微生物菌群培养组学在动物医学中的应用

实验室条件下可培养的微生物约占自然界中微生物总数的1%,这限制了人们对99%未知微生物的认识和利用,而研究表明,那些“不可培养的微生物”是可以被开发和利用的,未能被纯培养的微生物才是未知微生物的主体。微生物培养组学探索利用多种培养条件和长时间的培养,结合基质辅助激光解吸电离飞行时间质谱法（MALDI-TOF-MS）和16S核糖体RNA（rRNA）测序可以大规模鉴定各种微生物,同时利用全基因组测序和宏基因组测序手段对未知微生物进行深入分析。本文综述了国内外近年来微生物菌群培养组学在反刍动物胃肠道、禽类盲肠及家畜鼻腔微生物菌群研究中的最新进展,探讨将动物体内菌群培养组学方法应用于动物疾病防治领域的可行性。作为一个新兴的研究方法,尽管该培养组学还存在一些不够成熟的方面,但它的发展前景十分广阔,微生物菌群培养组学方法和其他研究方法的互补已经逐渐成为发展兽医微生物学新的突破口。     

鸡空肠和盲肠微生物区系的研究

应用２０日龄白来航小母鸡的肠道内容物,研究消化道部分区段微生物区系,分离鉴定出需氧菌和厌氧菌,并进行了毒力试验。结果表明,每克肠内容物含细菌的对数值（ｌｏｇｎ／ｇ）为：空肠中消化球菌９．６±０．２、双歧杆菌９．４±０．５、拟杆菌１０．６±０．６、葡萄球菌６．６±０．６、乳杆菌９．３、肠杆菌６．８±０．４;盲肠中葡萄球菌６．７±０．６、消化球菌９．８±０．３、双歧杆菌９．２±０．８、拟杆菌１０．２±０．２、棒状杆菌８．７±０．４、肠杆菌６．８±０．３、链球菌７．２±０．２、乳杆菌９．６±０．４、酵母菌４．２±１．４．     

合生素对肉用鹌鹑免疫功能和肠道菌群的影响

摘 要：本试验旨在研究日粮中添加合生素对肉用鹌鹑免疫功能和对肠道菌群的影响。将120只1日龄法国莎维麦脱肉鹌鹑随机分为对照组和合生素组,每组60只。对照组饲喂基础日粮,合生素组在基础日粮中添加0.05%合生素试验期35天。结果表明,合生素显著提高肉用鹌鹑明显提高了免疫器官指数、ND抗体水平和E-玫瑰花环的转化率,且28～35日龄差异显著（P<0.05）;明显的降低了空肠大肠杆菌和沙门氏菌的数量（P<0.05）,提高了乳酸杆菌的数量（P<0.05）。     

采用PCR-SSCP技术分析蚯蚓粪便的微生物群落结构

通过SDS/蛋白酶的方法抽提蚯蚓粪便微生物总DNA,以总DNA为模板,采用通用引物分别成功扩增细菌16S rRNA基因的V4-V5可变区,真菌18S rRNA基因的V8-V9可变区,应用单链构象多态性技术（SSCP）分析了蚯蚓粪便中不同空间层次的微生物种群的多样性。结果表明,在粪便的不同空间层次上呈现出明显的空间分布多样性,并且与纯蚯蚓粪微生物种群的多样性有很大的差别。为进一步的蚯蚓粪生态学功能研究提供了有益的指导。     

乳酸菌对牙鲆稚鱼养殖水体和肠道菌群的影响

在牙鲆稚鱼中投喂添加由单一鼠李糖乳杆菌P15制备的微生态制剂和黄霉素,在60d的投喂期内,采用平板计数法检测好氧性异养菌的总数、总弧菌数和乳酸菌的数量。结果表明,在投喂菌液和冻干菌粉后,养殖水体和牙鲆肠道的乳酸菌数均呈上升趋势,在30d后乳酸菌数量达到稳定并在肠道内定植。同时,由于乳酸菌的抑制作用,弧菌的数量下降,以肠道中的弧菌最明显。乳酸菌对养殖水体和牙鲆肠道的好氧性异养菌没任何影响。乳酸菌对养殖水体和肠道菌群的影响与抗生素具有相近的效果,表明乳酸菌作为饲料添加剂可以取代抗生素应用在牙鲆的养殖中。     

复合微生态制剂对断奶仔猪生长性能、腹泻率、免疫性能和肠道菌群的影响

本试验旨在研究复合微生态制剂对断奶仔猪生长性能、腹泻率、免疫性能和肠道菌群的影响。选取(30±2)日龄平均体重为(8.15±0.05)kg的莱芜黑猪断奶仔猪96头,随机分为4组,每组6个重复(公母各占1/2),每个重复4头猪。Ⅰ组为对照组,饲喂不添加任何益生菌和抗生素的基础饲粮;Ⅱ组为复合微生态制剂组,在基础饲粮中添加1.0‰肽菌素;Ⅲ组为抗生素组,在基础饲粮中添加500 mg/kg硫酸黏杆菌素;Ⅳ组为肽菌素和抗生素混用组,在基础饲粮中添加1.0‰肽菌素的同时添加200 mg/kg的喹乙醇和110 mg/kg的吉他霉素。预试期3 d,正试期28 d。结果表明,1)Ⅱ组平均日增重较Ⅰ、Ⅲ和Ⅳ组分别显著提高了13.16%、15.77%和10.66%(P0.05),F/G分别显著降低了8.54%、12.10%和6.50%(P0.05)。与Ⅰ组相比,Ⅱ、Ⅲ和Ⅳ组均显著降低了仔猪腹泻率(P0.05)。2)与Ⅰ和Ⅲ组相比,Ⅱ组肝脏指数和脾脏指数分别提高了12.13%、24.76%和14.60%、12.45%,血清免疫球蛋白G和免疫球蛋白A含量分别提高了49.73%、37.50%和61.05%、46.67%,且差异均显著(P0.05)。3)与Ⅰ和Ⅲ组相比,Ⅱ组显著降低了胃和十二指肠内容物的p H(P0.05),且显著提高了盲肠和结肠中乳酸杆菌数量(P0.05)。由此可知,本试验条件下,饲粮添加1.0‰复合微生态制剂肽菌素可改善断奶仔猪生长性能,降低腹泻率,促进免疫器官的发育,提高血清中免疫球蛋白含量,增强仔猪免疫力,降低胃和十二指肠内容物p H,改善肠道菌群平衡。     

