Vulnerability assessments in dairy cattle farms based on individual sensitivity to heat stress.

Climate change (CC) is expected to increase temperatures and the frequency of extreme weather events, which renewed interest in heat stress (HS) effects on dairy cattle farms. Resilience is a key concept that should be considered to better understand the dairy farms exposure to HS and to combat CC-related risks. Thus, this study aimed to investigate the aspects of HS vulnerability for Tunisian dairy cattle farming systems. Historical milk test-day records from official milk recording were merged with temperature and humidity data provided by public weather stations. Firstly, different models relying in two heat load indices were applied for HS exposure assessment. Secondly, broken line models were used to estimate HS thresholds, milk losses, and rates of decline of milk production associated with temperature-humidity index (THI) across parities. Thirdly, individual cow responses to HS estimated using random regression model were considered as key measures of dairy farming system sensitivity assessment to HS. Dairy farms are annually exposed for 5 months to high THI values above 72 in Tunisia. The tipping points, at which milk yield started to decline over parities with 3-day average THI, ranged between 65 and 67. The largest milk decline per unit of THI above threshold values was 0.135 ± 0.01 kg for multiparous cows. The milk losses estimated due to HS in the Holstein breed during the summer period (June to August) ranged between 110 and 142 kg/cow in north and south, respectively. A high HS sensitivity was proved especially in dairy farms characterized by large herd size and high milk production level. Hence, providing knowledge of dairy farms vulnerability to HS may provide the basis for developing strategies to reduce HS effects and plan for CC adaptation.

In vitro approach to evaluate the fermentation pattern of inulin-rich food in obese individuals.

Alterations of the gut microbiome have been associated with obesity and metabolic disorders. The gut microbiota can be influenced by the intake of dietary fibres with prebiotic properties, such as inulin-type fructans. The present study tested the hypothesis that obese individuals subjected for 12 weeks to an inulin-enriched v. inulin-poor diet have differential faecal fermentation patterns. The fermentation of cellulose and inulin hydrolysates of six different inulin-rich and inulin-poor vegetables of both groups was analysed in vitro on faecal inocula. The results showed that the microbiota from obese patients who received a fructan-rich diet for 3 weeks produces more gas and total SCFA compared with the microbiota taken from the same individuals before the treatment. Obese individuals fed with a low-fructan diet produce less gas and less SCFA compared with the treated group. The present study highlighted profound changes in microbiota fermentation capacity obtained by prebiotic intervention in obese individuals, which favours the production of specific bioactive metabolites.

Effect of dietary tannins on milk yield and composition, nitrogen partitioning and nitrogen use efficiency of lactating dairy cows: A meta-analysis.

Tannins are secondary plant compounds which have been extensively studied in order to improve the nitrogen use efficiency (NUE) of ruminants. A meta-analysis was performed of 58 in vivo experiments comparing milk yield, composition and nitrogen metabolism of lactating dairy cows fed diets with or without tannins. The meta-analysis shows that tannins have no impact on corrected milk yield, fat and protein content or NUE (p > .05). However, tannins reduce ruminal ammoniacal nitrogen (N) production by 16% (from 10.95 to 8.47 mg/dl on average), milk urea by 9% (from 15.82 to 14.03 mg/dl) and urinary N excretion (-11%; p < .05). This is compensated for by a lower apparent N digestibility (61.51% with dietary tannins compared to 66.17% without). The effect of tannin on N metabolism parameters increases with tannin dose (p < .05). The shift from urinary to faecal N may be beneficial for environment preservation, as urinary N induces more harmful emissions than faecal N. From a farmer's perspective, tannins seem unable to increase fat- and protein-corrected milk yield or reduce feed protein requirements and thus have no direct economic benefit. Potentially less costly than tannin extracts, forage or by-products naturally rich in tannins could still be useful to reduce the environmental impact of ruminant protein feeding.

Thermotolerance indicators related to production and physiological responses to heat stress of holstein cows.

