Calculations of apparent ruminal synthesis and intestinal absorption of biotin in dairy cows as influenced by the extraction method

Biotin is present in nature either free or as biocytin, which is only degraded under the action of a specific enzyme: biotinidase. This enzyme is not included in analytical assays generally used. A method for sample preparation using biotinidase was developed in our laboratory before analysis by ELISA. Three cows equipped with duodenal and ileal cannulae were used to compare the effects of methods of sample preparation on calculations of apparent ruminal synthesis and intestinal absorption of biotin. There was no apparent ruminal synthesis of biotin, no matter whether free or total biotin was measured (p = 0.84). Results also suggested that rumen microbes cannot utilize nor degrade biocytin present in the feed. Estimates of apparent intestinal absorption were influenced by the sample preparation method (p = 0.002). Analysis of free biotin caused an artefact, suggesting intestinal synthesis of this vitamin; whereas determination of total biotin concentrations showed that absorption was taking place in the small intestine.     

Intestinal absorption of biotin and biocytin in the rat

The uptake of biotin and biocytin was investigated in rat intestine using the everted sac technique. It has been shown that at biotin and biocytin concentrations !ess than 40 and 50 nM respectively, absorption proceeds by a saturable process, whereas at higher concentrations uptake by passive diffusion predominates. Fractionation of solublized brush border preparations indicates that biotinidase is the only protein which binds biotin in this preparation.     

Studies on the biotin flow at the duodenum of dairy cows fed diets with different concentrate levels and types of forages

Abstract

Biotin is involved in many vital metabolic pathways and must be provided for an efficient fermentation in the rumen, as well as for the intermediary metabolism of the host animal. Factors influencing ruminal biotin metabolism and output are widely unknown at present. Therefore, dairy cows fitted with permanent cannulas in the dorsal rumen and in the proximal duodenum were fed differently composed diets, and the biotin flow at the proximal duodenum was measured. The diets (on DM basis) consisted of 8.9 kg grass hay (Diet 1), 8.9 kg corn silage plus 2.0 kg concentrate (Diet 2), or 7.3 and 7.4 kg grass silage plus 10.0 kg concentrate (Diets 3 and 4). The concentrate in Diets 3 and 4 contained 87% wheat and corn grain, respectively. The cows were pre-fed the rations for 21 days. Thereafter duodenal digesta was sampled every two h for 5 days. Cr₂O₃ served as a flow marker and the microbial proportion of total nitrogen at the duodenum was estimated by near infrared spectroscopy (NIRS). The duodenal flow of biotin was not related to biotin intake, but to the amount of fermented organic matter (FOM) and the amount of microbial protein (Biotin [mg/d] = 0.518 · kg FOM?0.300; r = 0.85 and biotin [mg/d] = 0.012 · g microbial protein + 1.478; r = 0.84), irrespective of the composition of the diet fed. Mean daily biotin flow was 0.48 ± 0.11 mg/kg FOM without any systematic effect of diet composition. The ruminal biotin balance, calculated as the difference between biotin flow at the duodenum and biotin intake, was positive (1.4 – 2.0 mg/d) in cows fed the mixed roughage/concentrate diets and negative (?0.71 mg/d) when the pure hay diet was fed.     

Determination of biocytin

Biocytin (epsilon-N-[d-biotinyl]-L-lysine) is generally undetected in serum and other body fluids of normal healthy individuals in view of the ubiquitous distribution of biotinidase. It has been suggested that biocytin may be present in serum and urine of patients with inherited biotinidase deficiency. We have developed a noncompetitive assay for biocytin based on its interaction with avidin. Biocytin can be determined in biological samples containing both biotin and biocytin. Biotin from such samples is removed by an anion-exchange resin and the biocytin is determined by the avidin-binding assay. The effect of above-normal levels of biotin in the sample on the assay of biocytin is discussed.     

