Gut Sections Research Articles

Controlled Intestinal Microbiota Colonisation in Broilers under the Industrial Production System

The concept of designer microbiota in chicken is focused on early exposure of the hatchlings to pathogen-free microbiota inoculum, limiting the early access to harmful and pathogenic microorganisms, thus promoting colonisation of the gut with beneficial and natural poultry microbiota. In this study, we controlled colonisation of the intestine in broiler chickens in a large-scale industrial setting via at-hatch administration of a commercial product containing a highly diverse microbiota originating from the chicken caecum. The treatment significantly transformed the microbiota membership in the crop, proventriculus, jejunum and caecum and significantly altered the taxa abundance in the jejunum, jejunum mucosa, and caecum estimated using PERMANOVA and unweighted and weighted UniFrac distances, respectively. The treatment also improved the growth rate in chickens with no significant alteration in feed conversion ratio. A comparison of inoculum product microbiota structure revealed that the inoculum had the highest Shannon diversity index compared to all investigated gut sections, and the number of Observed Species second only to the caecal community. PCoA plots using weighted or unweighted UniFrac placed the inoculum samples together with the samples from the caecal origin.

A novel gut inflammatory rat model by laparotomic injection of peptidoglycan from Staphylococcus aureus.

Inflammatory bowel disease includes ulcerative colitis and Crohn's disease, and is globally increasing. An appropriate model system is required to dissect the disease pathogenesis and drugs screening for adequate treatment. In the present study, we established a novel model of gut inflammation by injecting peptidoglycan from Staphylococcus aureus using laparotomic procedure. For this, three different doses of peptidoglycan, i.e., 2.5, 5 and 10mg/kg body weight were used. The treatment effect was evaluated by studying the macrophage phagocytic function, spleen lymphocytes' proliferation and qRT-PCR for the assessment of peritoneal cells' gene expression. In addition, histological analysis of gut sections, gastric acidity, immunoglobulins and cytokines were assessed. There was significant increase in phagocytic activity in 10mg/kg body weight PGN group. A dose dependent increase in spleen lymphocytes' proliferation and a significant increase in total acid secretion in 5 and 10mg/kg body weight PGN treated rats were observed. In addition, a significant increment in TLR-2 and CD-14 mRNA expression in peritoneal cells, TNF-α, IL-6 and IFN-γ level and maximum distortion of gut architecture was observed in 10mg/kg body weight PGN group. Hence, peptidoglycan from S. aureus can be used for establishing the screening model to study the action and mechanism of anti-inflammatory food products and drugs.

Metatranscriptomic holobiont analysis of carbohydrate-active enzymes in the millipede Telodeinopus aoutii (Diplopoda, Spirostreptida)

