培菌白蚁菌圃和粪便微生物多样性分析

【背景】培菌白蚁是属于白蚁科的一类与鸡枞菌属真菌共生的高等白蚁,其与体内肠道微生物和体外菌圃微生物形成三维共生体系。【目的】分析培菌白蚁菌圃和粪便的微生物多样性,并与肠道微生物进行比较。【方法】通过Illumina MiSeq高通量测序方法对培菌白蚁菌圃和粪便样品进行细菌16S rRNA基因和真菌ITS测序分析。【结果】高通量测序获得培菌白蚁菌圃和粪便样品细菌和真菌的有效序列和OTU数目。5个样品细菌OTU数目在90-199之间,而真菌OTU在10-58之间,细菌的种类多样性明显大于真菌。不论是细菌还是真菌,粪便样品的OTU数目多于菌圃样品。经物种分类分析,菌圃样品主要优势细菌是变形菌门(Proteobacteria),其相对含量超过82.4%;其次是拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes);粪便样品中优势细菌为拟杆菌门,其次是变形菌门,粪便优势菌属为别样杆菌属和营发酵单胞菌属,这与培菌白蚁肠道菌多样性组成一致。培菌白蚁菌圃和粪便样品共生真菌主要为担子菌门(Basidiomycota)和子囊菌门(Ascomycota)。菌圃优势真菌为鸡枞菌属(Termitomyces),相对含量在51.83%以上,菌圃中还鉴定到炭角菌属(1%,Xylaria)。【结论】为今后培菌白蚁-体内外微生物共生关系研究以及微生物的分离培养提供了依据和参考。     

短尾猴粪便雌二醇和孕酮处理与 提取方法比较

非损伤性的粪便样本已在监测动物性激素水平中得到广泛应用,但粪便处理和激素的提取方法在不同物种中因其食性与代谢不同具有明显的物种特异性。本研究以野生和笼养两种环境下的短尾猴(Macacathibetana)为研究对象,采集粪便样品,利用放射免疫法(RIA)检测激素含量,比较不同粪便处理方式(湿便法、冷冻干燥粉碎法、冷冻干燥研筛法)和激素提取方法(乙醇加热法、甲醇乙醚法、乙醇丙酮法、甲醇乙醇法)的效果,探讨不同取食条件下短尾猴雌二醇(E2)和孕酮(P4)水平的最佳测定方法。粪便不同处理方法的比较中,冷冻干燥研筛法测得雌二醇和孕酮含量最高,冷冻干燥研筛法与冷冻干燥粉碎法在野生短尾猴中具有显著差异(P0.01),但在笼养短尾猴中无显著差异(P 0.05)。湿便法测得含量最低,与另两种处理方法均具有显著差异性(P 0.01)。激素提取方法的比较中,乙醇加热法提取雌二醇和孕酮含量最高,与甲醇乙醚法(P 0.01)、乙醇丙酮法(P 0.01)、甲醇乙醇法(P 0.01)之间均具有显著性差异。综合考虑激素平均含量、处理过程的简便性、抽提步骤的可行性,野生短尾猴粪便处理建议选用冷冻干燥研筛法,笼养短尾猴粪便由于冷冻干燥粉碎法操作简便且与冷冻干燥研筛法无显著差异,因此笼养短尾猴建议选用冷冻干燥粉碎法。野生与笼养短尾猴粪便激素的提取均建议采用乙醇加热法。     

Wild black-headed gulls (<Emphasis Type="Italic">Larus ridibundus</Emphasis>) as an environmental reservoir of <Emphasis Type="Italic">Salmonella</Emphasis> strains resistant to antimicrobial drugs

Salmonella were isolated from black-headed gulls (Larus ridibundus) in six locations in the Czech Republic from 1984 to 2005 (Chropyně and Nymburk in 1984–1986; Nové Mlyny, Bartošovice, and Hodonín in 1991–1994; and Nové Mlyny, Bartošovice, and Ostrava in 2005). Antimicrobial susceptibility was determined in 12 antimicrobial drugs using disk diffusion. Although 95% of Salmonella isolates (197 out of 207) were pansusceptible, the prevalences of resistance increased significantly from 1 (2%) out of 59 isolates in 1984–1986 and 3 (3%) out of 100 isolates in 1991–1994 to 6 (13%) out of 48 isolates in 2005. Furthermore, in 2005, two isolates were nalidixic acid-resistant and one isolate was multidrug-resistant Salmonella Typhimurium DT 104. These findings suggest that the occurrence of salmonellae in black-headed gulls depends to a large extent on the contamination where the gulls feed and possibly reflects the dissemination of these strains among farm animals and humans. Black-headed gulls may also become infected with resistant Salmonella and thus pose a potential risk of Salmonella contamination of surface water and animal feeds, and consequently dissemination.     

The impact of ecological variability on the reproductive endocrinology of wild female African elephants

Non-invasive endocrine methods enable investigation of the relationship between ecological variation and ovarian activity and how this impacts on demographic processes. The underlying physiological factors driving high variation in inter-calving intervals among multi-parous African elephants offer an interesting system for such an investigation. This study investigates the relationship between Normalized Differential Vegetation Index (NDVI), an ecosystem surrogate measure of primary productivity, and fecal progestin concentrations among wild female elephants. Matched fecal samples and behavioral data on reproductive activity were collected from 37 focal individuals during the two-year study. Linear mixed models were used to explore the relationship between fecal 5alpha-pregnane-3-ol-20-one concentrations and the independent variables of NDVI, calf sex, female age, gestation day, and time since last parturition. Among both non-pregnant and pregnant females, fecal 5alpha-pregnane-3-ol-20-one concentrations were significantly correlated with time-specific NDVI indicating a strong relationship between ecological conditions and endocrine activity regulating reproduction. In addition, the age of a female and time since her last parturition impacted hormone concentrations. These results indicate that the identification of an individual's reproductive status from a single hormone sample is possible, but difficult to achieve in practice since numerous independent factors, particularly season, impact fecal hormone concentrations. Regardless of season, however, fecal 5alpha-pregnane-3-ol-20-one concentrations below 1 microg/g were exclusively collected from non-pregnant females, which could be used as a threshold value to identify non-pregnant individuals. Collectively the information generated contributes to a better understanding of environmental regulation of reproductive endocrinology in wild elephant populations, information salient to the management and manipulation of population dynamics in this species.     