Influence of supplemental endoglucanase or xylanase on volatile fatty acid production from ruminant feed by ruminal in vitro cultures

Abstract

This study employed two commercial enzyme preparations to examine the effects of endoglucanase, xylanase or their combination on in vitro volatile fatty acid (VFA) production by ruminal microbial populations. Batch ruminal cultures were established with one of various feedstuffs or with a fescue hay-based diet and ruminal fluid from a heifer fed a 40% forage:60% concentrate diet. Addition of xylanase at 135 xylanase units (XU) per ml increased total VFA production from the fescue hay-based diet (44.3 vs. 57.2 mM, p < 0.05) without changing the acetate to propionate (A:P) ratio. Addition of endoglucanase at 2, 3, 4, and 5 carboxymethyl cellulase units (CMCU) per ml increased total VFA production from the fescue hay-based diet on average by 36% (p < 0.05). Addition of 3, 4 and 5 CMCU/ml also decreased (p < 0.05) the A:P ratio. The combined addition of xylanase (135 XU/ml) and endoglucanase (5 CMCU/ml) increased total VFA production from the fescue hay-based diet (40.9 vs. 61.5 mM, p < 0.05) and reduced the A:P ratio (3.4 vs. 1.5, p < 0.05). The effects of endoglucanase and xylanase supplementation on in vitro VFA production varied across the various substrates used. However, endoglucanase supplementation consistently reduced the A:P ratio with all substrates tested. The effects of the enzyme combination were generally greater than either enzyme alone. We conclude that endoglucanase and xylanase activities differ in their ability to affect ruminal VFA production, and endoglucanase but not xylanase, may improve fermentation efficiency by reducing the A:P ratio.     

Effect of different levels of concentrate on ruminal microorganisms and rumen fermentation in Nellore steers

The aim of this study was to investigate the effect of different dietary levels of concentrate on feed intake, digestibility, ruminal fermentation and microbial population in steers. Eight Nellore steers fitted with ruminal cannulas were used in a double 4 × 4 Latin square design experiment. The dietary treatments consist of four different proportions of concentrate to roughage: 30:70, 40:60, 60:40 and 80:20% in the dry matter, resulting in Diets 30, 40, 60 and 80, respectively. The roughage was corn silage, and the concentrate was composed of corn, soybean meal and urea. Apparent digestibility of organic matter and crude protein showed a linear association with concentrate proportion (p = 0.01), but the increased concentrate levels did not affect the digestibility of fibre. The lowest ruminal pH-values were observed in animals fed with Diet 80, remaining below pH 6.0 from 6 h after feeding, while in the other diets, the ruminal pH was below 6.0 not before 12 h after feeding. After feeding Diet 80, the ammonia concentration in the rumen was significantly the highest. Higher dietary concentrate levels resulted in a linear increase of propionic acid concentrations, a linear reduction of the ratio acetic acid to propionic acid (p < 0.01) and a linear increased synthesis of microbial nitrogen (p < 0.001). The predicted production of methane was lower in diets with greater amounts of concentrate (p = 0.032). The population of methanogens, R. flavefaciens and R. albus decreased with higher concentrate levels, while the population of S. ruminantium increased (p < 0.05). The results indicate that greater amounts of concentrate do not decrease ruminal pH-values as much as expected and inhibit some cellulolytic bacteria without impairing the dry matter intake and fibre digestibility in Nellore steers.     

Influence of feeding sunflower seed and meal protected against ruminal fermentation on ruminal fermentation,bacterial composition and in situ degradability in sheep

