Effect of biochanin A on the rumen microbial community of Holstein steers consuming a high fiber diet and subjected to a subacute acidosis challenge.

Subacute rumen acidosis (SARA) occurs when highly fermentable carbohydrates are introduced into the diet, decreasing pH and disturbing the microbial ecology of the rumen. Rumen amylolytic bacteria rapidly catabolize starch, fermentation acids accumulate in the rumen and reduce environmental pH. Historically, antibiotics (e.g., monensin, MON) have been used in the prevention and treatment of SARA. Biochanin A (BCA), an isoflavone produced by red clover (Trifolium pratense), mitigates changes associated with starch fermentation ex vivo. The objective of the study was to determine the effect of BCA on amylolytic bacteria and rumen pH during a SARA challenge. Twelve rumen fistulated steers were assigned to 1 of 4 treatments: HF CON (high fiber control), SARA CON, MON (200 mg d-1), or BCA (6 g d-1). The basal diet consisted of corn silage and dried distiller's grains ad libitum. The study consisted of a 2-wk adaptation, a 1-wk HF period, and an 8-d SARA challenge (d 1-4: 40% corn; d 5-8: 70% cracked corn). Samples for pH and enumeration were taken on the last day of each period (4 h). Amylolytic, cellulolytic, and amino acid/peptide-fermenting bacteria (APB) were enumerated. Enumeration data were normalized by log transformation and data were analyzed by repeated measures ANOVA using the MIXED procedure of SAS. The SARA challenge increased total amylolytics and APB, but decreased pH, cellulolytics, and in situ DMD of hay (P < 0.05). BCA treatment counteracted the pH, microbiological, and fermentative changes associated with SARA challenge (P < 0.05). Similar results were also observed with MON (P < 0.05). These results indicate that BCA may be an effective alternative to antibiotics for mitigating SARA in cattle production systems.

Evaluating the technological properties of lactic acid bacteria in Wagyu cattle milk.

This paper reveals the technological properties of lactic acid bacteria isolated from raw milk (colostrum and mature milk) of Wagyu cattle raised in Okayama Prefecture, Japan. Isolates were identified based on their physiological and biochemical characteristics as well as 16S rDNA sequence analysis. Streptococcus lutetiensis and Lactobacillus plantarum showed high acid and diacetyl-acetoin production in milk after 24 h of incubation at 40 and 30°C, respectively. These strains are thought to have potential for use as starter cultures and adjunct cultures for fermented dairy products.

Effects of feeding a live yeast on rumen fermentation and fiber degradability of tropical and subtropical forages.

BACKGROUND: The effect of live yeast Saccharomyces cerevisiae strain CNCM I-1077 (SC) on the ruminal degradability of different forages commonly found in dairy diets in South America was evaluated. We also assessed if SC supplementation interacts with forage group to affect ruminal fiber degradability. Four non-lactating rumen-cannulated Holstein cows were randomly assigned to two treatment sequences: Control-SC-Control or SC-Control-SC, in a switchback design, with three 30-day periods. Cows in the SC treatment were supplied with 1 × 1010 colony-forming units of yeast daily via rumen cannula. In situ degradability of dry matter (DM) and neutral detergent fiber (aNDF) was measured in 15 forages collected in South America. Forages were assigned to one of three groups: corn silages; tropical grasses (sugarcane silages and tropical grass silages); and temperate grasses and alfalfa (oat silages, ryegrass silages, alfalfa silage, and alfalfa hay). RESULTS: Cows supplemented with SC had higher (P = 0.05) counts of yeasts and lower (P = 0.03) concentration of lactate in rumen fluid. There was no interaction between forage group and yeast supplementation (P > 0.10) on in situ degradability. The SC increased DM (by 4.6%) and aNDF degradation (by 10.3%) at 24 h of incubation (P < 0.05). Metabolomics revealed that a chemical entity (C17 H29 N6 O3 , m/z 365.2284 [M + H]+ ) from the family of lipids and related molecules was suppressed in the rumen fluid of cows supplemented with SC. CONCLUSION: The SC supplementation improved DM and aNDF degradability regardless of the forage group. © 2021 Society of Chemical Industry.

Microbial community structure of the bovine rumen as affected by feeding cashew nut shell liquid, a methane-inhibiting and propionate-enhancing agent.

