Micronized fibres affect in vitro fermentation under normal buffered and osmotic stress conditions using porcine inocula

In this in vitro study, the modified Hohenheim gas test was used to determine fermentation activity and bacterial composition of pig's faecal microbial inoculum, when fermenting a standard pig diet with varying levels of crude protein (CP; 20, 24 and 28% CP), and supplemented with one of three fibre sources manufactured by micronization treatment. These were wheat envelopes (MWE), pea fibre (MPF) and lupine fibre (MLF). For comparison, inulin was used. As intestinal bacteria have to cope with varying osmotic conditions in their ecosystem, fermentation was performed under normal buffered and osmotic stress conditions. After 24 h of fermentation, total gas production and ammonia production were measured. In addition, the effect of MWE and inulin on short‐chain fatty acid (SCFA) production and numbers of total eubacteria, Lactobacillus spp., Bifidobacterium spp., Enterobacteriaceae, Enterococcus spp., Clostridium cluster XIVa and Clostridium cluster IV, were determined using quantitative real‐time PCR. Under normal buffered conditions, supplementation of MWE resulted in increased (p < 0.05) SCFA, acetic, propionic and valerianic acid production at CP levels of 20 and 28%. There was an increase (p < 0.05) in ammonia production for the micronized supplements, and for MWE an increased (p < 0.05) branched‐chain proportion was observed, possibly due to higher availability of protein for fermentation which was released during the micronization process. Osmotic stress conditions reduced (p < 0.05) total gas as well as total SCFA, acetic and propionic acid production for all treatments, while cell counts were increased (p < 0.05) for Bifidobacterium spp., Enterococcus spp. and Lactobacillus spp. Under normal buffered conditions in combination with 24 and 28% CP levels, lactobacilli were increased for MWE, compared to inulin (p < 0.05). In conclusion, micronized supplements such as MWE may beneficially modulate pigs' intestinal microbiota by increasing SCFA production in addition to a selective proliferation of lactobacilli.     

In vitro gas production of foliage from three browse tree species treated with different dose levels of exogenous fibrolytic enzymes

The aim of this study was to evaluate the effect of different dose levels of exogenous fibrolytic enzymes (EFE) on in vitro ruminal fermentation kinetics and energy utilization of foliages from three browse trees (Pithecellobium dulce, Heliocarpus velutinus and Guazuma ulmifolia). Mixture of EFE product was added to the leaves of the three browse tree species at three dose levels: 0 (control), 3.5 and 7.0 mg/g of DM. Chemical composition of the foliages, including plant secondary metabolites such as total phenolics (TP), saponins (SAP) and aqueous fraction (AF), was determined. In addition, in vitro assaying of ruminal gas production kinetics was determined for the three browse three foliages treated with EFE. P. dulce had the highest crude protein content (p < 0.05), whereas G. ulmifolia had the highest content of neutral detergent fibre and SAP (p < 0.05) and H. velutinus had the lowest content of TP (p < 0.05). The interaction between tree species and dose level of EFE was significant (p < 0.05) for gas production (GP) at 24 h of incubation, parameters b and c of the accumulated GP curve, short‐chain fatty acids (SCFA) and metabolizable energy (ME). The lowest (p < 0.01) extent of accumulated GP as well as the b and c values occurred in G. ulmifolia at 0 mg EFE/g DM. P. dulce had the highest (p < 0.05) values for ME and SCFA at the highest dose of EFE. Tree species and dose level had significant (p < 0.05) effects on all parameters describing in vitro ruminal fermentation kinetics and energy utilization. Addition of EFE improved the fermentation kinetics of the browse species considered in this study.     

