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Melatonin prevents nicotine-induced hepatotoxicity by modulating apoptosis and histopathological changes in rats. 褪黑素通过调节大鼠体内细胞凋亡和组织病理学变化,预防尼古丁诱导的肝中毒。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.152949
S A Şengül, I İçen Taşkın, F Aşır, A Eraslan Şakar, G Pektanç Şengül

Nicotine, the main toxic component of tobacco, directly or indirectly causes adverse effects on the liver metabolism. Melatonin, secreted by the pineal gland, has anti-apoptotic activity as well as antioxidant activity. The aim of this study was to reveal the antiapoptotic effects of melatonin in rats with experimentally induced chronic liver damage with nicotine. In this study, 32 male Wistar albino rats were divided into four groups: control, melatonin, nicotine and nicotine+melatonin. During the experiment, nicotine (1 mg/kg) and melatonin (10 mg/kg) were administered daily intraperitoneally for 56 days. At the end of the study, the liver tissues were taken for histopathological, immunohistochemical and molecular analysis. The administration of melatonin was determined to partially alleviate histopathological changes in the liver tissue induced by nicotine, such as hepatocyte degeneration, vascular dilatation and congestion, and leukocyte infiltration. It was observed that there was a significant decrease in Bax expression levels and a significant increase in Bcl-2 expression levels in the nicotine+melatonin group when compared to the injury group. On the other hand, it was determined that melatonin administration reduced the Bax/Bcl-2 ratio, which was significantly higher in the nicotine group compared to the other groups, to a level close to the control group. Additionally, as a result of immunohistochemical evaluation, it was observed that decreased Bax expression and increased Bcl-2 expression in hepatocytes in the nicotine+melatonin group were at a level close to the control group. Our results revealed that melatonin is a hepatoprotective and effective antioxidant by suppressing cell apoptosis and increasing the rate of healing after damage at both the immunohistochemical and molecular levels.

尼古丁是烟草的主要有毒成分,直接或间接地对肝脏代谢产生不良影响。褪黑素由松果体分泌,具有抗凋亡活性和抗氧化活性。本研究旨在探讨褪黑素对实验性慢性尼古丁肝损伤大鼠的抗凋亡作用。本研究将32只雄性Wistar白化大鼠分为对照组、褪黑素组、尼古丁组和尼古丁+褪黑素组。实验期间,每天腹腔注射尼古丁(1 mg/kg)和褪黑素(10 mg/kg),连续56 d。研究结束时,取肝组织进行组织病理学、免疫组化和分子分析。研究发现,褪黑素可部分缓解尼古丁引起的肝组织病理改变,如肝细胞变性、血管扩张充血、白细胞浸润等。我们观察到,与损伤组相比,尼古丁+褪黑素组Bax表达水平显著降低,Bcl-2表达水平显著升高。另一方面,研究人员确定,褪黑激素的服用降低了Bax/Bcl-2的比率,尼古丁组的Bax/Bcl-2的比率明显高于其他组,达到接近对照组的水平。此外,免疫组化评价发现,尼古丁+褪黑素组肝细胞Bax表达降低,Bcl-2表达升高,与对照组接近。我们的研究结果表明,褪黑素在免疫组织化学和分子水平上通过抑制细胞凋亡和提高损伤后的愈合速度,具有肝保护和有效的抗氧化作用。
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引用次数: 0
Virus neutralization test for confirmation of ELISA-positive SARS-CoV-2 antibodies in dogs. 犬中确认elisa阳性SARS-CoV-2抗体的病毒中和试验。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.152946
D Vujin, A Knežević, V Vračar, D Petrović, G Kozoderović, Lj Spasojević Kosić, V Lalošević, D Lalošević

