50 of them rapidly growing mycobacteria (RGM) (among which three-quarters were Mycobacterium chelonae, M. peregrinum, and M. fortuitum and the rest M. neoaurum, M. septicum, M. abscessus, M. mucogenicum, M. salmoniphilum, M saopaulense, and M. senegalense). The other 49 were slowly growing mycobacteria (SGM) isolates (among which only one was M. szulgai and the bulk M. marinum and M. gordonae). Minimum inhibitory concentrations for amikacin (AMK), kanamycin (KAN), tobramycin (TOB), doxycycline (DOX), ciprofloxacin (CIP), clarithromycin (CLR), sulfamethoxazole (SMX), isoniazid (INH) and rifampicin (RMP) were determined. Results The majority of the isolates were susceptible to KAN (95.95%: RGM 46.46% and SGM 49.49%), AMK (94.94%: RGM 45.45% and SGM 49.49%), CLR (83.83%: RGM 36.36% and SGM 47.47%), SMX (79.79%: RGM 30.30% and SMG 49.49%), CIP (65.65%: RGM 24.24% and SGM 41.41%), and DOX (55.55%: RGM 9.06% and SGM 46.46%). The majority were resistant to INH (98.98%: RGM 50.50% and SGM 48.48%) and RMP (96.96%: RGM 50.50% and SGM 46.46%). Conclusion The drug sensitivity of NTM varies from species to species. KAN, AMK, CLR and SMX were the most active against RGM isolates, and these same four plus DOX and CIP were the best drugs against SGM isolates.","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82534142","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
Comparison of Commercial Enzyme-linked Immunosorbent Assays for Diagnosis of Contagious Agalactia Caused By Mycoplasma Agalactiae 商用酶联免疫吸附法诊断无乳支原体致传染性无乳的比较
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0010
A. Sánchez, A. Contreras, MA Sánchez-Corral, Carmen Martínez-Nista, S. Collado, J. L. Sáez, O. Mínguez, C. de la Fe
Abstract Introduction Contagious agalactia (CA) is a disease affecting small ruminants with worldwide distribution and caused by several mycoplasmas, especially M. agalactiae. The main option for systematic diagnosis under monitoring control programmes is the enzyme-linked immunosorbent assay (ELISA) test. Material and Methods This study was designed to appraise the performance of two commercial indirect ELISA tests using M. agalactiae p48 protein and one using total protein, for antibody detection in small ruminants after natural infection with different M. agalactiae strains. We carried out the test evaluation using sera of confirmed M. agalactiae-positive goats with clinical signs. In addition, test agreement was assessed by kappa between the three commercial ELISA tests. Results All three ELISA tests showed high validity scores (Youden’s J: 72.9–84%). The sensitivity values for the P48 protein-based tests were 76.9% and 84.6%, and was 79% for the total protein-based test. The specificity of all tests was 100%. In addition, between the total protein-based ELISA test and the other two ELISA tests based on the P48 protein, the agreement was substantial (kappa: 0.762–0.763) and the agreement between the latter two tests was almost perfect (kappa: 0.93). Conclusion The validity parameters for all tests allowed their application for diagnostic purposes in lactating goats excreting M. agalactiae in milk and presenting clinical signs. The agreements show that any of these ELISA tests could be equally well used for diagnosis in programmes against CA.