A dose-response experiment evaluating the effects of oligofructose and inulin on nutrient digestibility, stool quality, and fecal protein catabolites in healthy adult dogs

In this experiment, three concentrations (0.3, 0.6, and 0.9% of diet, as-fed basis) of two fructans, oligofructose (OF) and inulin, were tested against a 0% supplemental fructan control. Seven ileal-cannulated adult female dogs were fed a meat-based, kibbled diet and assigned to treatments in a 7 x 7 Latin square design. Dietary supplementation of fructans had no effect on nutrient intakes or ileal digestibilities. Total-tract digestibilities of DM, OM, and CP decreased (P < 0.05) as a result of dietary OF and inulin supplementation. Dogs fed the control diet had a DM total-tract digestibility of 83.0%. The percentages of fecal DM for dogs fed the control and 0.3, 0.6, and 0.9% OF were 36.6, 33.3, 32.8, and 31.7%, respectively. When compared with the control, OF (P < 0.01) and inulin (P < 0.01) supplementation increased fecal ammonia concentrations. Higher fecal short-chain fatty acid (SCFA; P < 0.10) and isovalerate concentrations (P < 0.01) were noted for dogs fed both fructans. Total fecal SCFA for dogs fed the control diet and 0.3, 0.6, and 0.9% OF were 406.4, 529.9, 538.3, and 568.8 micromol/g of feces (DM basis), respectively. Dogs fed 0.3, 0.6, and 0.9% inulin had total fecal SCFA of 472.2, 468.8, and 471.5 micromol/g of feces (DM basis), respectively. Linear increases were observed in putrescine (P < 0.11), cadaverine (P < 0.07), spermidine (P < 0.12), and total amines (P < 0.05) in feces of dogs fed OF. Lower fecal phenol (P < 0.08) and total phenol (P < 0.04) concentrations occurred in dogs fed inulin, along with a linear decrease (P < 0.08) in total phenols with OF supplementation. Total fecal phenols for dogs fed the control, 0.3, 0.6, and 0.9% inulin were 3.03, 1.86, 1.97, and 2.23 micromol/g of feces (DM basis), respectively. Low-level dietary inclusion of inulin and OF positively affected indices known to be associated with gut health of the dog without seriously compromising nutrient digestibility or stool quality. Overall, the 0.9% OF treatment resulted in the best responses, including no adverse effect on nutrient intakes, ileal digestibilities, or stool quality, as well as increased fecal SCFA and decreased fecal phenols. The biological responses due to inulin were more variable.     

Effects of xylanase supplementation on growth performance,nutrient digestibility,blood parameters,fecal microbiota,fecal score and fecal noxious gas emission of weaning pigs fed corn‐soybean meal‐based diet

This study was conducted to evaluate the effects of xylanase supplementation on nutrient digestibility, growth performance, blood parameters, fecal microflora shedding, fecal score and fecal noxious gas emission of weaning pigs fed corn‐soybean meal based diet. A total of 150 weaning pigs with an average initial body weight (BW) of 7.85 ± 0.93 kg were randomly allocated to three treatments based on BW and sex (10 replicate pens with five pigs, two gilts and three barrows) were used in this 42‐day trial. Dietary treatments were: (1) CON, basal diet; (2) X1, basal diet +0.005% xylanase; (2) X2, basal diet +0.01% xylanase. The xylanase supplementation linearly increased (P < 0.05) average daily gain (ADG), and gain : feed ratio (G:F) from days 29 to 42 and the in overall period, dry matter, nitrogen and energy digestibility, and fecal Lactobacilli counts, and linearly decreased (P < 0.05) blood urea nitrogen (BUN) concentration, fecal NH₃ and H₂S emission. Additionally, at weeks 5 and 6, there was a linear decrease in fecal score with xylanase supplementation. In conclusion, dietary supplementation of xylanase improved growth performance, nutrient digestibility, shifted microbiota by increasing fecal Lactobacillus counts, decreased BUN concentration, fecal score, and fecal NH₃ and H₂S emission in weaning pigs.     