Constant antigenic changes, new variants and easy transmission of SARS-CoV-2 virus should acquire greater zoonotic attention and need to remain alert. In this retrospective study the aim was to analyze seropositivity to SARS-CoV-2 in dogs by commercial ELISA. The Virus neutralization test (VNT) was modified for the purpose of confirmation of SARS-CoV-2 antibodies in ELISA-positive dog sera. The sera were collected from 204 dogs from different veterinary clinics across Vojvodina Province, Serbia, during COVID-19 pandemic. For the screening of antibodies a commercial double multi-species antigen ELISA was used, followed by the VNT modified with SARS-CoV-2 as a confirmatory test. VNT was modified as "one step" test using local isolate of SARS-CoV-2 and the results were checked by cytopathic effect in cell culture on the 96-well microtiter plate. Obtained data have shown that 9 out of 204 dogs were positive by ELISA (4.4%), while 2 (0.97%) sera were doubtful. VNT confirmed 9 positive dogs, but 2 doubtful samples were negative, exhibiting the seroconversion to SARS-CoV-2 in 4.4% dogs from Vojvodina region during pandemic of COVID-19. VNT with SARS-CoV-2 helped to elucidate ELISA ambiguous results. The occurrence of antibodies to SARS-CoV-2 in dogs in this study during COVID-19 pandemic suggested the possibility of viral transmission to dogs, implicating the potential for zoonotic transmission. This was the first research on seroconversion to SARS-CoV-2 in dogs from the Province of Vojvodina, the northernmost part of Serbia.

SARS-CoV-2病毒的持续抗原变化、新变体和易传播应引起更大的人畜共患关注,并需要保持警惕。在这项回顾性研究中,目的是通过商业ELISA分析狗对SARS-CoV-2的血清阳性反应。对病毒中和试验(VNT)进行了改进,以确定elisa阳性犬血清中是否存在SARS-CoV-2抗体。在COVID-19大流行期间,从塞尔维亚伏伊伏丁那省不同兽医诊所的204只狗身上收集了血清。抗体的筛选采用商用双多种抗原ELISA法,然后用SARS-CoV-2修饰的VNT作为确认试验。采用SARS-CoV-2本地分离株将VNT改进为“一步法”,并在96孔微滴板上用细胞培养的细胞病变效应检验结果。获得的资料显示,204只狗中有9只(4.4%)ELISA阳性,2只(0.97%)血清可疑。VNT确认9只狗呈阳性,但2只可疑样本呈阴性,表明伏伊伏丁那地区在COVID-19大流行期间有4.4%的狗血清转化为SARS-CoV-2。带有SARS-CoV-2的VNT有助于阐明ELISA的不明确结果。在本研究中,在COVID-19大流行期间,狗中出现了SARS-CoV-2抗体,这表明病毒可能传播给狗,暗示人畜共患传播的可能性。这是对塞尔维亚最北部伏伊伏丁那省犬血清转化为SARS-CoV-2的首次研究。
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引用次数: 0
A comparative approach on the prophylactic impact of fermented beverages on acute ulcerative colitis in mouse model. 发酵饮料对小鼠急性溃疡性结肠炎预防作用的比较研究。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.152951
Z Erol, Ö Özmen

Acute ulcerative colitis is an inflammatory disease of the colon that is becoming increasingly prevalent. Yet, a growing body of evidence supports the efficacy of dietary interventions in preventing acute ulcerative colitis. Fermented beverages have been the focus of research in humans and animals for several years due to their potential to influence overall health functions with an emphasis on gut health. This research comprehensively explores the preventive effect of three fermented beverages (water kefir, dairy kefir, and kombucha) on acute ulcerative colitis in a CD-1 mouse model. Histopathological evaluation of the colon samples indicated that consumption of kombucha led to increased alleviation of the gross and histopathological lesions. Oral administration of kombucha positively affected overall intestinal microecological homeostasis by decreasing the coliform counts in this group contrasting the water and milk kefir groups. Moreover, physicochemical evaluation of the fermented beverages was conducted covering key parameters such as pH, acidity, total solids, radical scavenging activity and total phenolic content. Kombucha had the highest radical scavenging activity (85.61), total phenolic content (5.04 mg GAE/ mL), and total solids (0.70%), but the lowest pH (3.1) values. The findings from this research offer valuable insights into the distinct contribution of different fermented beverages on prevention of acute ulcerative colitis. Kombucha unravels a promising natural prevention approach for acute colitis, opening new perspectives for future research.

