Faecal parameters as biomarkers of the equine hindgut microbial ecosystem under dietary change

Faeces could be used for evaluating the balance of the equine hindgut microbial ecosystem, which would offer a practical method for assessing gut health and how this relates to disease. However, previous studies concluded that faeces microbial ecosystem was not representative of the proximal hindgut (caecum and ventral colon). This study aimed to evaluate if variations of the faecal microbial ecosystem were similar to those observed in the proximal hindgut. Six horses, fistulated in the caecum and right ventral (RV) colon, were subjected to a gradual change of diet, from a 100% hay (high fibre) diet (2.2 DM kg/day per 100 kg BW) to a 57% hay+43% barley (high starch) diet (0.8 DM kg/day per 100 kg BW hay and 0.6 DM kg/day per 100 kg BW barley). The two diets were iso-energetic and fed over a 3-week trial period. Samples of digesta from the caecum, RV colon and faeces were collected two times on the 10th and 20th day of the trial, for each diet to assess the microbial ecosystem parameters by both classical culture technics and biochemical methods. The variations observed in the caecal and colonic bacterial composition (increase in total anaerobic, amylolytic and lactate-utilizing and decrease in cellulolytic bacteria concentrations) and microbial activity (changes in volatile fatty acids concentrations and increase in lactate concentrations) demonstrated that the hay+barley diet caused changes in the hindgut microbial ecosystem. Similar variations were observed in the faecal microbial ecosystem. Feeding the hay+barley diet resulted in higher concentrations of faecal lipopolysaccharides. The functional bacterial group concentrations (cellulolytics, amylolytics and lactate utilizers) were significantly correlated between caecum and faeces and between colon and faeces. From analyses of the metabolites produced from microbial activity, only valerate concentration in the caecum and the proportion of propionate were significantly correlated with the same parameters in the faeces. Results of the principal component analysis performed between all the caecal/faecal and colonic/faecal parameters revealed that the total anaerobic and cellulolytic bacteria concentrations, as well as valerate, l-lactate and lipopolysaccharide concentrations were strongly correlated with several microbial parameters in the caecum (P<0.027; r>|0.45|) and in the colon (P<0.013; r>|0.50|). This demonstrated that faecal samples and their bacterial analyses could be used to represent caecum and RV colon hindgut microbial ecosystem in terms of variations during a change from a high-fibre to a high-starch diet, and thus could be markers of particular interest to diagnostic proximal hindgut microbial disturbances.     

Influence of diet on the structure and function of the bacterial hindgut community of crickets

The effect of diets varying in carbohydrate and protein content on the structure and function of the hindgut microbiota of crickets was evaluated by determining bacterial densities, fermentation activity, and guanine plus cytosine (G + C) profiles of the DNA extracted from the microbial hindgut community. DNA isolated from the gut community was fractionated and quantified according to G + C content as a comprehensive, coarse-level measure of the composition and structure of the community. The bacterial densities measured by direct counts were not significantly different among the four diets. The crickets were initially reared in the laboratory on cricket chow, which resulted in a hindgut community dominated by bacteria with a G + C content between 32% and 57%. Crickets shifted to an alfalfa diet showed a similar hindgut community G + C profile, although microbial populations with DNA between 35% and 45% G + C were more abundant in alfalfa- than chow-fed crickets. The apparent complexity of the gut community was reduced in crickets fed beet-pulp and protein-based diets compared to those fed chow and alfalfa, and was dominated by populations with a low percentage G + C content. Hindgut communities in crickets fed pulp and protein diets also showed a decrease in hydrogen and carbon dioxide production, suggesting that these diets affected the biochemical activity of the hindgut community. The protein-based diet resulted in a decrease in the rate of evolution of volatile fatty acids, while the ratio of butyrate production to acetate and propionate production was significantly higher in these crickets. Our results show the emergence of a new microbial community structure concomitant with changes in microbial biochemical activity due to shifts in the cricket's dietary regime.     

Layers of symbiosis--visualizing the termite hindgut microbial community

Jared Leadbetter takes us for a nature walk through the diversity of life resident in the termite hindgut--a microenvironment containing 250 different species found nowhere else on Earth. Jared reveals that the symbiosis exhibited by this system is multi-layered and involves not only a relationship between the termite and its gut inhabitants, but also involves a complex web of symbiosis among the gut microbes themselves.     