Fermentation Capacity of Fecal Microbial Inocula of Przewalski Horse,Kulan, and Chapman Zebra and Polysaccharide Hydrolytic Activities of Fecal Microbial Constituents (Ciliates and Bacteria) of Kulan and Chapman Zebra

We examined fermentation capacity of fecal microbial inocula of Przewalski horse (Equus ferus przewalskii), Asian wild ass - kulan (Equus hemionus hemionus), and Chapman zebra (Equus quagga chapmani) in vitro. Interactions of the substrates (amorphous cellulose, wheat straw, meadow hay, xylan from oat spelt, and ground barley grain) and type of fecal inocula in the gas volume and in vitro dry matter digestibility were detected in all substrates after 72 hours of fermentation in five replicates for each substrate and type of inocula. No effects of fecal inocula sources were detected on total short-chain fatty acids concentrations. No live fecal ciliate population was present in kulan feces. Complex ciliate populations in zebra feces and the number and genera resembled ciliates from the colon of horses. Fresh feces of kulan and zebra were fractionated by galvanotaxis and centrifugation to separate fecal ciliates and bacteria. Specific activities (μmol of reducing equivalents/mL min mg protein) of carboxymethyl cellulase (CM-cellulase), xylanase, α-amylase, and inulinase were measured in crude cell-free extract of fecal ciliates (zebra), fecal bacteria (zebra and kulan), and total fecal preparation (zebra and kulan). All examined specific enzymatic activities were present in zebra fecal samples. We were unable to measure the inulinase activity and CM-cellulase activity in kulan fecal samples. Zebra ciliates are actively involved in the digestion of plant storage (α-amylase, 0.53 ± 0.02; inulinase, 1.77 ± 0.01, specific activities) and structural polysaccharides (CM-cellulase, 0.4 ± 0.15; xylanase, 0.26 ± 0.06). For the first time, we measured inulinase activity in intestinal ciliates.     

Pharmacokinetics studies of enrofloxacin injectable in situ forming gel in dogs

The objective of this study was to evaluate the pharmacokinetic characteristics of enrofloxacin (ENR) injectable in situ gel we developed in dogs following a single intramuscular (i.m.) administration. Twelve healthy dogs were randomly divided into two groups (six dogs per group), then administrated a single 20 mg/kg body weight (b.w.) ENR injectable in situ gel and a single 5 mg/kg b.w. ENR conventional injection, respectively. High‐performance liquid chromatography (HPLC) was used to determine ENR plasma concentrations. The pharmacokinetic parameters of ENR injectable in situ gel and conventional injection in dogs are as follows: MRT (mean residence time) (45.59 ± 14.05) h verse (11.40 ± 1.64) h, AUC (area under the blood concentration vs. time curve) (28.66 ± 15.41) μg·h/mL verse (11.06 ± 3.90) μg·h/mL, c_max (maximal concentration) (1.59 ± 0.35) μg/mL verse (1.46 ± 0.07) μg/mL, t_max (time needed to reach c_max) (1.25 ± 1.37) h verse (1.40 ± 0.55) h, t_1/2λz (terminal elimination half‐life) (40.27 ± 17.79) h verse (10.32 ± 0.97) h. The results demonstrated that the in situ forming gel system could increase dosing interval of ENR and thus reduced dosing frequency during long‐term treatment. Therefore, the ENR injectable in situ gel seems to be worth popularizing in veterinary clinical application.     

Effect of different starch sources in a raw meat-based diet on fecal microbiome in dogs housed in a shelter

A dietary intervention study was assessed to determine if different sources of starch in homemade diets could significantly modify fecal microbiome of dogs. Twenty-seven adult dogs were enrolled and fed a diet based on a mixture of rice and pasta with fresh raw meat (CD). After 90 d, 8 dogs continued to receive CD diet, 10 dogs received a diet made of a raw meat and a complementary food with rice as the main source of starch (B1), and 9 dogs were fed a diet with the same raw meat and a complementary food with potato as the main source of starch (B2). Samples of feces were collected from each dog in the mornings at the beginning of the study and after 15 d and analyzed for pH, ammonia N (N–NH₃) and total N, short chain fatty acids (SCFA) and lactic acid. Relative abundance of fecal microbiota was assessed by sequencing and annotating the V3–V4 regions of the 16S rRNA. Total starch intake was similar between diets but differed in the in vitro rate digestion and in the resistant starch, which was higher in B2 than in B1 and CD diets. Dogs fed B2 diet showed lower (P < 0.05) N–NH₃ and pH but higher (P < 0.05) molar proportion of lactic acid. Linear discriminant analysis of the genera relative abundances indicated a significant (P < 0.01) increase of SMB53 genus at the end of the study in B1 diet and of Megamonas genus in B1 and B2 diets in comparison to CD diet. These results suggest that changes of starch source in a raw meat-based diet have limited effects on fecal microbiome in healthy dogs, but underline a high variability of microbiota among dogs.     