急性溃疡性结肠炎是一种越来越普遍的结肠炎症性疾病。然而,越来越多的证据支持饮食干预在预防急性溃疡性结肠炎方面的有效性。多年来,发酵饮料一直是人类和动物研究的焦点,因为它们有可能影响整体健康功能,尤其是肠道健康。本研究在CD-1小鼠模型中全面探讨了三种发酵饮料(水开菲尔、乳开菲尔和康普茶)对急性溃疡性结肠炎的预防作用。结肠样本的组织病理学评估表明,康普茶的消费导致增加减轻大体和组织病理学病变。与水和牛奶开菲尔组相比,口服康普茶通过减少大肠菌群计数对肠道微生态稳态产生积极影响。并对发酵饮料进行了理化评价,包括pH值、酸度、总固形物、自由基清除能力和总酚含量等关键参数。康普茶的自由基清除能力最高(85.61),总酚含量最高(5.04 mg GAE/ mL),总固结物含量最高(0.70%),但pH值最低(3.1)。本研究的发现为不同发酵饮料对预防急性溃疡性结肠炎的独特贡献提供了有价值的见解。康普茶揭示了一种有希望的自然预防急性结肠炎的方法,为未来的研究开辟了新的视角。
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引用次数: 0
Effects of caponization and age on the histology, lipid localization and fibre diameter in muscles of Rhode Island Red cockerels. 皂化和年龄对罗德岛红公鸡肌肉组织学、脂质定位和纤维直径的影响。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.152945
M Gesek, K Michalska, D Murawska

Poultry scientists are constantly studying different breeds of cockerels that would be suitable for capon meat production. Capon meat, although not yet very popular, is characterized by exceptional taste qualities that could appeal to many customers. Obtaining the appropriate palatability, structure and tenderness of capon meat is possible thanks to the reduction in androgen levels following the castration of roosters. Surgical or chemical castration affects the metabolism of fats, thus increasing their accumulation in the abdominal cavity, subcutaneous tissue and muscles. The main aim of our research was histological evaluation and analysis of the concentration and distribution of adipose tissue in muscles in Rhode Island Red cockerels and capons. In addition, we analysed the diameter of the pectoral muscle fibre. The experiment was performed on 200 Rhode Island Red cockerels; the testes were removed at 8 weeks of age. At 12, 16, 20, 24 and 28 weeks of age, 6 cockerels and 6 capons were slaughtered, and samples from the pectoral and thigh muscles were evaluated. Differences in the accumulation of adipose tissue with muscular atrophy (p<0.05) were observed in thigh muscles, with higher amounts in capons than in cockerels. All examined locations in the pectoral and thigh muscles of capons (around the blood vessels, in the perimysium, in the endomysium, and in the sarcoplasm) showed much higher concentrations of lipids compared to the levels in cockerels. The diameters of the pectoral muscle fibres were different (p<0.05) at 12 and 16 weeks of age, and the diameters of the giant fibres were different (p<0.05) at 12 and 20 weeks of age, with higher values in cockerels. The high concentration of lipids in the skeletal muscles of Rhode Island Red capons is impressive. These dual-purpose cockerel breeds can be a source of high-quality meat.

家禽科学家一直在研究不同品种的公鸡,以适合生产鸡肉。卡彭肉虽然还不是很受欢迎,但其独特的口感品质可能会吸引许多顾客。由于雄鸡去势后雄激素水平的降低,获得适当的适口性、结构和嫩度的阉鸡肉成为可能。手术或化学阉割会影响脂肪的代谢,从而增加脂肪在腹腔、皮下组织和肌肉中的积累。我们研究的主要目的是对罗德岛红公鸡和阉鸡肌肉中脂肪组织的浓度和分布进行组织学评价和分析。此外,我们还分析了胸肌纤维的直径。实验在200只罗德岛红小公鸡上进行;在8周龄时切除睾丸。分别于12、16、20、24和28周龄屠宰6只公鸡和6只阉公鸡,并对其胸肌和大腿肌样本进行评价。脂肪组织积累与肌肉萎缩的差异(p
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引用次数: 0
Non-invasive methods for diagnosing pregnancy in cows and their real value. 奶牛妊娠诊断的非侵入性方法及其真正价值。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.152957
J M Jaśkowski, G Wozniak, P Kaźmierczak