The uptake of macromolecular materials in the hindgut of viviparous rockfish embryos

Regions of the gut of near-term embryos and new-born larvae of the viviparous rockfish Sebastes taczanowskii were examined histologically and immunohistochemically in order to ascertain whether they possessed the potential to absorb intact macromolecules. Compared with the hindgut of larvae 24 h after parturition, that of near-term embryos (stage 31) was found to be markedly hypertrophied and to react positively when tested for the presence of eosinophilic materials and with antibodies raised against extracts of the rockfish yolk (ab.a-E). In addition, near-term embryos incubated in Ringer solution containing 1% bovine serum albumin appeared to accumulate the protein within the epithelium of the hindgut. Furthermore, as confirmed by immunodiffusion analysis, the ovarian fluid contained materials which reacted positively to ab.a-E. These results suggest that ingestion of intact macromolecules occurs during the late embryonic stages and that the yolk-like substances detected within the hindgut epithelium of such embryos may be derived from the bathing ovarian fluid.     

Evolutionary adaptations of ruminants and their potential relevance for modern production systems

Comparative physiology applies methods established in domestic animal science to a wider variety of species. This can lead to improved insight into evolutionary adaptations of domestic animals, by putting domestic species into a broader context. Examples include the variety of responses to seasonally fluctuating environments, different adaptations to heat and drought, and in particular adaptations to herbivory and various herbivore niches. Herbivores generally face the challenge that a high food intake compromises digestive efficiency (by reducing ingesta retention time and time available for selective feeding and for food comminution), and a variety of digestive strategies have evolved in response. Ruminants are very successful herbivores. They benefit from potential advantages of a forestomach without being constrained in their food intake as much as other foregut fermenters, because of their peculiar reticuloruminal sorting mechanism that retains food requiring further digestion but clears the forestomach of already digested material; the same mechanism also optimises food comminution. Wild ruminants vary widely in the degree to which their rumen contents 'stratify', with little stratification in 'moose-type' ruminants (which are mostly restricted to a browse niche) and a high degree of stratification into gas, particle and fluid layers in 'cattle-type' ruminants (which are more flexible as intermediate feeders and grazers). Yet all ruminants uniformly achieve efficient selective particle retention, suggesting that functions other than particle retention played an important role in the evolution of stratification-enhancing adaptations. One interesting emerging hypothesis is that the high fluid turnover observed in 'cattle-type' ruminants - which is a prerequisite for stratification - is an adaptation that not only leads to a shift of the sorting mechanism from the reticulum to the whole reticulo-rumen, but also optimises the harvest of microbial protein from the forestomach. Although potential benefits of this adaptation have not been quantified, the evidence for convergent evolution toward stratification suggests that they must be substantial. In modern production systems, the main way in which humans influence the efficiency of energy uptake is by manipulating diet quality. Selective breeding for conversion efficiency has resulted in notable differences between wild and domestic animals. With increased knowledge on the relevance of individual factors, that is fluid throughput through the reticulo-rumen, more specific selection parameters for breeding could be defined to increase productivity of domestic ruminants by continuing certain evolutionary trajectories.     

The relationship between forage cell wall content and voluntary food intake in mammalian herbivores

• 1 It is generally assumed that animals compensate for a declining diet quality with increasing food intake. Differences in the response to decreasing forage quality in herbivores have been postulated particularly between cattle (ruminants) and horses (hindgut fermenters). However, empirical tests for both assumptions in herbivorous mammals are rare.
• 2 We collected data on voluntary food intake in mammals on forage‐only diets and related this to dietary neutral detergent fibre (NDF) content, assuming a nonlinear correlation between these measurements. Generally, the paucity of corresponding data is striking.
• 3 Elephants and pandas showed very high food intakes that appeared unrelated to dietary fibre content. Only in small rodents, and possibly in rabbits, was an increase in food intake on forages of higher NDF content evident. In particular, other large herbivores, including horses, followed patterns of decreasing intake with increasing forage NDF, also observed in domestic cattle or sheep.
• 4 For large herbivores, empirical data therefore do not – so far – support the notion that intake is increased in response to declining diet quality. However, data are in accord with the assumption that most large herbivores have an anticipatory strategy of acquiring body reserves when high‐quality forage is available, and reducing food intake (and potentially metabolic losses) when only low‐quality forage is available.
• 5 Intake studies in which the influence of digestive strategy on food intake capacity is tested should be designed as long‐term studies that outlast an anticipatory strategy and force animals to ingest as much as possible.
• 6 We suggest that a colonic separation mechanism coupled with coprophagy, in order to minimize metabolic faecal losses, is necessary below a body size threshold where an anticipatory strategy (living off body reserves, migration) is not feasible. Future studies aimed at investigating fine‐scale differences, for example between equids and bovids, should focus on non‐domesticated species.

