Recreational sandboxes for children and dogs can be a source of epidemic ribotypes of Clostridium difficile

Different studies have suggested that the sand of public playgrounds could have a role in the transmission of infections, particularly in children. Furthermore, free access of pets and other animals to the playgrounds might increase such a risk. We studied the presence of Clostridium difficile in 20 pairs of sandboxes for children and dogs located in different playgrounds within the Madrid region (Spain). Clostridium difficile isolation was performed by enrichment and selective culture procedures. The genetic (ribotype and amplified fragment length polymorphism [AFLP]) diversity and antibiotic susceptibility of isolates was also studied. Overall, 52.5% (21/40) of samples were positive for the presence of C. difficile. Eight of the 20 available isolates belonged to the toxigenic ribotypes 014 (n = 5) and 106 (n = 2), both regarded as epidemic, and CD047 (n = 1). The other 12 isolates were non‐toxigenic, and belonged to ribotypes 009 (n = 5), 039 (n = 4), and 067, 151 and CD048 (one isolate each). Nevertheless, all isolates (even those of a same ribotype) were classified into different AFLP genotypes indicating non‐relatedness. In conclusion, our results revealed the presence of epidemic ribotypes of C. difficile in children's and dog's sandboxes located nearby, which constitutes a major health risk.     

牛舍空气微生物及向环境传播的研究

采用４ 台 Ａ Ｎ Ｄ Ｅ Ｒ Ｓ Ｅ Ｎ微生物空气样品收集器对一个犊牛舍内及环境空气细菌含量进行了测量, 需氧菌总数分别为８５３６ ～４６６９１ Ｃ Ｆ Ｕ／ｍ ３ 和２６４９ ～２４７７５ Ｃ Ｆ Ｕ／ｍ ３ , 厌氧菌浓度计３０１７ ～２４７７５ Ｃ Ｆ Ｕ／ｍ ３ 和６６１ ～４１２２ Ｃ Ｆ Ｕ／ ｍ３ 。根据 Ｈ Ｅ Ｓ Ｋ Ｅ Ｔ Ｈ （１９７４） 计算公式和 Ｋ Ｌ Ｕ Ｇ （１９６９） 气溶胶颗粒扩散模式, 得出犊牛舍菌源强度, 即向环境排放需氧菌为１１００００ ～１４００００ Ｃ Ｆ Ｕ／ｓ , 厌氧菌在２７０００ ～１７５０００ Ｃ Ｆ Ｕ／ｓ 之间。细菌浓度实际测量值与估算一致, 该公式和扩散模式适合于动物舍菌源强度和舍外环境不同距离空气微生物浓度的估算。距离犊牛舍１００ ｍ 下风处, 需氧菌高达６５０ ～８３９ Ｃ Ｆ Ｕ／ｍ ３ , 厌氧菌１６０ ～１０３４ 细菌／ｍ ３ , 各项均超过正常值１０ ～１５ 倍。     

Partial replacement of fishmeal with Clostridium autoethanogenum single‐cell protein in the diet for juvenile black sea bream (Acanthopagrus schlegelii)

Clostridium autoethanogenum protein (CAP) is a single‐cell protein derived from ethanol production and may have potential to become a substitute for fishmeal in aquafeeds. A 70‐day feeding trial was conducted with black sea bream (Acanthopagrus schlegelii) (mean initial weight 6.03 ± 0.09 g) to evaluate partial replacement of fishmeal with CAP in diets. Seven isonitrogenous and isoenergetic diets were formulated with graded levels of CAP (0, 4.85, 9.70, 14.55, 19.40, 38.80 and 58.20%) to replace fishmeal. The inclusion of CAP at all dietary levels tested did not significantly affect the growth performance (p > .05). Fish fed the CAP58.20% diet showed a significantly lower feeding rate, with significantly higher protein efficiency ratio and feed efficiency ratio compared with fish fed the other diets (p < .05). No statistical differences were found in dorsal muscle and whole‐body compositions. Total superoxide dismutase in serum of fish fed CAP58.20% diet was significantly lower compared with that of the control. Malondialdehyde, catalase, total antioxidant capacity and digestive enzyme activities revealed no significant differences among dietary treatments. Phosphorus retention efficiency significantly increased, and phosphorus discharge showed a downward trend with increasing CAP inclusion levels. In conclusion, the results indicated that CAP is a safe and effective alternative protein source, which can replace fishmeal in the diet of black sea bream up to 58.20%, without adverse effects on growth performance, antioxidation and digestive enzyme activity. This study has shown the potential of converting industrial waste into a high protein feed ingredient for aquafeeds.     

