Effects of harvest date and grass species on silage cell wall components and lactation performance of dairy cows.

This study evaluated the effect of harvest date and forage species on the concentration of hydroxycinnamic acids in silage and its relationship to dairy cow performance. Tall fescue and timothy were harvested at a regular date on June 27 and July 8, respectively, or at a late date on July 8 and 25, respectively, in the first regrowth. Forage was treated with a salt-based additive and ensiled in hard-pressed round bales. Forty-seven lactating dairy cows were used in a block design. Cows received 1 of 4 treatments: (1) tall fescue harvested at regular date (RTF), (2) timothy harvested at regular date (RTI), (3) tall fescue harvested at late date (LTF), and (4) timothy harvested at late date (LTI). Diets were formulated to have the same forage-to-concentrate ratio (46:54 on a dry matter basis). Harvesting at late date increased fiber components, but only for timothy, where LTI contained greater neutral detergent fiber, acid detergent fiber, and acid detergent lignin concentrations than the other silages. Concentrations of hydroxycinnamic acids were affected by forage species, where concentrations of esterified ferulic acid and p-coumaric acid were greater for tall fescue silages than for timothy silages. Cows fed the RTI diet showed the greatest intakes of dry matter, organic matter, and crude protein. Feeding diets containing timothy silages increased milk yield and energy-corrected milk yield compared with tall fescue diets when averaged over harvest dates. Cows fed the RTI diet had greater milk protein yield than cows fed the RTF and LTF diets, and milk lactose yield was greater for cows fed diets containing timothy silage compared with tall fescue silage when averaged over harvest dates. Cows fed the LTF diet showed greater urinary N excretion compared with the LTI diet, but RTI showed the lowest urinary N and urea N excretions when calculated as percent of N intake. Cows fed diets containing timothy silage excreted more uric acid than cows fed tall fescue diets. Allantoin excretion was greater for cows eating the RTI and LTI diets compared with cows eating the RTF diet. Cows fed the RTI diet had a greater estimated microbial N flow and a greater excretion of hippuric acid than the RTF and LTF groups. In conclusion, besides the effect of harvest date on increasing the fiber components of timothy, concentrations of hydroxycinnamic acids were mainly affected by forage species; consequently, milk production was only affected by forage species. This indicates that hydroxycinnamic acids, such as ferulic acid, which cross-links to glucuronoarabinoxylans, was a major factor regulating milk production of cows fed tall fescue- and timothy silage-based diets, where lower hydroxycinnamic acid concentrations were responsible for greater milk yield.

Silage review: Recent advances and future uses of silage additives.

Additives have been available for enhancing silage preservation for decades. This review covers research studies published since 2000 that have investigated the efficacy of silage additives. The review has been divided into 6 categories of additives: homofermentative lactic acid bacteria (LAB), obligate heterofermentative LAB, combination inoculants containing obligate heterofermentative LAB plus homofermentative LAB, other inoculants, chemicals, and enzymes. The homofermentative LAB rapidly decrease pH and increase lactic acid relative to other fermentation products, although a meta-analysis indicated no reduction in pH in corn, sorghum, and sugarcane silages relative to untreated silages. These additives resulted in higher milk production according to the meta-analysis by mechanisms that are still unclear. Lactobacillus buchneri is the dominant species used in obligate heterofermentative LAB silage additives. It slowly converts lactic acid to acetic acid and 1,2-propanediol during silo storage, improving aerobic stability while having no effect on animal productivity. Current research is focused on finding other species in the Lb. buchneri group capable of producing more rapid improvements in aerobic stability. Combination inoculants aim to provide the aerobic stability benefits of Lb. buchneri with the silage fermentation efficiency and animal productivity benefits of homofermentative LAB. Research indicates that these products are improving aerobic stability, but feeding studies are not yet sufficient to make conclusions about effects on animal performance. Novel non-LAB species have been studied as potential silage inoculants. Streptococcus bovis is a potential starter species within a homofermentative LAB inoculant. Propionibacterium and Bacillus species offer improved aerobic stability in some cases. Some yeast research has focused on inhibiting molds and other detrimental silage microorganisms, whereas other yeast research suggests that it may be possible to apply a direct-fed microbial strain at ensiling, have it survive ensiling, and multiply during feed out. Chemical additives traditionally have fallen in 2 groups. Formic acid causes direct acidification, suppressing clostridia and other undesired bacteria and improving protein preservation during ensiling. On the other hand, sorbic, benzoic, propionic, and acetic acids improve silage aerobic stability at feed out through direct inhibition of yeasts and molds. Current research has focused on various combinations of these chemicals to improve both aerobic stability and animal productivity. Enzyme additives have been added to forage primarily to breakdown plant cell walls at ensiling to improve silage fermentation by providing sugars for the LAB and to enhance the nutritive value of silage by increasing the digestibility of cell walls. Cellulase or hemicellulase mixtures have been more successful at the former than the latter. A new approach focused on Lb. buchneri producing ferulic acid esterase has also had mixed success in improving the efficiency of silage digestion. Another new enzyme approach is the application of proteases to corn silage to improve starch digestibility, but more research is needed to determine the feasibility. Future silage additives are expected to directly inhibit clostridia and other detrimental microorganisms, mitigate high mycotoxin levels on harvested forages during ensiling, enhance aerobic stability, improve cell wall digestibility, increase the efficiency of utilization of silage nitrogen by cattle, and increase the availability of starch to cattle.

