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.     

Ernährungsstudien in der Kälberfütterung

ABSTRACT

This in vitro study aimed to evaluate the effects of Yucca schidigera powder (YSP) and inulin (IN) on protein fermentation metabolites (short-chain fatty acids [SCFA] and branched-chain fatty acids, phenolic and indolic compounds, biogenic amines, ammonia and pH) by using faecal inocula from dogs fed either a low (L) or a high (H) protein diet (crude protein 201 or 377 g/kg as fed). Four treatments for each diet were evaluated in an in vitro batch culture system over 24 h: (1) control with no addition of substrates; (2) 4 g YSP/l; (3) 5 g IN/l; (4) 4 g YSP/l in combination with 5 g IN/l of faecal culture of dogs fed Diet L or H. Several changes in fermentation metabolites were analysed. Samples incubated with the faecal inocula of dogs fed Diet L produced higher concentrations of total SCFA, propionate (p = 0.001), acetate (p ≤ 0.001), d-lactate (p = 0.041) and indole (p = 0.003), whereas pH (p = 0.004) was decreased. Supplementation of IN increased the content of putrescine, d- and l-lactate, total SCFA, acetate, propionate, n-butyrate (p ≤ 0.001) and n-valerate (p = 0.003), while i-valerate, indole and pH (p ≤ 0.001) were reduced. Ammonia was lower (p = 0.013) in samples with faecal inocula from dogs fed Diet H and further reduced by the addition of IN (p ≤ 0.001). Samples with faecal inocula from dogs fed Diet L had a fewer quotient of ammonia and total SCFA (p = 0.040). Supplementation of YSP (p = 0.021), IN (p ≤ 0.001) and YSP in combination with IN (p = 0.047) led to a higher reduction of the quotient of ammonia and total SCFA. In conclusion, dietary protein concentration and the supplementation of IN resulted in a stimulation of fermentation while YSP appeared to have only minor effects.     

Effects of dietary inclusion of macaúba seed cake meal on performance,caecotrophy traits and in vitro evaluations for growing rabbits

The objective was to evaluate the inclusion of macaúba seed cake (MSC) meal in diets for growing rabbits by assessing their growth and slaughtering performance, haematological traits, nutritional contribution of caecotrophs, in vitro digestibility, degradability and fermentation parameters. A total of 88 rabbits were distributed to four groups with 22 animals each and fed diets containing 0, 50, 100 and 150 g/kg of MSC, respectively. The in vitro assays were conducted employing cecum inoculum on the same dietary treatments. The inclusion of MSC yielded a quadratic effect on in vitro dry matter digestibility (p < 0.001). The maximum amount of produced gas was raised linearly with the inclusion of MSC (p = 0.016). MSC linearly reduced several variables as the nutritional contribution of caecotrophs in dry matter (p = 0.017) and crude protein (p = 0.014), live weight at 51 d (p = 0.024), body weight gain (p = 0.039), average daily feed intake (ADFI) (p = 0.001) and feed conversion ratio (FCR) (p = 0.007) in the first period evaluated (30–50 d); furthermore the ADFI and FCR the second (51–72 d) and whole period (30–72 d) (p < 0.001). MSC addition caused a quadratic effect on white blood cells count (p = 0.026) and a linear decrease of eosinophils (p = 0.045). In conclusion, the inclusion of up to 150 g/kg of MSC improves the in vitro digestibility and fermentation potential of the diets, reflecting on the ADFI and FCR of the animals, although adverse effects are observed on the weight of the commercial carcass and nutritive contribution of the caecotrophs.     