摘要简介传染性无乳病(CA)是一种分布在世界各地的小反刍动物疾病,由几种支原体,尤其是无乳支原体引起。在监测控制规划下进行系统诊断的主要选择是酶联免疫吸附试验(ELISA)。材料与方法本研究采用两种商业化间接ELISA法分别检测无乳支原体p48蛋白和总蛋白对小反刍动物自然感染不同无乳支原体菌株后抗体的检测效果。我们使用临床症状确诊的无乳分枝杆菌阳性山羊的血清进行测试评估。此外,用kappa法评估了三种商用ELISA检测的一致性。结果3项酶联免疫吸附试验的效度均较高(约登J值:72.9 ~ 84%)。P48蛋白检测的敏感性分别为76.9%和84.6%,总蛋白检测的敏感性为79%。所有检查的特异性均为100%。此外,基于总蛋白的酶联免疫吸附试验与基于P48蛋白的其他两种酶联免疫吸附试验之间的一致性很好(kappa: 0.762-0.763),后两种酶联免疫吸附试验之间的一致性几乎是完美的(kappa: 0.93)。结论所有试验的效度参数均可用于诊断泌乳山羊在乳中排泄无乳乳杆菌并出现临床症状。这些协议表明,这些酶联免疫吸附试验中的任何一种都同样可以很好地用于诊断针对CA的规划。
{"title":"Comparison of Commercial Enzyme-linked Immunosorbent Assays for Diagnosis of Contagious Agalactia Caused By Mycoplasma Agalactiae","authors":"A. Sánchez, A. Contreras, MA Sánchez-Corral, Carmen Martínez-Nista, S. Collado, J. L. Sáez, O. Mínguez, C. de la Fe","doi":"10.2478/jvetres-2022-0010","DOIUrl":"https://doi.org/10.2478/jvetres-2022-0010","url":null,"abstract":"Abstract Introduction Contagious agalactia (CA) is a disease affecting small ruminants with worldwide distribution and caused by several mycoplasmas, especially M. agalactiae. The main option for systematic diagnosis under monitoring control programmes is the enzyme-linked immunosorbent assay (ELISA) test. Material and Methods This study was designed to appraise the performance of two commercial indirect ELISA tests using M. agalactiae p48 protein and one using total protein, for antibody detection in small ruminants after natural infection with different M. agalactiae strains. We carried out the test evaluation using sera of confirmed M. agalactiae-positive goats with clinical signs. In addition, test agreement was assessed by kappa between the three commercial ELISA tests. Results All three ELISA tests showed high validity scores (Youden’s J: 72.9–84%). The sensitivity values for the P48 protein-based tests were 76.9% and 84.6%, and was 79% for the total protein-based test. The specificity of all tests was 100%. In addition, between the total protein-based ELISA test and the other two ELISA tests based on the P48 protein, the agreement was substantial (kappa: 0.762–0.763) and the agreement between the latter two tests was almost perfect (kappa: 0.93). Conclusion The validity parameters for all tests allowed their application for diagnostic purposes in lactating goats excreting M. agalactiae in milk and presenting clinical signs. The agreements show that any of these ELISA tests could be equally well used for diagnosis in programmes against CA.","PeriodicalId":54685,"journal":{"name":"Onderstepoort Journal of Veterinary Research","volume":null,"pages":null},"PeriodicalIF":2.7,"publicationDate":"2022-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88653858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The Potential Risk of International Spread of Mycobacterium Bovis Associated with Movement of Alpacas 与羊驼活动有关的牛分枝杆菌国际传播的潜在风险
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0012
M. Krajewska-Wędzina, M. Miller, A. Didkowska, A. Kycko, Łukasz Radulski, M. Lipiec, M. Weiner
Abstract Introduction The study highlights the transboundary nature of tuberculosis (TB) in alpacas and the failure of current ante-mortem testing protocols (the tuberculin skin and Enferplex Camelid TB tests) to identify TB-free alpaca herds and individuals for export. Our research and the available literature indicate that the alpaca (Vicugna pacos) is extremely susceptible to Mycobacterium bovis infection, and that testing periodicity fails to take into account that animals do not manifest disease symptoms for a long time. The skin test failed to identify Mycobacterium bovis infection in two alpacas prior to their movement from the UK to Poland. The animals were purchased by a breeding centre in Poland, and were then shown at an international animal exhibition. The last owner of the alpacas before their deaths from TB bought the infected animals unwittingly in order to run rehabilitation activities with disabled children on his farm. Material and Methods Thoracic lymph node, lung and liver tissue samples obtained at necropsy were examined histopathologically after Ziehl–Neelsen staining. Tissue samples were homogenised and mycobacteria present there were cultured on Stonebrink’s medium during a 6-week incubation. A commercial test using polymorphism of the chromosomal direct repeat region provided species identification and additional identification was by spacer oligonucleotide typing and mycobacteria interspersed repetitive unit–variable number tandem repeat analysis with a gel electrophoresis protocol. Results The microbiological examination confirmed multiorgan TB caused by the SB0666 spoligotype of Mycobacterium bovis. Conclusion Due to the suboptimal performance of current diagnostic tests for TB in alpacas, there is a risk that infected animals may be moved unwittingly. A risk of TB spread associated with the international movement of alpacas is implied by this study.