Heat stress (HS) adversely influences dairy cattle welfare and productivity. This study aimed to investigate the effects of HS on production and physiological parameters of Holstein cows. Two experiments each lasted 6 weeks were conducted in four Tunisian farms, firstly during summer under HS (n = 80, THI = 77) and later during autumn under thermo-neutral (n = 80, THI = 54) conditions. Respiration rate (RR), skin temperature (ST), rectal temperature (RT) and milk yield were measured, and milk samples were collected on 2 days every week during each experimental period. Temperature and relative humidity were measured inside the barn to calculate the temperature-humidity index (THI). Mixed models were used to evaluate the effects of period and the relationships between THI and physiological and production traits. Reaction norm models were applied to quantify the individual responses of cows across the trajectory of THI during the HS period. A clustering methodology was developed to identify tolerant and sensitive cows to HS based on their slope for response of physiological and production traits during HS period. In summer, RR (61 breaths/min) and ST (37.7 °C) were 2.3- and 1.3-fold higher, whereas milk yield per milking was 24% lower compared with thermo-neutral conditions. Linear relationship between THI and RR, ST and RT was observed and showed increases by 2 breaths/min, 0.5 °C and 0.04 °C per increase in one THI unit, respectively. Inversely, milk, fat and protein yields showed a drop of 0.13 kg, 0.4 g and 0.3 g per milking per increase in one THI unit, respectively. Cows qualified to be heat tolerant by our work tended to have higher RR, ST, and RT and lower to almost no decay in milk yield compared to cows qualified to be heat sensitive. Specifically, RR could be used as a reliable indicator for thermotolerance. The results of this study deepen our understanding of different aspects of HS resilience.

Characterization of fructans and dietary fibre profiles in raw and steamed vegetables.

Dietary fibre (DF) has many positive effects on human health associated with its functionality in the gastrointestinal tract. These benefits vary according to the type of DF. Vegetables can be a natural source of DF in the diet. However, to provide adequate nutritional advice, the content and profile of their various DF types must be characterised. This study aimed to determine the DF profile of 29 vegetables cultivated in Wallonia (Belgium) and the impact of steaming on these profiles. Using a combination of enzymatic, gravimetric and chromatographic methods, fructans, total dietary fibre (TDF), low- and high-molecular-weight soluble dietary fibre (SDF), and insoluble dietary fibre (IDF) were analysed. Results show that the DF content varies considerably among the 29 investigated vegetable varieties and species, but the influence of steaming is limited to a shift from IDF to high-molecular-weight SDF for 18 of the 29 tested vegetables, while fructans are preserved with not actual reduction in the DP.

Screening and Characterization of Pediococcus acidilactici LC-9-1 toward Selection as a Potential Probiotic for Poultry with Antibacterial and Antioxidative Properties.

Growing interest has been focused on lactic acid bacteria as alternatives to antimicrobial growth promoters, which are characterized by the production of various functional metabolites, such as antimicrobial and antioxidants compounds. The present study was undertaken to evaluate a potential probiotic from the antioxidant perspective. LC-9-1, screened from the intestines of healthy animals, was revealed to be Pediococcus acidilactici on the basis of its morphological, biochemical, and molecular characteristics. The strain has excellent properties, including acid-production efficiency, antibacterial performance and antioxidant activity. The safety of the strain was also evaluated. Furthermore, the experiments in broiler chickens suggested that dietary LC-9-1 supplementation improved the growth performance and decreased the abdominal fat, and enhanced the antioxidant capability and intestinal innate immunity of broilers. Analysis of intestinal microbiota showed that a higher community diversity (Shannon index) was achieved. In addition to the significantly increased relative abundances of Pediococcus spp., beneficial genera such as Rothia spp. and Ruminococcus spp. were abundant, while opportunistic pathogens such as Escherichia-Shigella spp. were significantly reduced in LC-9-1-supplemented broilers. Collectively, such in-depth characterization and the available data will guide future efforts to develop next-generation probiotics, and LC-9-1 could be considered a potential strain for further utilization in direct-fed microbial or starter culture for fermentation.

Screening and characterization of Bacillus velezensis LB-Y-1 toward selection as a potential probiotic for poultry with multi-enzyme production property.