Seasonal changes in blood biochemical and endocrine responses of different indigenous goat breeds of tropical island agro-ecological environment

The objective of the current study was to observe the impact of two seasons viz. summer (February–May) and monsoon (June–August) on the blood biochemical and hormonal responses in different indigenous goats of tropical island agro-ecological environment maintained under extensive management system. Sixty animals of three different indigenous goat breeds were included in the study: Andaman local goat (AL, n = 20), Andaman local?×?Malabari (AL?×?M, n = 20), and Teressa goat (n = 20). Sixty serum samples (n = 10/season/breed) from the three groups of animals were analyzed. Study revealed that there was a significant increase (p ≤ 0.05) in serum total protein in Teressa goats during summer than monsoon. Serum albumin showed significant variation (p ≤ 0.05) between AL and AL?×?M during summer whereas significant variation of albumin (p ≤ 0.05) was observed between AL and Teressa, AL?×?M and Teressa during monsoon season. Significant differences in serum albumin (p ≤ 0.05) were also observed in AL and AL?×?M during summer and monsoon seasons, respectively. The serum cortisol levels were significantly higher (p ≤ 0.05) in AL goats during summer than in monsoon season. Thus, the study could able to establish the seasonal variation in biochemical and hormonal values of indigenous goat breeds in hot and humid tropical island environment.     

Equine pre-caecal and total tract digestibility of individual carbohydrate fractions and their effect on caecal pH response

The working hypothesis was that a minor postprandial caecal pH decline would affect apparent total tract digestibility (ATTD) of the fibre fraction in horses and, hence, that soluble fibre would amplify fermentation and consequently increase ATTD of fibre. This study was a 4 × 4 Latin Square design with a sequence of 17 days adaptation to the ration followed by 8 sampling days. The feed rations consisted of only timothy hay (Group H), hay plus molassed sugar beet pulp combined with either whole oats (Group OB) or barley (Group BB) and hay plus loose chaff based concentrate (Group M). Four horses fitted with permanent caecal cannulas and collection harnesses were used. A pH electrode with logger was inserted through the cannula and caecal pH was recorded at 1 min intervals for 8 h. The mobile nylon bag technique was used to quantify pre-caecal loss (PCL) of individual feedstuffs. Fibre was analysed as dietary fibre (DF), non-starch polysaccharides, soluble non-cellulosic polysaccharides (S-NCP), insoluble non-cellulosic polysaccharide (I-NCP) and neutral detergent fibre. The ATTD of the S-NCP fraction was above 0.8, which was 60% higher than for the I-NCP fraction. The PCL of starch were 0.98 (oats) and 0.75 (barley). The BB diet lowered (p < 0.001) postprandial caecal pH more than the other diets and a significant correlation was found between the lowest pH and ATTD of I-NCP (r = ?0.66; p = 0.005). In conclusion, this study successfully measured the in vivo digestibility of individual fibre fractions and found that S-NCP was more digestible than the I-NCP, and that a single meal of unprocessed barley was sufficient to decrease caecal pH to such an extent that the fibre digestibility of the whole diet was negatively affected.     

Addition of citrus pulp and apple pomace in diets for dogs: influence on fermentation kinetics,digestion, faecal characteristics and bacterial populations

Fermentation kinetics, digestibility, faecal characteristics and bacterial populations (aerobes, anaerobes, lactobacilli, lactic acid bacteria, enterococci, coliforms and clostridia) of dog food mixed with citrus pulp and apple pomace were evaluated. The in vitro gas production of a pre-digested dog food mixed with 0, 30, 50 and 70 g/kg dry matter (DM) of citrus pulp or apple pomace was measured, and also an experiment with dogs fed the same dog food with or without the addition of 70 g/kg of either fresh citrus pulp or apple pomace was conducted. Gas production increased linearly (p < 0.001) and quadratically (p < 0.001) as fibre levels augmented. The inclusion of fibre sources in the diets resulted in higher faecal output (p = 0.005) and defecation frequency (p < 0.001), and lower faecal pH (p < 0.001) and digestibility values (p < 0.01). Faecal consistencies and microbial populations did not differ among treatments. The addition of fresh citrus and apple was effective to stimulate the hindgut fermentation, but slightly depressed the digestion.     

