禁抗后仔猪细菌性腹泻的临床诊断与防治

为了促进食品安全和养猪业的健康发展，禁止在饲料中使用促生长抗生素势在必行。随着我国的饲料禁抗相关法规的推行，饲料企业被严格禁止在饲料生产过程中加入抗生素，这也导致了猪场疾病尤其是肠道疾病的高发，而仔猪肠道发育未完全，免疫系统未发育成熟，更容易发生严重的腹泻甚至死亡。文章对几种常见的仔猪细菌性腹泻疾病进行了回顾，病原包括大肠杆菌、沙门氏菌、魏氏梭菌，讨论了它们的临床症状与防控措施，为猪场在防控这些细菌性腹泻提供参考。     

Clostridium bifermentans and C. subterminale are associated with kiwifruit vine decline,known as moria,in Italy

Since 2012, a new pathogenic syndrome has frequently been observed in many areas of kiwifruit cultivation in Italy. The main symptoms include an initial withering of the leaves followed by a total and sudden collapse of plants, mainly occurring during summer. The withered leaves fall and the main and secondary feeder roots appear rotten, sometimes showing a reddish-brown discoloration. The disease, that affects both the green and yellow-fleshed cultivars, has been called kiwifruit vine decline and is locally known as moria. The syndrome has been found consistently associated with soil waterlogging, which frequently occurs either after the traditional agronomical practice of irrigating orchards through surface irrigation or after very heavy rainfall. So far, the role played by bacteria in this syndrome has not been investigated. In the present study, Clostridium spp. were isolated from both rotten roots and soils obtained from Italian kiwifruit orchards affected by the syndrome, indicating for the first time that anaerobic bacteria are able to cause damage to woody crops. C. bifermentans and C. subterminale incited symptoms in kiwifruit in both in vivo and in vitro pathogenicity tests. Soil waterlogging seems to potentially favour colonization of kiwifruit roots by anaerobic bacteria, probably because saturation of the soil can facilitate proliferation and persistence of these bacteria during long periods of the vegetative growth of the crop. The occurrence of anaerobic bacteria does not exclude the possibility that other microorganisms can play additional/synergic role(s) in causing the kiwifruit vine decline.     

黄芪多糖和丁酸梭菌对蛋雏鸭生长性能、血清生化指标的影响

本试验旨在研究日粮中添加黄芪多糖和丁酸梭菌及其复合剂对蛋雏鸭生长性能和血清生化指标的影响。选取1日龄健康绍兴公鸭600只，随机分为5组，每组6个重复，每个重复20只。Ⅰ组饲喂基础日粮（对照组），Ⅱ~Ⅴ组分别在基础日粮中添加40 mg/kg的杆菌肽锌（抗生素组）、800 mg/kg的黄芪多糖、250 mg/kg丁酸梭菌、800 mg/kg黄芪多糖+250 mg/kg丁酸梭菌复合剂，试验期为28 d。结果显示：①1~14日龄时，各组平均日增重（ADG）、平均日采食量（ADFI）和料重比（F/G）差异均不显著（P>0.05）。15~21日龄时，与Ⅰ、Ⅱ组相比，Ⅲ、Ⅳ、Ⅴ组ADG显著提高（P<0.05）、F/G显著降低（P<0.05）；22~28日龄时，Ⅲ、Ⅳ、Ⅴ组ADG显著高于Ⅰ组（P<0.05）；Ⅴ组ADFI显著高于其他组（P<0.05），Ⅲ、Ⅳ、Ⅴ组F/G显著低于Ⅰ、Ⅱ组（P<0.05）。1~28日龄时，与Ⅰ、Ⅱ组相比，Ⅲ、Ⅳ、Ⅴ组ADG显著提高（P<0.05）、F/G显著降低（P<0.05）。②试验第28天时，与Ⅰ组相比，Ⅴ组三碘甲状腺原氨酸（T3）含量显著提高（P<0.05）；Ⅲ、Ⅴ组甲状腺素（T4）、生长激素（GH）含量显著提高（P<0.05）；各组间血清皮质醇（CORT）含量无显著差异（P>0.05）；Ⅳ、Ⅴ组白细胞介素-2（IL-2）显著高于Ⅰ、Ⅱ组（P<0.05）。Ⅲ、Ⅴ组干扰素-γ（IFN-γ）显著高于其他组（P<0.05）。综上所述，日粮中添加黄芪多糖和丁酸梭菌可一定程度上提高蛋雏公鸭生长性能，改善血清生化指标，黄芪多糖和丁酸梭菌复合添加效果优于单独添加。     