The effect of cashew nut shell liquid (CNSL) feeding on bacterial and archaeal community of the bovine rumen was investigated by analyzing clone libraries targeting 16S rRNA genes, methyl-coenzyme reductase A-encoding genes (mcrA), and their respective transcripts. Rumen samples were collected from three non-lactating cows fed on a hay and concentrate diet with or without CNSL supplementation. DNA and complementary DNA (cDNA) libraries were generated for investigating rumen microbial communities. MiSeq analysis also was performed to understand more comprehensively the changes in the microbial community structures. Following CNSL supplementation, the number of operational taxonomical unit (OTU) and diversity indices of bacterial and archaeal community were decreased. Bacterial OTUs belonging to Proteobacteria, including Succinivibrio, occurred at a higher frequency with CNSL feeding, especially in cDNA libraries. The methanogenic archaeal community became dominated by Methanomicrobium. A bacterial community shift also was observed in the MiSeq data, indicating that CNSL increased the proportion of Succinivibrio and other genera known to be involved in propionate production. Methanogenic archaeal community shifts to increase Methanoplanus and to decrease Methanobrevibacter also were observed. Together, these results imply the occurrence of significant changes in rumen communities, not only for bacteria but also for methanogens, following CNSL feeding.

Milk production and composition efficiency as influenced by feeding Pennisetum purpureum cv. Mahasarakham with Tiliacora triandra, Diels pellet supplementation.

Ruminal fermentation efficiency has been shown to be closely related with milk production in dairy cows. This investigation aimed at the utilization of sweet grass and bamboo grass pellet supplementation on ruminal fermentation, feed utilization efficiency, milk quantity, and quality in lactating dairy cows. Four lactating Holstein Friesian crossbreds were randomly assigned in a 2 × 2 factorial arrangement in a 4 × 4 Latin square design to determine the effect of roughage sources and bamboo grass (Tiliacora triandra, Diels) pellet (BP) supplementation on voluntary feed intake, digestibility of nutrients, fermentation characteristics of the rumen, and milk quantity and quality. Sweet grass (SG) (Pennisetum purpureum cv. Mahasarakham) and rice straw (RS) were fed as roughage sources as the first factor, while the second factor was supplementation levels of BP (0 and 150 g/cow/day). The results revealed that SG (P < 0.01) and BP supplementation (P < 0.05) improved feed intake, digestibility of nutrients, especially roughage intake and digestibility of DM and NDF. Ruminal pH (P < 0.05), bacterial (P < 0.01), and fungal population (P < 0.01) were increased with SG feeding, enhancing the concentration of total VFAs (P < 0.01) and propionic acid (P < 0.01), while both SG and BP decreased methane production (P < 0.01). While milk yield (P < 0.01) and milk composition (P < 0.01), especially unsaturated fatty acids including those of conjugated linoleic acid (P < 0.001), were enhanced. Supplementation of BP containing bioactive compounds such as condensed tannins (CT) enhanced rumen bacterial population with increased total VFAs (P < 0.05) and propionic acid (P < 0.05) concentrations, while decreased methane production (P < 0.05). The findings of this study indicate that SG would be beneficial to improved rumen fermentation, feed utilization, and milk production of dairy cows, while bamboo grass pellet supplementation tended to additionally improve rumen fermentation and feed intake without negative effects on milk production.

Astragalus root extract improved average daily gain, immunity, antioxidant status and ruminal microbiota of early weaned yak calves.

BACKGROUND: Early weaning in yak calves is being attempted to improve yak reproduction rate. However, this has to be done with caution because of the high mortality rate of calves due to the lack of nutrients and the harsh environmental conditions. Twenty-four weaned male yak calves were used in a 60 day feeding trial in which astragalus root extract (ARE) was supplemented. They were assigned randomly to one of four dietary treatments (n = six per treatment) that differed in ARE level: 0 g kg-1 (control), ARE0 ; 20 g kg-1 , ARE20 ; 50 g kg-1 , ARE50; and 80 g kg-1 dry matter intake (DMI), ARE80 . RESULTS: Final bodyweight and average daily gain (ADG) were significantly higher and the DMI/ADG ratio was significantly lower in calves with ARE supplementation than control (ARE0 ) calves. Ruminal concentrations of acetate and propionate and serum concentration of superoxide dismutase in ARE80 calves were higher than in the other groups and serum concentration of insulin was higher in ARE80 calves than in ARE20 calves. Serum immunoglobulin G (IgG) and interleukin-2 (IL-2) concentrations in ARE-fed calves were higher than in controls. Serum tumor necrosis factor (TNF-α) concentration was higher in ARE50 and ARE80 groups than ARE0 calves and serum interleukin-6 (IL-6) concentration was higher in ARE80 than in ARE0 calves. Serum immunoglobulin A (IgA), IgG and immunoglobulin M (IgM) concentrations increased with age in ARE-fed calves. ARE supplementation increased the abundance of fiber degrading bacteria. CONCLUSION: ARE at a dosage of 5% to 8% DMI can be supplemented to early weaned yak calves to improve growth performance, antioxidant capacity and immunity. © 2020 Society of Chemical Industry.