In vitro fermentation of various carbohydrate-rich feed ingredients combined with chyme from pigs

Increased carbohydrate fermentation, compared with protein fermentation, could benefit gut health. In two in vitro experiments, the effect of carbohydrate-rich feed ingredients on fermentation characteristics of ileal chyme from pigs was assessed, using the cumulative gas production technique. Ingredients of the first experiment included gums, inulins, pectins, transgalacto-oligosaccharides, lactose and xylan. In the second experiment, a gum, pectin and transgalacto-oligosaccharides were added at different starting weights, to determine their effects on fermentation characteristics of chyme, in relation to differences in the carbohydrate concentrations. In comparison to fermentation of chyme alone, added carbohydrates led to higher total gas production (p < 0.05), faster maximum rate of gas production (except for xylan) (p < 0.05), and a decreased branched-chain fatty acids to straight chain fatty acids ratio (BCR) (p < 0.05). In the second experiment, for all carbohydrate ingredients, the BCR decreased with increasing starting weights (p < 0.05). If these supplemented dietary carbohydrates were to reach the terminal ileum of the living animal, carbohydrate fermentation in the large intestine could be stimulated, which is known to have beneficial effects on host health.     

Effect of a combination of inulin and polyphenol-containing adzuki bean extract on intestinal fermentation in vitro and in vivo

The effect of a combination of inulin (INU) and polyphenol-containing adzuki bean extract (AE) on intestinal fermentation was examined in vitro using fermenters for 48 h and in vivo using rats for 28 d. The total short-chain fatty acid concentrations in the fermenters were decreased by a combination of INU and AE, but the concentration in the INU + AE group was higher than the cellulose (CEL) and CEL + AE groups. The cecal propionate concentration was increased by a combination of INU and AE compared with their single supplement. The ammonia-nitrogen concentration in the fermenters and rat cecum was decreased by INU and AE. Cecal mucin levels were increased by INU and AE respectively. Therefore, our observations suggested that the combination of INU and AE might be a material of functional food that includes several healthy effects through intestinal fermentation.     

Effects of Enterococcus faecalis on egg production,egg quality and caecal microbiota of hens during the late laying period

This study was conducted to determine the effects of diet supplementation of laying hens with Enterococcus faecalis (EF) on egg production, egg quality and caecal microbiota. A total of 360 Hy-Line Brown laying hens (72 weeks old) were divided into three groups with four replicates of 30 birds each. The laying hens were fed with the basal diet (Control), the basal diet + 3.75 · 10⁸ cfu EF/kg (Group I) or the basal diet + 7.5 · 10⁸ cfu EF/kg (Group II). The experiment lasted for 45 d. Eggs and caecal samples were collected at the end of the experiment. Results showed that dietary supplementation with EF did not affect the average daily egg weight, cracked egg rate, mortality and egg quality. However, EF supplementation caused a significantly increased laying rate and decreased feed/egg ratio (p < 0.05). The differences in caecal microbiota between Group II and the Control were significant. The relative abundance of Verrucomicrobia and Cyanobacteria at the phylum level, Rikenellaceae, Christensenellaceae and Veillonellaceae at the family level, and the Faecalibacterium, Christensenellaceae R-7 group and Eubacterium coprostanoligenes group at the genus level changed significantly in Group II compared with the Control (p < 0.05). In conclusion, the tested dietary supplementations with EF improved product performance and affected the caecal microbial community structure of laying hens during the late laying period.     

Electrolytic extraction drives volatile fatty acid chain elongation through lactic acid and replaces chemical pH control in thin stillage fermentation

Background

Volatile fatty acids (VFA) are building blocks for the chemical industry. Sustainable, biological production is constrained by production and recovery costs, including the need for intensive pH correction. Membrane electrolysis has been developed as an in situ extraction technology tailored to the direct recovery of VFA from fermentation while stabilizing acidogenesis without caustic addition. A current applied across an anion exchange membrane reduces the fermentation broth (catholyte, water reduction: H₂O + e^? → ½ H₂ + OH^?) and drives carboxylate ions into a clean, concentrated VFA stream (anolyte, water oxidation: H₂O → 2e^? + 2 H⁺ + O₂).