     

Left-right asymmetry in the alimentary canal of the Drosophila embryo

In many animal groups, left-right (LR) asymmetry within the body is observed. The left and right sides of the body are generally defined with reference to the anterior-posterior (AP) and dorsal-ventral (DV) axes. In this study, we investigated whether LR asymmetry is solely dependent on the AP and DV polarities in Drosophila embryos. We focused on the proventriculus, a posterior part of the foregut, and the hindgut because LR asymmetry in these body parts is highly stable in normal embryos. In embryos with a fully reversed AP polarity, LR asymmetry in both the proventriculus and the hindgut was re-oriented in relation to the reversed AP polarity. This demonstrates that inversion of AP polarity does not affect LR asymmetry of these tissues, and implies that LR asymmetry is specified in relation to the AP and DV polarities. Our findings were not consistent with the alternative hypothesis that LR asymmetry is predetermined by maternal signals that localize asymmetrically along the LR axis in the oocyte and/or early embryo.     

Commensal bacilli inhibitory to mastitis pathogens isolated from the udder microbiota of healthy cows

AIMS: To isolate from the microbiota of the healthy cow udder commensal bacteria having antimicrobial activity against bovine mastitis pathogens, with a long-term view to their potential application as antimastitis probiotics. METHODS AND RESULTS: Bacterial isolates from four healthy cow udders were tested for inhibitory activity against three Gram-positive indicator bacteria. This led to the selection of nine broadly inhibitory strains. All were of the Bacillus genus and their antimicrobial activities, which appeared heterogeneous on the basis of their antibacterial spectra and heat susceptibilities, enabled grouping of the inhibitory bacilli into six different inhibitory profiles. All displayed strong in vitro activity against Gram-positive mastitis pathogens. Inhibitory bacilli were recovered from each of the 11 udder samples collected over 7 months from one of these cows and the isolates included representatives of all six inhibitory profiles. CONCLUSIONS: Bacilli present in the udder microbiota of healthy cows can produce a variety of broadly active inhibitors of Gram-positive bacteria, including potential mastitis pathogens. SIGNIFICANCE AND IMPACT OF THE STUDY: Inhibitor-producing strains of commensal Bacillus species have been identified, which may have the potential for use as possible antimastitis probiotics.     

Effect of environmental factors and influence of rumen and hindgut biogeography on bacterial communities in steers

Feces from cattle production are considered important sources of bacterial contamination of food and the environment. Little is known about the combined effects of arctic temperatures and fodder tannins on rumen and hindgut bacterial populations. Individual rumen liquor and rectal fecal samples from donor steers fed either alfalfa silage or sainfoin (Onobrychis viciifolia Scop.) silage and water ad libitum were collected weekly on the first three sampling days and fortnightly afterwards. The daily ambient temperatures were registered and averaged to weekly mean temperatures. Steers fed sainfoin silage had lower (P < 0.05) concentrations of branched-chain volatile fatty acids (VFA) than those fed alfalfa silage. All VFA concentrations were higher (P < 0.001) in rumen liquor samples than in fecal samples. The interaction of sample type and diet showed a significant effect (P < 0.05) on the proportions of the bacterial community that were from the phyla Proteobacteria and Verrucomicrobia. Ambient temperature had an indirect effect (P < 0.05) on the phylum Firmicutes, as it affected its proportional balance. The bacterial population diversity in samples appeared to decrease concurrently with the ambient temperature. The phylum Firmicutes explained the first principal component at 64.83 and 42.58% of the total variance in rumen liquor and fecal samples, respectively. The sample type had a larger effect on bacterial communities than diet and temperature. Certain bacterial populations seemed to be better adapted than others to environmentally adverse conditions, such as less access time to nutrients due to higher motility and rate of passage of digesta caused by extreme temperatures, or antimicrobials such as tannins, possibly due to an influence of their biogeographical location within the gut.     