Proper management of cattle reproduction has a major impact on the efficiency and profitability of dairy production. Ultrasound examination and transrectal palpation or the pregnancy-associated glycoprotein (PAG) test are currently the most commonly used methods for pregnancy diagnosis. However, alternative methods to those mentioned above are constantly being sought in order to minimise stress during the examination, the cost of veterinary services and to reduce the rate of errors in pregnancy diagnosis. Non-invasive methods of pregnancy diagnosis in cows are being improved, which include the barium chloride test, sulphuric acid, seed germination test, measurement of progesterone, interferon-tau or interferon-stimulated genes (ISGs), PAG, early pregnancy factor (EPF), estrone sulphate, thermography and electrocardiography. Over the past few decades, these methods have been extensively described. Some of these tests require blood, milk or urine for the diagnosis of pregnancy, while others require prolonged contact with the animal in order to take the appropriate measurements. Despite their advantages in terms of simplicity and lower cost compared with traditional methods of pregnancy diagnosis, they are sometimes problematic because of the difficulty of collecting material for testing. They allow the determination of a pregnancy without determining its age or pathology on the part of the development of the fetus and the reproductive system. They are also generally characterised by lower accuracy, sensitivity and specificity, which can have a negative impact on reproductive management and translate into the economics of dairy production. In the context of the above information, it appears that non-invasive methods of pregnancy diagnosis need to be further improved to minimise or eliminate the disadvantages cited.

牛的繁殖管理得当,对奶牛生产的效率和盈利能力有重大影响。超声检查和经直肠触诊或妊娠相关糖蛋白(PAG)试验是目前最常用的妊娠诊断方法。然而,为了尽量减少检查期间的压力、兽医服务的费用和减少怀孕诊断的错误率,正在不断寻求替代上述方法的方法。奶牛妊娠诊断的无创方法正在得到改进,其中包括氯化钡试验、硫酸、种子发芽试验、黄体酮测量、干扰素tau或干扰素刺激基因(ISGs)、PAG、早孕因子(EPF)、硫酸雌酮、热成像和心电图。在过去的几十年里,这些方法得到了广泛的描述。其中一些测试需要血液、乳汁或尿液来诊断是否怀孕,而另一些测试则需要与动物长时间接触,以便进行适当的测量。尽管与传统的妊娠诊断方法相比,它们具有简单和低成本的优势,但由于难以收集检测材料,它们有时会出现问题。它们可以在不确定其年龄或胎儿发育和生殖系统病理的情况下确定是否怀孕。它们通常还具有准确性、敏感性和特异性较低的特点,这可能对生殖管理产生负面影响,并转化为乳制品生产的经济性。在上述信息的背景下,似乎需要进一步改进非侵入性妊娠诊断方法,以尽量减少或消除所提到的缺点。
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引用次数: 0
TaqMan-based quantitative real-time polymerase chain reaction assay to detect porcine circovirus-like virus. 基于taqman的实时定量聚合酶链反应法检测猪圆环病毒样病毒。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.152954
Z R Yu, Y Shao, Z Chen, Y Zhang, F Y Cheng, H Liu, Z Y Wang, J Tu, X J Song, K Z Qi

The aim of this study was to develop a rapid, sensitive and highly specific TaqMan quantitative real-time polymerase chain reaction PCR (qPCR) assay for porcine circovirus-like virus (PCLV). The primers and probe were designed based on the conserved regions of the PCLV ORF4 gene. The assay has a good detection performance (y=-3.3257x+ 1.482, R2=0.9905), with a limit of detection of 10 copies, which was 100 times more sensitive than conventional PCR (cPCR). No cross-reactivity was observed with other common viruses. The intra- and inter-assay coefficients of variation were less than 1.25%. 36 fecal samples were analyzed using this method, detecting a positivity rate of 8.33% (3/36) that was higher than the cPCR detected. In summary, the established assay for PCLV detection has high specificity, sensitivity, and reproducibility and can be used as a tool for clinical diagnosis and epidemiological investigation.