Influence of feeding fresh colostrum from the dam or frozen colostrum from a pool on indicator gut microbes and the inflammatory response in neonatal calves.

The aim of this study was to evaluate the capacity of cells from colostrum to modulate the intestinal microbial colonization, the activity of the inflammatory response, and for their influence on the development of diarrheal disease in calves. Twenty calves were distributed into two groups: COL+ (n = 10) receiving fresh whole colostrum; COL- (n = 10) receiving pooled frozen colostrum, containing no viable cells. All assessments were made before colostrum intake (D0), the next day (D2), and weekly on the 7th (D7), 14th (D14), 21st (D21) and 28th (D28) day of age. Diarrhea was assessed using a fecal score, and the systemic inflammatory status was assessed using a combination of temperature, anemia, total serum iron level, total haptoglobin concentration and the need for systemic antimicrobial treatment. The number of indicator bacteria present in the fecal population was estimated using qPCR. However, COL- calves presented more frequent signs of systemic inflammatory response including, fever at D7 (P = 0.011); indicator haptoglobin levels on D7 and D14, and lower levels of iron on D7, D14. Anemia was detected more often in the COL- calves on D21 (P = 0.043) and D28 (P = 0.016). COL- calves had a 1.66 greater chance of having elevated haptoglobin and a 1.8 greater chance of needing treatment with antimicrobials than COL+. A lower number of DNA copies of Clostridium perfringens were detected in COL+ calves on D2 (P = 0.088) and D7 (P = 0.040). Similarly, a low number of DNA copies was observed for Escherichia coli and Lactobacillus spp. (P = 0.012) in the fecal samples of COL+ calves on D7.

Digestive disorders associated with the consumption of palm fiber (Elaeis guineensis) in feedlot cattle / Distúrbios digestivos associados ao consumo de fibra de dendê (Elaeis guineensis) em bovinos criados em regime de confinamento

Digestive disorders in cattle are associated with the breeding system and feed provided to the animals. Abomasal compaction is primarily related to the ingestion of forage with elevated levels of lignin, low quality, and difficult digestibility. In addition, the excess of fibrous food in the diet can lead to phytobezoars that may be responsible for intestinal obstruction disorders. This study aimed to describe pathological and clinical aspects of an outbreak of digestive disorders associated with the consumption of palm fiber (Elaeis guineensis). The outbreak struck a herd of 499 animals raised in a feedlot system after a change in diet that included an increase in the amount of palm fiber. Forty (8.01%) animals showed clinical signs such as fattening and regurgitation during rumination, and 21 (4.2%) animals died later. The cattle affected presented with apathy, emaciation, dehydration, distended abdomen, incomplete or absent ruminal movements, and congestive mucosa. Three animals were submitted to necropsy, and distended rumen and reticulum has a large amount of brownish liquid, long and tangled vegetable fibers with sand and stones. In two animals, the omasum had many rounded structures measuring approximately 5cm in diameter, made of vegetable fiber (phytobezoars). Abomasum of animals had similar material to the rumen, and one animal had compressed content. In two animals, dilatation was observed in the small intestine, and in the opening, the total obstruction of the lumen by phytobezoar was observed. During the follow-up of the slaughter of 76 cattle, 15 (19.7%) had phytobezoars of different sizes in the omasum and abomasum. The increased amount of oil palm fiber in animal feeding favored the occurrence of compression abomasum and intestinal obstruction phytobezoa, causing significant economic losses.(AU)

Alguns distúrbios digestivos em bovinos podem estar associados ao sistema de criação e alimentação dos animais. Entre estes estão à compactação de abomaso e a obstrução intestinal por fitobezoários, ambas relacionadas principalmente com a ingestão de alimentos com altos níveis de lignina e, consequentemente, de difícil digestibilidade. Neste trabalho são descritos os aspectos clínicos e patológicos de um surto de distúrbios digestivos em bovinos associados ao consumo de fibra de dendê (Elaeis guineensis). O surto acometeu um rebanho de 499 bovinos, criados em sistema de confinamento, após uma mudança na dieta que incluiu o aumento na quantidade de fibra de dendê. Após a mudança 40 animais (8,01%) apresentaram diarreia, distensão abdominal e regurgitação durante a ruminação e 21 animais (4,2%) morreram. Os bovinos examinados clinicamente apresentavam sinais de apatia, emagrecimento, desidratação, abdômen distendido, movimentos ruminais incompletos e ausentes, além de mucosas congestas. Três animais foram submetidos à necropsia e observou-se rúmen e reticulo distendidos e com grande quantidade de líquido acastanhado, fibras vegetais longas e emaranhadas e presença de areia e pedras. Em dois animais o omaso continha grande número de estruturas arredondadas medindo aproximadamente 5cm de diâmetro, constituídas de fibras vegetais (fitobezoários). No abomaso dos animais havia material semelhante ao do rúmen, sendo que um animal apresentou conteúdo compactado e um deles também tinha fitobezoários. Em dois animais foram observadas dilatação e obstrução total do lúmen do intestino delgado por fitobezoários. Durante o acompanhamento do abate de 76 bovinos, 15 (19,7%) apresentavam fitobezoários de diferentes tamanhos no abomaso e omaso. O aumento da quantidade de fibra de dendê na alimentação dos animais favoreceu a ocorrência de casos de compactação de abomaso e obstrução intestinal por fitobezoários, ocasionando perdas econômicas significativas.(AU)