As important decomposers of soil organic matter, millipedes contribute to lignocellulose decomposition and nutrient cycling. The degradation of lignocellulose requires the action of several carbohydrate-active enzymes (CAZymes) and, in most invertebrates, depends on the activity of mutualistic gut microorganisms. To address the question of the importance of the microbiota and endogenous (host) enzymes in digestive processes in millipedes, we analyzed metatranscriptomic data from the tropical millipedeTelodeinopus aoutiiat the holobiont level. Functional annotation included identification of expressed CAZymes (CAZy families and EC terms) in the host and its intestinal microbiota, foregut, midgut, and hindgut, compared to non-intestinal tissues. Most of the 175 CAZy families were expressed exclusively in the gut microbiota and more than 50% of these microbial families were expressed exclusively in the hindgut. The greatest diversity of expressed endogenous CAZymes from all gut sections was found in the midgut (77 families). Bacteria were the major microbial producers of CAZymes, Proteobacteria dominating in the midgut and Bacteriodetes with Firmicutes in the hindgut. The contribution of the eukaryotic microbiota to CAZymes production was negligible. Functional classification of expressed CAZy families confirmed a broad functional spectrum of CAZymes potentially expressed in the holobiont. Degradation of lignocellulose in the digestive tract of the millipedeT. aoutiidepends largely on bacterial enzymes expressed in the hindgut. Endogenous cellulases were not detected, except for the potentially cellulolytic family AA15, but an expression of cellulolytic enzymes of this family was not confirmed at the EC-number level. The midgut had the greatest diversity of expressed endogenous CAZymes, mainly amylases, indicating the importance of digesting α-glucosidases for the millipede. In contrast, bacterial lignocellulolytic enzymes are sparsely expressed here. The hindgut was the hotspot of microbial degradation of cellulose and hemicellulases. The gain of the millipede from the microbial lignocellulose degradation in the gut, and consequently the mutualistic status of the relationship between the millipede and its cellulolytic gut bacteria, depends on the ability of the millipede to take up microbial metabolites as nutrients through the hindgut wall. Enzymes expressed in the intestine can degrade all components of lignocellulose except lignin. Assuming that soil microbiota is partially degraded lignin in the millipede diet,T. aoutiican be considered a decomposer of soil organic matter relying primarily on its gut bacteria. The deposition of millipede fecal pellets containing an organic matter modified by the hindgut bacterial community could be of ecological significance.

Exploring the effect of a microencapsulated citrus essential oil on in vitro fermentation kinetics of pig gut microbiota.

Essential oils (EOs) have emerged as a potential alternative to antibiotics in pig breeding due to their antimicrobial properties. Citrus EOs, a common by-product of the orange juice industry, can be an interesting alternative from a financial perspective due to their huge offer in the global market. Thus, the effect of a citrus EO, and specifically different formulations of Brazilian Orange Terpenes (BOT), on pig gut microbiota was evaluated by means of an in vitro fermentation model simulating different sections of the pig gut (stomach, ileum, and colon). Treatments consisted in: BOT in its unprotected form (BOT, 1.85 and 3.70 mg/mL), microencapsulated BOT (MBOT, 3.50 and 7.00 mg/mL), colistin (2 μg/mL), and a control. BOT and MBOT altered in a similar way the total bacterial 16S rRNA gene copies in the stomach only from 18 h of incubation onwards, and no metabolite production in terms of short-chain fatty acids (SCFAs) was detected. In ileal and colonic fermentations, BOT and MBOT affected ileal and colonic microbiota in terms of total bacterial 16S rRNA gene copies, reduced phylogenetic diversity, and altered composition (p < 0.05) as evidenced by the significant reduction of certain bacterial taxa. However, more pronounced effects were found for MBOT, indicating its higher antimicrobial effects compared to the unprotected BOT, and suggesting that the antibacterial efficiency of the unprotected BOT was probably enhanced by microencapsulation. Furthermore, MBOT stimulated lactate production in ileal fermentations and greatly stimulated overall SCFA production in colonic fermentations. This indicates that besides the shifts in ileal and colonic microbiota by the delivered EO (BOT), the wall material of microcapsules (chitosan/modified starch) might have worked as an additional carbon source with prebiotic functioning, stimulating growth and metabolic activity (SCFAs) of colonic bacteria.

Representativeness of Fecal Microbiota Is Limited to Cecum and Colon in Domestic Yak

Gut microbiota are important for the health and adaptability of the domestic yak. Fecal microbiota are one of the most popular groups of microorganisms used to estimate the domestic yaks’ status, given the ease of obtaining fecal samples. However, because gut microbiota differ between gut sections, the representativeness of feces in microbiota is unclear in yak. To fill this gap, we compared the gut microbial diversities and functions of microbiota in the feces and seven other intestinal parts of domestic yaks based on 16S rRNA, including the rumen, duodenum, jejunum, ileum, cecum, colon, and rectum. The gut microbiota of eight intestinal parts showed significant differences at the beta-diversity level. However, there were no significant differences in the diversity and functions of microbiota between the feces and cecum and the feces and colon because of the digestive process. According to source-tracking analysis, most fecal microbiota originate from the cecum and colon. We speculated that the representativeness of fecal microbiota is limited to cecum and colon in domestic yak. Our study is the foundation of the use of fecal microbiota for animal husbandry research.

