Gastrointestinal microbiota play a crucial role in nutrient digestion, maintaining animal health and welfare. Various factors may affect microbial balance often leading to disturbances that may result in debilitating conditions such as colic and laminitis. The invention of next generation sequencing technologies and bioinformatics has provided valuable information on the effects of factors influencing equine gut microbiota. Among those factors are nutrition and management (e.g. diet, supplements, exercise), medical substances (e.g. antimicrobials, anthelmintics, anaesthetics), animal-related factors (breed and age), various pathological conditions (colitis, diarrhoea, colic, laminitis, equine gastric ulcer syndrome) as well as stress-related factors (transportation and weaning). The aim of this review is to assimilate current knowledge on equine microbiome studies, focusing on the effect of factors influencing equine gastrointestinal microbiota. Decrease in microbial diversity and richness leading to decrease in stability; decrease in Lachnospiraceae and Ruminococcaceae family members, which contribute to gut homeostasis; increase in Lactobacillus and Streptococcus; decrease in lactic acid utilising bacteria; decrease in butyrate-producing bacteria that have anti-inflammatory properties may all be considered as a negative change in equine gut microbiota. Shifts in Firmicutes and Bacteroidetes have often been observed in the literature in response to certain treatments or when describing healthy and unhealthy animals; however, these shifts are inconsistent. It is time to move forward and use the knowledge now acquired to start manipulating the microbiota of horses.

BackgroundMilk acute phase proteins (APP) have been identified and show promise as biomarkers of mastitis. However analysis of their profile in dairy cows from a production herd is necessary in order to confirm their benefits in mastitis diagnosis. The profiles of milk haptoglobin (Hp), mammary associated serum amyloid A3 (M-SAA3) and C-reactive protein (CRP) were determined in 54 composite milk (milk from all functional quarters of a cow’s udder collected in a common receptacle) samples (CMS) from a commercial dairy farm. Milk Hp was also determined in individual quarter milk (milk from a single udder quarter) samples (QMS) (n = 149) of the cows.An ELISA was developed and validated for the determination of milk Hp while commercial kits were used for M-SAA3 and CRP assay respectively. Composite milk APP results were compared with cow factors including parity, stage of lactation, percentage protein and fat as well as somatic cell counts (SCC).ResultsComposite milk Hp ranged from <0.4–55 μg/ml with a median of 3.5 μg/ml; composite milk M-SAA3 ranged from <0.6–50 μg/ml and had a median of 1.2 μg/ml, while CRP ranged from <1.80–173 ng/ml and had a median of 24.6 ng/ml. Significant correlations were found between composite SCC and Hp (P-value <0.009) as well as parity and Hp (P < 0.009), but not between M-SAA3 and SCC, M-SAA3 and Hp, M-SAA3 and CRP or M-SAA3 and parity. Milk CRP was correlated with % fat (P = 0.002) and % protein (P = 0.001) of the milk samples. The lack of correlation of SCC with the M-SAA3 and CRP could result from these APP being more sensitive to intra-mammary infection than SCC. Quarter milk Hp had a range of <0.4–420 μg/ml with a median value of 3.6 μg/ml, with 92 % of samples below 20 μg/ml.ConclusionBaseline values of Hp, M-SAA3 and CRP were established in composite milk from cows with normal SCC on the dairy farm. Parity was recognized as a possible confounding factor when diagnosing mastitis using Hp. The value of the APP, Hp, M-SAA3 and CRP as substitutes or to complement SCC in indicating udder inflammation, was demonstrated.

There is a need to further our understanding of the role that the equine hindgut ecosystem plays in digestive processes and diseases. The aim of the present study was to utilise the real-time PCR technique to determine the abundance of candidate cellulolytic (Ruminococcus flavefaciens; Fibrobacter succinogenes) and non-cellulolytic (Streptococcus bovis) bacteria in lumen contents from the caecum, ventral and dorsal colon, and rectum of healthy horses (n 14). Total DNA was extracted from frozen and lyophilised lumen contents, and PCR primers and Taqman w probes were designed based on 16S rDNA sequences for specific detection of candidate bacterial species. Overall, in frozen and lyophilised digesta, there were significantly (P, 0·01) fewer candidate bacteria in the caecum than the dorsal colon and rectum. In frozen digesta, candidate bacteria levels were similar between the ventral colon, dorsal colon and rectum, but in lyophilised digesta there were significantly (P,0·05) higher levels of bacteria in the dorsal colon and rectum. Frozen digesta contained disparate levels of candidate bacteria such that R. flavefaciens . F. succinogenes . S. bovis (P, 0·05), while in lyophilised digesta R. flavefaciens was present in significantly (P, 0·05) greater amounts than F. succinogenes and S. bovis. R. flavefaciens and F. succinogenes were abundant at significantly (P,0·05) greater levels in lyophilised digesta v. frozen digesta, with no difference in S. bovis levels. These data indicate that for these bacteria at least, faeces are a suitable model for studying the bacterial ecosystem within the equine colon. The present study also indicates that the preservation method of digesta affects levels of bacteria detected.