Analysis of the developing gut microbiota in young dairy calves-impact of colostrum microbiota and gut disturbances.

The aim of this study was to characterize the colostrum and fecal microbiota in calves and to investigate whether fecal microbiota composition was related to colostrum microbiota or factors associated with calf health. Colostrum samples were collected in buckets after hand milking of 76 calving cows from 38 smallholder dairy farms. Fecal samples were taken directly from the rectum of 76 calves at birth and at 14 days age. The bacterial community structure in colostrum and feces was analyzed by terminal restriction fragment length polymorphism for all samples, and the microbial composition was determined by 16S rRNA gene amplicon sequencing for a subset of the samples (8 colostrum, 40 fecal samples). There was a significant difference in fecal microbiota composition between day 0 and day 14 samples, but no associations between the microbiota and average daily gain, birth weight, or transfer of passive immunity. At 14 days of age, Faecalibacterium and Butyricicoccus were prevalent in higher relative abundances in the gut of healthy calves compared to calves with diarrhea that had been treated with antimicrobials. Colostrum showed great variation in composition of microbiota but no association to fecal microbiota. This study provides the first insights into the composition of colostrum and fecal microbiota of young dairy calves in southern Vietnam and can form the basis for future more detailed studies.

Rumen metabolism, omasal flow of nutrients, and microbial dynamics in lactating dairy cows fed fresh perennial ryegrass (Lolium perenne L.) not supplemented or supplemented with rolled barley grain.

The objective of this study was to evaluate the effect of rolled barley grain (RB) supplementation on rumen metabolism, omasal flow of nutrients, and microbial dynamics in lactating dairy cows fed fresh perennial ryegrass (Lolium perenne L.; PRG)-based diets. Ten ruminally cannulated Holstein cows averaging (mean ± standard deviation) 49 ± 23 d in milk and 513 ± 36 kg of body weight were assigned to 1 of 2 treatments in a switchback design. The treatment diets were PRG only (G) or PRG plus 3.5 kg of dry matter RB (G+RB). The study consisted of three 29-d periods where each period consisted of 21 d of diet adaptation and 8 d of data and sample collection. A double marker system was used to quantify nutrient flow entering the omasal canal along with labeled 15N-ammonium sulfate to measure bacterial, protozoal, and nonmicrobial N flow. Rumen evacuation techniques were used to determine nutrient and microbial pool size, allowing the calculation of fractional rates of digestion and microbial growth. There was no difference in daily milk yield or energy-corrected milk yield between treatments. Milk fat concentration and milk urea N decreased, whereas milk protein concentration increased in cows fed the G+RB diet. During the omasal sampling phase, dry matter intake was higher in cows fed the G+RB diet. Ruminal and total-tract neutral detergent fiber digestibility was lower in G+RB cows; however, no difference was observed in reticulorumen pH. The rumen pool size of fermentable carbohydrate was increased in cows fed the G+RB diet; however, the fractional rate of digestion was decreased. Flow of nonammonia N and bacterial N at the omasal canal increased in cows fed the G+RB diet compared with the G diet. Protozoa N flow was not different between diets; however, protozoa appeared to supply a much larger amount of microbial N and exhibited shorter generation time than previously considered. Feed N ruminal digestibility, corrected for microbial contribution, was similar for both treatments (88.4 and 89.0% for G and G+RB, respectively). In conclusion, RB supplementation did not benefit overall animal performance; however, it reduced ruminal neutral detergent fiber digestibility and increased bacterial N flow. The results demonstrate the large dependence of cows consuming PRG-based diets on microbial N as the main source of nonammonia N supply. Additional quantitative research is required to further describe the supply of nutrients and microbial dynamics in cows consuming PRG-based diets in an effort to determine most limiting nutrients.

Copper sulfate and sodium selenite lipid-microencapsulation modifies ruminal microbial fermentation in a dual-flow continuous-culture system.

