Ruminal Fistula Research Articles

Disturbances of Ruminal Microbiota and Liver Inflammation, Mediated by LPS and Histamine, in Dairy Cows Fed a High-Concentrate Diet.

The ecosystem of ruminal microbiota profoundly affects the health and milk production of dairy cows. High-concentrate diets are widely used in dairy farms and evoke a series of metabolic disorders. Several studies have reported the effects of high-concentrate diets on the ruminal microbiome, while the effect of changes in ruminal microbial flora, induced by high-concentrate diet feeding, on the liver of dairy cows has not been studied before. In this study, 12 mid-lactating Holstein Friesian cows (weight of 455 ± 28 kg; parities of 2.5 ± 0.5; starting milk yield of 31.59 ± 3.2 kg/d; DMI of 21.7 ± 1.1 kg/d; and a DIM at the start of the experiment of 135 ± 28 d) were fitted with ruminal fistulas, as well as with portal and hepatic vein catheters. All cows were randomly divided into 2 groups; then, they fed with low-concentrate diets (LC, concentrate: forage = 40:60) and high-concentrate diets (HC, concentrate: forage = 60:40) for 18 weeks. The forage sources were corn silage and alfalfa hay. After the cows of two groups were euthanized over two consecutive days, ruminal microbiota; the concentration of LPS in the rumen content; cecum content; the levels of blood and histamine in rumen fluid, blood, and the liver; the histopathological status of the rumen and cecum; and the inflammatory response of the liver were assessed in dairy cows under conditions of subacute ruminal acidosis (SARA). These conditions were caused by high-concentrate diet feeding. All data were analyzed using the independent t-test in SPSS. The results showed that high-concentrate diet feeding increased the concentration of LPS and histamine in the rumen and plasma of veins (p < 0.05). The abundance of Bacteroidetes at the phylum level, and of both Bacteroidetes and Saccharibacteria at the genus level, was decreased, while the abundance of Firmicutes at the phylum level and Oscillibacter at the genus level was increased by high-concentrate diet feeding. The decreased pH values of ruminal contents (LC = 6.02, HC = 5.90, p < 0.05) and the increased level of LPS in the rumen (LC = 4.921 × 105, HC = 7.855 × 105 EU/mL, p < 0.05) and cecum (LC = 11.960 × 105, HC = 13.115 × 105 EU/mL, p < 0.01) induced the histopathological destruction of the rumen and cecum, combined with the increased mRNA expression of IL-1β (p < 0.05). The histamine receptor H1R and the NF-κB signaling pathway were activated in the liver samples taken from the HC group. In conclusion, the elevated concentrations of LPS and histamine in the gut may be related to changes in the ruminal microbiota. LPS and histamine induced the inflammatory response in the ruminal epithelium, cecum epithelium, and liver. However, the cause-effect mechanism needs to be proved in future research. Our study offers a novel therapeutic strategy by manipulating ruminal microbiota and metabolism to decrease LPS and histamine release and to improve the health of dairy cows.

Supplementation of 5,6-Dimethylbenzimidazole and Cobalt in High-Concentrate Diet Improves the Ruminal Vitamin B12 Synthesis and Fermentation of Sheep

The objective of this experiment was to investigate the effects of 5,6-dimethylbenzimidazole (5,6-DMB) and cobalt (Co) on the ruminal vitamin B12 synthesis and fermentation parameters of sheep under high concentrate conditions. Twenty-four Kazakh rams (body weight = 39.23 ± 2.61 kg and 8 months old) fitted with permanent ruminal fistulas were randomly divided into four groups with six rams in each group. The control (CON) group was fed a basal ration with a concentrate-to-roughage ratio of 70:30, and the experimental groups (T60, T75 and T90) were fed a basal diet with 60 mg 5,6-DMB+ 0.25 mg Co, 75 mg 5,6-DMB+ 0.5 mg Co and 90 mg 5,6-DMB+ 0.75 mg Co supplied to each kilogram of basal diet, respectively. The experiment lasted for 26 days, with the first 14 days being an adaptation period to allow the sheep to adapt to the diet type and environment, and the second 12 days being a sample period. On 0, 7 and 12 d of the sample period, all sheep were weighed before the morning feed. Rumen fluid samples were collected from all sheep on the last 4 days of the sample period. The results showed that the ruminal vitamin B12 content was higher in trial groups than that of the CON group at 3 h after feeding (p &lt; 0.05). Rumen pH was higher in trial groups than in the CON group at 1, 3 and 5 h after feeding (p &lt; 0.05). The concentration of ruminal ammonia-N was significantly increased in trial groups when compared to the CON group (p &lt; 0.05) at 1 and 3 h after feeding. At 1 and 3 h after feeding, the concentration of propionate in trial groups was higher than that in the CON group (p &lt; 0.05). The rumen microbial protein content reached the highest value at 3 h after feeding, and all trial groups were higher than the CON group (p &lt; 0.05). Thus, the supplementation of 5,6-DMB and Co increased vitamin B12, propionate, ammonia-N and microbial protein contents and pH in the rumen of sheep, and the best results were obtained by the amounts of 75 mg/kg 5,6-DMB and 0.5 mg/kg Co, respectively.