本研究旨在建立一种快速、灵敏、高特异性的猪圆环病毒样病毒(PCLV) TaqMan实时定量PCR (qPCR)检测方法。根据PCLV ORF4基因的保守区设计引物和探针。该方法检测性能良好(y=-3.3257x+ 1.482, R2=0.9905),检出限为10份,灵敏度比常规PCR (cPCR)提高100倍。与其他常见病毒无交叉反应。试验内和试验间变异系数均小于1.25%。采用该方法对36份粪便标本进行了分析,阳性率为8.33%(3/36),高于cPCR检测结果。综上所述,所建立的PCLV检测方法具有较高的特异性、敏感性和重复性,可作为临床诊断和流行病学调查的工具。
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引用次数: 0
Pathogenicity and drug resistance characterization of Streptococcus agalactiae isolated from dairy cows. 奶牛无乳链球菌的致病性和耐药性研究。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.151748
L J Jiang, H R Liu, Z Y Liu, Q Li, Y C Wang, B W Tan

Streptococcus agalactiae, commonly known as S. agalactiae, is a critical zoonotic pathogen that significantly reduces milk yield and product quality and poses a significant risk to public health. Although S. agalactiae is increasingly recognised as a principal agent causing milkborne infections, research dedicated to this pathogen in dairy cattle has been less extensive than that of other pathogens. This study aimed to examine the antibiotic resistance profiles of S. agalactiae derived from dairy cows and assess its pathogenicity using validated in vivo models. The findings contribute essential scientific insights into the realm of environmental antibiotic resistance research. The resistance of S. agalactiae isolates to drugs was assessed using the broth microdilution technique. Additionally, PCR analysis was used to identify six important virulence genes. The study revealed that S. agalactiae was fully susceptible to streptomycin, meropenem, ciprofloxacin, clindamycin, cefquinome, and cloxacillin in general laboratory settings and within milk samples. However, among the antibiotics tested, tetracycline exhibited the highest level of resistance, with rates reaching 70%. Penicillin showed a resistance level of 50%, followed by doxycycline at 30%. Additionally, the resistance rates for apramycin and cefoxitin were both 20%, whereas florfenicol resistance was observed at a rate of 10%. All isolates of S. agalactiae carried the cfb gene. However, it is noteworthy that only one isolate possessed this gene exclusively, while the other nine isolates shared a uniform set of four additional virulence genes. The study highlighted the significant impact of these virulence factors on the pathogenic behaviour of S. agalactiae from dairy sources. This was demonstrated by the high mortality rates observed in experimental infections using Galleria mellonella (G. mellonella) larvae and mouse models. These findings contribute to understanding the relationship between the pathogenic properties of S. agalactiae and the virulence genes it carries.

无乳链球菌,俗称无乳链球菌,是一种重要的人畜共患病原体,可显著降低牛奶产量和产品质量,并对公众健康构成重大风险。尽管无乳链球菌越来越被认为是引起乳源性感染的主要病原体,但对奶牛中这种病原体的研究却没有对其他病原体的研究那么广泛。本研究旨在研究奶牛来源的无乳链球菌的抗生素耐药性特征,并通过验证的体内模型评估其致病性。这些发现为环境抗生素耐药性研究领域提供了重要的科学见解。采用肉汤微量稀释法测定了无乳链球菌对药物的耐药性。此外,PCR分析还鉴定了6个重要的毒力基因。研究表明,在一般实验室环境和牛奶样品中,无乳链球菌对链霉素、美罗培南、环丙沙星、克林霉素、头孢醌和氯西林完全敏感。然而,在所测试的抗生素中,四环素的耐药率最高,达到70%。青霉素的耐药水平为50%,其次是强力霉素的30%。此外,阿帕霉素和头孢西丁的耐药率均为20%,而氟苯尼考的耐药率为10%。所有的无乳链球菌都携带cfb基因。然而,值得注意的是,只有一个分离株独家拥有该基因,而其他9个分离株共享一套统一的4个额外的毒力基因。该研究强调了这些毒力因素对乳制品来源的无乳链球菌致病行为的重大影响。这一点可以通过在使用mellonella (G. mellonella)幼虫和小鼠模型的实验感染中观察到的高死亡率得到证实。这些发现有助于了解无乳链球菌的致病特性与其携带的毒力基因之间的关系。
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引用次数: 0
Cellular distribution of some intermediate filaments in the rat mammary gland during pregnancy, lactation and involution. 妊娠期、哺乳期和复归期大鼠乳腺某些中间纤维的细胞分布。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.152948
B Bayram, H Sağsöz, U Topaloğlu