Characterization of the nutritive value of tropical legume grains as alternative ingredients for small‐scale pork producers using in vitro enzymatic hydrolysis and fermentation*

In the tropic, the small‐scale pork production is negatively influenced by the low availability of high protein ingredients. The study aimed to compare the protein and starch hydrolysis as well as fibre fermentation of five tropical legume grains (Canavalia brasiliensis, CB; Lablab purpureus, LP; Vigna unguiculata, white WVU; pink PVU and red RVU) and a control (extruded full‐fat soybean (SB)), using an in vitro model that simulated digestion in the gastrointestinal tract of pigs. A sequential in vitro hydrolysis was carried out with pepsin (120 min) and pancreatin (240 min) to determine the degree of hydrolysis (DH) of protein and starch. The indigestible residue was fermented in vitro with pig faecal inoculum to compare the modelled kinetics of gas production over 72 h and the production of short‐chain fatty acids (SCFA). After 360 min of pepsin–pancreatin hydrolysis, SB and WVU had the highest protein hydrolysis (76% and 66%) and PVU and WVU the highest starch hydrolysis (70% and 64%) (p < 0.01). The in vitro fermentation of the indigestible residue of WVU resulted in the highest (482 ml/g DM; p < 0.001) and CB the lowest (335 ml/g DM) gas production. These data were consistent with the SCFA production. Butyrate, propionate and total SCFA were higher (or tended) for RVU and WVU when compared with CB and SB (p = 0.015–0.085). In conclusion, the high DH of protein and starch as well as the high gas and SCFA production obtained with raw WVU makes it an interesting alternative to SB as a feedstuff for swine nutrition in the tropic. Other legume grains (LP and CB) cannot be used by pigs in their raw form.     

Effects of tributyrin supplementation on ruminal microbial protein yield,fermentation characteristics and nutrients degradability in adult Small Tail ewes

Two trials were conducted to assess the effects of tributyrin (TB) supplementation on ruminal microbial protein yield and fermentation characteristics in adult sheep. In an in vitro trial, substrate was made to offer TB at 0, 2, 4, 6, and 8 g/kg on a dry matter (DM) basis and incubated for 48 hr. In an in vivo trial, 45 adult ewes were randomly assigned by initial body weight (55 ± 5 kg) to five treatments of nine animals over an 18‐day period. Total mixed ration was made to offer TB to ewes at 0, 2, 4, 6, and 8 g/kg on a DM basis. The in vitro trial showed that TB enhanced apparent degradation of DM (p = .009), crude protein (p < .001), neutral detergent fiber (p = .007) and acid detergent fiber (p = .010) and increased methanogenesis (p < .001), respectively. The in vivo trial showed that TB decreased DM intake (p < .001) and enhanced rumen microbial N synthesis (p < .001), respectively. Both in vitro and in vivo trials showed that TB increased total volatile fatty acid concentration and enhanced fibrolytic enzyme activity. The results indicated that TB might exert positive effects on microbial protein yield and fermentation in the rumen.     

Effects of supplementation of iodixanol to semen extender on quality and fertilization ability of frozen–thawed Thai native bull sperm

