Impact of chemical structure of flavanol monomers and condensed tannins on in vitro anthelmintic activity against bovine nematodes

Plants containing condensed tannins (CT) may have potential to control gastrointestinal nematodes (GIN) of cattle. The aim was to investigate the anthelmintic activities of four flavan-3-ols, two galloyl derivatives and 14 purified CT fractions, and to define which structural features of CT determine the anti-parasitic effects against the main cattle nematodes. We used in vitro tests targeting L1 larvae (feeding inhibition assay) and adults (motility assay) of Ostertagia ostertagi and Cooperia oncophora. In the larval feeding inhibition assay, O. ostertagi L1 were significantly more susceptible to all CT fractions than C. oncophora L1. The mean degree of polymerization of CT (i.e. average size) was the most important structural parameter: large CT reduced larval feeding more than small CT. The flavan-3-ols of prodelphinidin (PD)-type tannins had a stronger negative influence on parasite activity than the stereochemistry, i.e. cis- vs trans-configurations, or the presence of a gallate group. In contrast, for C. oncophora high reductions in the motility of larvae and adult worms were strongly related with a higher percentage of PDs within the CT fractions while there was no effect of size. Overall, the size and the percentage of PDs within CT seemed to be the most important parameters that influence anti-parasitic activity

In vitro rumen fermentation of diets with different types of condensed tannins derived from sainfoin (Onobrychis viciifolia Scop.) pellets and hazelnut (Corylus avellana L.) pericarps

The aim of this study was to evaluate the in vitro rumen fermentation parameters of diets including pellets of sainfoin pellets and/or hazelnut pericarps, which are two plant resources that containing different types of condensed tannins (CT) with contrasted structures, using a batch culture system during for 24 h. The treatments were a basal diet (control), the basal diet + pellets of dehydrated sainfoin (PS), the basal diet + freeze-dried hazelnut pericarps (HP), and the basal diet + PS + HP. The diets were adjusted to be isotannic (20 g/kg dry matter (DM), except for the control) and isoproteic (132 g/kg DM). Total gas and methane (CH4) productions were measured after 3.5 h and 24 h of incubation. At the end of incubation, pH, in vitro DM degradability (IVDMD) and the concentration of fermentation end-products in the medium were also measured. The CT structures of CT infrom PS and HP were very different: as PS showed a PD-dominant profilehad mostly prodelphinidins and HP showed a PC-dominant profile mostly procyanidins. After 24 h of incubation, the total gas and methane productions and IVDMD were greater for the basal diet than for the diet + HP and the diet + PS (P<0.05). The CH4 production increased significantly with the diet + HP in the presence of PEG, a compound CT-inactivating CTcompound (P<0.001), and tended to increase for the diet + PS (P<0.1). The volatile fatty acids (VFA) net productions were globally similar among treatments, while the NH3 concentration was lower for the diet + PS (with a significant PEG effect) than for the diets including HP, and was the highest for the basal diet. It was concluded that the inclusion of PS and HP in a basal diet results in lower rumen fermentability and that but their CT decreased CH4 production and protein degradability;, PS being were more efficient effective than HP for the latterreducing protein degradability

A mechanistic model of small intestinal starch digestion and glucose uptake in the cow

The high contribution of postruminal starch digestion (up to 50%) to total-tract starch digestion on energy-dense, starch-rich diets demands that limitations to small intestinal starch digestion be identified. A mechanistic model of the small intestine was described and evaluated with regard to its ability to simulate observations from abomasal carbohydrate infusions in the dairy cow. The 7 state variables represent starch, oligosaccharide, glucose, and pancreatic amylase in the intestinal lumen, oligosaccharide and glucose in the unstirred water layer at the intestinal wall, and intracellular glucose of the enterocyte. Enzymatic hydrolysis of starch was modeled as a 2-stage process involving the activity of pancreatic amylase in the lumen and of oligosaccharidase at the brush border of the enterocyte confined within the unstirred water layer. The Na+-dependent glucose transport into the enterocyte was represented along with a facilitative glucose transporter 2 transport system on the basolateral membrane. The small intestine is subdivided into 3 main sections, representing the duodenum, jejunum, and ileum for parameterization. Further subsections are defined between which continual digesta flow is represented. The model predicted nonstructural carbohydrate disappearance in the small intestine for cattle unadapted to duodenal infusion with a coefficient of determination of 0.92 and a root mean square prediction error of 25.4%. Simulation of glucose disappearance for mature Holstein heifers adapted to various levels of duodenal glucose infusion yielded a coefficient of determination of 0.81 and a root mean square prediction error of 38.6%. Analysis of model behavior identified limitations to the efficiency of small intestinal starch digestion with high levels of duodenal starch flow. Limitations to individual processes, particularly starch digestion in the proximal section of the intestine, can create asynchrony between starch hydrolysis and glucose uptake capacity