Substrate degradation pathways, conserved functions and community composition of the hindgut microbiota in the herbivorous marine fish Kyphosus sydneyanus

Symbiotic gut microbiota in the herbivorous marine fish Kyphosus sydneyanus play an important role in digestion by converting refractory algal carbohydrate into short-chain fatty acids. Here we characterised community composition using both 16S rRNA gene amplicon sequencing and shotgun-metagenome sequencing. Sequencing was carried out on lumen and mucosa samples (radial sections) from three axial sections taken from the hindgut of wild-caught fish. Both lumen and mucosa communities displayed distinct distributions along the hindgut, likely an effect of the differing selection pressures within these hindgut locations, as well as considerable variation among individual fish. In contrast, metagenomic sequences displayed a high level of functional similarity between individual fish and gut sections in the relative abundance of genes (based on sequencing depth) that encoded enzymes involved in algal-derived substrate degradation. These results suggest that the host gut environment selects for functional capacity in symbionts rather than taxonomic identity. Functional annotation of the enzymes encoded by the gut microbiota was carried out to infer the metabolic pathways used by the gut microbiota for the degradation of important dietary substrates: mannitol, alginate, laminarin, fucoidan and galactan (e.g. agar and carrageenan). This work provides the first evidence of the genomic potential of K. sydneyanus hindgut microbiota to convert highly refractory algal carbohydrates into metabolically useful short-chain fatty acids.

Investigation on the colonisation of Campylobacter strains in the pig intestine depending on available metabolites

Campylobacter (C.) spp. represent one of the most important causes for food-borne bacterial pathogen in humans worldwide. The aim of this study was to investigate metabolic requirements of two Campylobacter strains of different species based on substrate utilisation (in vitro). Based on these results, a correlation between the colonisation and the available substrates in different intestinal sections was recorded using an animal model. Campylobacter coli (ST-5777) and C. jejuni (ST-122) were used to inoculate 16 pigs, respectively, and one group of 16 pigs was used as control. The strains differed significantly in substrate utilisation - C. coli was able to metabolise various substrates (acetate, asparagine, serine, fucose, and propionate), while C. jejuni only utilised serine. Metabolomic analysis of intestinal content from different gut sections showed the presence of all previously tested metabolites, except for fucose. A significantly larger amount of glucose was found in the jejunum of those pigs infected with C. coli, while neither strain utilised it in vitro. The analysis of the intestinal contents revealed a very low proportion of Campylobacterales in the total microbiome, suggesting that the small percentage of the inoculated Campylobacter strains in the gut microflora of the animals is too low to cause differences between the control and infected groups in the composition of the metabolome. Nevertheless, knowledge of specific nutritional requirements of the pathogens combined with proof of different metabolites in the intestinal segments may provide clues about the site of colonisation in the host and improve our understanding of this zoonotic germ.

Morphology, microbiota, and metabolome along the intestinal tract of female turkeys

The global turkey industry is confronted with emerging challenges regarding health and welfare. Performance and disease resilience are directly linked to gut health. A clear definition of a healthy gut is a prerequisite to developing new strategies for improved gut health and, thus, general health, welfare and productivity. To date, detailed knowledge about gut health characteristics, especially during the critical fattening period, is still lacking for turkeys. Therefore, the goal of this study was to describe the morphology, microbiota, and metabolome along the intestinal tract of clinically healthy Salmonella- and Campylobacter-free commercial turkey hens throughout the fattening period from 7 to 10 wk posthatch, and obtain information on the stability of the investigated values over time. Feed changes were avoided directly preceding and during the investigation period. Investigation methods included histomorphometric measurement of intestinal villi and crypts, Illumina-sequencing for microbiota analysis, and proton nuclear magnetic resonance spectroscopy for metabolite identification and quantification. Overall, the study demonstrated a high repeatability across all 3 experiments and gut section differences observed coincided with their functions. It was demonstrated that gut maturation, defined by gut microbiota stability, is reached earlier in the ceca than any other intestinal section where morphological changes are ongoing throughout the fattening period. Therefore, the present study provides valuable information necessary to advise future studies on the development and implementation of measures to support gut maturation and establish a protective microbiota in commercial turkeys.