Undesirable interactions between trace mineral elements and ruminal contents may occur during digestion when mineral salts are supplemented. Antimicrobial effects of copper sulfate (CuSO4) may affect ruminal digestibility of nutrients when fed as a source of copper (Cu), while sodium selenite (Na2SeO3) may be reduced in the rumen to less available forms of selenium (Se). Our objective was to evaluate if protection of CuSO4 and Na2SeO3 by lipid-microencapsulation would induce changes on ruminal microbial fermentation. We used 8 fermentors in a dual-flow continuous-culture system in a 4 × 4 duplicated Latin square with a 2 × 2 factorial arrangement of treatments. Factors were CuSO4 protection (unprotected and protected by lipid-microencapsulation) and Na2SeO3 protection (unprotected and protected by lipid-microencapsulation). Treatments consisted of supplementation with 15 mg/kg of Cu and 0.3 mg/kg of Se from either unprotected or protected (lipid-microencapsulated) sources, as follows: (1) Control (unprotected CuSO4 + unprotected Na2SeO3); (2) Cu-P (protected CuSO4 + unprotected Na2SeO3); (3) Se-P (unprotected CuSO4 + protected Na2SeO3); (4) (Cu+Se)-P (protected CuSO4 + protected Na2SeO3). All diets had the same nutrient composition and fermentors were fed 106 g of dry matter/d. Each experimental period was 10 d (7 d of adaptation and 3 d for sample collections). Daily pooled samples of effluents were analyzed for pH, NH3-N, nutrient digestibility, and flows (g/d) of total N, NH3-N, nonammonia N (NAN), bacterial N, dietary N, and bacterial efficiency. Kinetics of volatile fatty acids was analyzed in samples collected daily at 0, 1, 2, 4, 6, and 8 h after feeding. Main effects of Cu protection, Se protection, and their interaction were tested for all response variables. Kinetics data were analyzed as repeated measures. Protection of Cu decreased acetate molar proportion, increased butyrate proportion, and tended to decrease acetate:propionate ratio in samples of kinetics, but did not modify nutrient digestibility. Protection of Se tended to decrease NH3-N concentration, NH3-N flow, and CP digestibility; and to increase flows of nonammonia N and dietary N. Our results indicate that protection of CuSO4 may increase butyrate concentration at expenses of acetate, while protection of Na2SeO3 tended to reduce ruminal degradation of N. Further research is needed to determine the effects of lipid-microencapsulation on intestinal absorption, tissue distribution of Cu and Se, and animal performance.

The effects of feeding high or low milk levels in early life on growth performance, fecal microbial count and metabolic and inflammatory status of Holstein female calves.

Gut microbial colonization and immune response may be affected by milk feeding method. The objective of this study was to determine the effects of feeding high or low volumes of milk on fecal bacterial count, inflammatory response, blood metabolites and growth performance of Holstein female calves. Colostrum-fed calves (n = 48) were randomly assigned to either high milk (HM; n = 24) or low milk (LM; n = 24) feeding groups. Low milk-fed calves were fed pasteurized whole milk at 10% of BW until weaning. In HM group, milk was offered to calves at 20% of BW for the first 3 weeks of life. Then, milk allowance was decreased gradually to reach 10% of BW on day 26 and remained constant until weaning on day 51. Calves were allowed free access to water and starter throughout the experiment. Body weight was measured weekly, and blood samples were taken on days 14, 28 and 57. Fecal samples were collected on days 7, 14 and 21 of age for the measurement of selected microbial species. By design, HM calves consumed more nutrients from milk during the first 3 weeks and they were heavier than LM calves on days 21, 56 and 98. High milk-fed calves had greater serum glucose and triglyceride levels on day 14 with no significant difference between groups on days 28 and 57. Blood urea nitrogen was higher in LM calves on day 14, but it was lower in HM calves on day 28. Calves in LM group had significantly greater blood tumor necrosis factor-α (TNF-α) than HM calves throughout the experiment. Serum amyloid A (SAA) concentration was higher in LM calves on day 14. However, HM calves showed higher levels of SAA at the time of weaning. Feeding high volumes of milk resulted in lower serum cortisol levels on days 14 and 28 but not at the time of weaning in HM calves compared to LM counterparts. Lactobacillus count was higher in feces sample of HM calves. Conversely, the numbers of Escherichia coli was greater in the feces of LM calves. Calves in HM group showed fewer days with fever and tended to have fewer days treated compared to LM group. In conclusion, feeding higher amounts of milk during the first 3 weeks of life improved gut microbiota, inflammation and health status and growth performance of Holstein dairy calves.

Antimicrobial Activity of Agarwood Oil Against Multiple-Drug-Resistant (MDR) Microbes of Clinical, Food and Environmental Origin.