摘要:本研究强调了羊驼结核病的跨界性,以及目前的死前检测方案(结核菌素皮肤和Enferplex Camelid结核病检测)在确定出口的无结核羊驼群和个体方面的失败。我们的研究和现有文献表明,羊驼(Vicugna pacos)对牛分枝杆菌感染非常敏感,而且检测的周期性未能考虑到动物在很长一段时间内没有表现出疾病症状。在两只羊驼从英国转移到波兰之前,皮肤测试未能确定牛分枝杆菌感染。这些动物是由波兰的一个繁殖中心购买的,然后在一个国际动物展览会上展出。羊驼死于结核病前的最后一位主人无意中购买了受感染的羊驼,以便在他的农场为残疾儿童开展康复活动。材料与方法尸检获得的胸淋巴结、肺、肝组织标本经Ziehl-Neelsen染色进行组织病理学检查。组织样本均质化,其中存在的分枝杆菌在Stonebrink培养基上培养6周。利用染色体直接重复序列区多态性进行的商业测试提供了物种鉴定,并通过间隔片段寡核苷酸分型和分枝杆菌穿插重复单位可变数串联重复序列凝胶电泳分析进行了额外鉴定。结果微生物学检查证实由牛分枝杆菌SB0666 spoligotype引起的多器官结核。结论由于目前羊驼结核病诊断测试的性能不理想,存在感染动物在不知情的情况下被转移的风险。这项研究暗示了与国际羊驼运动相关的结核病传播风险。
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引用次数: 4
In Vitro and in Vivo Activity of Lactobacillus Sakei L14 Strain Against Campylobacter Jejuni DC3 Strain 酒井乳杆菌L14对空肠弯曲杆菌DC3的体内外活性研究
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0015
J. Catacutan, Ma. Socorro Edden P. Subejano, Gil M Penuliar
Abstract Introduction Domestic poultry is a natural reservoir of Campylobacter, the host–pathogen interaction being predominantly asymptomatic. This study investigated whether chickens remain asymptomatic partly because of lactic acid bacteria (LAB). Material and Methods Campylobacter spp. and LAB were isolated from the gut of poultry chickens using enrichment and screening assays and were identified via rDNA sequencing. The C. jejuni DC3 isolate was grown in different cell-free supernatants (CFS) generated from a priority LAB isolate. An in vivo challenge involving the C. jejuni and LAB isolates using a chicken model was performed to confirm the in vitro findings. Results Twelve presumptive LAB isolates had anti-C. jejuni activity based on cross-streak and agar plug assays, with Lactobacillus sakei L14 isolate exhibiting the highest activity. Inhibition by L. sakei L14 CFS of the growth of C. jejuni occurred in a dose-dependent manner. Campylobacter jejuni DC3 inhibition was most evident in CFS harvested at 72 h and produced by co-culture with the pathogen. Neutralisation of the CFS abrogated the observed inhibition. Co-infection with C. jejuni DC3 and L. sakei L14 in vivo, however, failed to inhibit C. jejuni colonisation in chickens. Conclusion The results suggest that the anti-C. jejuni effect of L. sakei L14 in chickens may be due to mechanisms other than direct inhibition of growth.