Intermediate filaments (IFs) play a major role in determining and maintaining cell shape and anchoring intracellular organelles in place, in the tissues and organs of several species, starting from the early stages of development. This study was aimed at the immunohistochemical investigation of the presence, cellular localization and temporal distribution of the intermediate filaments keratin 8 (CK8), keratin 18 (CK18), keratin 19 (CK19), vimentin, desmin and laminin, all of which contribute to the formation of the cytoskeleton in the rat mammary gland during pregnancy, lactation and involution. On days 7, 14 and 21 of pregnancy (pregnancy period), on day 7 post-delivery (lactation period) and on day 7 post-weaning (involution period), under ketamine hydrochloride (Ketalar-Pfizer) (90 mg/kg) anesthesia, two mammary glands were fully excised from the abdominal region. It was determined that CK8 showed moderate immunoreactions in the alveolar and ductal epithelia, connective tissue and vascular endothelium of the rat mammary gland throughout pregnancy. On the 7th day of pregnancy, CK18 expression was absent in the alveolar and ductal epithelia, but was observed weakly in some connective tissue cells. Throughout pregnancy, lactation and involution, the alveolar and ductal epithelia of the rat mammary gland were determined to be negative for CK19. Desmin expression predominated in the mammary myoepithelium and vasculature throughout all three of the investigated periods. While vimentin was not expressed in any of the mammary tissue components during pregnancy and lactation, its moderate expression was observed in the alveolar and ductal epithelia during involution. The involution period was also characterized by the vimentin negativity of the myoepithelium, stroma, fat cells and blood vessels of the mammary gland. Throughout all three periods, laminin expression was strong in the alveolar and ductal epithelia, stromal and myoepithelial cells and blood vessels, and did not vary in strength between the investigated periods. These findings demonstrated that intermediate filaments showed cell- and tissue-specific expression patterns in the rat mammary gland under the effects of pregnancy, lactation and involution.

在一些物种的组织和器官中,中间丝(if)在决定和维持细胞形状和将胞内细胞器固定在适当位置上起着重要作用,从发育的早期开始。本研究旨在通过免疫组化方法研究大鼠乳腺细胞骨架在妊娠、哺乳期和复复期形成的中间丝角蛋白8 (CK8)、角蛋白18 (CK18)、角蛋白19 (CK19)、vimentin、desmin和层粘连蛋白的存在、细胞定位和时间分布。在妊娠第7、14、21天(妊娠期),分娩后第7天(哺乳期)和断奶后第7天(复复期),在盐酸氯胺酮(凯塔拉-辉瑞)(90 mg/kg)麻醉下,从腹部完全切除2个乳腺。结果表明,CK8在妊娠期间对大鼠乳腺的肺泡上皮、导管上皮、结缔组织和血管内皮均有中等程度的免疫反应。在妊娠第7天,CK18在肺泡和导管上皮中不表达,但在部分结缔组织细胞中表达较弱。在整个妊娠期、哺乳期和复归期,大鼠乳腺的肺泡上皮和导管上皮均检测到CK19阴性。在所有三个研究期间,Desmin的表达在乳腺肌上皮和脉管系统中占主导地位。在妊娠和哺乳期,波形蛋白在任何乳腺组织成分中均不表达,但在复旧期间,在肺泡和导管上皮中观察到其适度表达。复旧期还表现为乳腺肌上皮、间质、脂肪细胞和血管的波形蛋白阴性。在所有三个时期,层粘连蛋白在肺泡和导管上皮、基质和肌上皮细胞以及血管中的表达都很强烈,并且在不同时期的表达强度没有变化。这些结果表明,在妊娠、哺乳和复归的影响下,中间丝在大鼠乳腺中表现出细胞特异性和组织特异性的表达模式。
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引用次数: 0
Development of a capsid protein-based ELISA for the detection of PCV2 antibodies in swine serum. 基于衣壳蛋白的猪血清PCV2抗体ELISA检测方法的建立。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.151748
Y B Wang, P Li, Y C Gao, P F Hao, J W Feng, N Y Hu, J Cao, J H Hu, K Ding, L Wang