This study investigates the effects of iodixanol supplementation in varied concentrations to Tris egg yolk (TEY) extender on the quality and fertilization ability of frozen–thawed sperm of Thai native bulls. Each ejaculate was divided into four different groups, as follows: sperm were treated with TEY extender (control group) and TEY extender supplemented with three different concentrations of iodixanol (1.25%, 2.50% and 5.00%). Semen straws were frozen in liquid nitrogen vapor. After thawing, sperm motility characteristics, viability, plasma membrane integrity and acrosome integrity were determined. Also, frozen–thawed spermatozoa from all groups were used for in vitro fertilization and artificial insemination (AI) in natural estrus Thai native cows. The results showed that the post‐thaw quality of the 2.50% iodixanol group was superior to the other iodixanol groups (P < 0.05). However, iodixanol had no beneficial effect on post‐thaw sperm in vitro fertilization ability and pregnancy rate after AI (P > 0.05). It can be concluded that the supplementation of 2.50% iodixanol extender significantly improves the progressive motility, viability, plasma membrane integrity and acrosome integrity of cryopreserved semen from Thai native bulls, but it has no beneficial effect on in vitro fertilization ability and pregnancy rate after AI.     

Influence of dietary protein content and source on colonic fermentative activity in dogs differing in body size and digestive tolerance

Low-consistency, high-moisture feces have been observed in large dogs (Canis lupus familiaris), compared with small dogs, and particularly in sensitive breeds (e.g., German Shepherd dogs). The aim of this work was to determine if greater colonic protein fermentation is responsible for poorer fecal quality in large sensitive dogs. Twenty-seven bitches were allotted to 4 groups based on size and digestive sensitivity: small, medium, large tolerant, and large sensitive. Five experimental diets varying in protein source [highly digestible wheat gluten (WG) vs. medium digestible poultry meal (PM), and protein concentration from 21.4 to 21.6 (LP) to 38.2 to 39.2% CP (HP)] were tested. Diets were fed for 14 d and followed by a 12-d transition period. Digestive fermentation by-products were investigated in fresh stools [ammonia, phenol, indole, and short chain fatty acids including acetate, propionate, and butyrate (C2 to C4 SCFA), branched-chain fatty acids (BCFA), and valerate] and in urine (phenol and indole). Bacterial populations in feces were identified. The PM diets resulted in greater fecal concentrations of ammonia, BCFA, valerate, indole, and C2 to C4 SCFA than WG diets (P = 0.002, P < 0.001, P = 0.039, P = 0.003, and P = 0.012, respectively). Greater concentrations of ammonia, BCFA, and valerate were found in the feces of dogs fed HP compared with LP diets (P < 0.001, P < 0.001, and P = 0.012, respectively). The concentrations of ammonia, valerate, phenol, and indole in feces of large sensitive dogs were greater (P < 0.001, P < 0.001, P = 0.002, and P = 0.019, respectively) compared with the other groups. The Enterococcus populations were greater in feces of dogs fed with PMHP rather than WGLP diets (P = 0.006). Urinary phenol and indole excretion was greater when dogs were fed PM than WG diets (P < 0.001 and P = 0.038, respectively) and HP than LP diets (P = 0.001 and P = 0.087, respectively). Large sensitive dogs were prone to excrete a greater quantity of phenol in urine (P < 0.001). A diet formulated with highly digestible protein, such as WG, led to reduced concentrations of protein-based fermentation products in feces together with improved fecal quality in dogs, especially in large sensitive ones. Poor fecal quality in large sensitive dogs could be partly related to the pattern of protein fermentation in the hindgut.     

Effects of isoquinoline alkaloids from Macleaya cordata on physiological,immunological and inflammatory parameters in healthy beagles