Bacteriological and molecular typing of Clostridium perfringens strains isolated in retail beef in Beijing,China

Clostridium perfringens is an important zoonotic pathogen. This study was designed to explore the prevalence and toxin types of C. perfringens in retail beef collected from Beijing, China. Among 221 beef samples collected, 53 samples were positive for C. perfringens, resulting in the average prevalence as 23.98%. By toxin gene-based typing, the most C. perfringens strains belong to type A (96.23%, 51/53), only 2 strains were identified as type D. By a multi-locus sequence typing (MLST)-based analysis, a total of 36 sequence types (STs) were detected, and the most STs (n=30) represented just a single strain. These finding suggested that the prevalence of C. perfringens in retail beef in Beijing was considerably high and these bacteria displayed extreme diversity in genetics.     

丁酸梭菌和枯草芽孢杆菌对鸭肠道发育的影响

为研究饲粮中添加丁酸梭菌和枯草芽孢杆菌对育雏育成期高邮鸭肠道发育以及肠道组织形态、杯状细胞密度等的影响,选择1日龄体重相近的健康高邮鸭90只,随机分3组,每组3个重复,每个重复10只。对照组饲喂基础全价日粮,丁酸梭菌组添加丁酸梭菌活菌3×10⁵ CFU·g^-1,枯草芽孢杆菌组添加枯草芽孢杆菌活菌5×108 CFU·g-1,试验期60 d。结果表明,高邮鸭日粮中添加丁酸梭菌和枯草芽孢杆菌显著提高60日龄高邮鸭十二指肠、空肠和回肠重量(P <0.05),显著增加空肠长度(P <0.05)。丁酸梭菌添加组显著降低空肠和回肠的隐窝深度(P <0.05),显著增加回肠的黏膜厚度、肌层厚度和绒毛高度(P <0.05)。丁酸梭菌和枯草芽孢杆菌添加后提高高邮鸭十二指肠、空肠、回肠和盲肠杯状细胞密度(P <0.05);与对照组相比,丁酸梭菌添加组中十二指肠,空肠组织黏糖蛋白2编码基因MUC2m RNA表达量显著提高(P <0.05)。综上所述,高邮鸭自1日龄起日粮中添加丁酸梭菌和枯草芽孢杆菌后,改善其肠道显微结构,在全价日粮...     

健脾按摩法配合口服酪酸梭菌活菌散剂对小儿非感染性急性腹泻的干预效果

目的 观察健脾按摩法配合口服酪酸梭菌活菌散剂治疗小儿非感染性腹泻的干预效果。方法 选择我院门诊2012年3月～2015年9月期间收治113例非感染性腹泻患儿作为研究对象,对照组（35例）、单纯推拿组（38例）与观察组（40例）。对照组口服酪酸梭菌活菌散剂,单纯推拿组采取健脾按摩法,观察组口服酪酸梭菌活菌散剂联合健脾按摩法,比较三组治疗前后大便次数与大便性状症状积分、IL-6、IL-17、IgA、治愈率、总有效率、不良反应发生率。结果 对照组与单纯推拿组治愈率、总有效率比较无统计学意义;观察组治愈率、总有效率分别为82.5%、100.0%,高于对照组与单纯推拿组（均P<0.05）。三组不良反应发生率及治疗前大便次数、大便性状评分、IL-6、IL-17及IgA水平差异的两两比较无统计学意义;治疗后观察组各项症候评分、IL-6、IL-17水平低于对照组与单纯推拿组,IgA高于对照组与单纯推拿组（均P<0.05）。结论 健脾按摩法配合口服酪酸梭菌活菌散剂治疗小儿非感染性腹泻可调理胃肠菌群,健脾和胃、消食导滞,改善症状与机体消化功能与免疫功能,安全性与患儿接受度高。     