Divergent gut microbiota in two closely related house mouse subspecies under common garden conditions.

The gastrointestinal microbiota (GM) is considered an important component of the vertebrate holobiont. GM-host interactions influence the fitness of holobionts and are, therefore, an integral part of evolution. The house mouse is a prominent model for GM-host interactions, and evidence suggests a role for GM in mouse speciation. However, previous studies based on short 16S rRNA GM profiles of wild house mouse subspecies failed to detect GM divergence, which is a prerequisite for the inclusion of GM in Dobzhansky-Muller incompatibilities. Here, we used standard 16S rRNA GM profiling in two mouse subspecies, Mus musculus musculus and M. m. domesticus, including the intestinal mucosa and content of three gut sections (ileum, caecum, and colon). We reduced environmental variability by sampling GM in the offspring of wild mice bred under seminatural conditions. Although the breeding conditions allowed a contact between the subspecies, we found a clear differentiation of GM between them, in all three gut sections. Differentiation was mainly driven by several Helicobacters and two H. ganmani variants showed a signal of codivergence with their hosts. Helicobacters represent promising candidates for studying GM-host coadaptations and the fitness effects of their interactions.

Effect of Nosema ceranae infection and season on the gut bacteriome composition of the European honeybee (Apis mellifera)

Nosema ceranae is an intracellular parasite that infects honeybees’ gut altering the digestive functions; therefore, it has the potential of affecting the composition of the gut microbiome. In this work, individual bees of known age were sampled both in spring and autumn, and their digestive tracts were assessed for N. ceranae infection. Intestinal microbiome was assessed by sequencing the bacterial 16S rRNA gene in two different gut sections, the anterior section (AS; midgut and a half of ileum) and the posterior section (PS; second half of ileum and rectum). A preliminary analysis with a first batch of samples (n = 42) showed that AS samples had a higher potential to discriminate between infected and non-infected bees than PS samples. As a consequence, AS samples were selected for subsequent analyses. When analyzing the whole set of AS samples (n = 158) no changes in α- or β-diversity were observed between infected and non-infected bees. However, significant changes in the relative abundance of Proteobacteria and Firmicutes appeared when a subgroup of highly infected bees was compared to the group of non-infected bees. Seasonality and bees’ age had a significant impact in shaping the bacteriome structure and composition of the bees’ gut. Further research is needed to elucidate possible associations between the microbiome and N. ceranae infection in order to find efficient strategies for prevention of infections through modulation of bees’ microbiome.

Effect of Two Soybean Varieties Treated with Different Heat Intensities on Ileal and Caecal Microbiota in Broiler Chickens.