摘要简介家禽是弯曲杆菌的天然宿主,宿主-病原体相互作用主要是无症状的。本研究调查了乳酸菌(LAB)是否在一定程度上导致鸡无症状。材料与方法从家鸡肠道中分离到弯曲杆菌(Campylobacter spp)和乳酸菌(LAB),并通过rDNA测序对其进行鉴定。将C. jejuni DC3分离物培养在由优先LAB分离物产生的不同无细胞上清液(CFS)中。利用鸡模型对空肠梭菌和乳酸菌进行体内攻毒,以证实体外研究结果。结果12株推定乳酸菌具有抗- c抗体。通过交叉条纹法和琼脂塞法测定空肠活性,其中酒井乳杆菌L14的活性最高。酒井L14 CFS对空肠梭菌生长的抑制呈剂量依赖性。空肠弯曲杆菌DC3的抑制作用在72 h收获的CFS中最为明显,并与病原体共培养。中和CFS消除了观察到的抑制作用。然而,在鸡体内同时感染空肠梭菌DC3和L. sakai L14并不能抑制空肠梭菌在鸡体内的定殖。结论抗c。白井乳酸菌L14对鸡空肠的影响可能是由于其直接抑制生长以外的机制。
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引用次数: 0
Occurrence of Ornithobacterium Rhinotracheale in Polish Turkey Flocks 波兰火鸡禽群中鼻气管炎鸟杆菌的发生
IF 2.7 3区 农林科学 Q1 Veterinary Pub Date : 2022-03-01 DOI: 10.2478/jvetres-2022-0014
O. Kursa, G. Tomczyk, Anna Sawicka-Durkalec
Abstract Introduction Ornithobacterium rhinotracheale (ORT) causes significant economic losses to the poultry industry around the world. The bacterium often affects poultry as part of multiple infections causing very serious clinical signs that are usually not limited only to the respiratory system. This study’s main objective was the retrospective detection and identification of ORT in turkey flocks. Material and Methods ORT identification was performed in 6,225 samples taken from 133 different flocks between 2015 and 2020. Molecular methods were used, specifically real-time PCR and traditional PCR. We focused on partial 16S rRNA gene sequences of isolates, which were compared with sequences obtained from GenBank. The reaction products were analysed phylogenetically. Molecular methods indicating secondary infections was carried out, and the bacterial composition of the upper respiratory tract was 16S metasequenced for selected flocks to identify any other pathogens. Results The presence of ORT was detected in 30.83% of samples by real-time PCR and 28.57% by PCR. Phylogenetic analysis of the PCR products from the turkeys samples showed that their sequences resolved into two main genetic groups. Tests for the occurrence of secondary infections showed the presence of Mycoplasma gallisepticum and M. synoviae in some samples but the total absence of Bordetella avium. The upper respiratory tract in turkeys was dominated by two major phyla Firmicutes and Proteobacteria. At the genus level, the genera Ornithobacterium, Mycoplasma, Gallibacterium, Avibacterium, and Escherichia-Shigella were found which may include pathogenic bacteria that can cause clinical symptoms. Conclusion The results of the analysis of multiple infection carried out in flocks with respiratory signs are probably associated with outbreaks of ornithobacteriosis in turkey flocks in Poland.
摘要鼻气管鸟杆菌(Ornithobacterium rhinotracheale, ORT)给世界范围内的家禽业造成了巨大的经济损失。这种细菌经常作为多重感染的一部分影响家禽,引起非常严重的临床症状,通常不仅限于呼吸系统。本研究的主要目的是回顾性检测和鉴定火鸡群中的ORT。材料与方法2015 - 2020年对133个不同禽群的6225份样本进行了ORT鉴定。采用分子方法,特别是实时荧光定量PCR和传统荧光定量PCR。我们将重点放在分离株的部分16S rRNA基因序列上,并与从GenBank中获得的序列进行比较。对反应产物进行了系统发育分析。采用分子方法提示继发性感染,对选取的鸡群进行上呼吸道细菌组成16S序列分析,以确定是否存在其他病原体。结果实时荧光定量PCR检测ORT的检出率为30.83%,PCR检出率为28.57%。从火鸡样品中提取的PCR产物的系统发育分析表明,它们的序列可分为两个主要的遗传群。继发感染的检测显示,在一些样本中存在鸡毒支原体和滑膜支原体,但完全没有禽博德特拉菌。火鸡的上呼吸道主要由厚壁菌门和变形菌门两大门组成。在属水平上发现鸟杆菌属、支原体属、Gallibacterium属、Avibacterium属和Escherichia-Shigella属,其中可能包括可引起临床症状的致病菌。结论在有呼吸体征的鸡群中进行的多重感染分析结果可能与波兰火鸡群中鸟类杆菌病的暴发有关。