BACKGROUND AND OBJECTIVES: Multiple-Drug-Resistance (MDR) among bacteria is an imminent problem and alternative therapies are seen as a future abode. Agarwood Oil (AO) is described to possess antimicrobial activity besides many other medicinal utilities. This paper discusses the antimicrobial activity of AO on MDR and non-MDR strains of microbes of 69 genera isolated from clinical and non-clinical samples. METHODS AND RESULTS: In this study sensitivity of microbes was determined for conventional antimicrobials and AO using disc diffusion assay followed by determination of minimum inhibitory concentration (MIC) using agar well dilution assay. A total of 18.5% (522) strains were found sensitive to AO. Carbapenem resistant bacterial strains were more often (p, ≤0.01) resistant to antibiotics with 4.2 times more odds (99% CI, 2.99-5.90) of being MDR than carbapenem sensitive strains but no difference in their AO sensitivity was observed. However, MDR strains were more often (p, <0.001) resistant to AO than non-MDR strains. Bacteria isolated from dogs were more often sensitive to AO than those from buffaloes, human, horse, and cattle. On the other hand, bacteria from pigs were more often (p, ≤0.05) resistant to AO than bacteria from human, cattle, buffaloes, dogs, wild carnivores and birds. Oxidase positive Gram positive bacteria had 4.29 (95% CI, 2.94-6.27) times more odds to be AO sensitive than oxidase negative Gram negative bacteria. Bacillus species strains were the most sensitive bacteria to AO followed by strains of Streptococcus and Staphylococcus. The MIC of AO for different bacteria ranged from 0.01 mg/mL to > 2.56 mg/mL. CONCLUSION: The study concluded that MDR and AO resistance had a similar trend and AO may not be seen as a good antimicrobial agent against MDR strains.

Metabolic studies reveal that ruminal microbes of adult steers do not degrade rumen-protected or unprotected L-citrulline.

In vitro and in vivo experiments were conducted to determine the metabolism of rumen-protected or unprotected l-citrulline (Cit) plus l-glutamine (Gln) by ruminal microbes. In the in vitro experiment, whole ruminal fluid (3 mL, containing microorganisms) from steers was incubated at 37 ºC with 5 mM Cit plus 6 mM Gln (in a rumen-protected or unprotected form) for 0, 0.5, 2, or 4 h after which times 50 µL samples were collected for AA and ammonia analyses. In the in vivo experiment, at 0.5 h before and 0, 0.5, 1, 2, 4, and 6 h after cannulated adult steers consumed 0.56 kg dried-distillers' grain mixed with 70 g Cit plus 70 g Gln (in a rumen-protected or unprotected form), samples of ruminal fluid and jugular venous blood were obtained for AA analyses. Results from both in vitro and in vivo experiments demonstrated extensive hydrolysis of rumen-unprotected Gln into glutamate, but little degradation of the rumen-protected Gln or rumen-protected and unprotected Cit by ruminal microbes. Concentrations of Cit and arginine in the plasma of steers consuming rumen-protected or unprotected AA increased at 1 and 2 h after the meal, respectively, when compared with values at 0 h. Collectively, these novel findings indicate that ruminal microbes of adult steers do not degrade extracellular Cit in a rumen-protected or unprotected form. Our results refute the view that all dietary AAs are extensively catabolized by ruminal microorganisms and also have important implications for dietary supplementation with Cit to ruminants to enhance the concentration of arginine in their plasma and their productivity.

Branched-chain volatile fatty acids and folic acid accelerated the growth of Holstein dairy calves by stimulating nutrient digestion and rumen metabolism.

The combined addition of branched-chain volatile fatty acids (BCVFAs) and folic acid (FA) could improve growth performance and nutrient utilization by stimulating ruminal microbial growth and enzyme activity. This study was conducted to evaluate the effects of BCVFA and FA addition on growth performance, ruminal fermentation, nutrient digestibility, microbial enzyme activity, microflora and excretion of urinary purine derivatives (PDs) in calves. Thirty-six Chinese Holstein weaned calves (60 ± 5.4 days of age and 107 ± 4.7 kg of BW) were assigned to one of four groups in a randomized block design. Treatments were control (without additives), FA (with 10 mg FA/kg dietary DM), BCVFA (with 5 g BCVFA/kg dietary DM) and the combined addition of FA and BCVFA (10 mg/kg DM of FA and 5 g/kg DM of BCVFA). Supplements were hand-mixed into the top one-third of total mixed ration. Dietary concentrate to maize silage ratio was 50 : 50 on a DM basis. Dietary BCVFA or FA addition did not affect dry matter intake but increased average daily gain (ADG) and feed conversion efficiency. Ruminal pH and ammonia N were lower, and total volatile fatty acids (VFAs) concentration was higher for BCVFA or FA addition than for control. Dietary BCVFA or FA addition did not affect acetate proportion but decreased propionate proportion and increased acetate to propionate ratio. Total tract digestibility of DM, organic matter, CP and NDF was higher for BCVFA or FA addition than for control. Dietary BCVFA or FA addition increased activity of carboxymethyl cellulase and cellobiase, population of total bacteria, fungi, Ruminococcus albus, R. flavefaciens, Fibrobacter succinogenes and Prevotella ruminicola as well as total PD excretion. Ruminal xylanase, pectinase and protease activity and Butyrivibrio fibrisolvens population were increased by BCVFA addition, whereas population of protozoa and methanogens was increased by FA addition. The BCVFA × FA interaction was significant for acetate to propionate ratio, cellobiase activity and total PD excretion, and these variables increased more with FA addition in diet without BCVFA than in diet with BCVFA. The data indicated that supplementation with BCVFA or FA increased ADG, nutrient digestibility, ruminal total VFA concentration and microbial protein synthesis by stimulating ruminal microbial growth and enzyme activity in calves.