Bacillus spp. have gained increasing recognition as an option to use as antimicrobial growth promoters, which are characterized by producing various enzymes and antimicrobial compounds. The present study was undertaken to screen and evaluate a Bacillus strain with the multi-enzyme production property for poultry production. LB-Y-1, screened from the intestines of healthy animals, was revealed to be a Bacillus velezensis by the morphological, biochemical, and molecular characterization. The strain was screened out by a specific screening program, possessed excellent multi-enzyme production potential, including protease, cellulase, and phytase. Moreover, the strain also exhibited amylolytic and lipolytic activity in vitro. The dietary LB-Y-1 supplementation improved growth performance and tibia mineralization in chicken broilers, and increased serum albumin and serum total protein at 21 days of age (p < 0.05). Besides, LB-Y-1 enhanced the activity of serum alkaline phosphatase and digestive enzyme in broilers at 21 and 42 days of age (p < 0.05). Analysis of intestinal microbiota showed that a higher community richness (Chao1 index) and diversity (Shannon index) in the LB-Y-1 supplemented compared with the CON group. PCoA analysis showed that the community composition and structure were distinctly different between the CON and LB-Y-1 group. The beneficial genera such as Parasutterella and Rikenellaceae were abundant, while the opportunistic pathogen such as Escherichia-Shigella were reduced in the LB-Y-1 supplemented group (p < 0.05). Collectively, LB-Y-1 can be considered as a potential strain for further utilization in direct-fed microbial or starter culture for fermentation.

Effects of graded levels of xylo-oligosaccharides on growth performance, serum parameters, intestinal morphology, and intestinal barrier function in weaned piglets.

The objective of this study was to investigate the effects of xylo-oligosaccharides (XOSs) supplementation on growth performance, serum parameters, small intestinal morphology, intestinal mucosal integrity, and immune function in weaned piglets. A total of 240 weaned piglets with an average body weight (BW) of 8.82 ± 0.05 kg (28 d of age) were assigned randomly to four dietary treatments in a 28-d trial, including a control (CON) diet and three diets with XOS supplementation at the concentration of 100 (XOS100), 500 (XOS500), and 1,000 (XOS1000) mg/kg. There were four replicates per treatment with 15 pigs per pen. From day 1 to 14, there were no differences (P > 0.05) in average daily gain (ADG), average daily feed intake, and gain to feed ratio (G:F) during the different treatments. The different doses of XOSs showed a quadratic effect on BW on day 28, ADG, and G:F on day 1 to 28 of piglets (P < 0.05). From day 15 to 28, ADG of pigs fed the XOS500 diet was higher (P < 0.05) than pigs fed the CON diet. During the overall period (day 1 to 28), pigs fed the XOS500 diet had a higher BW, ADG, and G:F than pigs fed the CON diet (P < 0.05). In addition, compared with the CON group, the XOS500 group had significantly higher serum total antioxidant capacity, total superoxide dismutase and catalase levels, and lower malondialdehyde levels on days 14 and 28 (P < 0.05). The serum immunoglobulin G (IgG) concentration in the XOS500 group was also significantly higher compared with the CON group on days 14 and 28 (P < 0.05). However, serum immunoglobulin A and immunoglobulin M were not affected by the dietary treatments. Supplementation of XOS500 to the feed significantly increased the villus height (VH) and VH to crypt depth ratio in the jejunum and ileum in comparison with the CON and XOS1000 groups. Moreover, the XOS500 group significantly elevated the expression levels of occludin and zonula occludens protein-1 in the ileum compared with the CON group. The ileal interleukin (IL)-1ß, IL-8, and interferon (IFN)-γ mRNA expression levels in the XOS100 and XOS500 groups were markedly lower than in the CON group. In contrast, the ileal IL-10 mRNA expression levels were remarkably higher in the XOS500 than in the CON group. In conclusion, XOSs have a beneficial effect on growth performance by improving serum antioxidant defense system, serum IgG, small intestinal structure, and intestinal barrier function in weaned piglets.

Xylo-Oligosaccharides, Preparation and Application to Human and Animal Health: A Review.