The aim of the study was to investigate nutritional, physiological and immunological effects of a plant‐derived blend of isoquinoline alkaloids (Sangrovit® Extra) in healthy dogs. Two groups of healthy, adult beagles (N = 10) were tested in a cross‐over experiment, lasting two consecutive three‐week periods. The experimental group received 1.2 g additive/kg feed, according to the recommendation of 10–20 mg/kg live weight per day. The control group received the same feed without additive. Complete blood count, immunological parameters and amino acid concentrations in serum were assessed. Faeces were analysed for short‐chain fatty acids, lactate and ammonium; moreover, their quantity and consistency were determined. Neither feed intake, total apparent nutrient digestibility (crude protein and fat, organic matter, sodium, potassium) were affected by intake of the product. Lymphocyte and monocyte counts were slightly increased in both groups. Elevation was not treatment dependant. IgA, IgG, haptoglobin in serum and flow cytometric phenotyping of peripheral lymphocytes were not affected by alkaloids supplementation. Numerically greater methionine concentrations in blood serum occurred in the experimental group (p = 0.182). Quantity and consistency of faeces and ammonium concentration in faeces were not affected by the additive. Faecal concentrations of short‐chain organic acids differed between groups (acetic acid, % of total SCFA: control group 52.3 ± 5.2 vs. experimental group 57.1 ± 4.5, p = 0.042), lactate concentrations (d ‐, l ‐ and total) did not. Due to the shift of SCFA proportions in faeces, an effect of isoquinoline alkaloids (IQs) on the metabolic activity of intestinal microbiota is probable. In conclusion, the addition of IQs in the given dose was well tolerated and did not have adverse effects in healthy dogs.     

Detection and isolation of Toxoplasma gondii from fresh semen of naturally infected dogs in Southern Brazil

The aim of this study was to isolate Toxoplasma gondii and determine the viability of the parasite in fresh semen samples of clinically healthy adult dogs naturally infected. Eleven seropositive dogs with T. gondii IgG antibodies from southern Brazil were selected to confirm the presence and viability of T. gondii in fresh semen samples using in vitro isolation in Vero cell culture, polymerase chain reaction (PCR) and sequencing analysis. The presence of viable T. gondii was confirmed by in vitro isolation and PCR in five semen samples. The ITS1 region of the isolated protozoa (TG S4) was amplified and sequenced. The nucleotide sequence obtained was 99% compatible with the T. gondii DNA sequences stored in the GenBank. It has been shown that T. gondii tachyzoites may be isolated in vitro from fresh semen samples of clinically healthy dogs seropositive for T. gondii.     

Effect of cellobiose supplementation on in vitro fermentation activity and bacterial numbers of porcine inocula

In this in vitro study, the modified Hohenheim gas test (HGT) was applied to determine fermentation activity and bacterial composition of pig's faecal microbial inoculum using different concentrations of cellobiose. Incubation procedures included normal buffered and osmotic stress conditions (elevated medium salinity). After 24 hr of fermentation, production of gas, ammonia and short‐chain fatty acid (SCFA) was measured, and the gene copy numbers of total bacteria, Lactobacillus spp., Bifidobacterium spp., Roseburia spp., Clostridium Cluster IV spp. and Enterobacteriaceae were analysed using real‐time polymerase chain reaction. There was a significant reduction in gas production after 24 hr when comparing osmotic stress conditions with normal buffered conditions. Under osmotic stress, increasing cellobiose concentrations linearly increased gas production (p < .001), while ammonia, acetic acid and isobutyric acid concentrations decreased (p < .001, p = .012, p = .035 respectively). Under normal buffered conditions, Roseburia spp. gene copies linearly increased with increasing cellobiose concentrations (p = .048). Lactobacillus spp. and Bifidobacterium spp. numbers were higher under osmotic stress (p < .001) compared to normal conditions. Results might point towards a positive impact of cellobiose supplementation on gut health especially under osmotic stress conditions.     

Dietary protein level affects nutrient digestibility and ileal microbiota structure in growing pigs

This study aimed to determine whether dietary protein content influences pig health as indicated by ileal microbiota structure and coefficients of total tract apparent digestibility (CTTAD) of nutrients. Seventy‐two gilts, with an initial body weight of 29.9 ± 1.5 kg, were used in this 42‐day feeding study. Pigs were randomly allotted to one of three dietary treatments of corn‐soybean meal contained 14, 16 or 18% crude protein (CP). As dietary CP content decreased, the CTTAD of most essential amino acids (AAs), except for arginine and histidine, increased linearly, while those of most nonessential AAs decreased linearly. The concentration of total short‐chain fatty acids (SCFA) was higher in pigs fed the diet with 14% CP content than others. Ileal microbiota structure was changed by dietary treatments. In particular, at the phylum level, the relative abundance of Tenericutes in ileal digesta decreased as the dietary protein content reduced, while that of cyanobacteria increased. At the genus level, the relative abundance of Weeksella, Phaseolus acutifolius, Slackia, Sulfurimonas and Aerococcus showed significant differences among the three dietary treatments. In conclusion, ileal microbiota structure was changed by dietary protein content. Moderate reduction of protein intake can benefit gut health by enhancing the gut microbial fermentation and SCFA formation.     

