Identification of stool proteins in C57BL/6J mice by two-dimensional gel electrophoresis and MALDI-TOF mass spectrometry

Gastrointestinal disease is a major cause of mortality in humans and animals, and the detection of disease-associated protein in stool is an established diagnostic method in this context. Yet, no data currently exists about the protein composition of mammalian faeces. Using a newly developed two-dimensional (2D) gel method, 28 of the most abundant proteins in murine faeces were identified. Mammalian faeces contains protein from multiple species (from the individual, from gastrointestinal bacteria, from food, etc.). Yet, it was found that the majority of mouse stool proteins were of mouse origin, with a minority of proteins being derived from food (in particular soybean glycinin and conglycinin) and bacteria (flagellin). Most mouse proteins were proteases and saccharidases derived from the exocrine pancreas. In addition, two unexpected mouse proteins were identified: one was a newly described mucin-like protein from intestinal goblet cells (FcγBP); the other was the secreted form of carbonic anhydrase (type VI) from salivary gland. The data suggest that 2D analysis of faecal protein is likely to provide meaningful information about the physiological stage of the gastrointestinal tract. Compared with studies based on biopsies, faecal protein analysis may reduce the number of laboratory animals, and might also allow quicker bridging from animal studies to humans, where biopsy material is more difficult to obtain and is less relevant for general practice use.     

人工养殖条件下三种锦蛇的粪便微生物多样性

采用高通量测序技术,分析比较了实验室人工饲养的王锦蛇(Elaphecarinata)、棕黑锦蛇(E.schrenckii)和赤峰锦蛇(E. anomala)3种锦蛇粪便样本的微生物多样性。结果显示,在送检的粪便样本中,Shannon多样性指数(P 0.05)和Simpson多样性指数(P 0.05)在王锦蛇和棕黑锦蛇之间有显著性差异,赤峰锦蛇与前两者均没有差异;ace与chao1指数在3种锦蛇之间没有显著性差异。共检测出8个菌门,厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)和变形菌门(Proteobacteria)是3种锦蛇共享菌门。在属水平上3种锦蛇的优势菌群明显不同,志贺氏菌属(Shigella)、梭状芽胞杆菌属(Clostridium_sensu_stricto_11)及拟杆菌属(Bacteroides)分别是王锦蛇、棕黑锦蛇和赤峰锦蛇粪便样本中相对丰度最高的菌属。3种锦蛇粪便微生物多样性存在着明显的差异,但其是否可以作为区分这3种锦蛇的依据还需要进一步研究。     

The suitability of a faecal suspension of sheep as inocula for the estimation of utilizable crude protein of feeds by in vitro incubation

The objective of this review is to outline those parts of modelling approaches in pig production which are not highly developed; these are the partitioning of protein and lipid accretion in different anatomical body parts. The authors introduce present models with a critical evaluation and draw some conlusions for further developments. Based on present knowledge this paper demonstrates the process of protein and fat accretion in different body compartments in pigs and influencing factors. A further aim is to assist in the conceptual development of a new pig model, which is more detailed, precise and accurate than currently available models. Exsisting models are generally deficient with regard to the translation of lipid and protein gain into lean and fatty tissue. Only assumed values for this translation have been used so far and the concepts underlying these values are not well understood. Therefore, it may be appropriate to develop a compartimental model to predict protein and fat deposition in growing and fattening pigs. With this new approach the model can supply sufficiently the changing consumer demands regarding to the possibility of meat quality prediction.     