Effect of concentrate feeding sequence on equine hindgut fermentation parameters

Feeding a diet with a small amount of hay and a high proportion of concentrate given in large meals entails a risk of colic to horses as this can impact the hindgut microbial ecosystem. To counteract this potential negative effect, one feeding strategy is to modify the distribution sequence of concentrate and hay. The purpose of the present study was to assess the impact of feeding the concentrate meal before or with the hay meal on fermentative parameters postprandial variations in caecum and right ventral colon contents and faeces. We focused on the comparison of pH value and volatile fatty acid (VFA) concentrations between the three compartments. Four horses fitted with two cannulas in the caecum and right ventral colon were fed a 100% hay diet (H) for a 3-week period, and then a 40% concentrate–60% hay diet for two consecutive periods of 3 weeks each. Concentrate and hay were offered simultaneously in two equal meals at 0800 and 1700 h (HC1) during one period, or separately in two equal meals at 0800 and 1600 h for the concentrate and at 1000 and 1730 h for hay (HC2) during the second one. During the 2^nd week of each diet period (H, HC1, HC2), caecal, right ventral colonic and faecal samples were collected before (0) and at 2, 4, 6 and 8 h after the morning meal. pH values and VFA concentrations were measured. Adding concentrate to the hay was associated with a decrease of pH value in the caecum, right ventral colon and faeces and an increase of faecal VFA concentrations (P<0.01). An impact of the concentrate feeding sequence was observed on caecal pH and faecal propionate concentration (P<0.001) but none on the other parameters investigated. In the caecum and right ventral colon, pH values decreased and VFA concentrations increased within the range of 4 to 8 h after the morning meal. Our data suggested that the variation of faecal pH could be an indicator of those of the caecal and right ventral colonic pH with a delay of 2 h. The present study showed that hay and commercial pelleted could be safely distributed simultaneously, thus making easier every day horse feeding.     

Production of methyl mercury in the gut of the Australian termite Mastotermes darwiniensis

Animals are often exposed to or can ingest heavy metals along with their food. Therefore, we tested whether the hindgut microbiota of Mastotermes darwiniensis possesses the capability to form methyl mercury. The termite M. darwiniensis (Isoptera) was fed with saw dust containing different concentrations of inorganic mercury. Methyl mercury was determined by purge‐and‐trap capillary gas chromatography‐atom fluorescence spectrometry (CGC‐AFS) using ethyl mercury chloride as the internal standard. Total mercury concentrations were determined in the termite tissue by inductive coupled plasma‐isotope dilution mass spectrometry (ICP‐IDMS) after microwave‐assisted digestion. The obtained results showed in vivo methyl mercury production in terrestrial insects for the first time. Desulfovibrio intestinalis isolated from M. darwiniensis was identified as a biomethylating species of the intestinal microbiota.     

The role of volumetric feedback from the hindgut in the regulation of meal size in fifth-instar Locusta migratoria nymphs

ABSTRACT. Behavioural studies show that fullness of the hindgut influences the size of meals taken by fifth-instar Locusta migratoria L. nymphs, with the relevant information coming from the ileum and passing via the terminal abdominal ganglion. Morphological and electrophysiological studies suggest that stretch receptors on the posterior ileum, associated with branches of the rectal nerves, register extension of the ileal fold and probably also an increase in ileal volume. A system in which the volumetric feedbacks from both the crop and hindgut interact in the regulation of meal size is postulated.     

Ultrastructure of the hindgut of Drosophila larvae, with special reference to the domains identified by specific gene expression patterns

The hindgut of Drosophila larvae consists of nine domains that have been distinguished by specific gene expression patterns. In the present study, we examined the ultrastructure of the hindgut of Drosophila larvae, with special reference to the domains, in order to determine whether or not the domains are morphologically distinct functional units. Each domain showed specific ultrastructural features that suggested specific corresponding functions. According to the morphological features, terms are proposed for each domain: the imaginal ring; the "pylorus," which has a thick cuticular layer and well-developed sphincter muscles; the "large intestine," which occupies a major middle portion of the hindgut and has a unique dorsal and ventral subdivision; "border cells," which delineate the anterior and posterior borders of the large intestine and the border between the dorsal and ventral domains of the large intestine; and the "rectum," which is situated at the posterior end of the hindgut and has a thick cuticular layer and sphincter muscles. The morphological features indicate that the large intestine has active absorptive activities. The domains, which have been distinguished by gene expressions, were demonstrated to be functional tissue units of the gut.