Simple SummarySoybeans are an essential part of today’s poultry nutrition diets because of their high protein content and quality. To ensure optimum digestibility in monogastric animals, soybeans need to be thermally processed. As required heat intensities depend on individual soybean properties, the emergence of highly heterogenic soybean batches is a challenge for adequate processing conditions. Molecular changes occurring during heat treatment can alter the microbial communities colonizing the animals’ guts. Gut microbiota is of great importance for both its host animal’s performance and health. To investigate the effect of heat treatment and soybean variety on the chickens’ microbiota, two soybean varieties were selected, treated at two different heat intensities and subjected to a feeding trial. DNA was then extracted and sequenced to identify different bacterial populations in the digesta of certain gut sections. Results showed that both the soybean variety and the applied heat treatment affected the abundance of certain bacterial species in the gut of chickens, but no effect on the taxonomy level of family or genus appeared. This underlines the sensitivity and reactivity of the highly complex microbial community to apparently small dietary differences.Soybean products are of high importance for the protein supply of poultry. Heat treatment of soybeans is essential to ensure optimal digestibility because of intrinsic antinutritive factors typical for this feed category. However, excessive treatment promotes the Maillard reaction and reduces protein digestibility. Furthermore, Europe’s efforts are to decrease dependence on imports of soybean products and enlarge local production. This process will include an increase in the variability of soybean batches, posing great challenges to adequate processing conditions. Intrinsic soybean properties plus heat treatment intensity might be able to modulate the gut microbiota, which is of crucial importance for an animal’s health and performance. To assess the influence of heat treatment and soybean variety on gut microbiota, 2 soybean cakes from 2 varieties were processed at 110 °C or 120 °C and subsequently fed to 336 one-day-old broiler chickens. After 36 days, the animals were slaughtered, and the digesta of the ileum and caecum was collected. Next, 16S rRNA amplicon sequencing of the extracted DNA revealed a high discrepancy between gut sections, but there were no differences between male and female birds. Significant differences attributed to the different soybean varieties and heat intensity were detected for certain bacterial taxa. However, no effect on specific families or genera appeared. In conclusion, the results indicated the potential of processing conditions and soybean variety as microbiota-modulating factors.

Trust your guts? The effect of gut section on diet composition and impact of Mus musculus on islands using metabarcoding.

DNA metabarcoding is widely used to characterize the diet of species, and it becomes very relevant for biodiversity conservation, allowing the understanding of trophic chains and the impact of invasive species. The need for cost‐effective biodiversity monitoring methods fostered advances in this technique. One question that arises is which sample type provides a better diet representation.Therefore, with this study, we intended to evaluate if there were differences in diet estimates according to the section of the gastrointestinal tract analysed and which section(s) provided the best diet representation. Additionally, we intended to infer the ecological/economic impacts of an invader as a model of the potential effects in an originally mammal‐free ecosystem.We examined the gut contents of the house mouse Mus musculus introduced to Cabo Verde, considering three sections: stomach, small intestine, and large intestine. We applied a DNA‐metabarcoding approach using two genetic markers, one specific for plants and another for invertebrates.We showed that this invader consumed 131 taxa (73 plants and 58 invertebrates). We obtained significant differences in the composition of two of the three sections, with a higher incidence of invertebrates in the stomach and plants in the intestines. This may be due to stomach inhibitors acting on plants and/or to faster absorption of soft‐body invertebrates compared to the plant fibers in the intestines. We verified that the impact of this invader in the ecosystem is predominantly negative, as at least 50% of the ingested items were native, endemic, or economically important taxa, and only 19% of the diet items were exotics.Overall, results showed the need to analyse only two gastrointestinal tract sections to obtain robust diet data, increasing the cost‐effectiveness of the method. Furthermore, by uncovering the native taxa most frequently preyed on by mice, this DNA‐metabarcoding approach allowed us to evaluate efficiently which are at the highest risk.

Distinct microbiota composition and fermentation products indicate functional compartmentalization in the hindgut of a marine herbivorous fish.