Efficacy of a single oral dose of a live bivalent E. coli vaccine against post-weaning diarrhea due to F4 and F18-positive enterotoxigenic E. coli.

F4- and F18-positive enterotoxigenic E. coli strains (F4-ETEC and F18-ETEC) are important causes of post-weaning diarrhea (PWD) in pigs. F4 (antigenic variant ac) and F18 (ab and ac) fimbriae are major antigens that play an important role in the early stages of infection. Herein, the efficacy of a live oral vaccine consisting of two non-pathogenic E. coli strains, one F4ac- and one F18ac-positive, was evaluated using F4ac-ETEC and F18ab-ETEC challenge models. A randomized, masked, placebo-controlled, block design, parallel-group confirmatory study with two different vaccination-challenge intervals (7 and 21 days) was conducted for each challenge model. The vaccine was administered in one dose, to ≥18-day-old piglets via drinking water. Efficacy was assessed by evaluating diarrhea, clinical observations, weight gain and fecal shedding of F4-ETEC or F18-ETEC. Anti-F4 and anti-F18 immunoglobulins in blood were measured. The vaccination resulted in significant reductions in clinical PWD and fecal shedding of F4-ETEC and F18-ETEC after the 7- and 21-day-post-vaccination heterologous challenges, except for after the 21-day-post-vaccination F4-ETEC challenge, when the clinical PWD was too mild to demonstrate efficacy. A significant reduction of mortality and weight loss by vaccination were observed following the F18-ETEC challenge. The 7-day protection was associated with induction of anti-F4 and anti-F18 IgM, whereas the 21-day protection was mainly associated with anti-F4 and anti-F18 IgA. The 7-day onset and 21-day duration of protection induced by this vaccine administered once in drinking water to pigs of at least 18days of age were confirmed by protection against F4-ETEC and F18-ETEC, and induction of F4 and F18-specific immunity. Cross protection of the vaccine against F18ab-E. coli was demonstrated for both the 7- and 21-day F18-ETEC challenges.

Description and evaluation of a net energy intake model as a function of dietary chewing index.