Effect of supplementation of rice bran and fumarate alone or in combination on in vitro rumen fermentation,methanogenesis and methanogens

This study investigated the effect of fumarate (FUM) and rice bran (RB), alone and together, on in vitro rumen fermentation, methanogenesis and methanogens. In vitro incubation was performed with six media that were either unsupplemented (control) or supplemented with 10% RB, 5 mmol/L FUM, 10% RB + 5 mmol/L FUM, 10 mmol/L FUM, or 10% RB + 10 mmol/L FUM. Methane (CH₄) production, dry matter digestibility, CH₄ per digested dry matter, total short‐chain fatty acid (SCFA) production, proportion of SCFA, acetate : proprionate ratio, production of NH₃‐N, and population density of rumen microbes were determined. Supplementation with 10% RB + 10 mmol/L FUM yielded a 36% decrease in CH₄ production compared to the control. Supplementation of FUM, in the presence or absence of RB, provided increases in total SCFA production and propionate proportion up to 61% and 31%, respectively. Total bacteria, methanogens and protozoa populations were significantly (P < 0.05) decreased with the 10% RB + 10 mmol/L FUM supplementation. The effect of anti‐methanogenesis of FUM was enhanced by the addition of RB. Notably, the CH₄ production attenuation was achieved by 10% RB + 10 mmol/L FUM without reduction of digestibility or of ruminal fermentation.     

Aspergillus awamori‐fermented mung bean seed coats enhance the antioxidant and immune responses of weaned pigs

The potential benefits of Aspergillus‐fermented mung bean seed coats (FMSC) for weaned pigs remain unexplored. Both in vitro and in vivo experiments were employed to evaluate the potential of FMSC supplement on the growth, antioxidant and immune responses of weaned pigs. The total polyphenols and DPPH scavenging capability of ethanol extract of FMSC exhibited a greater (p < 0.01) increase than those of pre‐fermentation. With the addition of the polyphenol of FMSC extract, an increase in phagocytosis by neutrophils and proliferation of peripheral blood mononuclear cells (PBMC) were found. However, these observations were significantly inhibited (p < 0.05) in those activated cells. Next, 96 weaned pigs were allotted with a randomized complete block design into four dietary treatments, including 0 (control), 600, 1200 or 1800 mg/kg FMSC in a corn–soya bean meal basal diet for a 35‐day trial. The pigs were injected with swine enzootic pneumonia (SEP) vaccines at day 3 and day 21 respectively. The results showed that dietary treatment failed to affect growth performance or serum SEP titre. The diet supplemented with 600–1800 mg/kg FMSC decreased faecal lactoferrin on day 21 and increased plasma trolox equivalent antioxidant capacity (TEAC) and erythrocytes catalase activity, as well as decreased (p < 0.01) plasma malondialdehyde (MDA) concentration on day 35. Diet supplementation of 1800 mg/kg FMSC increased phagocytosis by neutrophils and PBMC proliferation induced by pokeweed mitogen (PWM). However, the polymorphonuclear leucocytes (PMN)‐positive respiratory burst cells were decreased in the supplementation of 1200 or 1800 mg/kg FMSC respectively. In addition, the serum haptoglobin concentration was decreased in the supplementation with 1200 mg/kg FMSC. Taken together, FMSC enriches polyphenols with antioxidative and immune modulated properties. After feeding FMSC, an improvement in antioxidative capability and immunocompetence was found, implying that FMSC could provide as a feed additive at optimal level 1200 mg/kg for weaned pigs.     