Many marine herbivorous fishes harbour diverse microbial communities in the hindgut that can play important roles in host health and nutrition. Kyphosus sydneyanus is a temperate marine herbivorous fish that feeds predominantly on brown seaweeds. We employed 16S rRNA gene amplicon sequencing and gas chromatography to characterize microbial communities and their metabolites in different hindgut regions of six K. sydneyanus. Measurements were confined to three distal sections of the intestine, labelled III, IV and V from anterior to posterior. A total of 625 operational taxonomic units from 20 phyla and 123 genera were obtained. Bacteroidota, Firmicutes and Proteobacteria were the major phyla in mean relative abundance, which varied along the gut. Firmicutes (76%) was the most dominant group in section III, whereas Bacteroidota (69.3%) dominated section V. Total short‐chain fatty acid (SCFA) concentration was highest in sections IV and V, confirming active fermentation in these two most distal sections. The abundance of Bacteroidota correlated with propionate concentration in section V, while Firmicutes positively correlated with formate in sections III and IV. Acetate levels were highest in sections IV and V, which correlated with abundance of Bacteroidota. Despite differences in gut microbial community composition, SCFA profiles were consistent between individual fish in the different hindgut regions of K. sydneyanus, although proportions of SCFAs differed among gut sections. These findings demonstrate functional compartmentalization of the hindgut microbial community, highlighting the need for regional sampling when interpreting overall microbiome function. These results support previous work suggesting that hindgut microbiota in marine herbivorous fish are important to nutrition in some host species by converting dietary carbohydrates into metabolically useful SCFAs.

Analysis of enterotoxigenic effect of Staphylococcus aureus and Staphylococcus epidermidis enterotoxins C and L on mice

Pathogenic properties of orthologues to S. aureus staphylococcal enterotoxin C (SEC) and staphylococcal enterotoxin L (SEL) produced by S. epidermidis are largely unexplored. We assessed the enteropathogenic effects of S. epidermidis SECepi and SELepi and S. aureus SEC3 and SEL after oral administration to Balb/c mice. Intestinal sections from SE-treated mice were analyzed histopathologically. The T cell lineage markers (αβ and γδ TCR CD3, CD4, CD8), T-cell activation marker CD69 and proliferation-related marker CD71 were assessed in intraepithelial lymphocytes (IEL), mesenteric lymph nodes (MLN) and spleens (SPL). Serum concentrations of SEC and SEL were determined. Ortologous S. epidermidis and S. aureus SEs exerted a number of common histopathological changes in the mouse gut. Atrophy, generation of villi gap and edema of the villi were the most prominent effects of SE treatment observed in mouse gut sections. The most marked effect of ortologous S. epidermidis and S. aureus SEs on the number of goblet cells, crypt depth and villi height was noted in the mice duodenum and jejunum. We indicate early changes of TCRαβ CD4-CD8a+ T and TCRαβ CD4+CD8a+ T cells in response to both S. aureus and S. epidermidis SEs. Upon the treatment with SEs, markers of T cell activation and proliferation were upregulated in both αβ and γδ T cell populations derived from IEL and MLN. We demonstrated that S. epidermidis-encoded SEs applied via oral route exert pathological changes in mice gut similarly to S. aureus-encoded SEs. For the first time we indicated that SEL co-produced together with SEC by both S. aureus and S. epidermidis induces some elements of mice gut immune response and contributes to gastrointestinal tract damage. Our results indicate the potential involvement of CoNS-encoded enterotoxins in the pathogenesis of SFP.

Effect of different dietary fibre sources on the zootechnical performance, feeding behaviour and intestinal physiology of growing and finishing pigs

The aim of this study was to determine the effect of different sources of fibre in the diets of fattening pigs on performance, feeding behaviour and intestinal physiology. A total of 60 barrows and gilts (initial body weight 28.4 ± 0.4 kg) were allotted to four dietary treatments: control (CON), lignocellulose (LC), mycelium (MYC) and corn gluten feed (CGF). Diets were calculated to provide balanced available nutrient contents. Including MYC in the diet resulted in an increased average daily gain (P &amp;lt; 0.05) compared to CON and CGF, and improved gain to feed ratio (P &amp;lt; 0.05) compared to LC. Pigs in CON (P &amp;lt; 0.05) ate the fewest but largest meals, whereas treatment CGF (P &amp;lt; 0.05) showed the opposite effect, resulting in the same daily feeder occupation time. Regarding intestinal physiology, in ileum, no differences were observed between the contents of short chain fatty acids (SCFA), lactic acid and biogenic amines. In the colon, MYC showed an increased concentration of acetic acid (P &amp;lt; 0.05) as well as the total content of SCFA (P &amp;lt; 0.05), compared to LC and CGF. Distinct fermentation profiles of ammonia were recorded in ileal and colonic digesta, although contents remained below harmful concentrations. Morphometrical measurements showed differences between the fibre sources LC and MYC, as well as LC and the CON in all investigated gut sections. These results provide evidence that the inclusion of specific dietary fibre sources/contents can positively influence the gut morphology and performance of pigs. However, further studies are needed regarding the mode of action and physico-chemical characteristics of the different fibre sources as a precondition for their successful application in pig diets.