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引用次数: 2
Antibody response to Raboral VR-G® oral rabies vaccine in captive and free-ranging black-backed jackals (Canis mesomelas). 人工饲养和放养的黑背豺对 Raboral VR-G® 口服狂犬病疫苗的抗体反应。
IF 1.5 3区 农林科学 Q2 VETERINARY SCIENCES Pub Date : 2022-02-10 DOI: 10.4102/ojvr.v89i1.1975
Katja N Koeppel, Peter Geertsma, Brian F Kuhn, Ockert L Van Schalkwyk, Peter N Thompson

Rabies is a zoonotic disease that remains endemic in large parts of southern Africa because of its persistence in wildlife and domestic dog vectors. The black-backed jackals (Canis mesomelas) is primarily the wildlife vector responsible for rabies outbreaks in northern parts of South Africa. Two trials were carried out to investigate antibody responses to the oral rabies vaccine Raboral V-RG® in black-backed jackals under captive and free-ranging conditions. In captive jackals 10/12 (83%; 95% confidence interval [CI]: 52% - 98%), seroconverted after single oral vaccination. Nine captive jackals had protective antibody titres ( 0.5 IU/mL) at 4 weeks (median: 2.1 IU/mL; inter quartile range [IQR]: 0.6-5.7) and 10 jackals had at 12 weeks (median: 3.5 IU/mL; IQR: 1.5-8.3) and three maintained antibody titres for up to 48 weeks (median: 3.4 IU/mL; IQR: 2.0-6.3). Four sites were baited with Raboral V-RG® vaccine for wild jackals, using fishmeal polymer and chicken heads. Baits were distributed by hand or from vehicle at three sites in north-eastern South Africa, with an average baiting density of 4.4 baits/km2 and at one site in central South Africa, at 0.12 baits/km2. This resulted in protective antibody titres in 3/11 jackals (27%; 95% Cl: 6-61) trapped between 3 and 12 months after baiting in north-eastern South Africa, compared with 4/7 jackals (57%; 95% Cl: 18-90) trapped after 3-18 months in central South Africa. This study shows the potential utility of oral rabies vaccination for the control of wildlife-associated rabies in north-eastern and central South Africa, but extensive studies with wider distribution of bait are needed to assess its potential impact on rabies control in wild jackals.

狂犬病是一种人畜共患病,由于其在野生动物和家犬载体中的持续存在,在南部非洲大部分地区仍然是地方病。黑背豺(Canis mesomelas)是南非北部地区爆发狂犬病的主要野生动物病媒。我们进行了两项试验,调查黑背豺在圈养和放养条件下对口服狂犬病疫苗 Raboral V-RG® 的抗体反应。在圈养豺中,10/12(83%;95% 置信区间 [CI]:52% - 98%)只在口服一次疫苗后出现血清转换。9 只人工饲养的豺在 4 周时获得了保护性抗体滴度(0.5 IU/mL)(中位数:2.1 IU/mL;四分位数间距 [IQR]:0.6-5.7),10 只豺在 12 周时获得了保护性抗体滴度(中位数:3.5 IU/mL;IQR:1.5-8.3),3 只豺的抗体滴度维持了 48 周(中位数:3.4 IU/mL;IQR:2.0-6.3)。在四个地点使用鱼粉聚合物和鸡头作为诱饵,诱杀野豺的 Raboral V-RG® 疫苗。在南非东北部的三个地点用人工或车辆投放毒饵,平均投饵密度为每平方公里 4.4 个毒饵,在南非中部的一个地点,平均投饵密度为每平方公里 0.12 个毒饵。在南非东北部,诱饵投放后 3 至 12 个月内被诱捕的 3/11 只豺(27%;95% Cl:6-61)获得了保护性抗体滴度,而在南非中部,诱饵投放后 3 至 18 个月内被诱捕的 4/7 只豺(57%;95% Cl:18-90)获得了保护性抗体滴度。这项研究表明,口服狂犬病疫苗对南非东北部和中部野生动物相关狂犬病的控制具有潜在作用,但还需要进行更广泛的诱饵分布研究,以评估其对野生豺狂犬病控制的潜在影响。
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引用次数: 0
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Onderstepoort Journal of Veterinary Research
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