Xylo-oligosaccharides (XOS) are considered as functional oligosaccharides and have great prebiotic potential. XOS are the degraded products of xylan prepared via chemical, physical or enzymatic degradation. They are mainly composed of xylose units linked by ß-1, 4 bonds. XOS not only exhibit some specific physicochemical properties such as excellent water solubility and high temperature resistance, but also have a variety of functional biological activities including anti-inflammation, antioxidative, antitumor, antimicrobial properties and so on. Numerous studies have revealed in the recent decades that XOS can be applied to many food and feed products and exert their nutritional benefits. XOS have also been demonstrated to reduce the occurrence of human health-related diseases, improve the growth and resistance to diseases of animals. These effects open a new perspective on XOS potential applications for human consumption and animal production. Herein, this review aims to provide a general overview of preparation methods for XOS, and will also discuss the current application of XOS to human and animal health field.

Effects of Xylo-Oligosaccharides on Growth and Gut Microbiota as Potential Replacements for Antibiotic in Weaning Piglets.

Xylo-oligosaccharides (XOS) is a well-known kind of oligosaccharide and extensively applied as a prebiotic. The objective of this study was to investigate the effect of XOS supplementation substituting chlortetracycline (CTC) on growth, gut morphology, gut microbiota, and hindgut short chain fatty acid (SCFA) contents of weaning piglets. A total of 180 weaned piglets were randomly allocated to three treatments for 28 days, as follows: control group (basal diet, CON), basal diet with 500 mg/kg (XOS500) XOS, and positive control (basal diet with 100 mg/kg CTC). Compared with the CON group, the piglets in the XOS500 group improved body weight (BW) on days 28, average daily gain (ADG) and reduced feed: gain ratio during days 1-28 (P < 0.05). The XOS500 supplementation increased Villus height and Villus height: Crypt depth ratio in the ileum (P < 0.05). Villus Height: Crypt Depth of the ileum was also increased in the CTC treatment group (P < 0.05). Meanwhile, the XOS500 supplementation increased significantly the numbers of goblet cells in the crypt of the cecum. High-throughput 16S rRNA gene sequencing revealed distinct differences in microbial compositions between the ileum and cecum. XOS500 supplementation significantly increased the bacterial diversity. However, CTC treatment markedly reduced the microbial diversity (P < 0.05). Meanwhile, XOS500 supplementation in the diet significantly increased the abundance of Lactobacillus genus compared to the CON and CTC group in the ileum and cecum (P < 0.01), whereas the level of Clostridium_sensu_stricto_1, Escherichia-Shigella, and Terrisporobacter genus in the XOS500 group were markedly lower than the CON and CTC group (P < 0.05). In addition, dietary supplementation with XOS500 significantly increased the total short-chain fatty acids, propionate and butyrate concentrations and decreased the acetate concentration compared to the CON group in the cecum (P < 0.05). In summary, dietary supplemented with XOS500 could enhance specific beneficial microbiota abundance and decrease harmful microbiota abundance to maintain the structure of the intestinal morphology and improve growth performance of weaned piglets. Thus, XOS may potentially function as an alternative to in-feed antibiotics in weaned piglets in modern husbandry.

The Walloon farmers position differently their ideal dairy production system between a global-based intensive and a local-based extensive model of farm.

Dairy farming systems are evolving. This study presents dairy producers' perceptions of their ideal future farm (IFF) to ensure revenue, and attempts to determine the reasons for this choice, the environmental aspects related to this choice, the proximity between the current farm and the IFF and the requirements for reaching this IFF. Just before the end of the European milk quota, a total of 245 Walloon dairy producers answered a survey about the characteristics of their IFF and other socio-environmental-economic information. A multiple correspondence analysis (MCA) was carried out using seven characteristics of the IFF (intensive vs. extensive, specialised vs. diversified, strongly vs. weakly based on new technologies, managed by a group of managers vs. an independent farmer, employed vs. familial workforce, local vs. global market, standard vs. quality-differentiated production) to observe the relationships between them. Based on the main contributors to the second dimension of the MCA, this axis was defined as an IFF gradient between the local-based extensive (LBE) producers (26%) and the global-based intensive (GBI) producers (46%). The differences of IFF gradient between modalities of categorical variables were estimated using generalised linear models. Pearson correlations were calculated between the scores on the IFF gradient and quantitative variables. Finally, frequencies of IFF characteristics and the corresponding characteristic for the current situation were calculated to determine the percentages of "unhappy" producers. Some reasons for the choice of IFF by the producers have been highlighted in this study. Environmental initiatives were more valued by LBE than GBI producers. Low similarity was observed between the current farm situation of the respondents and their IFF choice. LBE and GBI producers differed significantly regarding domains of formation (technical and bureaucratic vs. transformation and diversification respectively) and paths of formation (non-market vs. market respectively). Two kinds of farming systems were considered by dairy producers and some socioeconomic and environmental components differed between them.