Previously, a linear relationship has been found between net energy intake (NEI) and dietary chewing index (CI) of the diet for different types of cattle. Therefore, we propose to generalize and calibrate this relationship into a new model for direct prediction of NEI by dairy cows from CI values (CINE; min/MJ of NE). Furthermore, we studied the forage-to-concentrate substitution rate in this new NEI model. To calibrate the model on a diverse set of situations, we built a database of mean intake from 14 production experiments with a total of 986 primi- and multiparous lactating dairy cows of different breeds fed 136 different diets ad libitum. The NEI were estimated by the Nordic feed evaluation system. The CINE value of diets was estimated from the intake of concentrate, intake of forage neutral detergent fiber (NDFf), particle length of forage, indigestible NDFf/NDFf, body weight, NDFf/body weight, and the content of NE in DM. We show that the slope values in this regression are proportional to the squared intercepts, giving the nonlinear equation NEI=NEI0-k×NEI0a×CINE, where the parameter k represents the decline in NEI with the increasing CINE of the diet and a was estimated to have a value of 2, implying a constant maximum daily chewing time. The intercept NEI0 in the regression of NEI on CINE may be interpreted as metabolic net energy intake capacity of the cows fed without physical constraints on intake. Based on experimental data, the maximum chewing time was estimated as 1/(4 × k). The NEI0 values were parameterized as a linear function of metabolic body size, energy-corrected milk yield (kg/d), days in milk, and days in milk squared. Prediction accuracy was evaluated by mean square prediction error (MSPE) and its decomposition into central tendency, regression, and disturbance, across and within experiments on independent data from 19 experiments including 812 primi- and multiparous lactating dairy cows of different breeds fed 80 different diets ad libitum. The NEI model predicted NEI with an MSPE of 8% of observed, and across the 19 experiments the error central tendency, error regression, and error disturbance were 4.2, 40.6, and 84.9% of MSPE, respectively. The described intake model implies a variable forage-to-concentrate substitution rate as a nonlinear function of NEI0, CINE of forage, and supplementation of concentrate.

Variations in automatically recorded rumination time as explained by variations in intake of dietary fractions and milk production, and between-cow variation.

Individual recording of rumination time (RT) is now possible in commercial dairy herds, through development of a microphone-based sensor, which is able to record RT by the sound of rumination activity. The objectives of this study were to examine the relationship between daily RT and intakes of different dietary fractions, the relationship between RT in minutes per kilogram of dry matter intake (DMI) and milk production, and to examine the variation in RT within and between mid-lactating dairy cows. Data from 3 production trials were used in which a total of 27 different diets were fed. The data contained 761, 290, and 203 daily recordings of RT, milk yield, milk components, DMI, and intake of dietary fractions recorded on 29, 26, and 24 Holstein and Swedish Red cows from trials 1, 2, and 3, respectively. The dietary fractions included forage neutral detergent fiber (NDF), concentrate NDF, crude protein, sugar, starch, and the remaining fraction represented by organic matter--(forage NDF+concentrate NDF+crude protein+sugar+starch). The relationship between the dietary fractions and RT was analyzed in 2 steps. In step 1, the dietary fractions, which were significantly related to RT, were selected and simultaneously checked for multicollinearity between the dietary components; in step 2, a multivariate model, including the effect of repeated measurements, the main effect of the selected dietary fractions from step 1, random effects of cow(trial) and trial, and information on breed, days in milk, and parity was used to analyze the relationship between RT and the selected dietary fractions. Relationships between RT in minutes per kilogram of DMI and milk yield and milk components were analyzed, using the same multivariate model as in step 2. Approximately 32% of the variation in daily RT could be explained by variations in intakes of the dietary fractions, whereas 48% of the total variation in RT was accounted for by individual variations between cows. Intakes of forage NDF and starch were positively related to daily RT, whereas intakes of sugar and the remaining fraction were negatively related to daily RT. Rumination time in minutes per kilogram of DMI was negatively related to milk yield and protein percentage, but positively related to milk fat percentage.

Status of vitamins E and A and ß-carotene and health in organic dairy cows fed a diet without synthetic vitamins.