Low emission feed : opportunities to mitigate enteric methane production of dairy cows

As global demand for high-quality food originating from animal production is expected to rise due to an increasing human population and consumer income level, the expected role of ruminants in meeting this demand brings multiple challenges. Ruminant production needs to adapt to environmental changes and, at the same time, reduce its impact on the environment. Ruminants production systems have a major impact on the environment through the emission of greenhouse gases such as methane (CH4), nitrous oxide and carbon dioxide. Microbial fermentation of feeds in the gastrointestinal tract, known as enteric fermentation, is the main source of CH4 emissions from dairy production. Enteric CH4 emission is strongly related to the amount of feed fermented in the rumen, which depends on feed intake, feed composition and rumen fermentation conditions associated to the intrinsic characteristics of these feeds and the characteristics of the whole diet. Important gaps in knowledge remain however. The prime aim of this thesis was to investigate the effects of various feeding strategies to mitigate enteric CH4 emissions of dairy cows. First experiment was conducted to investigate the effects of type and level of starch in the concentrate. Inclusion of a high level (53%) of starch in the concentrate that accounted for 40% of the total mixed ration dry matter (DM) produced lower CH4 per unit of estimated rumen fermentable organic matter (eRFOM) than a low level (27% of DM) of starch (43.1 vs. 46.9 g/kg of eRFOM). Methane production per kg of eRFOM also was lower for diets based on rapidly fermentable starch (gelatinized maize grain) compared to diets based on slowly fermentable starch (native maize grain) (42.6 vs. 47.4 g/kg of eRFOM). However, inclusion of 53% of starch in the concentrate from both types of starch did not affect CH4 emission intensity (CH4 Ei) (CH4 emission per kg of fat- and protein-corrected milk; FPCM). In a subsequent experiment, maize silage was prepared from whole-plant maize harvested at a very early (25% DM), early (28% DM), medium (32% DM) and late (40% DM) stage of maturity and fed to dairy cows as an alternative to concentrate as starch source. Diet consisted of (on DM basis) 75% maize silage, 20% concentrate and 5% wheat straw. Increasing harvest maturity of maize silage linearly decreased CH4 yield (21.7, 23.0, 21.0 and 20.1 g/kg of DM intake) and CH4 emission as a fraction of gross energy intake (6.3, 6.7, 6.3 and 6.0%). Methane Ei tended to decrease linearly with maturity (13.0, 13.4, 13.2 and 12.1 g/kg FPCM). In another experiment grass silage as roughage source was tested. This experiment was designed to investigate the effects of N fertilisation of grassland and maturity of grass at cutting on CH4 emission in dairy cows. Two N fertilisation rates (65 vs. 150 kg of N/ha) were examined in combination with three stages of grass maturity (early, 28 days of regrowth; mid, 41 days of regrowth; and late, 62 days of regrowth). Diet contained 80:20 ratio (on DM basis) of grass silage (mainly ryegrass) and concentrate. Dry matter intake decreased with N fertilisation and maturity, and FPCM decreased with maturity but was unaffected by N fertilisation. Methane Ei (mean 15.0 g/kg of FPCM) increased by 31% and CH4 per unit digestible OM intake (mean 33.1 g/kg of DOMI) increased by 15% with increasing maturity. Methane yield (mean 23.5 g/kg of DM intake) and CH4 as a fraction of gross energy intake (mean 7%) increased by 7 and 9% with maturity, respectively, which implies an increased loss of dietary energy with progressing grass maturity. Rate of N fertilisation had no effect on CH4 Ei and CH4 yield. Despite the importance of in vitro gas production technique for evaluating feeds, in vitro study as a stand-alone approach was considered inadequate to fully evaluate the potential effect of feeds and rumen fermentation modifiers on CH4 production, because in vitro studies are frequently performed separately rather than in parallel with in vivo studies. To test this hypothesis, both in vitro and in vivo CH4 measurements were measured simultaneously using cows in the first experiment that were fed (and adapted to) the same dietary material used as a substrate for in vitro incubation, as donor for microbial inoculum. It was found that 24-h in vitro CH4 (mL/g of incubated organic matter) correlated well with in vivo CH4 when expressed per unit of eRFOM (R2 = 0.54), but not when expressed per unit of organic matter ingested (R2 = 0.04). In the same experiment, results showed that incubation of the same substrate with rumen inocula obtained from donor cows adapted to different diets produced a variable amount of CH4 suggesting that it is important to consider the diet of the donor animal when collecting rumen inocula for in vitro incubation. Even though the in vitro technique has limitations to represent in vivo conditions, it is useful for screening of large sets of animal feeds or feed additives to be used as a CH4 mitigation strategy. In this thesis, two in vitro experiments were conducted to examine the effects of variation in structural composition of condensed tannins (CT) in sainfoin accessions collected from across the world on CH4 production, and CT extracts obtained from a selected sainfoin accessions on CH4 production. Results revealed substantial variation among CT in their effect on in vitro CH4 production and this variation was attributed to differences in chemical structure of CT. Condensed tannins evaluated in this thesis showed to have potential to reduce in vitro CH4 production, but require further investigations to fully evaluate their in vivo effects. In conclusion, results from the research work conducted in this thesis show that changes in the basal diet of dairy cows and in roughage production management can substantially reduce the amount of enteric CH4 produced and thereby influence the impact of dairy production on the environment