Adhesion and growth inhibitory effect of chicken egg yolk antibody (IgY) on Salmonella enterica serovars Enteritidis and Typhimurium in vitro.

The protective effects of powder preparation of egg yolk immunoglobulin Y (IgY), specific to Salmonella Enteritidis and Salmonella Typhimurium outer membrane proteins (OMP), against these two Salmonella sp. serovars were investigated in vitro in two different assays: adhesion-prevention and growth-inhibition. The adhesion-prevention assay was conducted using polarized monolayers of the human intestinal epithelial Caco-2 cell line. First, the conditions of Salmonella adherence to Caco-2 cells were optimized, and interferences of bacteria with the transepithelial electrical resistance (TER) of fully differentiated Caco-2 cell monolayers and the lactate dehydrogenase release upon exposure of the cells to Salmonella were evaluated. Both Salmonella sp. serovars were able to adhere to Caco-2 cells and decreased TER. Results from the adhesion-prevention assay demonstrated that specific IgY reduced the decrease in TER of the infected Caco-2 cell monolayers and blocked the Salmonella sp. adhesion in a concentration-dependent manner (p < 0.05). Nonspecific IgY also exhibited an inhibitory effect on these two parameters, but to a lesser extent than that of the specific IgY (p < 0.05). The protective effect of nonspecific IgY could be attributed to the low-density lipoprotein component of the water-soluble fraction of egg yolks that may not have been eliminated during ultrafiltration. The growth-inhibition assay revealed that specific IgY had an inhibitory effect on the bacterial growth, markedly during the late exponential phase, whereas nonspecific IgY failed to do so. Taken together, these results suggest that the in vitro growth inhibitory effect of specific IgY on Salmonella spp. resulted from the specific binding activity of these IgY to Salmonella sp. OMP. Passive immunization with Salmonella sp. OMP-specific IgY could thus be useful to prevent Salmonella colonization in broiler chickens and the subsequent carcass contamination during processing.

The use of inulin and wheat bran only during the starter period or during the entire rearing life of broilers: effects on growth performance, small intestinal maturation, and cecal microbial colonization until slaughter age.

Inulin and wheat bran were added to broiler diets during the starter period or during the entire rearing period to investigate whether the effects of using these ingredients remained until slaughter age. Diets containing no inulin and no wheat bran (CON), 2% inulin (IN), 10% wheat bran (WB), or 2% inulin + 10% wheat bran (IN+WB) were provided until day 11. Thereafter, each dietary treatment was further divided into a continued diet with supplementation or a control diet, resulting in 7 groups (CON, IN/IN, IN/CON, WB/WB, WB/CON, IN+WB/IN+WB, or IN+WB/CON). On day 40, 12 chickens per group were euthanized. The IN/IN group increased the cecal molar ratio of butyrate but had a lower relative abundance of Lactobacillus (P < 0.05). Additionally, the cecal molar ratio of propionate was higher in the IN/CON group compared to the IN/IN group (P = 0.034). The WB/CON group had the best results on BW and feed conversion ratio (FCR) (P < 0.05). Only the cecal molar ratio of iso-butyrate was higher in the WB/WB group (P = 0.013). Moreover, compared to the CON group, both WB/WB and WB/CON groups reduced the relative abundances of Bifidobacterium and Escherichia coli, and only the WB/WB group reduced the relative abundance of Enterobacteriaceae (P < 0.05). Both IN+WB/IN+WB and IN+WB/CON groups increased BW until day 21 and lowered the relative abundance of Bifidobacterium (P < 0.05). The IN+WB/IN+WB group increased the cecal molar ratio of butyrate but reduced the molar ratio of propionate with a higher relative abundance of Enterobacteriaceae (P < 0.05). In conclusion, the lack of positive effects induced by inulin might be explained by the dose being too high. The beneficial effects on BW, FCR, and microbiota induced by wheat bran during the starter period were lasting when supplementation was stopped, suggesting that wheat bran could be a favorable ingredient during the starter period.