Results

In this study, we fermented thin stillage to generate a mixed VFA extract without chemical pH control. Membrane electrolysis (0.1 A, 3.22 ± 0.60 V) extracted 28 ± 6 % of carboxylates generated per day (on a carbon basis) and completely replaced caustic control of pH, with no impact on the total carboxylate production amount or rate. Hydrogen generated from the applied current shifted the fermentation outcome from predominantly C2 and C3 VFA (64 ± 3 % of the total VFA present in the control) to majority of C4 to C6 (70 ± 12 % in the experiment), with identical proportions in the VFA acid extract. A strain related to Megasphaera elsdenii (maximum abundance of 57 %), a bacteria capable of producing mid-chain VFA at a high rate, was enriched by the applied current, alongside a stable community of Lactobacillus spp. (10 %), enabling chain elongation of VFA through lactic acid. A conversion of 30 ± 5 % VFA produced per sCOD fed (60 ± 10 % of the reactive fraction) was achieved, with a 50 ± 6 % reduction in suspended solids likely by electro-coagulation.

Conclusions

VFA can be extracted directly from a fermentation broth by membrane electrolysis. The electrolytic water reduction products are utilized in the fermentation: OH^? is used for pH control without added chemicals, and H₂ is metabolized by species such as Megasphaera elsdenii to produce greater value, more reduced VFA. Electro-fermentation displays promise for generating added value chemical co-products from biorefinery sidestreams and wastes.

     

Seasonal variation in in vitro immune activity of milk leukocytes in elite and non-elite crossbred cows of Indian sub-tropical semi-arid climate

Immunity of mammary gland in terms of in vitro activity of milk leukocytes has been evaluated during hot-humid, summer, and winter season in elite (n = 10) and non-elite (n = 10) crossbred cows. Milk samples were collected from all the cows throughout the year at 15-day interval. Milk somatic cell counts (SCC) and differential leukocyte counts (DLC) were evaluated microscopically. Milk neutrophils, macrophages, and lymphocytes were isolated and cultured in vitro. In vitro PI of milk neutrophils and macrophages was evaluated by colorimetric NBT (nitro-blue tetrazolium) reductive assay. Mitogen-induced milk lymphocyte blastogenic response was measured by colorimetric MTT (thiazolyl blue tetrazolium bromide) assay. Milk SCC was found to be significantly (p < 0.01) higher in elite cows compared to non-elite cows irrespective of season. There was significant (p < 0.05) increase in milk SCC during hot-humid season compared to winter season in both the group of the cows. There was no significant difference between group and season in terms of DLC. In vitro phagocytic index of elite cows was significantly (p < 0.01) higher than non-elite cows. The phagocytic index was significantly (p < 0.01) decreased in summer and hot-humid season compared to winter season in both the group of animals. Macrophages isolated from elite cows having significantly (p < 0.01) lower phagocytic index than non-elite cows which significantly (p < 0.01) decreased during summer and hot-humid season compared to winter. In vitro milk lymphocyte proliferative response was significantly (p < 0.01) lower in elite cows. Activity of B-lymphocytes decreased significantly (p < 0.01) during summer and hot-humid season than winter, but activity of T-lymphocytes remains unaltered during different seasons. In conclusion, the mammary immunity in terms of in vitro activity of milk leukocytes is compromised during summer and hot-humid season in elite crossbred cows; therefore, better care and management should be taken in high-yielding cows during summer and hot-humid season to minimize intramammary infections.     

Levan production by Zymomonas mobilis cells. Attached to plaited spheres

In this work, an immobilization method for polymer-levan production by a non-flocculating Z mobilis culture was developed. The extent of cell attachment to the stainless steel wire surface, culture growth and product synthesis were described. It was established that during short-term passive immobilization of non-flocculation Z mobilis cells on a stainless steel wire surface, sufficient amounts of biomass for proper levan and ethano fermentation could not be obtained. Adherence of cells was improved by pressing the paste-like biomass within stainless steel spheres knitted from wire with subsequent dehydration. Biomass fixed in metal spheres was used for repeated batch fermentation of levan. The activation period of cells within wire spheres (WS) was 48 h in duration. During this time, cell growth stabilized at production levels of ethanol and levan of Q_eth = 1.238 g/l × h and q_eth = 0.47 g/l × h; Q_eth = 0.526 g/l × h and q_eth = 0.20 g/l × h. Five stable fermentation cycles were realized using one wire sphere inoculum, and maintaining a stable ratio of 2.4 of biomass suspended in the medium to biomass fixed in the sphere. Using fixed Z mobilis biomass in the WS, the total amount of inoculum could be reduced for batch fermentation. Large plaited wire spheres with biomass may have potential in fermentation in viscous systems, including levan production.     