Effect of 2-hydroxy-4-(methylthio) butanoate (HMTBa) supplementation on rumen bacterial populations in dairy cows when exposed to diets with risk for milk fat depression.

Diet-induced milk fat depression (MFD) is a condition marked by a reduction in milk fat yield experimentally achieved by increasing dietary unsaturated fatty acids and fermentable carbohydrates. 2-Hydroxy-4-(methylthio) butanoate (HMTBa) is a methionine analog observed to reduce diet-induced MFD in dairy cows. We hypothesize that the reduction in diet-induced MFD by HMTBa is due to changes in the rumen microbiota. To test this, 22 high-producing cannulated Holstein dairy cows were placed into 2 groups using a randomized block design and assigned to either control or HMTBa supplementation (0.1% of diet dry matter). All cows were then exposed to 3 different diets with a low risk (32% neutral detergent fiber, no added oil; fed d 1 to 7), a moderate risk (29% neutral detergent fiber and 0.75% soybean oil; fed d 8 to 24), or a high risk (29% neutral detergent fiber and 1.5% soybean oil; fed d 25 to 28) for diet-induced MFD. Rumen samples were collected on d 0, 14, 24, and 28, extracted for DNA, PCR-amplified for the V1-V2 region of the 16S rRNA gene, sequenced on an Illumina MiSeq (Illumina, San Diego, CA), and subjected to bacterial diversity analysis using the QIIME pipeline. The α diversity estimates (species richness and Shannon diversity) were decreased in the control group compared with the HMTBa group. Bacterial community composition also differed between control and HMTBa groups based on both weighted UniFrac (relative abundance of commonly detected bacteria) and unweighted UniFrac (presence/absence) distances. Within the HMTBa group, no differences were observed in bacterial community composition between d 0 and d 14, 24, and 28; however, in the control group, d 0 samples were different from d 14, 24, and 28. Certain bacterial genera including Dialister, Megasphaera, Lachnospira, and Sharpea were increased in the control group compared with the HMTBa group. Interestingly, these genera were positively correlated with milk fat trans-10,cis-12 conjugated linoleic acid and trans-10 C18:1, fatty acid isomers associated with biohydrogenation-induced MFD. It can be concluded that diet-induced MFD is accompanied by significant alterations in the rumen bacterial community and that HMTBa supplementation reduces these microbial perturbations.

Associative effects of wet distiller's grains plus solubles and tannin-rich peanut skin supplementation on in vitro rumen fermentation, greenhouse gas emissions, and microbial changes1.

Two sets of in vitro rumen fermentation experiments were conducted to determine effects of diets that included wet distiller's grains plus solubles (WDGS) and tannin-rich peanut skin (PS) on the in vitro digestibility, greenhouse gas (GHG) and other gas emissions, fermentation rate, and microbial changes. The objectives were to assess associative effects of various levels of PS or WDGS on the in vitro digestibility, GHG and other gas emissions, fermentation rate, and microbial changes in the rumen. All gases were collected using an ANKOM Gas Production system for methane (CH4), carbon dioxide (CO2), nitrous oxide (N2O), and hydrogen sulfide (H2S) analyses. Cumulative ruminal gas production was determined using 250 mL ANKOM sampling bottles containing 50 mL of ruminal fluid (pH 5.8), 40 mL of artificial saliva (pH 6.8), and 6 g of mixed diets after a maximum of 24 h of incubation. Fermenters were flushed with CO2 gas and held at 39 °C in a shaking incubator for 24 h. Triplicate quantitative real-time polymerase chain reaction (qPCR) analyses were conducted to determine microbial diversity. When WDGS was supplied in the diet, in the absence of PS, cumulative CH4 production increased (P < 0.05) with 40% WDGS. In the presence of PS, production of CH4 was reduced but the reduction was less at 40% WDGS. In the presence of PS, ruminal lactate, succinate, and acetate/propionate (A/P) ratio tended to be less with a WDGS interaction (P < 0.01). In the presence of PS and with 40% WDGS, average populations of Bacteroidetes, total methanogens, Methanobrevibacter sp. AbM4, and total protozoa were less. The population of total methanogens (R2 = 0.57; P < 0.01), Firmicutes (R2 = 0.46: P < 0.05), and Firmicutes/Bacteroidetes (F/B) ratio (R2 = 0.46; P < 0.03) were strongly correlated with ruminal CH4 production. Therefore, there was an associative effect of tannin-rich PS and WDGS, which suppressed methanogenesis both directly and indirectly by modifying populations of ruminal methanogens.

