Effect of glyphosate contaminated feed on rumen fermentation parameters and in sacco degradation of grass hay and corn grain

Abstract Four rumen fistulated wethers were used to investigate the effect of glyphosate contaminated feed on rumen fermentation. The rations were based on corn silage, urea and a vitamin-mineral premix, either in the absence or presence of 0.77 g glyphosate per kg DM. Furthermore, rations were fed either with or without aromatic amino acid supplementation. During four periods of 28 days, sheep received each of the four dietary treatments according to a Latin square. After 14 days of adaptation rumen fermentation parameters (pH, ammonia, volatile fatty acids) were measured on day 15 over a five-hour period after the morning feeding. The remaining 13 days served for in sacco degradation studies with grass hay and corn grain. Ammonia (NH3) and pH of rumen fluid were within the normal range for all dietary treatments (NH3: 9.1-32.3 mmol x l(- l), pH: 6.2-6.7). Neither rumen fermentation parameters nor in sacco DM and NDF degradation of incubated feedstuffs were significantly affected by glyphosate, with or without aromatic amino acid supplementation. Kinetic profiles of the in sacco dry matter and NDF degradation of grass hay were almost identical for the dietary treatments.     

Effect of glyphosate contaminated feed on rumen fermentation parameters and in sacco degradation of grass hay and corn grain

Four rumen fistulated wethers were used to investigate the effect of glyphosate contaminated feed on rumen fermentation. The rations were based on corn silage, urea and a vitamin-mineral premix, either in the absence or presence of 0.77?g glyphosate per kg DM. Furthermore, rations were fed either with or without aromatic amino acid supplementation. During four periods of 28 days, sheep received each of the four dietary treatments according to a Latin square. After 14 days of adaptation rumen fermentation parameters (pH, ammonia, volatile fatty acids) were measured on day 15 over a five-hour period after the morning feeding. The remaining 13 days served for in sacco degradation studies with grass hay and corn grain. Ammonia (NH₃) and pH of rumen fluid were within the normal range for all dietary treatments (NH₃: 9.1 – 32.3?mmol·l^???1, pH: 6.2 – 6.7). Neither rumen fermentation parameters nor in sacco DM and NDF degradation of incubated feedstuffs were significantly affected by glyphosate, with or without aromatic amino acid supplementation. Kinetic profiles of the in sacco dry matter and NDF degradation of grass hay were almost identical for the dietary treatments.     

Effects of vitreousness and particle size of maize grain on ruminal and intestinal in sacco degradation of dry matter,starch and nitrogen

We assessed the effects of vitreousness and particle size of maize grain on ruminal and intestinal in sacco degradation of dry matter, starch and nitrogen. Six maize grain (Zea mays) genotypes characterized by differing vitreousness (proportion of vitreous in total endosperm) were ground (3-mm screen; Gr, ground particles, mean particle size (MPS): 526 μm) and cracked with a roller mill using two gap width settings (CS, cracked small particles, MPS: 1360 μm; CL, cracked large particles, MPS: 2380 μm). The ruminal and intestinal in sacco degradation of dry matter, starch and nitrogen was measured on three dry Holstein cows, fitted with rumen, proximal duodenum and terminal ileum cannulas, fed maize silage ad libitum twice daily. The ruminal starch degradability and intestinal digestibility differed among genotypes (P<0.001) and decreased as particle size increased (P<0.001). For the same particle size, starch ruminal degradability decreased (P<0.05) and intestinal digestibility decreased (P<0.002) with vitreousness. Particle size and vitreousness of maize grain are efficient factors for manipulating the amount of starch escaping rumen degradation, but may be limiting for the amount of starch digested in the small intestine.     

Validation of in sacco method: influence of sampling site,nylon bag or rumen contents,on fibrolytic activity of solid-associated microorganisms

Three ruminally cannulated cows, fed twice daily with a 70:30 forage:concentrate diet, were used to investigate the differences in fibrolytic activity of solid-associated microorganisms between nylon bags and rumen contents. Two different grass hays (regrowth and late harvested) were incubated in ruminal nylon bags. After 2 h or 23 h incubation time, pH was measured in bags and rumen contents, and enzymes of solid-associated microorganisms were extracted from bag residues and surrounding digesta by grinding, freezing, defrosting and sonication. Xylanase, avicelase, β-D-xylosidase and β-D-glucosidase activities were measured. Activities were lower in bag residues than in rumen digesta, and differences were greater after 2 h than after 23 h incubation time. Causes of these differences are discussed. For each incubation time and each enzyme, the differences in solid-associated microorganisms activities between rumen and bags contents were independent of the quality of hay in the bag. Thus the lower fibrolytic activity inside the bags may account for an underestimation of in vivo ruminal fiber degradation by the in sacco method, but this underestimation may be similar whatever the nature and content of forage cell walls.     