Formation of volatile organic compounds during the fermentation of maize as affected by sealing time and silage additive use

ABSTRACT

The study investigated the production of volatile organic compounds during the fermentation of maize containing 26.8% dry matter (DM). Forage was ensiled without additive or treated with 2 ml/kg of a chemical silage additive (SA) containing per litre 257 g sodium benzoate, 134 g potassium sorbate and 57 g ammonium propionate, and either sealed immediately or with a delay of 24 h. During the fermentation process, DM-losses, fermentation pattern (including ethyl lactate [EL] and ethyl acetate [EA]) and yeast numbers were determined. Delayed sealing and no SA resulted in highest DM losses with significant interactions between sealing time (ST) and SA on all sampling days (p < 0.001). The effects on organic acid production were variable depending on storage length. Ethanol production was affected by ST and SA, but promptly sealed silage treated with SA had consistently the lowest concentrations. Higher ethanol content during fermentation was associated with higher DM losses, as reflected by a strongly linear, positive relationship (R² = 0.70, p < 0.001). Compared with promptly sealed silage, the counts of yeasts were higher after delayed sealing during the first 7 d of storage (p < 0.001). Moreover, SA reduced yeast numbers compared with untreated silage (p < 0.01). EL concentrations increased throughout storage, whereas EA acetate accumulation was very rapid and intense already during the early stages of fermentation and peaked on d 34. The differences in concentrations and accumulation pattern between EL and EA, especially during the early fermentation phases, make evident that their synthesis was facilitated by different pathways and reactions, respectively.     

Characterisation of a phenomenological model for commercial pneumatic muscle actuators

This study focuses on the parameter characterisation of a three-element phenomenological model for commercially available pneumatic muscle actuators (PMAs). This model consists of a spring, damping and contractile element arranged in parallel. Data collected from static loading, contraction and relaxation experiments were fitted to theoretical solutions of the governing equation for the three-element model resulting in prediction profiles for the spring, damping and contractile force coefficient. For the spring coefficient, K N/mm, the following relationships were found: K = 32.7 ? 0.0321P for 150 ≤ P ≤ 314 kPa and K = 17 + 0.0179P for 314 ≤ P ≤ 550 kPa. For the damping coefficient, B Ns/mm, the following relationship was found during contraction: B = 2.90 for 150 ≤ P ≤ 550 kPa. During relaxation, B = 1.57 for 150 ≤ P ≤ 372 kPa and B = 0.311 + 0.00338P for 372 ≤ P ≤ 550. The following relationship for the contractile force coefficient, F _ce N, was also determined: F _ce = 2.91P+44.6 for 150 ≤ P ≤ 550 kPa. The model was then validated by reasonably predicting the response of the PMA to a triangular wave input in pressure under a constant load on a dynamic test station.     

Resveratrol affects in vitro rumen fermentation,methane production and prokaryotic community composition in a time- and diet-specific manner

This study aimed to investigate the effect of resveratrol on methane production, rumen fermentation and microbial composition under high-concentrate (HC) and high-forage (HF) diets using the in vitro fermentation system. A total of 25 mg of resveratrol was supplemented into 300 mg of either HC or HF diet. Methane production, total volatile fatty acid (VFA) concentration, molar proportion of VFA, metabolites of resveratrol and prokaryotic community composition were measured after 12 and 24 h of in vitro fermentation. Resveratrol reduced methane production (ml per mg of dry matter degraded) by 41% and 60% under both HC and HF diets (P < 0.001), respectively, and this result could be associated with the lower abundance of Methanobrevibacter (P < 0.001) in response to resveratrol. The molar proportion of propionate was significantly higher in the resveratrol group only under the HC diet (P = 0.045). The relative abundance of 10 bacterial genera was affected by the three-way interaction of treatment, diet and time (P < 0.05). Resveratrol was partly converted to dihydroresveratrol after 24 h of fermentation, and its degradation could be associated with microbes belonging to the order Coriobacteriales. Our results suggest that multiple factors (e.g. diet and time) should be considered in animal experiments to test the effect of polyphenol or other plant extracts on rumen fermentation, methane emission and microbial composition.     