ABSTRACT

The effects of treating sunflower seed (SS) and meal (SM), as well as of a mixture of both feeds (SSM; 45:55) with a solution of malic acid (1 M; 400 ml/kg feed) and heating for protection against ruminal degradation were studied. Four rumen-fistulated sheep were fed two mixed diets composed of oat hay and concentrate (40:60) and differing only in the concentrate, that contained either a mixture of untreated SS and SM (control diet) or treated SS and SM (MAH diet). A crossover design with two 24-d experimental periods was used, and each period included 10 d of diet adaptation, 9 d for in situ incubations of SS, SM and SSM, and 5 d for measuring ruminal fermentation characteristics and rumen emptying. From day 6 onwards a solution of (¹⁵NH₄)₂SO₄ was continuously infused into the rumen of each sheep to label ruminal bacteria. Feeding the MAH diet did not affect either ruminal pH or concentrations of total volatile fatty acids and NH₃-N, but decreased (p ≤ 0.01) the molar proportions of acetate and propionate and increased those of butyrate (p< 0.001). Organic matter and lipid contents of ruminal bacteria were lower whereas both N content and ¹⁵N enrichment were greater (p ≤ 0.05) in MAH-fed sheep. The in situ effective degradability (ED) of different fractions of SS, SM and SSM were calculated from the ruminal rates of particle comminution and passage, and values were corrected for microbial contamination. The MAH treatment decreased the ED of most fractions for all feeds and increased the supply of by-pass crude protein (CP) by 19.1% and 120% for SS and SM, respectively, and that of fat by 34% for SS. The MAH treatment also increased the in vitro intestinal digestibility of the by-pass CP for both SS (from 60.1% to 75.4%) and SM (from 83.2% to 91.0%). The simultaneous heating of both feeds (SSM) reinforced the protective effect of the MAH treatment and increased the by-pass CP without altering its intestinal digestibility, increasing the intestinally digested CP content by 16.8% compared with the value estimated from the results obtained for MAH-treated SS and SM incubated independently. These results indicate that the MAH treatment was effective to protect sunflower protein against rumen degradation and increased its intestinal digestibility.     

Purification of biotinidase from human plasma and its activity on biotinyl peptides

Biotinidase catalyzes the hydrolysis of N epsilon-biotinyllysine (biocytin) to form biotin and free lysine. The enzyme has been purified 4800-fold from outdated human plasma and was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis to have a molecular weight of (76 +/- 2) X 10(3). The same molecular weight was found by molecular sieve chromatography under nondenaturing conditions, indicating biotinidase is a monomer. This value is in contrast to a molecular weight of 115 000 determined by Pispa [Pispa, J. (1965) Ann. Med. Exp. Biol. Fenn., Suppl. 5, 5-39] with an impure biotinidase. The Km for biocytin was 6.2 X 10(-6) M, and biotinidase was found to be sensitive to phenylmethanesulfonamide and iodoacetamide in agreement with earlier studies by Knappe and co-workers [Knappe, J., Brümmer, W., & Bierderbick, K. (1963) Biochem. Z. 338, 599-613], who suggested that serine hydroxyl groups and sulfhydryl groups are essential for enzymatic activity. The specificity of biotinidase was examined by using synthetic and natural biotinyl peptides isolated by specific proteolytic cleavage of the biotinyl subunit of transcarboxylase. It was found that the rate of hydrolysis of biocytin was 83-fold higher than that found for biotin-containing peptides 5-13 residues in length. Removal of methionine from either side of the conserved region around the biocytin did not greatly alter the rate of cleavage. Increasing the peptide to 65-123 residues in length decreased the rate 1200-fold compared to that of biocytin.(ABSTRACT TRUNCATED AT 250 WORDS)     

Comparison of a xylanase and a complex of non starch polysaccharide-degrading enzymes with regard to performance and bacterial metabolism in weaned piglets