Influence of a Fusarium culmorum inoculation of wheat on the progression of mycotoxin accumulation, ingredient concentrations and ruminal in sacco dry matter degradation of wheat residues

The Fusarium head blight (FHB)-susceptible winter wheat cv. Ritmo was inoculated with spores of Fusarium culmorum at the beginning of full blossom. Samples of whole wheat plants were taken once weekly from anthesis until harvest and subsequently fractionated into straw, glumes and spindles, which were examined for deoxynivalenol (DON) and zearalenone (ZON). Additionally, the content of crude protein (CP) and non-starch polysaccharides (NSP) was scrutinized. Synthesis of the Fusarium toxins DON and ZON generally differed in terms of date of formation and concentration. Final mean DON concentrations of 37.5, 28.1 and 5.0 mg/kg DM were measured in glumes, spindles and straw, respectively, at the time of harvest. At this time, maximal mean ZON concentrations of 587, 396 and 275 microg/kg DM in spindles, glumes and straw, respectively, were determined. Moreover, Fusarium infected wheat residues contained higher CP but lower NSP contents at the last three sampling dates. In addition, collective samples of wheat straw and chaff were taken to investigate the effect of the Fusarium contamination on their in sacco DM degradation in dairy cows. Samples were analysed for mycotoxins and selected quality parameters. The dried and milled collective samples of straw and chaff were weighed into nylon bags and subjected to ruminal incubation for 4, 8, 16, 24, 48, 72, 96 and 120 h in two dairy cows equipped with a permanent rumen cannula. Marked differences in level of mycotoxin contamination as well as in ingredient composition between the variants of straw and chaff were detected. Moreover, after 120 h rumen incubation the in sacco DM degradation of inoculated straw and chaff were lower compared to the accordant controls. The soluble fraction was increased in inoculated samples, whereas a diminishment in the potentially degradable but insoluble fraction was more pronounced. Thereby, a decrease in the potential degradability was obtained for inoculated straw and even if less pronounced for chaff compared to the non inoculated corresponding controls. In conclusion, infection with F. culmorum of wheat involves an increased risk of mycotoxin contamination in straw. Also, a Fusarium infection may have an impact on chemical composition and may result in Fusarium growth-related modifications of host cell wall components.     

Proteolytic activity of ruminal digesta during the feeding cycle in sheep receiving grass hay/concentrate or maize silage/concentrate diets

Proteolytic activity was measured by the digestion of 14C-labelled casein in digesta removed from the rumen of four sheep receiving a grass hay/concentrate diet and four sheep receiving a maize silage/concentrate diet. Samples were removed immediately before feeding and at 2-h intervals after feeding up to 12 h. Animals on both diets produced similar proteolytic activities (1.83 (S.D. 0.41) and 2.14 (S.D. 0.61) mg 14C-casein hydrolysed (ml ruminal fluid)-1 h-1 with the maize silage- and grass hay-based diets, respectively). Time after feeding had no effect on proteolytic activity, but between-animal variation was consistent and highly significant, with the highest-activity animals having activities 64 and 74% higher than the lowest-activity animals on the two diets, respectively.     

Effect of whole-plant corn silage treated with lignocellulose-degrading bacteria on growth performance,rumen fermentation,and rumen microflora in sheep

Lignification of cellulose limits the effective utilisation of fibre in plant cell wall. Lignocellulose-degrading bacteria secrete enzymes that decompose lignin and have the potential to improve fibre digestibility. Therefore, this study aimed to investigate the effect of whole-plant corn silage inoculated with lignocellulose-degrading bacteria on the growth performance, rumen fermentation, and rumen microbiome in sheep. Twelve 2-month-old male hybrid sheep (Dorper ♂ × small-tailed Han ♀) were randomly assigned into two dietary groups (n = 6): (1) untreated whole-plant corn silage (WPCS) and (2) WPCS inoculated with bacterial inoculant (WPCSB). Whole-plant corn silage inoculated with bacterial inoculant had higher in situ NDF digestibility than WPCS. Sheep in the WPCSB group had significantly higher average daily gain, DM intake, and feed conversion rate than those in the WPCS group (P < 0.05). Furthermore, higher volatile fatty acid concentrations were detected in WPCSB rumen samples, leading to lower ruminal pH (P < 0.05). The WPCSB group showed higher abundance of Bacteroidetes and lower abundance of Firmicutes in the rumen microbiome than the WPCS group (P < 0.05). Multiple differential genera were identified, with Prevotella being the most dominant genus and more abundant in WPCSB samples. Moreover, the enriched functional attributes, including those associated with glycolysis/gluconeogenesis and citrate cycle, were more actively expressed in the WPCSB samples than in the WPCS samples. Additionally, certain glucoside hydrolases that hydrolyse the side chains of hemicelluloses and pectins were also actively expressed in the WPCSB microbiome. These findings suggested that WPCSB increased NDF digestibility in three ways: (1) by increasing the relative abundance of the most abundant genera, (2) by recruiting more functional features involved in glycolysis/gluconeogenesis and citrate cycle pathways, and (3) by increasing the relative abundance and/or expression activity of the glucoside hydrolases involved in hemicellulose and pectin metabolism. Our findings provide novel insights into the microbial mechanisms underlying improvement in the growth performance of sheep/ruminants. However, the biological mechanisms cannot be fully elucidated using only metagenomics tools; therefore, a combined multi-omics approach will be used in subsequent studies.     