OPTIMIZATION OF SOLID-STATE FERMENTATION FOR PHYTASE PRODUCTION BY Thermomyces lanuginosus USING RESPONSE SURFACE METHODOLOGY

A strain of Thermomyces lanuginosus, isolated from hot spring water in Turkey, was studied for optimization of phytase production using solid-state fermentation. Effects on fermentation of different production parameters such as substrate type, moisture, culture time, and inoculum size were investigated using a one-factor-at-a-time approach. Central composite design (CCD) of response surface methodology was applied for the optimization of four factors (culture temperature, initial pH, aeration area, age of seeding culture) that were affecting phytase production by Thermomyces lanuginosus in rice bran. Maximum phytase activity was achieved by using rice bran. The optimum levels of variables that supported maximum enzyme activity were moisture 70%, culture time 7 days, inoculum size 40%, culture temperature 55°C, initial pH 7.5, aeration area 30%, age of seeding culture 5 days, sucrose 1%, and ZnSO₄ 2.5 mM. An overall 10.83-fold enhancement in phytase activity (0.30 to 3.248 U) was attained due to the optimization.     

Production of a highly glucose tolerant β-glucosidase by Paecilomyces variotii MG3: optimization of fermentation conditions using Plackett–Burman and Box–Behnken experimental designs

β-Glucosidases (β-d-glucoside glucohydrolase, 3.2.1.21) are a group of enzymes mainly involved in the hydrolysis of β-glycosidic bonds connecting carbohydrate residues in different classes of β-d glycosides. During cellulose degradation they convert cellobiose and cellooligosaccharides produced by the endo and exoglucanases to glucose. Most of the microbial β-glucosidases are inhibited by glucose. This limits their application in commercial scale cellulose degradation ventures. Solid state fermentation production of a highly glucose tolerant β-glucosidase by a novel isolate of Paecilomyces was optimized using a two step statistical experiment design. In the first step which employed a Plackett–Burman design, the effects of parameters such as moisture, temperature, pH, inoculum concentration, incubation time and different concentrations of (NH₄)₂SO₄, KH₂PO₄, NaCl, peptone and cellobiose were evaluated. The parameters with significant influence on the process were selected and fine tuned in the second step using a Box–Behnken design. The model obtained was validated and a peptone concentration of 2 g/l, inoculum concentration of 1.2 × 10⁶ spores/ml and an incubation period of 96 h were found to be optimum for the maximum production of the enzyme. The optimization resulted in a doubling of the enzyme production by the fungus.     

Dietary carbohydrates affect caecal fermentation and modify nitrogen excretion patterns in rats II. Studies with diets differing in protein quality

In 2 two‐factorial experiments, each conducted on 80 growing male rats, the effects of substituting 10% raw potato starch (PS), pectins (PEC), or cellulose (CEL) for wheat starch (WS) and the addition of tannic acid to WS (WSTA) were studied using diets differing in protein quality. Casein unsupplemented or supplemented with DL‐methionine and gluten unsupplemented or supplemented with lysine, methionine and tryptophan were used as protein sources in Experiment 1 and 2, respectively. Parameters indicative of caecal fermentation intensity (pH, acetic, propionic and butyric acid contents, digesta and tissue weight) and of protein metabolism (urea blood concentration, faecal and urinary nitrogen excretion) were determined. Ten‐day balance experiments were preceded by a 10‐day adaption period to respective carbohydrates given in a diet containing balanced protein.

In both experiments the type of carbohydrates affected the caecal concentration of individual and total SCFA and other parameters of fermentation intensity. Pectins and potato starch were fermented more intensively than cellulose. Faecal N excretion was increased by all carbohydrates substituted for cereal starch, and by tannic acid. Urinary excretion was greater on CEL than on PEC and WSTA containing casein and on other diets containing gluten. In both experiments urinary N excretion was the lowest on PEC diets. Protein quality had the greatest effect on apparent biological value and net protein utilization but all indices of protein utilization were also affected by carbohydrates. It is concluded that not only the amount of N excreted in faeces but also in urine is affected by the type and fermentability of carbohydrates.     