Abstract

Weaned piglets were fed a wheat based diet either non-supplemented or supplemented with a multi-enzyme preparation or a xylanase mono-enzyme preparation, respectively. Both enzyme preparations increased live weight gain nonsignificantly, but only animals of the xylanase group showed a trend (p = 0.076) for an improved feed conversion. Only precaecal digestibility of total amino acids was improved significantly when the mono-enzyme preparation was added. Improvements of digestibility of crude fat, crude protein and starch did not reach the significance level. Both enzyme preparations reduced jejunal viscosity, however viscosity in the colon was only reduced by the mono-enzyme preparation. Both enzymes significantly reduced Lactobacillus spp. cell numbers as well as bacterial metabolites in the stomach and showed similar nonsignificant modifications in jejunum contents except for acetate in the mono-enzyme group. Total jejunal bile acids were unchanged. Compared to the control, the ratio of the main conjugated to the main deconjugated bile acid was significantly higher in the mono-enzyme group. This study has shown that the mono- and multi-enzyme preparation can lead to improved performance in wheat based diets for piglets. Like in poultry, the main mode of action seems to be the reduction of small intestinal viscosity. However, the generation of fermentable carbohydrates by the multi-enzyme preparation may mask beneficial effects on performance due to the development of an active bile acid deconjugating microbiota in the small intestine.     

Effects of the supplementation of distillers’ grape residues on ruminal degradability,whole tract digestibility and nitrogen metabolism in sheep

ABSTRACT

Five ruminally and duodenally cannulated Kazakh male lambs (30 ± 2.75 kg) maintained singly in a metabolic cage were used in a 5 × 5 Latin square experiment to investigate the effect of supplementing a ration with five different levels of distillers’ grape residue (DGR) on ruminal degradability, whole tract digestibility and nitrogen (N) metabolism of growing lambs. The rations were isoenergetic and isonitrogenous and contained 0, 3.85, 7.70, 11.55 and 15.41% DGR (DM basis). Each experimental period lasted for 18 d: 10 d for adaptation to the dietary treatment and 8 d for faecal, urinary, ruminal and duodenal digesta sample collections. The outflow rate of ruminal digesta increased (p = 0.032) linearly with the increased level of dietary neutral detergent fibre content, caused by the supplementation of DGR. As a result, the effective degradability of dry matter and crude protein decreased significantly with the treatments. Although the dietary intake of N, duodenal flow of total N, and the endogenous N at the duodenum were not affected by experimental treatments, N fractions in the digesta were altered. Ruminal microbial N decreased (p < 0.01) linearly; in contrast, ruminal un-degradable protein increased linearly (p < 0.01) in response to the increased addition of DGR. Although there was no significant difference in faecal N among treatments, N retention was increased linearly (p = 0.014), owing to the remarkable reduction (p = 0.016) of urinary N excretion with an increasing level of DGR. The results indicate that the DGR has some potential benefits of increasing the supply of bypass protein and of improving the utilisation efficiency of N for sheep. Therefore, the supplementation of DGR in ruminant feeding is recommended at levels not exceeding 10% of the diet.     

Association of Per3 length polymorphism with susceptibility of Alzheimer disease (AD) in Chinese population

Objectives: The association of Per3 length polymorphism with susceptibility of Alzheimer Disease (AD) was examined in the present study. Methods: This study was constructed using the case-control method and investigated the association of Per3 length polymorphism with susceptibility of AD. Genotypes of APOE and Per3 length were determined by a PCR restriction fragment length polymorphism detection method. Results: In this study, we gathered 130 unrelated AD patients and 188 controls in performing an analysis the association of Per3 length polymorphism with susceptibility of AD. In the whole sample or APOE ε4 non-carriers, an increased prevalence of five repeat homozygotes of Per3 length in AD patients had significant higher than that in controls (in the whole sample: χ² = 7.261,= 0.0176; in APOE ε4 non-carriers: χ² = 6.086, p = 0.030). And, among APOE ε4 carriers, an increased prevalence of five repeat homozygotes of Per3 length in AD patients had also significant higher than that in controls (χ² = 3.893, p = 0.0319). Conclusions: Among APOE ε4 non-carriers, five repeat homozygotes of Per3 length was associated with a high susceptibility of AD among APOE ε4 carriers and non-carriers.     