Nutraceuticals: An Alternative Strategy for the Use of Antimicrobials.

Livestock industries strive to improve the health of their animals and, in the future, they are going to be required to do this with a continued reduction in antimicrobial use. Nutraceuticals represent a group of compounds that may help fill that void because they exert some health benefits when supplemented to livestock. This review is focused on the mechanisms of action, specifically related to the immune responses and health of ruminants. The nutraceutical classes discussed include probiotics, prebiotics, phytonutrients (essential oils and spices), and polyunsaturated fatty acids.

Decoding multifarious role of cow dung bacteria in mobilization of zinc fractions along with growth promotion of C. annuum L.

Zinc is one of the micronutrients, required by all types of crops. About 10-100ppm of zinc is present in soil which is generally immobile. The cow dung sustains all life and being practice since aeons. Exploitation of cow dung bacteria can mobilize nutrients besides contributing in sustainable agriculture. Therefore, to examine mobilization of Zn, cow dung is used as a source of bacteria. The objectives of the present study were to isolate an array of bacteria from cow dung and to characterize them for their Zn (ZnO and ZnCO3) mobilization ability in addition to establish the optimum conditions for dissolution of zinc. A total of seventy bacterial isolates have been screened for Zn mobilization. Out of which most potent (CDK15 and CDK25) were selected to study the effect of various parameters viz. pH, temperature and concentration of Zn. These parameters were assessed qualitatively in diverse growth medium and quantitatively using Atomic absorption spectroscopy. Optimum pH and temperature for mobilization was recorded at pH 5 (ZnO) and 37 °C (ZnCO3) by CDK25, whereas, optimum zinc concentration for mobilization was recorded at 0.05% (ZnO) by CDK15. Maximum amount of Zn solubilized was recorded by CDK25 in ZnO (20ppm). Considering the abilities of most potent bacterial isolates with reference to P-mobilization and growth promoting traits, pot culture assay of C. annuum L. was carried out. The findings of which conclude that, bacterium CDK25 (Bacillus megaterium) could be exploited for factors viz. nutrient management of Zn, growth promoting agent, and Zn augmentation in soil.

Towards a rapid identification and a novel proteomic analysis for dermatophytes from human and animal dermatophytosis.

BACKGROUND: Since accurate identification of dermatophyte species is essential for epidemiological studies and implementing antifungal treatment, overcoming limitations of conventional diagnostics is a fruitful subject. OBJECTIVES AND METHODS: In this study, we investigated real-time polymerase chain reaction(q-PCR), matrix-assisted laser desorption/ionisation time of flight mass spectrometry (MALDI-TOF MS) and nano-electrospray ionisation mass spectrometry (nano-ESI-MS) to detect and identify the most frequently isolated dermatophytes from human and animal dermatophytosis in comparison with conventional methods. RESULTS: Among 200 samples, the identified species were Microsporum canis (78.22%), Trichophyton verrucosum (10.89%) and T. mentagrophytes (5.94%). Q-PCR assay displayed great execution attributes for dermatophytes detection and identification. Using MALDI-TOF MS, M. canis, but none of T. violacium, T. verrucosum or T. mentagrophytes, could be identified. Nano-ESI-MS accurately identified all species. The potential virulence attributes of secreted proteases were anticipated and compared between species. Secreted endoproteases belonging to families/subfamilies of metalloproteases, subtilisins and aspartic protease were detected. The analysed exoproteases are aminopeptidases, dipeptidyl peptidases and carboxypeptidases. Microsporum canis have three immunogenic proteins, siderophore iron transporter mirB, protease inhibitors, plasma membrane proteolipid 3 and annexin. CONCLUSION: In essence, q-PCR, MALDI-TOF MS and nano-ESI-MS assays are very nearly defeating difficulties of dermatophytes detection and identification, thereby, supplement or supplant conventional diagnosis of dermatophytosis.