Effect of different levels of concentrate allowances on rumen fluid pH,nutrient digestion,nitrogen retention and growth performance of weaner lambs

This experiment was conducted for 90 d to assess the effect of feeding graded levels of concentrate allowance on rumen fermentation characteristics, performance and nutrient utilisation of weaner lambs on restricted or high concentrate allowance using 60 weaner lambs of initial average live weight of 13.90 kg BW in a randomized design. The experimental treatments were 15 or 25 g kg⁻¹ BW or ad libitum concentrate allowance. Roughage source which contained Khejri (Prosopis cineratia) and Siris (Albizia lebback) leaves in 50:50 ratio was offered ad libitum to all the animals. Lambs supplemented with 15 g or ad libitum concentrate had similar dry matter intake (4.2 kg/100 kg BW) but significantly (p < 0.01) lower than 25 g concentrate supplemented group (4.9 kg/100 kg BW). Organic matter and CP intakes increased with increasing concentrate supplementation. Apparent digestibilities of dry matter, organic matter, CP, NDF, ADF and cellulose were significantly (p < 0.01) higher in ad libitum concentrate supplemented than 15 and 25 g concentrate supplemented lambs. Daily ME intake was significantly (p < 0.05) higher in 25 g and ad libitum concentrate supplemented lambs while ME intake kg⁻¹ gain was lower in ad libitum concentrate supplemented lambs (57 MJ kg⁻¹ gain) than those supplemented with 15 or 25 g concentrate (91 MJ kg⁻¹ gain). Generally, average daily gain increased with increasing levels of concentrate supplementation. Ad libitum concentrate supplemented lambs had significantly (p < 0.01) higher daily gains (151 g) than 15 and 25 g concentrate supplemented lambs (77 and 98 g, respectively). Feed efficiency was similar for 15 and 25 g concentrate supplemented lambs but significantly (p < 0.01) lower than the ad libitum concentrate supplemented lambs. All animals were in positive N-balance and the N-balance increased with increasing concentrate supplementation. Mean rumen fluid pH was significantly (6.6, p < 0.01) lower in ad libitum concentrate supplemented lambs compared to 15 or 25 g concentrate fed lambs (6.9). Rumen NH₃-N and total-N-concentrations peaked at 3 h post-feeding. Optimum rumen fluid pH, better nutrient digestibilities, higher N-retention improved growth by 49% of ad libitum concentrate fed lambs.     

Temporal fermentation and microbial community dynamics in rumens of sheep grazing a ryegrass-based pasture offered either in the morning or in the afternoon

Eight ruminally-fistulated wethers were used to examine the temporal effects of afternoon (PM; 1600h) v. morning (AM; 0800 h) allocation of fresh spring herbage from a perennial ryegrass (Lolium perenne L.)-based pasture on fermentation and microbial community dynamics. Herbage chemical composition was minimally affected by time of allocation, but daily mean ammonia concentrations were greater for the PM group. The 24-h pattern of ruminal fermentation (i.e. time of sampling relative to time of allocation), however, varied considerably for all fermentation variables (P⩽0.001). Most notably amongst ruminal fermentation characteristics, ammonia concentrations showed a substantial temporal variation; concentrations of ammonia were 1.7-, 2.0- and 2.2-fold greater in rumens of PM wethers at 4, 6 and 8h after allocation, respectively, compared with AM wethers. The relative abundances of archaeal and ciliate protozoal taxa were similar across allocation groups. In contrast, the relative abundances of members of the rumen bacterial community, like Prevotella 1 (P=0.04), Bacteroidales RF16 group (P=0.005) and Fibrobacter spp. (P=0.008) were greater for the AM group, whereas the relative abundance of Kandleria spp. was greater (P=0.04) for the PM group. Of these taxa, only Prevotella 1 (P=0.04) and Kandleria (P<0.001) showed a significant interaction between time of allocation and time of sampling relative to feed allocation. Relative abundances of Prevotella 1 were greater at 2h (P=0.05), 4h (P=0.003) and 6h (P=0.01) after AM allocation of new herbage, whereas relative abundances of Kandleria were greater at 2h (P=0.003) and 4h (P<0.001) after PM allocation. The early post-allocation rise in ammonia concentrations in PM rumens occurred simultaneously with sharp increases in the relative abundance of Kandleria spp. and with a decline in the relative abundance of Prevotella. All measures of fermentation and most microbial community composition data showed highly dynamic changes in concentrations and genus abundances, respectively, with substantial temporal changes occurring within the first 8h of allocating a new strip of herbage. The dynamic changes in the relative abundances of certain bacterial groups, in synchrony with a substantial diurnal variation in ammonia concentrations, has potential effects on the efficiency by which N is utilised by the grazing ruminant.     