Time courses of changes of para-, meta-, and ortho-tyrosine in septic patients: A pilot study

Objectives: Sepsis is associated with oxidative stress. Due to oxidative stress, three tyrosine isoforms, para-, meta-, and ortho-tyrosine (p-, m-, and o-Tyr), can be formed non-enzymatically in smaller amounts. p-Tyr is mainly formed physiologically in the kidneys through the activity of the phenylalanine hydroxylase enzyme. The three tyrosine isoforms may undergo different renal handling.

Methods: Twenty septic patients were involved in the study and 25 healthy individuals served as controls. Blood and urine levels of p-, m-, and o-Tyr were measured on admission and four consecutive days.

Results: Serum m-Tyr levels were higher in septic patients than in controls on days 2 (P = 0.031) and 3 (P = 0.035). Serum p-Tyr levels were lower in the cases than in controls on days 1 (P = 0.005) and 2 (P = 0.040), and subsequently normalized due to a day-by-day elevation (P = 0.002). The tendency of urinary m-Tyr concentration was decreasing (P = 0.041), while that of urinary p-Tyr concentration was increasing (P = 0.001). Fractional excretion of m-Tyr (FE_m-Tyr) showed a decreasing tendency (P = 0.009), and was, on all days, higher than FE_p-Tyr, which remained near-normal, less than 4%. Procalcitonin showed significant correlation with FE_m-Tyr (r = 0.454; P < 0.001).

Discussion: Our data suggest that the oxidative stress marker m-Tyr and physiologic p-Tyr may be handled differently in septic patients. The excretion of m-Tyr correlates with inflammation. m-Tyr may be actively secreted or produced in the kidney in some patients, whereas the decreased serum level of p-Tyr is a consequence of diminished renal production and not of renal loss.     

Effects of soybean oil and dietary copper levels on nutrient digestion,ruminal fermentation,enzyme activity,microflora and microbial protein synthesis in dairy bulls

ABSTRACT

The study evaluated the effects of soybean oil (SO) and dietary copper levels on nutrient digestion, ruminal fermentation, enzyme activity, microflora and microbial protein synthesis in dairy bulls. Eight Holstein rumen-cannulated bulls (14 ± 0.2 months of age and 326 ± 8.9 kg of body weight) were allocated into a replicated 4 × 4 Latin square design in a 2 × 2 factorial arrangement with factors being 0 or 40 g/kg dietary dry matter (DM) of SO and 0 or 7.68 mg/kg DM of Cu from copper sulphate (CS). The basal diet contained per kg DM 500 g of corn silage, 500 g of concentrate, 28 g of ether extract (EE) and 7.5 mg of Cu. The SO × CS interaction was significant (p < 0.05) for ruminal propionate proportion and acetate to propionate ratio. Dietary SO addition increased (p < 0.05) intake and total tract digestibility of EE but did not affect average daily gain (ADG) of bulls. Dietary CS addition did not affect nutrient intake but increased (p < 0.05) ADG and total tract digestibility of DM, organic matter, crude protein and neutral detergent fibre. Ruminal pH was not affected by treatments. Dietary SO addition did not affect ruminal total volatile fatty acids (VFA) concentration, decreased (p < 0.05) acetate proportion and ammonia N and increased (p < 0.05) propionate proportion. Dietary CS addition did not affect ammonia N, increased (p < 0.05) total VFA concentration and acetate proportion and decreased (p < 0.05) propionate proportion. Acetate to propionate ratio decreased (p < 0.05) with SO addition and increased (p < 0.05) with CS addition. Dietary SO addition decreased (p < 0.05) activity of carboxymethyl cellulase, cellobiase and xylanase as well as population of fungi, protozoa, methanogens, Ruminococcus albus and R. flavefaciens but increased (p < 0.05) α-amylase activity and population of Prevotella ruminicola and Ruminobacter amylophilus. Dietary CS addition increased (p < 0.05) activity of cellulolytic enzyme and protease as well as population of total bacteria, fungi, protozoa, methanogens, primary cellulolytic and proteolytic bacteria. Microbial protein synthesis was unchanged with SO addition but increased (p < 0.05) with CS addition. The results indicated that the addition of CS promoted nutrient digestion and ruminal fermentation by stimulating microbial growth and enzyme activity but did not relieve the negative effects of SO addition on ruminal fermentation in dairy bulls.     