Influence of adding fibrolytic enzymes on the ruminal fermentation of date palm by-products.

This study was conducted in order to assess the influence of four doses (0, 0.5, 1, and 2 mg (g dry matter) - 1 of commercial fibrolytic enzymes (MAXFIBER-I ® , SHAUMANN GmbH, Wahlstedt, Germany) on in vitro fermentation of date palm (Phoenix dactylifera) by-products: date kernels, wasted dates, floral stems, and palm fronds. Rumen contents were obtained from two non-lactating Holstein cows. Enzyme supplementation to by-products was carried out 12 h prior to incubation. Compared to the control, the enzymatic supplementation quadratically increased the extent but not the gas production rate of date kernel fermentation. Indeed, the potential gas production increased notably by 14.8 % with the lowest enzymes dose following recorded gas production after 48, 72, and 96 h of incubation. The estimated organic-matter digestibility, metabolisable energy, and total volatile fatty acids in the incubation fluid tended to be increased with the lowest dose by 7.8 %, 8.4 %, and 13.9 % respectively. For the wasted dates, this feed additive tended to linearly increase the gas production rate of fermentation with the highest dose. On the other hand, this supplementation had no effect on the ruminal fermentation of the floral stems and palm fronds. The exogenous fibrolytic enzymes were more effective on fibrous but not on lignified date palm by-products.

Early Feeding Regime of Waste Milk, Milk, and Milk Replacer for Calves Has Different Effects on Rumen Fermentation and the Bacterial Community.

We investigated the effects of different types of early feeding on rumen fermentation parameters and the bacterial community in calves. Fifty-four Holstein calves were assigned to three treatments and fed whole milk (M), pasteurized waste milk (WM), or milk replacer (MR). Male calves were slaughtered at the age of two months to measure the stomach masses. The female calves were followed for six months to determine the body weight, blood indices, rumen fermentation, and ruminal bacterial community. At the age of two months, the average daily gain was lower, but the concentration of total volatile fatty acids was greater in the MR group. Starter intake and stomach mass were lower, but the isovalerate molar proportion was greater in the WM group. The blood indices and ruminal bacterial community of the WM group differed from those of the other groups. At the age of six months, the ruminal propionate molar proportion was lower, but the ruminal pH and acetate/propionate ratio were greater in the MR group. In conclusion, calves fed WM had different rumen fermentation and bacterial community during the weaning period, whereas feeding MR produced a long-lasting effect on the rumen environment.

The effect of inulin and wheat bran on intestinal health and microbiota in the early life of broiler chickens.

Inulin and wheat bran were added to the starter diets of broiler chickens to investigate the potential of these ingredients to improve the host's health and growth performance, as well as the underlying mechanisms of their effects. A total of 960 1-day-old chicks were assigned to 4 treatments: control (CON), 2% inulin (IN), 10% wheat bran (WB), and 10% wheat bran +2% inulin (WB+IN). On day 11, 6 chicks per treatment were euthanized. A general linear model procedure with Tukey's multiple range test was performed to compare a series of parameters between treatments. The WB-containing treatments improved BW on day 7, day 11, day 35, and BW gain until day 11 (P < 0.05), but only the WB+IN treatment showed a lower feed conversion ratio than the CON treatment (P = 0.011). Furthermore, the WB+IN treatment showed the highest villus height in the jejunum and ileum (P < 0.05), and the highest jejunal ratio villus height/crypt depth (P = 0.035). The concentration of acetate in the ceca was higher in the CON treatment compared to the IN treatment (P = 0.040). The IN treatment increased the concentration (P = 0.003) and ratio (P = 0.004) of iso-butyrate compared to the WB+IN and the CON treatments (P < 0.05). A clustering result exhibited similar intestinal microbiota profiles in the chicks receiving the IN and the WB+IN diets (P > 0.05), but these profiles were different from those found in chicks receiving the WB and the CON diets (P < 0.05). In conclusion, wheat bran and the combination of wheat bran and inulin ameliorated the growth performance and gut morphology of the starter chicks, which resulted in a higher BW until day 35. Inulin, on the other hand, had a greater ability to influence the microbiota profile. The beneficial results found in relation to BW and gut morphology during the starter period suggested a synergistic effect of inulin and wheat bran.