Effects of Dietary Non-Digestible Oligosaccharides on Microbial Characteristics of Ileal Chyme and Faeces in Weaner Pigs

Fructooligosaccharides (FOS) and transgalactooligosaccharides (TOS), which are non-digestible oligosaccharides (NDO), were included at 10 and 40g/kg in an NDO-free control diet at the expense of purified cellulose. Each of the 5 diets was fed to 4 weaner pigs and microbial characteristics of their ileal chyme and faeces were assessed. The NDO-pigs had lower ileal pH than the control pigs. Dietary NDO did not affect the ileal volatile fatty acid concentration, though FOS-pigs had a higher concentration of lactic acid and relatively more iso-valeric acid and less acetic acid than TOS-pigs. The NDO-pigs had lower ileal aerobic bacterial counts than the control pigs, whilst the FOS-pigs had a larger ileal anaerobic bacterial counts than the TOS-pigs. The NDO-pigs had an higher faecal pH and their faecal volatile fatty acid pool contained relatively more iso-butyric acid and iso-valeric acid than the control pigs. The TOS-pigs tended to have higher faecal anaerobic bacterial counts and had a smaller concentration of faecal volatile fatty acid than the FOS-pigs. We concluded that whilst effects at the faecal level may have been partly due to a reduced intake of cellulose, dietary NDO can exert precaecal prebiotic effects in weaner pigs.     

Non-invasive methods for identifying oocysts of Sarcocystis spp. from definitive hosts

Because the excreted sporocysts and/or oocysts of various species of Sarcocystis may not be discriminated morphologically, we sought to validate a diagnostic technique based on variation in the 18S rDNA sequence. Oocysts and/or sporocysts from three taxa of Sarcocystis were collected from human, feline, and canine definitive hosts that had fed upon meats infected with the muscle cysts of Sarcocystis hominis, Sarcocystis fusiformis and a species of Sarcocystis from water buffalo that could not be distinguished from Sarcocystis cruzi. Using a new collection method employing filter paper, these excreted oocysts and sporocysts were subjected to DNA extraction, as were the corresponding muscle cysts. Methods employing PCR–RFLP and DNA sequencing of a partial 18S rDNA gene (ssrRNA) sequence were then used to successfully distinguish among the three taxa. The same, unique restriction digestion pattern characterizes the tissue cysts and oocysts and/or sporocysts of each parasite taxon. The technique makes possible amplification and identification of species specific gene sequences based on DNA extracted from as few as 7 excreted sporocysts (the equivalent of 3 and 1/2 oocysts) from freshly prepared material, or as few as 50 sporocysts from feces samples that had been stored in potassium dichromate (K₂Cr₂O₇) for as long as 6 years. This represents the first report using molecular diagnostic procedures to diagnose oocysts of Sarcocystis in faecal samples, describing a valuable new tool for studying the epidemiology of various Sarcocystis species.     

Testing the potential seed availability in dung samples: comparison of two seedling emergence methods

To optimize the estimation of species composition and viable seed content of herbivore faeces and to make different approaches comparable, two seedling emergence methods are evaluated. The Ter Heerdt method (TH) employs concentrated samples, potentially increasing and accelerating seedling emergence, as shown for soil samples (95% of all seedlings emerged within 6 weeks). Samples are kept under controlled conditions (glasshouse or climate room). Secondly, a common garden method (CG) using unconcentrated samples so that seasonal changes could fulfill the germination requirements of a broad species spectrum (experiment duration approximately 15 months) was applied. The methods were tested by the use of sheep faeces samples, collected during a six-day grazing period in a threatened dry grassland (Allio-Stipetum capillatae).Both methods proved largely similar in species composition (QS=0.81) and viable seed content (QS=0.69). More species (e.g. monocotyls) and a higher seedling emergence of hard-seeded species (Fabaceae and Cistaceae) were found in the CG method. Besides a higher emergence of some small-seeded winter annuals, few other species emerged exclusively by use of the TH method. Nevertheless, all species detected by only one method were found in low individual numbers (4).Depending on research interest and availability of space and time, the most appropriate method can be chosen. If the main focus is on the species composition, unconcentrated faeces samples can be studied by CG. In case the overall viable seed content is more important and/or a shorter time period is available, TH serves as a suitable alternative.     