1. The objective of this study was to investigate the effect of supplementing broiler diets with xylanase or xylo-oligosaccharide (XOS) on growth performance, the concentration of non-starch polysaccharide (NSP) hydrolysis products in the ileum and concentration of short chain fatty acids (SCFA) in the caeca of broiler chickens. 2. In total, 500 male Ross 308 broilers were used in this 29-day (d) study. The treatments were organised into a 2 × 2 plus 1 factorial arrangement consisting of two additives (xylanase or XOS) at two levels (low or high) plus a control treatment with no additives. This gave five treatments with 100 birds in each treatment group. The diets were slightly deficient in protein by 20 g/kg and energy by 1 MJ/kg. 3. On d 14 and 28, two birds per pen were euthanised, the caeca content collected and analysed for short chain fatty acid (SCFA) concentration. On d 29, six birds per pen were euthanised and ileal digesta were collected and analysed for the concentration of NSP fractions. 4. On d 14, caecal acetic acid, iso-butyric acid, iso-valeric acid, n-valeric acid and total SCFA concentrations were significantly greater (P ≤ 0.05) when diets were supplemented with XOS compared with xylanase. 5. Ileal concentration of arabinose, galactose and glucuronic acid (GlucA2) were significantly greater (P ≤ 0.05) in the insoluble NSP fraction when diets were supplemented with a high level of xylanase, compared with the control treatment. Ileal concentration of fructose was significantly greater (P ≤ 0.05) in the water soluble NSP when a high level of xylanase or low level of XOS were included in the diet compared with the control. 6. It was concluded that xylanase and XOS had similar effects on NSP concentration and SCFA in the caeca, although there was little effect on performance. This observation demonstrated further benefits of xylanase supplementation in wheat-based broiler diets beyond digesta viscosity reduction and the release of extra nutrients.

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Gut microbiota have been associated with health, disease and behaviour in several species and are an important link in gut-brain axis communication. Diet plays a key role in affecting the composition of gut microbiota. In horses, high-starch diets alter the hindgut microbiota. High-starch diets are also associated with increased behavioural reactivity in horses. These changes in microbiota and behaviour may be associated. This study compares the faecal microbiota and behaviour of 10 naïve ponies. A cross-over design was used with experimental groups fed high-starch (HS) or high-fibre (HF) diets. Results showed that ponies were more reactive and less settled when being fed the HS diet compared to the HF diet. Irrespective of diet, the bacterial profile was dominated by two main phyla, Firmicutes, closely followed by Bacteroidetes. However, at lower taxonomic levels multivariate analysis of 16S rRNA gene sequencing data showed diet affected faecal microbial community structure. The abundance of 85 OTUs differed significantly related to diet. Correlative relationships exist between dietary induced alterations to faecal microbiota and behaviour. Results demonstrate a clear link between diet, faecal microbial community composition and behaviour. Dietary induced alterations to gut microbiota play a role in affecting the behaviour of the host.

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The neurotransmitters/neuromodulators galanin (GAL) and galanin-like peptide (GALP) are known to operate through three G proteincoupled receptors, GALR1, GALR2 and GALR3. The aim of this study was to investigate changes in expression of mRNA for galanin, GALP and GALR1-3 in the hypothalamus and pituitary gland, of male and female sheep, to determine how expression changed in association with growth and the attainment of reproductive competence. Tissue samples from the hypothalami and pituitary glands were analysed from late foetal and pre-pubertal lambs and adult sheep. Although mRNA for galanin and GALR1-3 was present in both tissues, at all ages and in both genders, quantification of GALP mRNA was not possible due to its low levels of expression. mRNA expression for both galanin and its receptors was seen to change significantly in both tissues as a function of age. Specifically, hypothalamic galanin mRNA expression increased with age in the male, but decreased with age in the female pituitary gland. mRNA expression for all receptors increased between foetal and pre-pubertal age groups and decreased significantly between pre-pubertal and adult animals. The results indicate that the expression of mRNA for galanin and its receptors changes dynamically with age and those significant differences exist with regard to tissue type and gender. These changes suggest that galaninergic neuroendocrine systems could be involved in the regulation of ovine growth and or the development of reproductive competence. The roles played by these systems in the sheep, however, may differ from other species, in particular the neuroendocrine link between nutrition and reproduction and GALR1's role in pituitary signalling.