Differences in Ureolytic Bacterial Composition between the Rumen Digesta and Rumen Wall Based on ureC Gene Classification.

Ureolytic bacteria are key organisms in the rumen producing urease enzymes to catalyze the breakdown of urea to ammonia for the synthesis of microbial protein. However, little is known about the diversity and distribution of rumen ureolytic microorganisms. The urease gene (ureC) has been the target gene of choice for analysis of the urea-degrading microorganisms in various environments. In this study, we investigated the predominant ureC genes of the ureolytic bacteria in the rumen of dairy cows using high-throughput sequencing. Six dairy cows with rumen fistulas were assigned to a two-period cross-over trial. A control group (n = 3) were fed a total mixed ration without urea and the treatment group (n = 3) were fed rations plus 180 g urea per cow per day at three separate times. Rumen bacterial samples from liquid and solid digesta and rumen wall fractions were collected for ureC gene amplification and sequencing using Miseq. The wall-adherent bacteria (WAB) had a distinct ureolytic bacterial profile compared to the solid-adherent bacteria (SAB) and liquid-associated bacteria (LAB) but more than 55% of the ureC sequences did not affiliate with any known taxonomically assigned urease genes. Diversity analysis of the ureC genes showed that the Shannon and Chao1 indices for the rumen WAB was lower than those observed for the SAB and LAB (P < 0.01). The most abundant ureC genes were affiliated with Methylococcaceae, Clostridiaceae, Paenibacillaceae, Helicobacteraceae, and Methylophilaceae families. Compared with the rumen LAB and SAB, relative abundance of the OTUs affiliated with Methylophilus and Marinobacter genera were significantly higher (P < 0.05) in the WAB. Supplementation with urea did not alter the composition of the detected ureolytic bacteria. This study has identified significant populations of ureolytic WAB representing genera that have not been recognized or studied previously in the rumen. The taxonomic classification of rumen ureC genes in the dairy cow indicates that the majority of ureolytic bacteria are yet to be identified. This survey has expanded our knowledge of ureC gene information relating to the rumen ureolytic microbial community, and provides a basis for obtaining regulatory targets of ureolytic bacteria to moderate urea hydrolysis in the rumen.

Illumina Sequencing Approach to Characterize Thiamine Metabolism Related Bacteria and the Impacts of Thiamine Supplementation on Ruminal Microbiota in Dairy Cows Fed High-Grain Diets.