Feeding nitrate and docosahexaenoic acid affects enteric methane production and milk fatty acid composition in lactating dairy cows

An experiment was conducted to study potential interaction between the effects of feeding nitrate and docosahexaenoic acid (DHA; C22:6 n-3) on enteric CH4 production and performance of lactating dairy cows. Twenty-eight lactating Holstein dairy cows were grouped into 7 blocks of 4 cows. Within blocks, cows were randomly assigned to 1 of 4 treatments: control (CON; urea as alternative nonprotein N source to nitrate), NO3 [21 g of nitrate/kg of dry matter (DM)], DHA (3 g of DHA/kg of DM and urea as alternative nonprotein N source to nitrate), or NO3 + DHA (21 g of nitrate/kg of DM and 3 g of DHA/kg of DM, respectively). Cows were fed a total mixed ration consisting of 21% grass silage, 49% corn silage, and 30% concentrates on a DM basis. Feed additives were included in the concentrates. Cows assigned to a treatment including nitrate were gradually adapted to the treatment dose of nitrate over a period of 21 d during which no DHA was fed. The experimental period lasted 17 d, and CH4 production was measured during the last 5 d in climate respiration chambers. Cows produced on average 363, 263, 369, and 298 g of CH4/d on CON, NO3, DHA, and NO3 + DHA treatments, respectively, and a tendency for a nitrate × DHA interaction effect was found where the CH4-mitigating effect of nitrate decreased when combined with DHA. This tendency was not obtained for CH4 production relative to dry matter intake (DMI) or to fat- and protein corrected milk (FPCM). The NO3 treatment decreased CH4 production irrespective of the unit in which it was expressed, whereas DHA did not affect CH4 production per kilogram of DMI, but resulted in a higher CH4 production per kilogram of fat- and protein-corrected milk (FPCM) production. The FPCM production (27.9, 24.7, 24.2, and 23.8 kg/d for CON, NO3, DHA, and NO3 + DHA, respectively) was lower for DHA-fed cows because of decreased milk fat concentration. The proportion of saturated fatty acids in milk fat was decreased by DHA, and the proportion of polyunsaturated fatty acids was increased by both nitrate and DHA. Milk protein concentration was lower for nitrate-fed cows. In conclusion, nitrate but not DHA decreased enteric CH4 production and no interaction effects were found on CH4 production per kilogram of DMI or per kilogram of FPCM