Death of a neonatal lamb due to Clostridium perfringens type B in New Zealand

ABSTRACT

Case history and clinical findings: A flock of 20 sheep was kept within three paddocks on a single property. None of the animals in the flock had been vaccinated against any disease for at least three years. Abdominal bloating and haemorrhagic diarrhoea were observed in Lamb 1 at 24 hours-of-age. The lamb subsequently died within an hour of the onset of clinical signs. Lamb 2 was 3-days-old when observed to be recumbent with opisthotonus. The lamb was treated with dextrose, vitamins B1 and B12, and penicillin G, but died 4 hours later.

Pathological findings: Examination of Lamb 1 revealed markedly increased gas within the peritoneum and within dilated loops of intestine. The intestines were dark red and contained large quantities of haemorrhagic fluid. Histology of the intestines revealed peracute mucosal necrosis with minimal accompanying inflammation. The intestinal lumen contained cell debris, haemorrhage, and myriad large Gram-positive bacilli. The intestines of Lamb 2 did not appear bloated or reddened. However, multiple fibrin clots were visible within the pericardial sac. Histopathological examination revealed small foci of necrosis within the mucosa of the distal intestine. The necrotic foci were often associated with large numbers of large Gram-positive bacilli.

Immunohistochemsitry and molecular biology: Intestinal samples from Lamb 1 were processed for Clostridium perfringens immunohistochemistry, which revealed large numbers of intralesional, positively immunostained rods. Fragments corresponding to the expected sizes for genes encoding alpha, beta, and epsilon C. perfringens typing toxins were amplified by PCR from DNA extracted from formalin-fixed sections of intestine.

Diagnosis: Lamb dysentery due to C. perfringens type B.

Clinical relevance: C. perfringens bacteria have a worldwide distribution, but disease due to C. perfringens type B has only been diagnosed in a small number of countries and has never been reported in New Zealand or Australia. C. perfringens type B produce both beta toxin and epsilon toxins, therefore both haemorrhagic enteritis and systemic vascular damage can develop. As many animals are exposed to C. perfringens without developing disease, there must be additional unknown factors that resulted in disease in these particular sheep. Vaccines that specifically protect against C. perfringens type B are available and may be recommended for use in smaller non-commercial flocks, as in the present case.     

肉毒毒素重组抗原表达系统研究进展

肉毒中毒是肉毒杆菌引起的一种致死性的神经麻痹性疾病,应对内毒中毒的最有效方法是预防或疫苗接种,肉毒毒素Hc段是研制亚单位疫苗的首选.通过基因重组技术能够将肉毒毒素Hc片段在大肠杆菌或酵母表达系统中进行大规模表达.阐述了这2种表达载体在肉毒毒素亚单位疫苗的研制及发酵生产情况.     

规模化养鸡场A型产气荚膜梭菌的分离鉴定及其对海南霉素敏感性的测定

为探究海南霉素对产气荚膜梭菌的敏感性,在某大型养鸡场采集了209份鸡肠道样本,采用选择性培养基和MALDI-TOF质谱仪微生物鉴定仪鉴定,并对鉴定的菌株进行分型。采用琼脂稀释法测定了海南霉素、恩拉霉素、盐霉素、丁酸甘油酯对临床分离的产气荚膜梭菌的抗菌活性,以期为当前限抗禁抗后养殖场选择合适的药物控制产气荚膜梭菌提供理论依据。试验结果表明：共分离出61株产气荚膜梭菌,分离率为29.2%;通过多重PCR对所分离的产气荚膜梭菌进行分型,结果分离株均为A型产气荚膜梭菌;4种药物均有不同程度的抑菌效果,其中海南霉素与恩拉霉素对鸡源产气荚膜梭菌的抑菌效果最好,且MIC值范围均为0.004～0.06μg/mL。海南霉素具有很好的抗梭菌效果。     