产雌马酚梭菌C1对小鸡沙门氏菌攻毒保护效果

部分梭菌对机体的免疫系统具有明显的调节作用,可开发成新型益生菌。由于鸡盲肠内容物中梭菌含量较高,从小鸡肓肠内容物中分离培养功能性梭状芽孢杆菌,并在小鸡中初步研究其对沙门氏菌攻毒的保护效果。该研究首先采用梭菌选择性培养基,从小鸡肓肠内容物中分离获得了一株可以将大豆素转化为雌马酚的革兰氏阳性纤维状梭菌C1,以四环素为对照,研究梭菌C1对小鸡沙门氏菌攻毒的保护作用。结果表明,梭菌C1组和四环素组均具有降低肓肠内容物中沙门氏菌活菌数和促进小鸡生长的功能,推测梭菌C1具有开发为四环素替代品的潜能。虽然较高浓度的S 雌马酚（5 mg·mL-1）在体外对沙门氏菌有一定的抑制作用,梭菌C1也具有将大豆素转化为雌马酚的能力,但梭菌C1在体外对沙门氏菌的抑制作用并不明显,其益生机制还有待进一步研究。     

鸡源高产淀粉酶丁酸梭菌的分离鉴定

为了获得产淀粉酶的丁酸梭菌,本研究从鸡小肠表面黏液中进行丁酸梭菌的分离与筛选,首先对样品进行80℃水浴10 min除去非芽孢菌后,然后接种到TSN培养基进行菌株的分离与筛选。对分离到的菌株进行菌落形态、显微形态初步观察,然后对疑似丁酸梭菌的菌株进行产酸能力和淀粉酶酶活检测,最后对既产酸又高产淀粉酶的菌株进行生理生化鉴定和16S rDNA序列分析。结果表明分离到一株革兰氏阳性厌氧菌C.B1,菌体呈短杆状,能形成圆形或椭圆形芽孢,生理生化结果也符合丁酸梭菌的基本特征,16S rDNA序列长度为1450 bp,与丁酸梭菌的同源性高达99%以上,因此确定该分离菌株为丁酸梭菌,为进一步开发新的微生态制剂奠定了基础。     

产气荚膜梭菌α毒素基因的克隆、表达及其抗血清的制备

本试验通过设计并合成1对引物,PCR扩增B型产气荚膜梭菌C58-2株α毒素完整成熟肽序列,并将其插入到pGEM-T Easy载体中,构建克隆载体pGEM-T-α。对克隆载体pGEM-T-α进行EcoRⅠ和Hind Ⅲ的双酶切,将得到的1125 bp片段以正确的阅读框架定向克隆于pET-28a(+)中,然后将重组质粒转化大肠杆菌BL21(DE3)plys宿主菌中,37 ℃、1.0 mmol/L IPTG诱导该片段获得良好表达。经SDS-PAGE分析,其表达的蛋白约为46.1 ku,与预期大小一致。Western blotting结果显示,该重组α毒素蛋白可被A型产气荚膜梭菌定型血清识别,表明该重组α毒素蛋白具备与天然毒素相似的反应原性。重组α毒素蛋白在菌液上清、超声波裂解上清和超声波裂解沉淀中均有分布,且以包涵体表达为主,表明重组α毒素蛋白可同时在胞外、周质和胞浆表达。小鼠毒力试验结果表明,重组α毒素蛋白不具有毒性。毒素—抗毒素中和试验结果表明,该抗血清具有α毒素特异性。以重组α毒素蛋白作为抗原免疫家兔制备血清,效价测定结果表明每1 mL重组α毒素蛋白抗血清可以中和100 MLD的A型毒素。     

艰难梭菌毒素致病机理的研究进展

艰难梭菌是一种革兰氏阳性厌氧芽胞梭菌,是人类肠道感染的主要致病菌,其主要致病因素为毒素A(肠毒素)和毒素B(细胞毒素).毒素A引发细胞损伤后,毒素B即可侵入肠黏膜,引起细胞病变,导致一系列与感染相关的临床表现.同时,艰难梭菌毒素也是引起猪、鸡等畜禽发生腹泻的重要因素,因此探讨艰难梭菌毒素对机体的损伤作用,有利于揭示艰难梭菌的致病机理,为其防控提供理论依据.