A high-concentrate diet induces colonic inflammation and barrier damage in Hu sheep

Long-term feeding of a high-concentrate diet can induce subacute ruminal acidosis (SARA) in ruminants, which further leads to systemic inflammatory response. However, few studies have examined the effects of feeding a high-concentrate diet on the hindgut of ruminants. The purpose of this study was to investigate the effects of a high-concentrate diet on the composition of gut microbiota in colonic contents, inflammatory response, and barrier damage in the colon tissue of ruminants. A total of 12 healthy multiparous lactating Hu sheep were randomly allotted into the following 2 groups: a high-concentrate (HC) group (concentrate:forage = 7:3) and a low-concentrate (LC) group (concentrate:forage = 3:7). All sheep were fitted with ruminal fistulas. The formal feeding experiment lasted for 8 wk. After the feeding experiment, rumen fluid, portal vein blood, hepatic vein blood, colonic contents, and colon tissue samples were collected. The results showed that feeding the HC diet induced SARA in Hu sheep and significantly reduced pH in the colonic contents. The abundances of Firmicutes, Verrucomicrobiota, and Actinobacteriota decreased significantly, whereas those of Bacteroidota, Spirochaetota, and Fibrobacterota significantly increased in colonic contents. At the genus level, the relative abundances of 29 genera were significantly altered depending on the different type of diets. Analysis of the 10 bacterial genera with high relative abundance revealed that feeding the HC diet significantly reduced the abundance of UCG-005, Christensenellaceae R-7 group, UCG-010-norank, Monoglobus, [Eubacterium] coprostanoligenes group_norank, and Alistipes, whereas the abundances of Rikenellaceae RC9 gut group, Treponema, Bacteroides, and Prevotella increased. Compared with the LC group, feeding the HC diet significantly increased the concentration of LPS in rumen fluid, portal vein blood, hepatic vein blood, and colonic contents, and significantly upregulated the mRNA expression levels of proinflammatory cytokines in colon tissue, including TNF-α, IL-1β, IL-6, and IL-8, indicating the occurrence of inflammatory response in the colon tissue. In addition, the structure of colonic epithelial cells was loose, the intercellular space became larger, epithelial cells were exfoliated, and the mRNA and protein abundances of ZO-1, occludin, claudin-1, claudin-3, and claudin-4 were significantly decreased in the HC group, which was consistent with the results of immunohistochemistry. Furthermore, feeding the HC diet increased the ratios of DNA methylation and chromatin compaction in the promoter regions of occludin and claudin-1, which in turn inhibited their transcriptional expression. Therefore, the present study demonstrated that feeding an HC diet induced SARA in Hu sheep, altered the composition and structure of the microbial community in the colonic contents, induced an inflammatory response, and disrupted the intestinal mucosal barrier in the colonic tissue.

Effect of forage type on swallowed bolus mass and a method for counting swallows in dairy cattle

Dry matter intake (DMI) is a primary determinant of milk production in grazing dairy cows and an ability to measure the DMI of individual cows would allow herd managers to formulate supplementary rations that consider the amount of nutrients ingested from grass. The 2 related aims of this experiment were to define the mean number of swallowed boli and mass of the swallowed boli in Holstein-Friesian dairy cattle offered a variety of forages commonly fed in the dairy industry of southeastern Australia, and to evaluate 2 indirect methods for counting the number of swallows. Twelve ruminally-fistulated, lactating Holstein-Friesian cows were randomly assigned to 3 replicated 4 × 4 Latin square designs and offered 4 forages: fresh chicory (FC), fresh perennial ryegrass (RP), alfalfa hay (AH), and perennial ryegrass silage (RS). The experiment was conducted over 28 d with each of 4 periods consisting of 7 d with 3 d of measurement. Forage diets were offered to individual cows following the partial evacuation of the rumen. The first 20 min after forage was offered constituted the measurement period, during which all swallowed boli were manually captured by samplers who placed their hand through the ruminal fistula and over the cardia entrance of the rumen of each cow. Concurrently, microphones and video cameras were used for the indirect measurement of swallows. The average swallowed bolus mass overall was 17.4 g dry matter (DM) per bolus with the lowest mass observed in cows offered FC (8.9 g DM/bolus), followed by RP (14.9 g DM/bolus), compared with cows offered AH (23.6 g DM/bolus) and RS (22.3 g DM/bolus). The swallowing rate was greater in cows offered FC (78 swallows/20 min) than in cows offered RP, AH, and RS (62.3 swallows/20 min). The audio recording method showed greater concordance (Lin's concordance correlation coefficient = 0.90) with the physical capturing of the boli through the rumen, than the video recording method did (Lin's concordance correlation coefficient = 0.54). It is concluded that the mass of the swallowed boli is related to forage type and that using a microphone attached to the cow's forehead can provide an accurate measure of the number of swallows when verified against the actual number of swallows counted by manual interception of the boli at the rumen cardia.

Silage additives improve fermentation quality, aerobic stability and rumen degradation in mixed silage composed of amaranth and corn straw.