Antagonistic intestinal microflora produces antimicrobial substance inhibitory to pseudomonas species and other spoilage organisms

Chicken intestine harbors a vast number of bacterial strains. In the present study, antimicrobial substance produced by lactic acid bacteria (LAB) isolated from the gastrointestinal tract of healthy chicken was detected, characterized, and purified. Based on 16S rRNA sequencing, the bacteria were identified as Lactobacillus plantarum vN. The antimicrobial substance produced by this bacterium was designated vN-1 and exhibited a broad-spectrum of activity against many important pathogenic and spoilage microorganisms, including Pseudomonas aeruginosa, Staphylococcus aureus, Micrococcus luteus, Salmonella Typhimurium, and Erwinia amylovova. vN-1 was determined to be thermostable, insensitive to pH values ranging from 2.0 to 8.0, resistant to various organic solvents and to enzymatic inactivation. The inhibition kinetics displayed a bactericidal mode of action. This study revealed an antimicrobial substance with low molecular mass of less than 1 kDa as determined by ultrafiltration and having features not previously reported for LAB isolated from chicken intestines. The detection of this antimicrobial substance addresses an important aspect of biotechnological control agents of spoilage caused by Pseudomonas spp. and promises the possibility for preservation of refrigerated poultry meat

Effects of grass silage quality and level of feed intake on enteric methane production in lactating dairy cows

The objective of this study was to determine the effect of level of feed intake and quality of ryegrass silage as well as their interaction on enteric methane (CH4) emission from dairy cows. In a randomized block design, 56 lactating dairy cows received a diet of grass silage, corn silage, and a compound feed meal (70:10:20 on DM basis). Treatments consisted of 4 grass silage qualities prepared from grass harvested from leafy through late heading stage, and offered to dairy cows at 96 ± 2.4 (mean ± SEM) days in milk (namely, high intake) and 217 ± 2.4 d in milk (namely, low intake). Grass silage CP content varied between 124 and 286 g/kg of DM, and NDF content between 365 and 546 g/kg of DM. After 12 d of adaptation, enteric CH4 production of cows was measured in open-circuit climate-controlled respiration chambers for 5 d. No interaction between DMI and grass quality on CH4 emission, or on milk production, diet digestibility, and energy, and N retention was found (P ≥ 0.17). Cows had a greater DMI (16.6 vs. 15.5 kg/d; SEM 0.46) and greater fat- and protein-corrected milk (FPCM) yield (29.9 vs. 25.4 kg/d; SEM 1.24) at high than low intake (both P ≤ 0.001). Apparent total-tract nutrient digestibility was not affected (P ≥ 0.08) by DMI level. Total enteric CH4 production (346 ± 10.9 g/d) was not affected (P = 0.15) by DMI level. A small, significant (P = 0.025) decrease at high compared with low intake occurred for CH4 yield (21.8 ± 0.59 g/kg of DMI; −4%). Methane emission intensity (12.8 ± 0.56 g/kg of FPCM; −12%) was considerably smaller (P ≤ 0.001) at high intake as a result of greater milk yields realized in early lactation. As grass quality decreased from leafy through late heading stage, FPCM yield and apparent total-tract OM digestibility declined (−12%; P ≤ 0.015), whereas total CH4 production (+13%), CH4 yield (+21%), and CH4 emission intensity (+28%) increased (P ≤ 0.001). Our results suggest that improving grass silage quality by cutting grass at an earlier stage considerably reduces enteric CH4 emissions from dairy cows, independent of DMI. In contrast, losses of N in manure increased for the earlier cut grass silage treatments. The small increase in DMI at high intake was associated with a small to moderate reduction in CH4 emission per unit of DMI and GE intake. This study confirmed that enteric CH4 emissions from dairy cows at distinct levels of feed intake depend on the nutritive value and chemical composition of the grass silage