Intraruminal infusion of n-butyric acid induces an increase of ruminal papillae size independent of IGF-1 system in castrated bulls

Abstract

The objective of this study was to explore morphological alterations of rumen papillae induced by n-butyric acid in relation to the insulin-like growth factor (IGF) system in adult castrated bulls. Three animals fitted with rumen cannula were fed twice daily at a low and high nutritional level (LL and HL), i.e., at 1.1 × maintenance (M) and 1.6 × M, respectively. Diets contained artificial dried grass and concentrate (74:26 and 52:48). Bulls received no (B0) or daily intraruminal infusions of 500 g n-butyric acid (B500) over 14 d. The infusion started 1 h after the morning feeding (9:00) and lasted for 3.5 h. Thus, four treatments (B0LL, B500LL, B0HL, and B500HL) were compared. Blood and rumen mucosa samples from the atrium ruminis were taken at the last day of each period. Length, width and surface of rumen papillae were greater (p < 0.001) in B0HL than in B0LL. Treatment with n-butyric acid resulted in an increase of the papillae surface of 20 – 40% (p = 0.047) for both nutritional levels as compared to periods without n-butyric acid treatments. The higher nutritional level and intraruminal n-butyric acid infusion induced epithelial cell death. The percentage of proliferative cells was doubled by n-butyric acid treatment. The mRNA of IGF-1 and IGF type 1 receptor (IGF-1R), as well as IGF-1R binding capacity were unaffected by butyric acid treatments. The abundance of IGF-1 mRNA tended to be lower (p = 0.1) and IGF-1R abundance was lower (p = 0.03) in response to the HL. The plasma IGF-1 concentration was lower with butyric acid treatment (p < 0.01), but was unaffected by the nutritional level. In conclusion, under described experimental preconditions of daily short-time intraruminal n-butyric acid infusion alterations of rumen papillae morphology is not mediated by ruminal IGF type 1 receptor and by local IGF-1 expression in papillae in castrated bulls.     

Effects of dietary addition of cellulase and a Saccharomyces cerevisiae fermentation product on nutrient digestibility,rumen fermentation and enteric methane emissions in growing goats

This study was designed to assess the effectiveness of dietary cellulase (243 U/g, derived from Neocallimastix patriciarum) and a Saccharomyces cerevisiae fermentation product (yeast product) on ruminal fermentation characteristics, enteric methane (CH₄) emissions and methanogenic community in growing goats. The experiment was conducted in a 5 × 5 Latin square design using five Xiangdong black wether goats. The treatments included a Control and two levels of cellulase (0.8 g and 1.6 g/kg dry matter intake (DMI), i.e. 194 U/kg and 389 U/kg DMI, respectively) crossed over with two levels (6 g or 12 g/kg DMI) of the yeast product. There were no significant differences regarding feed intake, apparent digestibility of organic matter, neutral detergent fibre and acid detergent fibre among all the treatments. In comparison with the Control, the ruminal ammonia N concentration was decreased (p = 0.001) by cellulase and yeast product addition. The activities of carboxymethylcellulase and xylanase were decreased after cellulase addition. Moreover, dietary cellulase and yeast product addition led to a significant reduction (p < 0.05) of enteric CH₄ emissions although the diversity and copy numbers of methanogens among treatments were not dissimilar. The present results indicate that the combination of cellulase and yeast fermentation product can reduce the production of CH₄ energy and mitigate the enteric CH₄ emissions to a certain degree.     