Comparison of nutritional and antinutritional traits among different species (Lupinus albus L., Lupinus luteus L., Lupinus angustifolius L.) and varieties of lupin seeds

In order to promote the use of lupin in pig nutrition, in this research the nutritional characteristics (i.e. dietary fibre, alkaloid and fatty acid profile) and the in vitro gas production of 12 lupin varieties grown in the Mediterranean basin and belonging to three lupin species (Lupinus albus, Lupinus angustifolius and Lupinus luteus) were assessed. Four varieties of L. albus (Asfer, Lublanc, Lutteur and Multitalia) were grown in South Campania. Three varieties of L. luteus (Dukat, Mister and Taper), three of L. angustifolius (Jindalee, Sonet and Wonga) and two of L. albus (Rosetta and Luxor) were grown in Eastern Sicily. Lupinus albus varieties showed interesting nutritional and dietetic characteristics (i.e. high protein and low fibre content); the lipid fraction, rather elevated, is well represented by monounsaturated fatty acids (544 g/kg), whereas saturated fatty acids (SFAs) are less represented (167 g/kg) and the n‐3/n‐6 ratio (0.510) is the most favourable. Lupinus luteus varieties presented the most remarkable dietetic aspects, in terms of polyunsaturated fatty acid (PUFA) content (569 g/kg), n‐6 PUFA series (490 g/kg), UFA/SFA (5.24) and PUFA/SFA (3.56) ratios and atherogenic (0.059) and thrombogenic (0.100) indices and very low alkaloid content (1.07 mg per 100 g). Lupinus angustifolius varieties showed the least interesting nutritional and dietetic characteristics: low protein and fat content, high fibre level, high SFA amount (248 g/kg) and the lowest favourable nutritional indices (IA: 0.164 and IT: 0.334). Regarding the fermentation process, in L. albus, the tendency to increase the rate of gas production during the early stages of fermentation suggests that the high presence of alkaloids did not affect the in vitro degradability, production of short‐chain fatty acids and fermentation process, probably due to their concentration and/or water solubility. Lupinus angustifolius and L. luteus showed intermediate and slightly worse in vitro fermentation patterns respectively. From a nutritional and dietetic point of view, lupin may represent an interesting alternative to soya bean in pig feeding.     

Effect of Ca Ionophore On Blastocyst Production Following Intracytoplasmic Sperm Injection in Caprine Oocytes

The aim of the present investigation was to study the effect of calcium ionophore activation on blastocyst production following intracytoplasmic sperm injection (ICSI) in in vitro‐matured Caprine oocytes. A total of 470 in vitro‐matured oocytes were selected and randomly divided in to three groups. Cumulus oocyte complexes (COCs) recovered by slicing the Caprine ovaries were matured in TCM199 supplemented with 10% foetal bovine serum (FBS) + 10% follicular fluid + FSH (5 μg/ml) + LH (10 μg/ml) + estradiol (1 μg/ml) + EGF (10 ng/ml) + BSA (3 mg/ml) for 27 h in humidified atmosphere at 38.5°C with 5% CO₂ in CO₂ incubator. After 27 h of culture, selected COCs (n = 470) were separated from cumulus cells by treating with 0.1% hyaluronidase enzyme and passing repeatedly through a fine pipette and randomly divided into three groups. In group 1, (n = 168) matured oocytes were injected with injection micropipette without sperm as control. In group 2, (n = 152) capacitated spermatozoa were injected into cytoplasm of in vitro‐matured oocytes through injection micropipette. In group 3, (n = 150) capacitated spermatozoa were injected into cytoplasm of in vitro‐matured oocytes through injection micropipette and then activated with 5 μm Ca ionophore for 5 min. The oocytes of all groups were then culture in RVCL media for embryo development. The cleavage rate was observed after 48–72 h of injection. The cleavage rate and blastocyst production in group 1, 2 and 3 were 0.00 and 0.00, 18.42 and 3.57 and 61.33% and 16.30%, respectively. The result indicated that mechanical activation failed to induce cleavage in in vitro‐matured Caprine oocytes, whereas chemical activation of intracytoplasmic sperm‐injected in vitro‐matured Caprine oocytes showed significantly higher cleavage rate and blastocyst production as compare to non‐activated oocytes.