Deep,subsurface microflora after excavation respiration and biomass and its potential role in degradation of fossil organic matter

Three types of Miocene claystones (amorphous, lamellar, and transitional) were aseptically sampled from depths of 30 m and 150 m below the soil surface. Respiration of these sediments was measured under conditions that prevented inoculation by other microorganisms not indigenous to the claystones in situ. Microbial respiration was higher in lamellar than amorphous claystones and was not affected by sampling depth. During cultivation, microbial biomass (as indicated by PLFA) significantly increased. Microbial biomass after cultivation was significantly higher in sediments from 30 m than from 150 m depth. Both microbial respiration and biomass increased after glucose addition.     

Effect of wheat hay particle size and replacement of wheat hay with wheat silage on rumen pH,rumination and digestibility in ruminally cannulated non-lactating cows

This study examined the effects on intake, diurnal rumen pH changes, rumination and digestibility of feeding ruminally cannulated non-lactating cows in a Latin square design (four cows×four periods) with four total mixed rations (TMRs) typical for lactating cows. TMRs were based on: long wheat hay or short wheat hay, wheat silage or wheat silage+1.5% NaHCO₃ buffer, as the sole roughage source (30% of TMR dry matter (DM)). The level of physically effective NDF remaining above the 8 mm screen (peNDF) was similar in the long hay and silage-based TMRs (9.45% to 9.64% of DM) and lower in the short hay TMR (7.47% of DM). The four TMRs were offered individually at 95% of ad libitum intake to avoid orts within 24 h. Cows fed long hay consumed less DM than the short hay and silage groups (9.6 v. 10.5 and 10.8 kg/day, respectively) and sorted against large hay particles at 12 h post-feeding. Under the limitations of this study (non-lactating cows fed at restricted intake) short hay TMR prevented sorting within 12 h post-feeding, encouraged rumination per kg peNDF ingested, and had higher average rumen pH (6.24), whereas preventing sub acute ruminal acidosis (SARA, defined as pH<5.8 for at least 5 h/day). In contrast, the long hay and silage-based groups were under SARA. In vitro methane production of rumen fluid was higher in the hay-fed cows than in their silage-fed counterparts, and in all treatments lower at 1 h pre-feeding than at 6 h post-feeding. In vivo DM and NDF digestibility were similar for the short hay and silage TMRs, and higher than those of the long hay TMR. Under the conditions of this study, addition of 1.5% buffer to the wheat silage TMR had no effect on intake, rumen pH, creation of SARA and digestibility.     

The effect of rumen ciliates on chitinolytic activity,chitin content and the number of fungal zoospores in the rumen fluid of sheep

The objective of this study was to investigate the effect of selected protozoa on the degradation and concentration of chitin and the numbers of fungal zoospores in the rumen fluid of sheep. Three adult ewes were fed a hay-concentrate diet, defaunated, then monofaunated with Entodinium caudatum or Diploplastron affine alone and refaunated with natural rumen fauna. The average density of the protozoa population varied from 6.1 · 10⁴ (D. affine) to 42.2 · 10⁴ cells/ml rumen fluid (natural rumen fauna). The inoculation of protozoa in the rumen of defaunated sheep increased the total activity of chitinolytic enzymes from 2.9 to 3.6 μmol N-acetylglucosamine/g dry matter (DM) of rumen fluid per min, the chitin concentration from 6.3 to 7.2 mg/g DM of rumen fluid and the number of fungal zoospores from 8.1 to 10.9 · 10⁵ cells/ml rumen fluid. All examined indices showed diurnal variations. Ciliate population density was highest immediately prior to feeding and lowest at 4 h thereafter. The opposite effects were observed for the numbers of fungal zoospores, the chitin concentration and chitinolytic activity. Furthermore, it was found that chitin from zoospores may account for up to 95% of total microbial chitin in the rumen fluid of sheep. In summary, the examined ciliate species showed the ability of chitin degradation as well as a positive influence on the development of the ruminal fungal population.