Effects of quebracho tannin extract (Schinopsis balansae Engl.) and activated charcoal on nitrogen balance,rumen microbial protein synthesis and faecal composition of growing Boer goats

Under irrigated arid conditions, organic fertiliser rich in slowly decomposable nitrogen (N) and carbon (C) is needed for soil fertility maintenance. Feeding ruminants with condensed tannins will lower ruminal protein degradation, reduce urinary N excretion and might increase the faecal fraction of slowly decomposable N. Supplementation with activated charcoal (AC) might enrich manure with slowly degrading C. Therefore, we investigated the effects of feeding quebracho tannin extract (QTE) and AC on the N balance of goats, the efficiency of microbial protein synthesis in the rumen (EMPS) and the composition of faeces. The feeding trial comprised three periods; in each period, 12 male Boer goats (28 ± 3.9 kg live weight) were assigned to six treatments: a Control diet (per kg diet 500 g grass hay and 500 g concentrate) and to further five treatments the Control diet was supplemented with QTE (20 g and 40 g/kg; diets QTE2 and QTE4, respectively), with AC (15 g and 30 g/kg, diets AC1.5 and AC3.0, respectively) and a mixture of QTE (20 g/kg) plus AC (15 g/kg) (diet QTEAC). In addition to the N balance, EMPS was calculated from daily excretions of purine derivatives, and the composition of faecal N was determined. There was no effect of QTE and AC supplementation on the intake of organic matter (OM), N and fibre, but apparent total tract digestibility of OM was reduced (p = 0.035). Feeding QTE induced a shift in N excretion from urine to faeces (p ≤ 0.001) without altering N retention. Total N excretion tended to decrease with QTE treatments (p = 0.053), but EMPS was not different between treatments. Faecal C excretion was higher in QTE and AC treatments (p = 0.001) compared with the Control, while the composition of faecal N differed only in concentration of undigested dietary N (p = 0.001). The results demonstrate that QTE can be included into diets of goats up to 40 g/kg, without affecting N utilisation, but simultaneously increasing the excretion of slowly decomposable N and C fractions. Feeding AC up to 30 g/kg of the diet increases slowly degradable faecal C concentration, without negative effects on N metabolism of goats.     

Effects of DNP on the cell surface properties of marine bacteria and its implication for adhesion to surfaces

Abstract

The effect of 2, 4-dinitrophenol (DNP) on the extracelluar polysaccharides (EPS), cell surface charge, and the hydrophobicity of six marine bacterial cultures was studied, and its influence on attachment of these bacteria to glass and polystyrene was evaluated. DNP treatment did not influence cell surface charge and EPS production, but had a significant effect on hydrophobicity of both hydrophilic (p = 0.05) and hydrophobic (p = 0.01) cultures. Significant reduction in the attachment of all the six cultures to glass (p = 0.02) and polystyrene (p = 0.03) was observed after DNP treatment. Moreover, hydrophobicity but not the cell surface charge or EPS production influenced bacterial cell attachment to glass and polystyrene. From this study, it was evident that DNP treatment influenced bacterial cell surface hydrophobicity, which in turn, reduced bacterial adhesion to surfaces.     

Effects of betaine and condensed molasses solubles on nitrogen balance and nutrient digestibility in piglets fed diets deficient in methionine and low in compatible osmolytes

Abstract

A balance experiment was carried out to investigate the effects of betaine monohydrate (BET) or betaine derived from condensed molasses solubles (CMS) as a substitute for methionine and choline on nitrogen (N) balance and total tract nutrient digestibility in weaned piglets. The experiment included four treatments with 32 barrows with an average initial body weight (BW) of 13.5 kg. The supplementation of DL-methionine and choline (positive control = PC) to the basal diet, which was deficient in methionine and low in compatible osmolytes in the form of betaine or its precursor choline (negative control = NC) resulted in a significant increase in N retention of 0.8 g/d. The substitution of DL-methionine and choline with BET or CMS did not affect N retention compared to the PC and the NC treatment either. Feeding the PC diet increased the digestibilities of organic matter, NDF, ADF, NFE, crude ash, Ca, P, methionine, tryptophan and cystine by 1.9%, 7.3%, 9.7%, 1.1%, 6.3%, 13.9%, 7.7%, 15.9%, 4.3% and 2.8%, respectively, and tended (p < 0.20) to increase the digestibilities of most other amino acids by 1.6 – 3.4%. Digestibility of CP, EE (HCl), Mg and Na was 3.1% (p = 0.09), 5.1% (p = 0.09), 5.1% (p = 0.06) and 3.3% (p = 0.17) higher, respectively, when compared to the NC treatment. BET and CMS supplementation increased most nutrient digestibilities in the same magnitude as for the PC treatment. In summary, the supplementation of betaine, originating from different sources, to a diet with low contents of compatible osmolytes increased in particular the fermentation of fibre and enhanced mineral absorption. The supplementation of the NC with DL-methionine was more efficient in improving N retention than the replacement of DL-methionine by betaine originating from BET or CMS.     