Intestinal ciliate composition found in the feces of the Turk rahvan horse Equus caballus, Linnaeus 1758

Species composition and distribution of large intestinal ciliates were investigated in the feces from 15 Turk rahvan horses, living in the vicinity of Izmir, Turkey. Twenty-two ciliate genera consisting of 36 species were identified. This is the first report on intestinal ciliates in Turk rahvan horses and no previously unknown species were observed. The mean number of ciliates was 14.2 ± 13.9 × 10⁴ cells ml⁻¹ of feces and the mean number of ciliate species per host was 9.9 ± 7.1. No ciliates were observed in 2 horses. Bundleia and Blepharocorys were considered to be the major genera since these ciliates were constantly found in high proportions. In contrast, Paraisotricha, Didesmis and Gassovskiella were only observed at low frequencies. The ciliates found in this survey had almost the same characteristics as those described in previous reports, suggesting that there was no significant geographic variation in the intestinal ciliate fauna of equids.     

Prediction of total fecal output in sheep fed silage using the Captec chrome controlled-release capsule

Six Finnish wethers (average weight, 52.3 kg) were used to determine the effectiveness of a controlled-release Cr₂O₃ bolus and the internal marker acid insoluble ash (AIA) for predicting fecal output. Wethers were fed grass silage for ad libitum intake. Each wether was dosed orally with one continuous-release Cr₂O₃ bolus (CRC) on the first day of the experiment. Chronology of the trial was as follows: Day 0 to 14, marker adaptation period; Day 15 to 21, total fecal collection; Day 22 to 23, fecal grab sampling every 4 h; Day 25 to 35, once-daily fecal grab sampling. Accuracy of marker-estimated fecal output derived from each method was compared within a marker with total fecal collection values using a paired t-test. Concentrations of AIA and Cr varied (P < 0.10) between sampling times. Accuracy of fecal output estimates was better with the controlled-release Cr₂O₃ bolus (r = 0.97; P = 0.03) than with AIA (r = −0.08; P = 0.89). Grab samples taken once produced reliable (r = 0.96; P = 0.02) estimates of fecal output of silage-fed wethers using the controlled-release Cr₂O₃ bolus. The controlled-release Cr₂O₃ bolus is a better marker than acid-insoluble ash for predicting fecal output of silage-fed wethers.     

Destruction of Salmonella typhimurium, Escherichia coli O157:H7 and Listeria monocytogenes in chicken manure by drying and/or gassing with ammonia

Escherichia coli O157:H7 and Listeria monocytogenes were able to grow for a period of 2 days in fresh chicken manure at 20 degrees C with a resulting 1-2 log units increase in CFU; Salmonella typhimurium remained stable. Prolongation of the storage time to 6 days resulted in a 1-2 log decreases of S. typhimurium compared to the initial count and a 3-4 log decrease of E. coli O157:H7; the number of L. monocytogenes did not decrease below the initial. These changes were accompanied by an increase in pH and accumulation of ammonia in the manure. The destruction of the three microorganisms was greatly increased by drying the manure to a moisture content of 10% followed by exposure to ammonia gas in an amount of 1% of the manure wet weight; S. typhimurium and E. coli O157:H7 were reduced by 8 log units, L. monocytogenes by 4.     

人工饲养幼猴粪便的正常菌群研究

目的通过对10只幼猴粪便正常菌群的测定,了解幼猴肠道正常菌群的分布情况,为猕猴肠道疾病的治疗和微生态制剂的开发提供一定理论依据.方法用微生态学检测方法,对10只健康幼猴粪便中正常菌群(9种细菌)进行定性、定量分析.结果在健康幼猴肠道中,双歧杆菌(10.02±0.25)、类杆菌(10.00±0.09)、韦荣球菌(9.96±0.31)、肠球菌(9.98±0.43)和梭杆菌(9.90±0.24)为优势菌群,而肠杆菌(7.32±1.49)、葡萄球菌(5.74±0.49)和酵母菌(5.42±1.47)含量则较少.结论该实验测出健康幼猴粪便内正常菌群值可作为健康幼猴肠道内正常菌群值的参考.     

逆转录套式PCR检测粪便样本中的SARS冠状病毒

为了观察SARS冠状病毒在SARS患者粪便中的存在规律,建立了检测SARS冠状病毒RNA的逆转录-聚合酶链反应(RT-PCR)方法,并应用该方法检测了241份SARS患者粪便样本。部分PCR产物应用测序技术进行验证。RT-PCR的灵敏度为10^-10稀释度的病毒原液(原液为10^8TCID50／ml)。241份粪便样本的总体检出率为24．1％(58／241),其中发病后的前10d和20d的检出率均为50．0％。随着发病时间的延长,阳性检出率呈下降趋势。应用RT-PCR从粪便中检测SARS冠状病毒是可行的,在发病50d以后仍有17．0％左右的阳性检出率,提示SARS恢复期患者具有排毒的可能性,给后续的卫生防疫措施提供了一定的参考数据。