The objective of this research was to investigate effects of different additives on the fermentation quality, aerobic stability and rumen degradation of mixed silage composed of amaranth and corn straw. The mixture ratio of amaranth to corn straw was 78%: 22%. Three additives were selected in this study and five groups were as follows: control group (CON, without additive), lactic acid bacteria group (LAB, 5 mg/kg, Lactobacillus plantarum ≥ 1.6×1010 CFU/g and L. buchneri ≥ 4.0×109 CFU/g), glucose group (GLU, 30 g/kg), cellulase group (CEL, 2 mg/kg) and lactic acid bacteria, glucose and cellulase group (LGC, added at the same levels as in individual group). The period of ensiling was 60 days. Fermentation quality, chemical composition and aerobic stability of mixed silage were analyzed. Four cows with permanent ruminal fistula were selected as experimental animals. Nylon bag technique was used to study rumen degradation characteristic of dry matter (DM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) of mixed silage. Compared with CON group, the addition of different silage additives could improve mixed silage quality of amaranth and corn straw to some extent. Combining three additives significantly increased (P < 0.05) the DM, CP and lactic acid contents, whereas decreased (P < 0.05) the ADF and NDF contents as well as pH and ammonia nitrogen/total nitrogen. Moreover, the aerobic stability and rumen degradation of DM, CP and NDF were significantly improved (P < 0.05) in LGC group when compared to other groups. In conclusion, the combined addition of lactic acid bacteria, glucose and cellulase increased DM, CP and lactic acid contents as well as lactic acid bacteria count, decreased NDF and ADF contents and aerobic bacteria and mold counts, improved aerobic stability and rumen degradation of amaranth and corn straw mixed silage.

Comparative Study of the Nutritional Value and Degradation Characteristics of Amaranth Hay in the Rumen of Goats at Different Growth Stages.

The objective of this study was to investigate the rumen degradation characteristics of grain amaranth hay (Amaranthus hypochondriacus) at four different growth stages. The aim of this study was to evaluate the nutritional value of grain amaranth hay at different growth stages by chemical composition, in vivo digestibility, and in situ degradability. Three Boer goats with permanent ruminal fistulas were selected in this study. Amaranthus hay at four different growth stages (squaring stage (SS), initial bloom stage (IS), full-bloom stage (FS) and mature stage (MS)) was crushed and placed into nylon bags. Each sample was set up with three replicates, and two parallel samples were set up in fistulas at each time point. The rumen degradation rates of dry matter (DM), crude protein (CP), neutral detergent fibre (NDF) and acid detergent fibre (ADF) were determined at 0, 6, 12, 24, 36, 48 and 72 h. The results were as follows: (1) The concentration of CP in SS was the highest and was significantly higher than that in other stages (p < 0.05), whereas the contents of NDF and ADF gradually increased with the extension of the growing period and reached a maximum in MS; (2)The degradation of CP in the rumen at 72 h of SS and IS was more than 80%. Compared with other stages, the effective degradability of CP was highest in SS (p < 0.05) and reached 87.05% at 72 h, and the degradation rate was the lowest in MS; and (3) The effective degradability of NDF in IS was the highest (p < 0.05) and reached 69.326% at 72 h. The effective degradability of ADF in MS was the highest (p < 0.05) and reached 65.728% at 72 h. The effective degradability of DM and CP in SS was the highest. In conclusion, among the four stages, IS was superior in chemical composition and rumen degradability characteristics.

Connection between Circadian Rhythm and Rumen Digestibility of Concentrate and Roughage in Sheep

This study investigated the effects of circadian rhythm on rumen nutrient digestibility using the nylon bag method and the digestibility characteristics of dietary nutrients during the daytime and at night. It also presents modification suggestions for evaluating the nutritional value of raw feed materials. The rumen nutrient digestibility was measured and investigated for correlation with circadian rhythm using the nylon bag method. We established a sheep-feeding program to determine the differences in rumen nutrient digestibility between the daytime and nighttime. The digestibility of 18 standard feed samples was investigated in 6 Hu sheep with ruminal fistulas (body weight: 33.59 ± 0.39 kg, 12 months old). Samples were incubated in the rumen for three periods: daytime (12 h, 6:00 AM to 6:00 PM), night (12 h, 6:00 PM to 6:00 AM), and the entire day (24 h, 6:00 AM to 6:00 AM). The activities of enzymes amylase, lipase and cellulase were higher during the day than at night. The rumen digestibility of organic matter was significantly higher during the daytime than at night compared to the digestibilities of dry matter (DM), crude protein (CP), and ether extract (EE). Among them, the rumen digestibility of DM and CP was higher, but the digestibility of EE was lower during the daytime than at night in cereal feed fed sheep. The rumen digestibility of DM was higher in roughage-fed sheep but lower in rice straw- and corn silage-fed sheep during the day than at night. Circadian rhythms correlate strongly with the activities of major digestive enzymes in the rumen and the rumen digestibility of nutrients. Moreover, the nylon bag method needs to be modified and improved to ascertain the appropriate time for placing the nylon bag in the rumen and thus establish a unified procedure.