Effect of soybean oil availabilities on rumen biohydrogenation and duodenal flow of fatty acids in beef cattle fed a diet with crude glycerine

Soybean oil with different ruminal availability (whole soybeans (WS), soybean oil (SO) and calcium salts (CS)) was used to evaluate the fatty acid (FA) intake, rumen biohydrogenation (BH) and duodenal flow of FA in Nellore steers fed diets with crude glycerine (CG). Eight castrated Nellore steers were fitted with a ruminal and duodenal silicone cannula, and distributed in a double, simultaneous, Latin square 4 × 4 design with four diets and four experimental periods. Concentrates contained ground maize, urea, mineral salts, CG (100 g/kg DM) and soybean products with different availability of soybean oil: (1) no additional fat (CO), (2) WS, (3) SO or (4) CS. Fat supplementation was fixed to obtain 50 g ether extract/kg DM. Experimental treatments had no effect on DM intake, DM duodenal flow or ruminal turnover rate of C:16 FA. However, fat addition increased C:18 and turnover rates of total FA rumen (p < 0.05). CS resulted in lower C:18 turnover rates and lower ruminal BH of monounsaturated and unsaturated FA (UFA) than WS (p < 0.05). SO resulted in a greater duodenal flow of C18:0 (stearic acid), C18:1t-11 (vaccenic acid) and saturated FA than the WS and CS diets (p < 0.05). CS resulted in a higher duodenal flow of C18:3n-3 (linolenic acid) than WS (p < 0.05). The association of CG and calcium salts in Nellore steers was the best nutritional strategy to increase duodenal flow of healthier UFA, which may increase the deposition of these FA in meat. However, SO associated with CG association increased the duodenal flow of vaccenic acid, which is main precursor of endogenous synthesis of conjugated linoleic acids in tissues.     

Effects of fibre concentration of diets consisting of hay and slowly degradable concentrate on ruminal fermentation and digesta particle size in mid-lactation dairy cows

The objective of this study was to investigate the intestinal microbiota of growing kittens fed moderate- or high-protein diets using DNA-based qualitative and quantitative techniques. Kittens were weaned to a high-protein (HP; n = 7) or moderate-protein (MP; n = 10) diet at 8 weeks of age. Fresh faecal samples were collected at 8, 12, and 16 weeks of age. DNA was extracted and quantitative PCR used to quantify Bifidobacterium, Lactobacillus, Clostridium perfringens, and Escherichia coli concentrations. Denaturing gradient gel electrophoresis was performed to create a dendrogram and unrooted trees using Bionumerics 5.0 to identify similarity due to litter, age, or diet. Kittens fed HP had lower (p = 0.02) Bifidobacteria and Lactobacillus counts than MP-fed kittens. E. coli was lower (p = 0.02) in HP-fed kittens and tended to be affected by age (p = 0.09). Kittens were clustered by litter at 8 weeks of age, and then clustered by diet at 12 and 16 weeks of age. Our data suggest that faecal microbiota of growing kittens change after weaning and that dietary protein concentration affects E. coli, Bifidobacterium, and Lactobacillus populations. The relevance of these data in terms of intestinal health and disease remain to be determined and justifies further study.