The requirements of thiamine in adult ruminants are mainly met by ruminal bacterial synthesis, and thiamine deficiencies will occur when dairy cows overfed with high grain diet. However, there is limited knowledge with regard to the ruminal thiamine synthesis bacteria, and whether thiamine deficiency is related to the altered bacterial community by high grain diet is still unclear. To explore thiamine synthesis bacteria and the response of ruminal microbiota to high grain feeding and thiamine supplementation, six rumen-cannulated Holstein cows were randomly assigned into a replicated 3 × 3 Latin square design trial. Three treatments were control diet (CON, 20% dietary starch, DM basis), high grain diet (HG, 33.2% dietary starch, DM basis) and high grain diet supplemented with 180 mg thiamine/kg DMI (HG+T). On day 21 of each period, rumen content samples were collected at 3 h postfeeding. Ruminal thiamine concentration was detected by high performance liquid chromatography. The microbiota composition was determined using Illumina MiSeq sequencing of 16S rRNA gene. Cows receiving thiamine supplementation had greater ruminal pH value, acetate and thiamine content in the rumen. Principal coordinate analysis and similarity analysis indicated that HG feeding and thiamine supplementation caused a strong shift in bacterial composition and structure in the rumen. At the genus level, compared with CON group, the relative abundances of 19 genera were significantly changed by HG feeding. Thiamine supplementation increased the abundance of cellulolytic bacteria including Bacteroides, Ruminococcus 1, Pyramidobacter, Succinivibrio, and Ruminobacter, and their increases enhanced the fiber degradation and ruminal acetate production in HG+T group. Christensenellaceae R7, Lachnospira, Succiniclasticum, and Ruminococcaceae NK4A214 exhibited a negative response to thiamine supplementation. Moreover, correlation analysis revealed that ruminal thiamine concentration was positively correlated with Bacteroides, Ruminococcus 1, Ruminobacter, Pyramidobacter, and Fibrobacter. Taken together, we concluded that Bacteroides, Ruminococcus 1, Ruminobacter, Pyramidobacter, and Fibrobacter in rumen content may be associated with thiamine synthesis or thiamine is required for their growth and metabolism. In addition, thiamine supplementation can potentially improve rumen function, as indicated by greater numbers of cellulolytic bacteria within the rumen. These findings facilitate understanding of bacterial thiamine synthesis within rumen and thiamine's function in dairy cows.

Cooking Has Variable Effects on the Fermentability in the Large Intestine of the Fraction of Meats, Grain Legumes, and Insects That Is Resistant to Digestion in the Small Intestine in an in Vitro Model of the Pig's Gastrointestinal Tract.

This study aimed to evaluate the fermentation in the large intestine of indigestible dietary protein sources from animal, insect, and plant origin using an in vitro model of the pig's gastrointestinal tract. Protein sources were used raw and after a cooking treatment. Results showed that the category of the ingredient (meats, insects, or grain legumes) exerts a stronger impact on enzymatic digestibility, fermentation patterns, and bacterial metabolites such as short-chain fatty acids (SCFA) and hydrogen sulfide (H2S) than the cooking treatment. The digestibility and the fermentation characteristics of insects were more affected by the cooking procedure than the other categories. Per gram of consumed food, ingredients from animal origin, namely, meats and insects, were associated with fewer fermentation end-products (gas, H2S, SCFA) than ingredients from plant origin, which is related to their higher small intestinal digestibility.

Insights into Abundant Rumen Ureolytic Bacterial Community Using Rumen Simulation System.

Urea, a non-protein nitrogen for dairy cows, is rapidly hydrolyzed to ammonia by urease produced by ureolytic bacteria in the rumen, and the ammonia is used as nitrogen for rumen bacterial growth. However, there is limited knowledge with regard to the ureolytic bacteria community in the rumen. To explore the ruminal ureolytic bacterial community, urea, or acetohydroxamic acid (AHA, an inhibitor of urea hydrolysis) were supplemented into the rumen simulation systems. The bacterial 16S rRNA genes were sequenced by Miseq high-throughput sequencing and used to reveal the ureoltyic bacteria by comparing different treatments. The results revealed that urea supplementation significantly increased the ammonia concentration, and AHA addition inhibited urea hydrolysis. Urea supplementation significantly increased the richness of bacterial community and the proportion of ureC genes. The composition of bacterial community following urea or AHA supplementation showed no significant difference compared to the groups without supplementation. The abundance of Bacillus and unclassified Succinivibrionaceae increased significantly following urea supplementation. Pseudomonas, Haemophilus, Neisseria, Streptococcus, and Actinomyces exhibited a positive response to urea supplementation and a negative response to AHA addition. Results retrieved from the NCBI protein database and publications confirmed that the representative bacteria in these genera mentioned above had urease genes or urease activities. Therefore, the rumen ureolytic bacteria were abundant in the genera of Pseudomonas, Haemophilus, Neisseria, Streptococcus, Actinomyces, Bacillus, and unclassified Succinivibrionaceae. Insights into abundant rumen ureolytic bacteria provide the regulation targets to mitigate urea hydrolysis and increase efficiency of urea nitrogen utilization in ruminants.