Porcine circovirus type 2 (PCV2) is the major causative agent of postweaning multisystemic wasting syndrome which leads to significant economic losses in the global swine industry. In China, there is a widespread dissemination of PCV2 infection in the pig population. Serological diagnosis of the disease is considered as an effective control measure. Here, we developed a capsid protein (Cap)-based enzyme-linked immunosorbent assay (Cap-ELISA) for the detection of PCV2 antibodies in swine serum using a nuclear localization signal-truncated capsid protein produced in Escherichia coli. The Cap protein was expressed as water-soluble and purified using nickel-nitrilotriacetic acid (Ni-NTA) chromatography. After the optimization of the working conditions of the Cap-ELISA using chessboard titrations, a total of 649 serum samples were tested using the Cap-ELISA and a commercial ELISA kit. The diagnostic sensitivity (DSN), diagnostic specificity (DSP) and accuracy of the Cap-ELISA were determined to be 96.7%, 94.1% and 99.5%, respectively. Cross-reactivity analysis indicated that the Cap-ELISA was PCV2-specific and possessed no cross-reactions with antibodies against other common swine pathogens including porcine circovirus type 1 (PCV1), porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), porcine parvovirus (PPV), foot and mouth disease virus (FMDV), porcine epidemic diarrhea virus (PEDV) and pseudorabies virus (PRV). Repeatability of the experiment showed that Cap-ELISA was highly repeatable with the intra- and inter-plate coefficients of variation less than 10%. Hence, the Cap-ELISA has the potential for the swine industry to monitor PCV2 epidemiology and to evaluate PCV2 vaccine efficacy.

猪圆环病毒2型(PCV2)是断奶后多系统消耗综合征的主要病原体,给全球养猪业造成重大经济损失。在中国,PCV2感染在猪群中广泛传播。血清学诊断被认为是有效的控制措施。在这里,我们开发了一种基于衣壳蛋白(Cap)的酶联免疫吸附试验(Cap- elisa),利用大肠杆菌生产的核定位信号截断的衣壳蛋白检测猪血清中的PCV2抗体。Cap蛋白以水溶性形式表达,采用镍-硝基三乙酸(Ni-NTA)层析纯化。在棋盘滴定法优化Cap-ELISA的工作条件后,使用Cap-ELISA和商用ELISA试剂盒对649份血清样本进行检测。Cap-ELISA的诊断灵敏度(DSN)、诊断特异性(DSP)和准确性分别为96.7%、94.1%和99.5%。交叉反应性分析表明,Cap-ELISA对猪圆环病毒1型(PCV1)、猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)、猪细小病毒(PPV)、口蹄疫病毒(FMDV)、猪流行性腹泻病毒(PEDV)和伪狂犬病毒(PRV)等常见猪病原体的抗体均无交叉反应。实验重复性表明,Cap-ELISA重复性高,板内和板间变异系数均小于10%。因此,Cap-ELISA具有监测PCV2流行病学和评估PCV2疫苗疗效的潜力。
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引用次数: 0
Carriage of antimicrobial resistance genes in Escherichia coli of bovine origin. 牛源大肠杆菌耐药基因的携带。
Pub Date : 2024-12-01 DOI: 10.24425/pjvs.2024.151749
G Ozbey, E S Tanriverdi, M N Acik, R Kalin, B Otlu, F Zigo