白马雪山黑白仰鼻猴(Rhinopithecus bieti)垂直迁移

2000-2001年间,在白马雪山国家级自然保护区南任村附近,通过在四个山坡每隔海拔100米的样带(宽5.0米)内搜集黑白仰鼻猴的粪便来研究猴群不同季节的垂直迁移模式。同时,通过在每个地区山脊上和山沟内搜集2.5米宽垂直样带内的粪便,来研究猴群不同季节对山脊和山沟的选择性。结果表明:猴群在海拔3500-4300米的区域活动。虽然猴群全年集中利用3900-4200米的林带;但是具有季节性差异:夏季最高(4200米),依次是秋季(4100米)和冬季(4000米),夏季最低(3900米)。另外,冬季山沟中的粪便多于山脊说明猴群在沟中停留时间长,这与沟中少风且温度高有关。不同海拔带上的粪便密度和松萝量间正相关,这意味着食物资源的垂直分布是影响猴群垂直迁移的重要因素。春季,猴群会下到低海拔采食嫩芽/叶,而冬季则下到低海拔处躲避首次大雪,这很可能导致猴群集中利用第二个海拔带。     

Faecal pellets of Reeves’ muntjac,<Emphasis Type="Italic"> Muntiacus reevesi</Emphasis>: defecation rate,decomposition period,size and weight

Captive muntjac fed on vegetation had a daily defecation rate of 7 to 8.2.pellet groups. Even in the cold month of February 97.6% of pellets decayed within 80 days but for 8 other months the range was 4 to 23 days. Emphasis is given to the need for site-specific information when decay rate is required to assess population size from dung counts. The morphology, dimensions, weight and group size of pellets are described as an aid to distinguish them from the faeces of other cervids.     

Experimentally elevated testosterone increases status signalling in male Greylag geese (<Emphasis Type="Italic">Anser anser</Emphasis>)

Testosterone modulates male vertebrates sexual and social behaviour. We experimentally investigated the testosterone-sensitive behaviours in male greylag geese (Anser anser) by implanting silastic tubes containing crystalline testosterone during the mating season (February; 5 implanted and 5 control males) and in the early winter (November; 7 and 7). Focal animals were part of a semi-tame, unrestrained flock with fully intact social relationships. Excreted testosterone and corticosterone immunoreactive metabolites (TM, BM) were determined by enzyme immunoassay. Individual faecal samples and behavioural protocols were collected daily over a period of 5 weeks, including 1 control week before implantation. In February, no significant behavioural effects of the supplemental testosterone were observed, which may be due to the naturally occurring high systemic androgen levels in spring. In November, however, implanted males had higher TM excretion rates and performed status signalling behaviour (beak up) more frequently than control males. No differences between implanted and control males were found with respect to BM, agonistic interactions or vigilance behaviour. Furthermore, during the second week after implantation, TM positively correlated with the frequency of beak up of implanted males, whilst their female partners were attacked with lower latency by other members of the flock than the females of control males. Hence, status signalling in greylag ganders seems to be testosterone-sensitive year-long and inappropriate status signalling of males may draw attacks towards their females.     

A new 2CTAB/PCI method improves DNA amplification success from faeces of Mediterranean (Barbary macaques) and tropical (lowland gorillas) primates

Animal genomic DNA extracts of sufficient quality to address questions about population biology or behavioural ecology can be obtained from faeces when adequate extraction procedures are used. The presence of PCR inhibitors in extract products appears generally the main factor limiting DNA amplification success. We compared DNA amplification success from faeces of a tropical primate (western lowland gorilla, Gorilla g. gorilla) and a Mediterranean primate (Barbary macaque, Macaca sylvanus) between a standardized extraction technique widely used in animals (QIAamp^® stool kit), a technique mainly used in plant species (CTAB) and a new protocol (2CTAB/PCI). Amplification success varied from 51% to 97%, the highest success being reached with the 2CTAB/PCI protocol in both species.