凝结芽孢杆菌对感染产气荚膜梭菌肉鸡生长性能、肠道病变及免疫器官指数的影响

本试验旨在研究日粮中添加凝结芽孢杆菌对感染产气荚膜梭菌肉鸡生长性能、肠道病变及免疫器官指数的影响。选用312只1日龄科宝500肉鸡公雏,随机分成4个处理,每个处理6个重复,每个重复13只鸡。采用2×2双因子完全随机设计,产气荚膜梭菌分为无感染产气荚膜梭菌组和感染产气荚膜梭菌组,日粮中凝结芽孢杆菌水平分别为0和400mg/kg。试验期35d。结果表明,感染产气荚膜梭菌后15~35日龄肉鸡平均日增重显著降低(P0.05),28日龄肠道病变严重(P0.01),35日龄法氏囊指数极显著降低(P0.01);日粮中添加凝结芽孢杆菌可显著提高15~35日龄肉鸡平均日增重(P0.05),显著降低料重比(P0.05),能缓解感染导致的肠道损伤(P0.05),28日龄肉鸡胸腺指数极显著提高(P0.01),35日龄法氏囊指数极显著提高(P0.01)。肠道病变结果分析表明,日粮中添加凝结芽孢杆菌与感染产气荚膜梭菌有显著交互作用(P0.05)。综上所述,日粮中添加凝结芽孢杆菌能够提高感染产气荚膜梭菌肉鸡生长性能及免疫器官指数,增强动物免疫能力,并能缓解由于产气荚膜梭菌感染而导致的不利影响。     

Cloning,Expression of Clostridium perfringens α-toxin Gene and Preparation of its Antisera

One pair of primers had been designed and synthesized based on the α-toxin gene of Clostridium perfringens.The complete α-toxin gene fragment was amplified by polymerase chain reaction (PCR), and then was cloned into pGEM-T Easy vector to construct pGEM-T-α.Digested with EcoRⅠ and Hind Ⅲ, a fragment of 1125 bp was cloned into the expression plasmid vector pET-28a(+).The recombinant plasmid was transformed into the BL21(DE3)plys and induced by 1.0 mmol/L IPTG at 37 ℃.The expression product was found to be 46.1 ku as expected one identified by SDS-PAGE, and confirmed by Western blotting with Clostridium perfringens type A antisera, indicating similar reactivity with native α-toxin.Recombinant α-toxin protein was simultaneously found in culture supernatant, postsonic supertanant and inclusion bodies, most protein was expressed in inclusion bodies, which indicated recombinant α-toxin protein was expressed in the extracellular, periplasm and cytoplasm.Recombinant α-toxin protein in postsonic supertanant could not make mice die, indicating its non-toxicity.Toxin-antitoxin neutralization test showed that antisera of recombinant α-toxin protein were specific to α-toxin.Upon immunization of rabbit with the recombinant α-toxin protein, antisera with high antibody titer neutralizing 100 MLD toxin per 1 mL were prepared.     

Preventive effects of Clostridium butyricum on gastric ulceration induced by pylorus ligature in mice

AIM: To investigate the preventive effects of Clostridium butyricum (C. butyricum) on the type of pylorus ligated gastric ulcer (GU) in mice and the underlying mechanisms. METHODS: ICR mice were randomly divided into 4 groups: sham operation group, model group, C. butyricum pretreatment group and omeprazole pretreatment group. Gastric pyloric ligation was adopted to establish GU model in mice. The gastric juice was collected to measure the content of gastric free mucus, the pH of gastric juice and the activity of pepsin. The gastric tissues were collected for routine HE staining to observe the pathological changes. The content of glycogen was detected by PAS staining. The protein expression of Bax and Bcl-2 in the gastric mucosa was also assessed by immunohistochemical staining. RESULTS: The HE and PAS staining showed that the C. butyricum pretreatment obviously attenuated the mucosa lesion induced by ligation. Compared with model group, the pH of gastric juice was significantly raised. The activity of pepsin fell off in C. butyricum group, which was lower than that in omeprazole group. In comparison with model group, the content of gastric free mucus was dramatically increased and PAS staining showed a significant rise in C. butyricum group, but not in omeprazole group. The protein expression of Bax was decreased and the protein expression of Bcl-2 was upgraded in C. butyricum group than those in model group. CONCLUSION: C. butyricum protects gastric mucosa against the challenge of pylorus ligation in mice and its mechanism may be related to inhibiting gastric acid secretion and the activation of pepsin, increasing the production of gastric free mucus, strengthening the expression of bcl-2 gene and inhibiting the expression of bax gene.