Role of diet-microbiota interactions in precision nutrition of the chicken: facts, gaps, and new concepts

In the intestine, host-derived factors are genetically hardwired and difficult to modulate. However, the intestinal microbiome is more plastic and can be readily modulated by dietary factors. Further, it is becoming more apparent that the microbiome can potentially impact poultry physiology by participating in digestion, the absorption of nutrients, shaping of the mucosal immune response, energy homeostasis, and the synthesis or modulation of several potential bioactive metabolites. These activities are dependent on the quantity and quality of the microbiota alongside its metabolic potential, which are dictated in large part by diet. Thus, diet-induced microbiota alterations may be harnessed to induce changes in host physiology, including disease development and progression. In this regard, the gut microbiome is malleable and renders the gut microbiome a candidate ‘organ’ for the possibility of precision nutrition to induce precision microbiomics—the use of the gut microbiome as a biomarker to predict responsiveness to specific dietary constituents to generate precision diets and interventions for optimal poultry performance and health. However, it is vital to identify the causal relationships and mechanisms by which dietary components and additives affect the gut microbiome which then ultimately influence avian physiology. Further, an improved understanding of the spatial and functional relationships between the different sections of the avian gut and their regional microbiota will provide a better understanding of the role of the diet in regulating the intestinal microbiome.

Macronutrients and essential amino acids on digestive process of the freshwater teleost Matrinxã

The objective of this work was to evaluate the effect of macronutrients and essential amino acids on digestive process of the freshwater teleost Matrinxã (Brycon amazonicus). Juveniles were fed with diets containing starch plus free amino acids or oil plus free amino acids for 15 days. These fish were compared with others fed with diets containing starch or oil without addition of free amino acids. After the experimental span, 12 fish from each treatment were randomly sampled to collect stomach, pyloric cecum, anterior and posterior intestine for assaying digestive enzymes activity. Increase of gastric proteolysis due to dietary amino acids were observed. Amylolytic, proteolytic and lipolytic activities in intestine sections were also positive related to dietary amino acids. However, proteolytic and lipolytic activities in pyloric cecum were not responsive to dietary changes. Moreover, the absence of starch in the diets resulted in decrease of amylolysis, and very low levels of oil did not change the lipolytic activity. In conclusion, activities of amylase, protease and lipase of Matrinxãare selectively responsive to addition of free essential amino acids concerning the gut section.

The flow of non-starch polysaccharides along the gastrointestinal tract of broiler chickens fed either a wheat- or maize-based diet.