Histamine promotes adhesion of neutrophils by inhibition of autophagy in dairy cows with subacute ruminal acidosis.

Subacute ruminal acidosis (SARA), a common digestive disease in dairy cows, is accompanied by systemic inflammation and high concentrations of histamine in blood. Histamine-induced neutrophil adhesion may play an important role in the systemic inflammation experienced by cows during SARA. Autophagy, an intracellular degradation system, regulates recycling of membrane-associated integrin and may be involved in histamine-induced adhesion of bovine neutrophils. In the present study, 20 multiparous mid-lactation cows (average body weight 486 ± 24 kg) fitted with ruminal fistula were assigned to a control group (n = 10) or a SARA group (n = 10). We induced SARA by feeding different combinations of wheat-barley pellets and chopped alfalfa hay for 8 wk; SARA was defined as a ruminal pH <5.6 for at least 3 h/d. Blood samples were collected in wk 8. Compared with controls, SARA cows had greater serum concentrations of tumor necrosis factor-α, IL-6, IL-1β, lipopolysaccharide (LPS)-binding protein, haptoglobin, and serum amyloid A. Serum concentrations of these proinflammatory factors had strong positive correlations with the concentration of serum histamine and LPS. In ex vivo adhesion experiments, the number of adherent neutrophils was greater in the SARA group. Additionally, membrane protein abundance of adhesion molecules such as integrin α-L precursor (CD11a) and integrin α-M precursor (CD11b) was greater in neutrophils of the SARA group, confirming enhanced adhesion ability. Neutrophils of SARA cows had greater number of autophagosomes, greater protein abundance of autophagy substrate sequestosome-1 (p62), and higher ratio of microtubule associated proteins 1A/1B light chain 3 (LC3)-II to LC3-I, indicating congestion during the late phase of autophagy flux. For in vitro experiments, neutrophils isolated from control cows were incubated with 0.4 endotoxin units (EU)/mL LPS or 7 μM histamine for 0, 1, 2, and 4 h, respectively. We detected linear and quadratic effects for the number of adherent neutrophils after histamine treatment with a peak response at 2 h, whereas no significant effect was detected after LPS treatment. Membrane protein abundance of CD11a and CD11b was greater after histamine treatment, suggesting that it may have an inhibitory effect on the degradation of adhesion molecules. The greater abundance of p62, higher ratio of LC3-II to LC3-I, and increased co-localization between CD11b and LC3 after histamine treatment indicated that recycling of adhesion molecules and autophagy flux were blocked. These effects were not aggravated further in the presence of chloroquine, a specific inhibitor of the late phase of autophagy flux. Overall, our results revealed that histamine increases adhesion of neutrophils by inhibiting autophagy in dairy cows with SARA.

Near-infrared calibration models for estimating volatile fatty acids and methane production from in vitro rumen fermentation of different total mixed rations

Volatile fatty acids (VFA) and methane (CH4) are the major products of rumen fermentation. The VFA are considered an energy source for the animal and rumen microbiota, and CH4 (which is released by eructation) is considered an energy loss. Quantification of these fermentation products is fundamental for the evaluation of feeds and diets, and provides important information regarding the use of nutrients by ruminants. Near-infrared (NIR) spectroscopy is increasingly used for the evaluation of animal feeds because it is rapid, nondestructive, noninvasive, and inexpensive; does not require reagents; and the results are reproducible. The aim of this study was to develop NIR calibration models for estimating the production of VFA (acetic, propionic, butyric, valeric, isovaleric, and isobutyric acids), total gas, and CH4 using in vitro gas production tests with buffered rumen inoculum throughout fermentation. Fifty-four total mixed rations (TMRs) were examined, and rumen fluid was manually collected from 2 dry Holstein dairy cows that had ruminal fistulas and were fed at maintenance energy levels. Then, 30 mL of buffered rumen fluid was incubated in bottles with ~220 mg of TMR. The total gas, VFA, and CH4 were measured after 2, 5, 9, 24, 30, 48, and 72 h of rumen incubation for each TMR. The VFA were measured on 32 randomly selected TMR. In particular, 7 bottles were used for each TMR, one for each incubation time. Methane was measured in the headspace and VFA were measured in the buffered rumen fluid. The bottles were considered experimental units for calibration purposes. The production of CH4 was quantified from the bottle headspaces by gas chromatography, and total gas production was measured using a pressure transducer at each incubation time. Two aliquots of the fermented liquids were sampled by opening the bottles at each incubation time, and (1) the concentrations of VFA were determined by gas chromatography or (2) spectra were obtained from Fourier-transform NIR spectroscopy. The data were randomly divided into calibration and validation data sets. The average concentrations of acetic acid (45.30 ± 11.92 and 43.86 ± 11.93 mmol/L), propionic acid (14.97 ± 6.08 and 14.38 ± 6.56 mmol/L), butyric acid (8.47 ± 3.47 and 8.65 ± 3.79 mmol/L), total gas (111.34 ± 81.90 and 116.46 ± 82.44 mL/g of organic matter), and CH4 (9.65 ± 9.45 and 10.35 ± 9.33 mmol/L) were similar in the 2 data sets. The best calibration models were retained based on the coefficient of determination (R2) and the ratio of prediction to deviation (RPD). The R2 values for prediction of VFA ranged from 0.69 (RPD = 3.28) for valeric acid to 0.94 (RPD = 4.20) for acetic acid. The models also provided good predictions of CH4 (R2 = 0.89, RPD = 3.05) and cumulative gas production (R2 = 0.91, RPD = 3.30). The models described here precisely and accurately estimated the production of CH4 and VFA during in vitro rumen fermentation tests. Validations at additional laboratories may provide more robust calibrations.