The present study aimed to search for the presence of the plasmid-mediated antimicrobial resistance genes in 106 Escherichia coli (E. coli) isolates from a total of 240 fresh fecal samples collected from 12 private cattle farms in Bingol province of East Turkey from November 2021 to January 2022. In those colistin-resistant E. coli (mcr-1 to -9), the major carbapenemase (blaOXA-48, blaNDM-1, blaIMP, blaVIM, and blaKPC), β-lactamase (blaTEM-1, blaCTX-M and blaSHV-1) and OXA-48 like β- lactamase (blaOXA-162, blaOXA-163, blaOXA-181, blaOXA-204 and blaOXA-232) resistance genes were searched for determined a multiplex polymerase chain reaction (PCR) method and Next-generation sequencing (NGS) - PCR Amplicons with Nanopore Technology. Only the mcr-4 gene was found in one isolate and the remaining genes (mcr-1-9) were not shown in all E. coli isolates from cattle. The minimal inhibitory concentration (MIC) to colistin was detected in mcr-4 positive E. coli isolates using broth microdilution. We assessed the antimicrobial susceptibilities of mcr-4 positive E. coli isolates using the Kirby-Bauer disk diffusion method. E coli isolate was detected as negative for carbapenemase and OXA-48 like β-lactamase resistance genes and positive for β-lactamase. In addition, E. coli isolates carrying mcr-4 were more resistant to colistin. Antimicrobial susceptibility testing using the disk diffusion assay indicated that all 106 E. coli isolates (100%) were sensitive to AMK, 105 E. coli isolates (99.1%) exhibited sensitivity to imipenem, meropenem and doripenem, and 1 E. coli isolate (0.9%) had intermediate resistance to imipenem, meropenem and doripenem; It was observed that all strains (100%) were resistant to cefotaxime. E. coli isolates are resistant to ampicillin (95.3%), amoxicillin/clavulanic acid (95.3%), cefepime (14.2%), cefixime (19.8%), cephalexin (74.5%), gentamicin (42.5%), kanamycin (37.7%), streptomycin (69.8%), tetracycline (80.2%), ciprofloxacin (60.4%), norfloxacin (13.2%), chloramphenicol (59.4%) and trimethoprim/sulfamethoxazole (68.9%). When we investigated the sequence in the Blast database, the genome of the E. coli isolate indicated high similarity with the mcr-4 sequences. To our knowledge, this is the first report investigating on the mcr-4 gene in E. coli identified from cattle in Turkey. Our results highlighted that cattle might be a potential risk in transmitting mcr genes.

本研究旨在从2021年11月至2022年1月从东土耳其Bingol省12个私人牛场收集的240份新鲜粪便样本中分离出106株大肠杆菌(E. coli),寻找质粒介导的抗微生物药物耐药性基因的存在。在耐粘菌素大肠杆菌(mcr-1 ~ -9)中,利用多重聚合酶链式反应(PCR)方法和纳米孔技术新一代测序(NGS) - PCR扩增子,寻找主要的碳青霉烯酶(blaOXA-48、blaNDM-1、blaIMP、blaVIM和blaKPC)、β-内酰胺酶(blaem -1、blaCTX-M和blaSHV-1)和OXA-48样β-内酰胺酶(blaOXA-162、blaOXA-163、blaOXA-181、blaOXA-204和blaOXA-232)耐药基因。仅在一个分离株中发现了mcr-4基因,其余基因(mcr-1-9)未在所有牛大肠杆菌分离株中发现。采用肉汤微量稀释法检测mcr-4阳性大肠杆菌分离株对粘菌素的最低抑制浓度(MIC)。采用Kirby-Bauer圆盘扩散法对mcr-4阳性大肠杆菌分离株进行了抗菌药物敏感性评价。大肠杆菌碳青霉烯酶和OXA-48样β-内酰胺酶耐药基因均阴性,β-内酰胺酶阳性。此外,携带mcr-4的大肠杆菌分离株对粘菌素的耐药性更强。药敏试验结果表明,106株大肠杆菌(100%)对AMK敏感,105株(99.1%)对亚胺培南、美罗培南、多利培南敏感,1株(0.9%)对亚胺培南、美罗培南、多利培南耐药;所有菌株(100%)均对头孢噻肟耐药。大肠杆菌对氨苄西林(95.3%)、阿莫西林/克拉维酸(95.3%)、头孢吡肟(14.2%)、头孢克肟(19.8%)、头孢氨苄(74.5%)、庆大霉素(42.5%)、卡那霉素(37.7%)、链霉素(69.8%)、四环素(80.2%)、环丙沙星(60.4%)、诺氟沙星(13.2%)、氯霉素(59.4%)、甲氧苄啶/磺胺甲恶唑(68.9%)耐药。当我们研究Blast数据库中的序列时,大肠杆菌分离物的基因组显示与mcr-4序列高度相似。据我们所知,这是对土耳其牛大肠杆菌中mcr-4基因进行调查的第一份报告。我们的结果强调,牛可能是传播mcr基因的潜在风险。
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引用次数: 0
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Polish journal of veterinary sciences
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