Alteration in the in vitro activity of milk leukocytes during different parity in high yielding cross-bred cows

In vitro activity of milk leukocytes (viz. neutrophils, lymphocytes and macrophages) was evaluated in forty-eight (48) clinically healthy high-yielding cross-bred cows of mid-lactation stage (100–200 days of lactation), divided into four groups namely 1st parity (n = 12), 2nd parity (n = 12), 3rd parity (n = 12) and 4th and above parity (n = 12). Milk samples were taken (250 ml/cow) were taken. Milk somatic cell counts (SCC) and differential leukocyte counts (DLC) were performed microscopically. In vitro phagocytic index (PI) of milk neutrophils and macrophages was evaluated by colorimetric nitro blue tetrazolium reductive assay. Mitogen-induced milk lymphocyte blastogenic response was measured by colorimetric MTT (tetrazolium) assay after isolation of the milk leukocytes by density gradient centrifugation. Milk SCC differed significantly (p < 0.01) between different parity. Cows of 4 and above parity showed significantly (p < 0.01) higher milk SCC compared to primiparous cows. There was no significant difference in milk DLC during different parities in high-yielding cross-bred cows. There was a significant (p < 0.01) variation in lymphocyte blastogenesis amongst parity. The highest value of lymphocyte blastogenesis was seen at 3rd parity, whereas lowest value was obtained in the cows of both 1st and 4th or above parity. PI of milk neutrophils did not differ significantly between parity. PI of milk macrophages was significantly (p < 0.01) higher in 3rd parity and lower (p < 0.01) in 1st and 4th parities. The study indicated that depressed activity of milk lymphocytes and macropages was lower and SCC was higher in the cows of 4th and above parity indicating more mammary stress and hence susceptible to udder infection and mastitis. Therefore, better care and managemental interventions should be taken around these periods.     

猪不同肠段微生物体外培养对蛋氨酸的代谢特性

【目的】旨在通过微生物体外发酵技术,以回肠微生物为参照,研究猪盲肠及结肠微生物对在小肠微生物中代谢率较低的蛋氨酸的代谢特性。【方法】采集4头健康100 kg左右杜×长×大杂交猪的盲肠、结肠与回肠食糜作为接种物,分别接种于10 mmol/L蛋氨酸的培养基中,37°C体外培养24 h。分别设含蛋基酸溶液和含各肠段食糜接种物的空白对照组。【结果】(1)不同肠段微生物以蛋氨酸为底物体外发酵,盲肠组蛋氨酸消失率(21.9%)显著高于结肠组(16.7%)与回肠组(16.3%)(P0.05)。盲肠组总SCFA量显著高于结肠与回肠组(P0.05),伴随着p H值下降程度最高;盲肠组MCP产量也显著高于结肠与回肠组(P0.05);在产气量与NH3-N浓度上,盲肠组与结肠组均显著低于回肠组(P0.05)。(2)以蛋氨酸为底物体外发酵,门水平上,总菌、厚壁菌门含量在各肠段组间无显著差异(P0.05),拟杆菌门含量在盲肠组最高;与不加蛋氨酸底物的对照组比较,三个肠段试验组总菌、厚壁菌门含量均显著高于对照组(P0.05),而拟杆菌门含量在试验组与对照组间差异不显著(P0.05)。属水平上,盲肠组和结肠组大肠杆菌属数量显著低于回肠组(P0.05),而柔嫩梭菌属和梭菌XIV属数量在盲肠组和结肠组均高于回肠组;各肠段组间双歧杆菌数量无显著差异(P0.05)。【结论】以蛋氨酸为底物,体外培养猪盲肠微生物对蛋氨酸代谢率高于回肠微生物,伴随着其他发酵参数的变化,并且发酵产生更多的菌体蛋白。相比于回肠微生物发酵,大肠微生物发酵后,柔嫩梭菌属和梭菌XIV属数量较高,而大肠杆菌属数量较低。