PSVI-1 Evaluation of the synergistic effect of iron and an aqueous extract of oak bark on rumen microbiome of cattle

Abstract The search for new substances, alternatives to feed antibiotics in animal husbandry, is relevant in connection with the development of antibiotic resistance of pathogenic microorganisms. Substances of plant and mineral origin including nanoforms are of particular interest. The experiment studied the synergistic effect of iron (Fe; d = 80 nm) and an aqueous extract of oak bark (Ex; 1:10) on the microbiome of ruminal fluid of bulls (age 14 months, n = 5). Group I - control; Group II - Ex (0.6 ml / kg / lm); Group III - Fe (0.1 mg/kg/FM) with Ex in the same dosage. The collection of ruminal fluid was performed through a chronic ruminal fistula. Metagenomic analysis of cicatricial fluid was performed on a MiSeq device (Illumina, USA). Analysis of the data showed that the Bacteria domain is more influenced by Ex, there is a decrease by 27.0% (P ≤ 0.05), while in group III by 3.0% relative to the control. At the phylum level, Bacteroidetes and Firmicutes had the greatest importance, in group I their ratio was 39.3 and 54.7%, in group II - 37.7 and 56.4%, in group III - 46.1 and 47.0% of the total the number of bacteria. The use of Ex increases the amount of Bacilli by 14.7 and 13.4 (P ≤ 0.05) % relative to groups I and III. The introduction of the composition Ex and Fe increases Lachnospiraceae - 15.0% and Ruminococcaceae - 8.1% of the total number of microorganisms and reduces Streptococcus relative to other groups by 1.3 and 14.9% (P ≤ 0.05). Thus, the combined use of Ex and Fe affects the numerical value of microorganisms, as well as their ratio in the rumen through inhibition. At the same time, the data obtained require further research, including in vivo experiments. This research was performed with financial support from the RSF (20-16-00088).

Combined effects of 3-nitrooxypropanol and canola oil supplementation on methane emissions, rumen fermentation and biohydrogenation, and total tract digestibility in beef cattle.

The individual and combined effects of 3-nitrooxypropanol (3-NOP) and canola oil (OIL) supplementation on enteric methane (CH4) and hydrogen (H2) emissions, rumen fermentation and biohydrogenation, and total tract nutrient digestibility were investigated in beef cattle. Eight beef heifers (mean body weight ± SD, 732 ± 43 kg) with ruminal fistulas were used in a replicated 4 × 4 Latin square with a 2 (with and without 3-NOP) × 2 (with and without OIL) arrangement of treatments and 28-d periods (13 d adaption and 15 d measurements). The four treatments were: control (no 3-NOP, no OIL), 3-NOP (200 mg/kg dry matter [DM]), OIL (50 g/kg DM), and 3-NOP (200 mg/kg DM) plus OIL (50 g/kg DM). Animals were fed restrictively (7.6 kg DM/d) a basal diet of 900 g/kg DM barley silage and 100 g/kg DM supplement. 3-NOP and OIL decreased (P < 0.01) CH4 yield (g/kg DM intake) by 31.6% and 27.4%, respectively, with no 3-NOP × OIL interaction (P = 0.85). Feeding 3-NOP plus OIL decreased CH4 yield by 51% compared with control. There was a 3-NOP × OIL interaction (P = 0.02) for H2 yield (g/kg DM intake); the increase in H2 yield (P < 0.01) due to 3-NOP was less when it was combined with OIL. There were 3-NOP × OIL interactions for molar percentages of acetate and propionate (P < 0.01); individually, 3-NOP and OIL decreased acetate and increased propionate percentages with no further effect when supplemented together. 3-NOP slightly increased crude protein (P = 0.02) and starch (P = 0.01) digestibilities, while OIL decreased the digestibilities of DM (P < 0.01) and neutral detergent fiber (P < 0.01) with no interactions (P = 0.15 and 0.10, respectively). 3-NOP and OIL increased (P = 0.04 and P < 0.01, respectively) saturated fatty acid concentration in rumen fluid, with no interaction effect. Interactions for ruminal trans-monounsaturated fatty acids (t-MUFA) concentration and percentage were observed (P = 0.02 and P < 0.01); 3-NOP had no effect on t-MUFA concentration and percentage, while OIL increased the concentration (P < 0.01) and percentage (P < 0.01) of t-MUFA but to a lesser extent when combined with 3-NOP. In conclusion, the CH4-mitigating effects of 3-NOP and OIL were independent and incremental. Supplementing ruminant diets with a combination of 3-NOP and OIL may help mitigate CH4 emissions, but the decrease in total tract digestibility due to OIL may decrease animal performance and needs further investigation.