The present study characterised the types and amounts of non-starch polysaccharides (NSP) remaining undigested along the gastrointestinal tract (GIT) of broiler chickens offered a typical wheat- or maize-based diet. One-day old Cobb 500 mixed-sex chicks were assigned to 24 pens, with 10 birds/pen and 12 pens/treatment. Birds were offered the experimental diets in 3 phases (starter, day 0 to 10; grower, day 11 to 24 and finisher, day 25 to 35). Excreta and digesta samples from the crop, gizzard, duodenum, jejunum, ileum and caeca were collected at 12 and 35 days of age, and analysed for the NSP flow. The wheat-based diet contained higher levels of soluble NSP than the maize-based diet, whereas insoluble NSP levels were similar between the 2 diets. Detailed analysis of NSP constituents revealed that arabinoxylans were the primary NSP in the wheat-based diet, mostly in insoluble form. Pectins were the predominant NSP in the maize-based diet, followed by arabinoxylans. Overall, birds offered the wheat-based diet presented higher levels of soluble NSP remaining in all gut sections compared to birds offered the maize-based diet, at both 12 and 35 days of age (P < 0.050). Accumulation of insoluble NSP in the gizzard was noted in birds fed both diets, but was more pronounced in birds offered the maize-based diet compared to the wheat-based diet, at both 12 and 35 days of age (P < 0.001). The present study highlights marked differences in the amounts and types of NSP delivered to the different gut sections when feeding wheat-compared to maize-based diets, particularly in the gizzard and the lower GIT of birds.

Detoxification, Hydrogen Sulphide Metabolism and Wound Healing Are the Main Functions That Differentiate Caecum Protein Expression from Ileum of Week-Old Chicken

Sections of chicken gut differ in many aspects, e.g., the passage of digesta (continuous vs. discontinuous), the concentration of oxygen, and the density of colonising microbiota. Using an unbiased LC-MS/MS protocol, we compared protein expression in 18 ileal and 57 caecal tissue samples that originated from 7-day old ISA brown chickens. We found that proteins specific to the ileum were either structural (e.g., 3 actin isoforms, villin, or myosin 1A), or those required for nutrient digestion (e.g., sucrose isomaltase, maltase–glucoamylase, peptidase D) and absorption (e.g., fatty acid-binding protein 2 and 6 or bile acid–CoA:amino acid N-acyltransferase). On the other hand, proteins characteristic of the caecum were involved in sensing and limiting the consequences of oxidative stress (e.g., thioredoxin, peroxiredoxin 6), cell adhesion, and motility associated with wound healing (e.g., fibronectin 1, desmoyokin). These mechanisms are coupled with the activation of mechanisms suppressing the inflammatory response (galectin 1). Rather prominent were also expressions of proteins linked to hydrogen sulphide metabolism in caecum represented by cystathionin beta synthase, selenium-binding protein 1, mercaptopyruvate sulphurtransferase, and thiosulphate sulphurtransferase. Higher mRNA expression of nuclear factor, erythroid 2-like 2, the main oxidative stress transcriptional factor in caecum, further supported our observations.

Characteristics of Biotin lipoyl attachment and 2-oxoacid dehydrogenase acyltransferase of the parasitic nematode Haemonchus contortus and its modulatory functions on goat PBMCs in vitro.

Biotin lipoyl attachment and 2-oxoacid dehydrogenase acyltransferase (BLAODA), as an essential excretion of Haemonchus contortus (HcESPs), was identified to have antigenic functions. T helper-9 (Th9) cells secrete interleukin (IL)-9, a signature cytokine associated with tumour immunology, allergy and autoimmunity. Nonetheless, the understanding of modulatory functions of BLAODA on Th9 and other immune cells is limited. In this study, the BLAODA gene was cloned, and the recombinant (r) protein of BLAODA (rHcBLAODA) was expressed and immunoblotting was performed. The results revealed that HcBLAODA gene was successfully cloned and rHcBLAODA protein was expressed. The localization of rHcBLAODA was confirmed on the surface of gut sections from adult H. contortus. The rHcBLAODA protein capability to react precisely with anti-H. contortus antibodies were confirmed by immunoblotting and immunofluorescence assay (IFA). Further functional analysis showed that interaction of rHcBLAODA with host cells significantly enhanced Th9 cells generation, IL-9 expression, nitric oxide production and cell apoptosis while suppressing the cells proliferation and cells migration depending on the concentration. Overall, these findings suggest that rHcBLAODA protein could modulate the host immune response by inducing Th9 cells to secrete IL-9 cytokine in vitro.