Suppression of ovarian progesterone production in dairy cows using an implant of GnRH-agonist (deslorelin) for the purpose of evaluating progesterone metabolism

OBJECTIVE: To evaluate the potential of an implant of a GnRH-agonist (deslorelin) to create a progesterone free animal suitable for studying progesterone (P4) metabolism in intact cows by measuring blood P4 and faecal P4 metabolites. METHODS: Experiment 1: Eighteen non-lactating cycling Holstein-Friesian cows, 4 to 7 years old, were allocated to one of three groups to study plasma P4 concentrations preceding an intravaginal insert. These groups comprised: i) a deslorelin group (GnRH-agonist implanted); ii) a PGF group receiving two injections of prostaglandin (PGF2alpha) 12 days apart; and, iii) an ovariectomised (OVX) group. An intravaginal device (CIDR) was inserted into the vagina of each animal and left in place for 11 days. Plasma P4 concentrations were measured during the study period. Experiment 2: Twelve non-lactating cycling Holstein-Friesian cows, 4 to 7 years old, were allocated to two groups: i) a deslorelin group (GnRH-agonist implanted); and ii) an ovariectomised group. Plasma P4 and faecal P4 metabolites (20-oxo-pregnanes, 20alpha-OH and 20beta-OH) were monitored for a period of 5 weeks. RESULTS: Experiment 1: Average plasma P4 concentration did not differ between the three groups (1.28, 1.43 and 1.55 ng/mL for deslorelin, OVX and PGF cows, respectively, P = 0.8) during the period of supplementation. Experiment 2: There was no difference in plasma P4 (mean plasma P4 < 0.02 ng/mL, P = 0.9) and faecal P4 metabolites between deslorelin and OVX cows 2 weeks after the implantation (P = 0.7). CONCLUSIONS: These data showed that a GnRH-agonist (deslorelin) implant may be used as an alternative to ovariectomy to create a progesterone free animal suitable for studying the metabolism of administered P4.     

饲喂乳杆菌雏鸡对鸡白痢菌感染的影响

雏鸡饲喂乳杆菌培养物后,人工感染鸡白痢沙门氏菌,观察雏鸡盲肠主要正常菌群定植及雏鸡抵抗力情况。研究结果表明：人工感染鸡白痢沙门氏菌后,饲喂乳杆菌组同未饲喂乳杆菌组相比,雏鸡盲肠内正常菌群有一定变化,大肠杆菌数明显降低（Ｐ＜００１）,双歧杆菌和乳杆菌数明显增加（Ｐ＜００１）,而且死亡率降低２０％。     

辐照对瓜实蝇遗传区性品系成虫肠道微生物的影响

为研究辐照对瓜实蝇肠道微生物群落多样性和结构组成的影响,以100 Gy137Cs辐照来自瓜实蝇遗传区性品系(GSS)的5、8、11和13日龄的雌、雄成虫,并采用Illumina MiSeq测序技术分别对其肠道菌群进行测序分析。结果表明,瓜实蝇雌雄成虫的肠道中存在19个主要的菌属,包括17个属名已知,1个属名未知和1个属不可培养。其中,17个已知属分别属于4个细菌门,即变形菌门(Proteobacteria)(占总量的50%~98%)、厚壁菌门(Firmicutes)(占总量的2%~47%)、拟杆菌门(Bacteroidetes)(占总量的1%~16%)和1个至今微生物界还未确定的稀少菌门(unclassified rare phyla)(总量1%)。此外,辐照降低了瓜实蝇肠道中的微生物多样性以及优势菌的丰富度,且这种影响与瓜实蝇的性别和成虫日龄无关。本研究结果为评估和修复辐照对瓜实蝇的损伤提供了参考。     

Effect of Daidzein on Ileum Microflora Biodiversity in Hy-Line Variety Brown Layers

Daidzein is always added into poultry feed to make the production performance and immunity of poultry better. In this study, a total of 600 40-week-old Hy-Line variety brown layers were randomized into five groups and fed with a corn-soybean-mixed basal diet supplement with 0, 10, 50, 100, and 500 mg · kg^-1 daidzein, respectively. Then, two PCR-based typing methods（RAPD-PCR and ERIC-PCR） were combined to analyze the ileum content and explore the changes of ileum microflora biodiversity. The results of RAPD-PCR and ERIC-PCR showed that bands under 10 mg · kg^-1 and 50 mg · kg^-1 were the most, and their similarity was the largest. Bands under 500 mg · kg^-1 were the least and similarity with other groups was the minimum. Ileum microflora biodiversity under 10 mg · kg^-1 or 50 mg · kg^-1 was richer than that under 500 mg · kg^-1. A corn-soybean-mixed basal diet supplement with 10 mg · kg^-1 to 50 mg · kg^-1 of daidzein might be beneficial to Hy-Line variety brown layers intestinal bacteria.