PSXIV-23 Characterization of microbial communities associated with the rumen lining, digesta and rumen fluid from beef cattle consuming a high energy diet using 16S rRNA gene amplicon sequencing

Abstract The true etiology of liver abscesses is not well known. Therefore, the objective of this study was to characterize the microbial communities in the rumen lining, digesta, and rumen fluid from beef cattle consuming a high energy diet, using 16S rRNA gene amplicon sequencing. Twelve crossbred feedlot steers (450 ±10 kg; ~ 3.0 years of age) fitted with ruminal fistulas, consuming a high energy finishing diet (1.43 NEg, Mcal/kg DM) for 21 d were utilized in this experiment. Microbial DNA from three regions within the rumen [rumen lining (ventral/lateral), digesta (geometric center of the rumen), and rumen fluid] was extracted and the V4 region of the 16S rRNA gene was amplified and sequenced. Across all sample regions, bacterial sequences were classified into 34 phyla, 76 classes, 143 orders, and 254 families. Bacteroidetes and Firmicutes were the predominant phyla present across all samples. The relative abundance of Bacteroidetes detected in rumen fluid was lesser (P &amp;lt; 0.05) when compared to bacteria sampled from the rumen lining and digesta. In contrast, the relative abundance of Firmicutes were greater (P &amp;lt; 0.05) in rumen fluid and the rumen lining when compared to digesta samples. There are very few publications describing the complex community of the rumen microbiome. To our knowledge this is the first publication categorizing microbial populations in three distinct locations within the rumen using next generation sequencing in feedlot cattle.

Evaluation of Hair Characteristics and Animal Age on the Impact of Hair Cortisol Concentration in Feedlot Steers.

Hair cortisol is a novel biomarker of chronic stress. The objective of this study was to determine the effect of hair color and length, as well as, animal age on hair cortisol concentration in beef feedlot steers. Nineteen beef crossbred steers used for nutrition research and housed in a small feedlot setting were used for this study. Seven of the steers (680 ± 4.5 kg; ~9 years of age) were fitted with ruminal fistulas and duodenal cannulas. The other 12 steers (473 ± 3.1 kg; ~2.5 years of age) were fitted with only ruminal fistulas. Hair samples from each steer were collected throughout a period of 6 weeks from six different areas and analyzed for cortisol concentrations. One pre-determined area was shaved each week for 5 weeks (Weeks 1–5). During week 6, all five, previously shaved areas and an additional area was shaved to collect hair samples of various lengths. Hair length was recorded prior to the collection of each hair sample. Only data from the last week (Week 6) of collection were included in the analyses. Steers were categorized into one of three groups: old with black hair (OB, n = 3); old with white hair (OW, n = 3); young with black hair (YB, n = 12). Older steers exhibited greater hair cortisol concentrations than younger steers (P < 0.001). Hair cortisol concentration was not impacted by duration of growth (P = 0.33). Cortisol concentrations exhibited a weak, positive correlation with hair length (r = 0.33, P-value = 0.01). The average hair growth per week of beef steers in the winter months was calculated to be 0.90 mm. Further research should be performed to improve our understanding of the effect of hair characteristics, sampling methodologies and analysis techniques on hair cortisol concentrations.

Effects of oregano essential oil on the ruminal pH and microbial population of sheep.

Oregano essential oil (OEO), which has antimicrobial properties, may be used for altering the ruminal pH and microbial populations of sheep, as observed by the altered volatile fatty acid patterns. To further elucidate the effects of OEO on ruminal pH and microbial populations of sheep, 3 German merino sheep × local sheep crossbred rams with permanent ruminal fistulas were randomly assigned to a 3 × 3 Latin square design with 12-d periods. The treatments were as follows: control (CON); OEO4: OEO supplied at 4 g•d-1; and OEO7: OEO supplied at 7 g•d-1. Starting on day 11, rumen fluid was collected at 0 h, and at 4, 8, 12, 24 and 48 h after supplying OEO, and then pH values of rumen fluid were immediately measured. The abundance of microbial populations was determined by using qPCR. The ruminal pH values were similar among the sheep from all treatments. The abundance of ruminal fungi was higher for the sheep supplied OEO7 compared with the sheep supplied CON and OEO4, especially at 4 and 12 h. The abundance of ruminal protozoa decreased with supplied OEO, indicating that OEO could inhibit the protozoa. The abundance of the total ruminal bacteria was similar for the sheep from all treatments, but R. flavefaciens, R. albus and F. succinogenes increased in the sheep supplied OEO4 compared with those in the sheep supplied CON, however, the sheep supplied OEO7 had higher abundances of R. flavefaciens than the sheep supplied CON. These results demonstrated that supplying OEO to sheep did not affect the ruminal pH but could shift the rumen microbial population to one with less protozoa. Supplying OEO can preferentially enhance the growth of certain rumen microbial populations, but the shifts were influenced by the supply rate. Therefore, supplying low amount (i.e. 4 g•d-1) of OEO could have positive effects on ruminal microbial populations, whereas supplying elevated doses of OEO could be detrimental to those same ruminal microbial populations.

A study on comparative feeding value of corn flakes according to temperature and retention time in the pressurized steam chamber.

This study aimed to investigate the effects of temperature and retention time of the pressurized steam chamber on the ruminal fermentation characteristics and nutrient degradability of corn flakes in three Korean native Hanwoo cows and three Holstein cows implanted with a ruminal fistula. Corn kernels were categorized into 13 groups based on the chamber temperature (range, 100°C–116°C) and retention time (range, 700–950 s). The pH value was lowest in T1 regardless of breed. Propionate concentration was the highest in T2 (p < 0.05). Total-volatile fatty acid (VFA) concentration was slightly but not significantly greater in T2 than in other conditions. Dry matter (p < 0.05), starch, and crude protein (p < 0.05) degradability were the highest in T1. At different incubation times and with different breeds, dry matter, starch, and crude protein degradability of corn flakes were the highest in T1. Thus, the present results indicate that the optimal temperature and retention time of the pressurized steam chamber should be 100°C–105°C and 700–720 s.

Técnicas quirúrgicas para ruminotomía con fistulación ruminal en cabras

El presente trabajo formó parte del proyecto de investigación Calidad Nutricional, degradabilidad ruminal y digestibilidad fecal de Moringa oleifera. Se fistularon caprinos machos criollos de 24 a 36 meses, peso promedio 35kg, evaluados clínicamente sin presentar síntomas patológicos. Previo a la cirugía las cabras fueron sometidas a ayuno de sólidos y líquidos, en el pre operatorio se utilizó Meloxican, Penicilina G Procaína, acepromacina y Ketamina, además se realizó una sedación regional con epidural craneal y bloqueos de los nervios paravertebrales izquierdo T13, L1, L2 con lidocaína al 2 %. La ruminotomía es la apertura del rumen donde se aplicaron dos técnicas de fistulación ruminal, en ambas se colocó al animal de cubito lateral izquierdo para incidir en la zona del ijar, superado el plano subcutáneo divulsionando las fibras de los músculos oblicuo abdominal externo, interno y el transverso abdominal, peritoneo para luego exponer el rumen e incidir en él y colocar la fistula ruminal fijándola con una sutura de jareta para introducirlo a la cavidad abdominal en ambas técnicas. En la primera, se unieron con hilos de sutura superficie del rumen, el peritoneo y los músculos mencionados para posterior cierre de la piel. En la segunda técnica solo se dejó que los músculos se replieguen a su posición inicial finalmente, se hizo acercamiento subcutáneo y sutura de piel con hilo nylon. A los 7 días postcirugía se determinó que los procesos de cicatrización y adherencia de los músculos y el rumen fueron similares en ambas técnicas.

Dose-response effect of crude extracts produced by actinobacteria on in vitro rumen fermentation

Actinobacteria have been researched as a source that produces crude extracts, which contain bioactive compounds able to act as antimicrobial agents. The present investigation evaluated the dose-response effect of two crude extracts, obtained at Caatinga rhizosphere (Caat) and Rhizophora mangle (AMC), on in vitro ruminal fermentation by: cumulative gas production, digestibility of dry (IVDMD) and organic matter (IVOMD), and short-chain fatty acids concentration (SCFA). Three multiparous Holstein dairy cows with ruminal fistula were used as the inoculum donors and fed a basal diet consisting of corn silage, soybean meal, urea, ground corn and mineral supplement. Ruminal fluid samples were incubated in glass bottles with 1 g of the dried and milled diet, a buffer solution, and the crude extracts evaluated in four doses (0.3, 0.6, 0.9 and 1.20 mg/10 mL inoculum) in a randomized block design, and the donators were considered as blocks with random effects. Additionally, negative controls were used. The results were expressed as average values based on triplicate analyses. Decreased cumulative gas production was observed according to linear dose response at 24, 48 and 72 h of incubation with the addition of Caat extract. The IVOMD showed a linear decrease at 72 h of incubation with dose Caat inclusion. Furthermore, the inclusion of Caat extract linearly reduced butyric and isovaleric acid concentrations, as well as acetate:propionate ratio. Finally, the Caat inclusion increased the propionic acid concentration in comparison to AMC extract. However, the inclusion of AMC extract did not affect any of the analyzed variables at the used doses. The Caat extract could be used as a modulator of in vitro ruminal fermentation, since it reduced acetate:propionate ratio and cumulative gas production.

Effects of adding mannan oligosaccharides on digestibility and metabolism of nutrients, ruminal fermentation parameters, immunity, and antioxidant capacity of sheep.

The purpose of this study was to investigate the effects of adding mannan-oligosaccharides (MOS) on the following parameters in sheep: digestibility and retention rate of nutrients, ruminal fermentation, immunity, and antioxidant capacity. Twelve healthy crossbred wethers (Suffolk ♂ × Small tail Han-yang ♀) with external ruminal fistula and similar body weights (28.04 ± 2.07 kg) were fed individually four treatments, three repeats of each treatment. The wethers diets were supplemental MOS at 0%, 1.2%, 1.6%, and 2.0%·kg-1 of basal diet (as fed basis). The experiment lasted 17 d, including 10 d of acclimation and 7 d of formal experimentation. The results showed that MOS did not influence the apparent digestibility and retention rate of nutrients, ruminal fermentation, and immunity or concentration of serum nitric oxide and activity of serum nitric oxide synthase (P ≥ 0.07). However, the apparent digestibility of neutral detergent fiber and acid detergent fiber at MOS supplementation rates of 1.6% and 2.0% both tended to be greater than the control group (P ≤ 0.103). There was also moderate evidence that MOS might increase the nitrogen retention rate (P = 0.082). MOS increased the antioxidant ability of sheep (P ≤ 0.018), especially at a dose of 1.6%: an increase in activity of total superoxide dismutase (P = 0.007), glutathione peroxidase (P = 0.018) and total antioxidant capacity (P < 0.001), and a decrease in concentration of malondialdehyde (P < 0.001) were found. The results indicated that in sheep MOS improved fiber digestion, N retention and some antioxidant abilities, but these effects may be too small to improve health and performance.

Productive Response of Dairy Cows Fed with Different Levels of Totally Mixed Ration and Pasture

Forty multiparous Holstein cows in early lactation (66 ± 19 days) were assigned to one of four treatments that consisted in different ratios of total mixed rations (TMR) and pasture at 100% TMR (T0), 75:25 (T1) 50:50 (T2) and 25:75 (T3) over 9 weeks in autumn-winter. Measures of rumen parameters and digestion were performed on 4 additional Holstein cows in late lactation (287 days postpartum) fitted with permanent ruminal fistulae and producing 22.6 (±5.4) kg milk in a 4 × 4 Latin Square design. In T1 to T3 cows were taken to the grazing plot once they finished the pre-planned TMR intake and pasture was offered at 2 times the expected forage DM intake. Milk was analyzed for chemical composition, milk fatty acid (FA) profile and antioxidant compounds. The cows were individually weighed and body condition score (BCS) was determined. After the morning milking, blood samples were taken every 2 weeks and plasma was analyzed for glucose, urea, non-esterified fatty acids (NEFA), insulin, growth hormone (GH) and insulin-like growth factor (IGF-I). Herbage mass in pre grazing strips of pasture averaged 2540 ± 343 kg DM/ha. As TMR intake increased, production variables increased linearly excepting milk fat (3.88 g/100g) and milk protein (3.43 g/100g) contents that were not affected. Milk yield (kg/cow/d) resulted maximal in T0 (34.2) and linearly decreased (p 3-N concentration resulted higher in treatments with higher proportion of pasture. Kinetic parameters of DM and NDF digestion from pasture or corn silage were not affected. Milk FA profile and milk antioxidant quality showed unfavorably changes as TMR increased by a decrease in rumenic and linolenic acids and content of antioxidant vitamins, without effect on the atherogenicity index. Results suggested a depressing effect of the pasture on total DM and energy intake probably explained by qualitative deficiencies chemical composition of the forage and/or factors that affect animal behavior that may induce losses in body condition of high yielding dairy cows.

The rumen microbial metaproteome as revealed by SDS-PAGE

BackgroundRuminal digestion is carried out by large numbers of bacteria, archaea, protozoa and fungi. Understanding the microbiota is important because ruminal fermentation dictates the efficiency of feed utilisation by the animal and is also responsible for major emissions of the greenhouse gas, methane. Recent metagenomic and metatranscriptomic studies have helped to elucidate many features of the composition and activity of the microbiota. The metaproteome provides complementary information to these other –omics technologies. The aim of this study was to explore the metaproteome of bovine and ovine ruminal digesta using 2D SDS-PAGE.ResultsDigesta samples were taken via ruminal fistulae and by gastric intubation, or at slaughter, and stored in glycerol at −80 °C. A protein extraction protocol was developed to maximise yield and representativeness of the protein content. The proteome of ruminal digesta taken from dairy cows fed a high concentrate diet was dominated by a few very highly expressed proteins, which were identified by LC-MS/MS to be structural proteins, such as actin and α- and β-tubulins, derived from ciliate protozoa. Removal of protozoa from digesta before extraction of proteins revealed the prokaryotic metaproteome, which was dominated by enzymes involved in glycolysis, such as glyceraldehyde-3-phosphate dehydrogenase, phosphoenolpyruvate carboxykinase, phosphoglycerate kinase and triosephosphate isomerase. The enzymes were predominantly from the Firmicutes and Bacteroidetes phyla. Enzymes from methanogenic archaea were also abundant, consistent with the importance of methane formation in the rumen. Gels from samples from dairy cows fed a high proportion of grass silage were consistently obscured by co-staining of humic compounds. Samples from beef cattle and fattening lambs receiving a predominantly concentrate diet produced clearer gels, but the pattern of spots was inconsistent between samples, making comparisons difficult.ConclusionThis work demonstrated for the first time that 2D-PAGE reveals key structural proteins and enzymes in the rumen microbial community, despite its high complexity, and that taxonomic information can be deduced from the analysis. However, technical issues associated with feed material contamination, which affects the reproducibility of electrophoresis of different samples, limits its value.