Synthetic vitamin supplementation is not consistent with organic production, so it is important to investigate whether dairy cows can maintain their health and production without synthetic vitamins being added to their diet. In basic dairy cow diets, provitamin A (ß-carotene) and vitamin E are mainly found in pasture and in grass and legume silages, but the concentrations are highly variable. This study compared the vitamin status and health of cows without synthetic vitamin supplementation (NSV group) with control cows (CON group) fed synthetic vitamins according to Swedish recommendations (600 IU of vitamin E and 80,000 IU of vitamin A per cow per day) to investigate whether dairy cows can fulfill their requirements of vitamins A and E without supplementation with synthetic vitamins. Vitamin concentrations in blood plasma and milk, health, fertility, milk yield, and milk composition were measured in Swedish Holstein cows (n=28) during 2 complete lactations. All cows were fed a 100% organic diet containing grass-legume silage, cold-pressed rapeseed cake, peas, cereal grains, and minerals. Blood samples were collected from each cow 3 wk before expected calving, at calving, and 3 wk, 3 to 5 mo, and 7 to 9 mo after calving. Samples of colostrum were taken and milk samples were collected 4d after calving and at the same time as the 3 blood samplings after calving. The only difference in vitamin status between groups was found in colostrum in yr 1, when CON cows tended to have a higher concentration of α-tocopherol, and their ß-carotene concentration was higher compared with NSV cows. The NSV cows tended to have more cases of mastitis than CON cows in yr 2. Within the NSV group, fewer cows were healthy and more cases of mastitis were observed in yr 2 than in yr 1. The groups did not differ in production parameters. In conclusion, the vitamin status in blood and milk of the studied cows indicated that cows in organic dairy production can fulfill their requirements of vitamins A and E without any supplementation of synthetic vitamins, except at the time around calving, when the requirements are high. However, the impaired health of NSV cows in yr 2 may indicate a long-term negative health effect in cows fed no synthetic vitamins.

Effect of stage of maturity of grass at harvest on intake, chewing activity and distribution of particle size in faeces from pregnant ewes.

This study was conducted to investigate the effect of stage of maturity at harvest on the intake of grass silage, eating and ruminating activity and the distribution of faecal particle size in ewes during late pregnancy. A total of 18 Swedish Finull x Dorset 85 ± 8 kg (mean 6 ± s.d.) ewes bearing twins were randomly assigned to three dietary treatments 6 weeks before lambing. The treatments included ad libitum feeding with early harvested (EH), medium harvested (MH) or late harvested (LH) primary-growth grass silage with 45%, 58% and 63% NDF on a dry matter (DM) basis, respectively. Intake and chewing activity were recorded and faeces were sampled over 4 continuous days for each individual ewe. The faeces samples were washed in nylon bags, freeze dried and sieved with pore sizes from 2.4mm to 0.1 mm; particles less than 0.1mm in size were also collected. Subsamples of each sieving fraction were scanned and the dimensions of the individual particles in each sieving fraction were measured by image analysis. In addition, the number of particles longer than 7 mm was counted from the particles retained on a sieve with a pore size of 2.4mm using a simple wet sieving technique. The time spent eating and ruminating per kg of DM intake was affected by the stage of maturity at harvest; it was shorter in ewes fed EH compared with ewes fed MH and LH ( P < 0.05). In comparison with feeding LH, feeding EH resulted in the retention of a larger proportion of particles in the lower and upper sieve fractions (< 0.2mm and > 1 mm, respectively, P < 0.01), a smaller mean particle size ( P < 0.05) and a smaller mean particle width in faeces ( P < 0.01). The results from the simple wet sieving technique confirmed the results from dry sieving and image analysis, showing a higher number of large particles in faeces from ewes fed the EH compared with the ewes fed the MH and LH (P < 0.001). In conclusion, the distribution of faecal particle size might be considered as a footprint of the characteristics of forage fibre eaten by ewes.

Intake and digestion of whole-crop barley and wheat silages by dairy heifers.

The effect of maturity at harvest on the digestibility and intake of large bale silage made from whole-crop barley and wheat when fed to growing heifers was evaluated. Two crops of spring barley (Hordeum distichum cv. Filippa and Kinnan) and 1 of winter wheat (Triticum aestivum cv. Olevin) were harvested at the heading, milk, and dough stages of maturity. The silage was fed to 36 dairy heifers in a balanced crossover experiment with 3 periods and 9 treatments (diets based on 3 crops and 3 stages of maturity), organized into 6 pairs of 3 × 3 Latin squares. No clear relationship was observed between intake and stage of maturity of whole-crop cereal silage, but intake was positively correlated to silage DM content (P < 0.001, r = 0.46) and negatively correlated to NDF content (P < 0.001, r = -0.42). Organic matter digestibility decreased between the heading and milk stages of maturity in all crops (P < 0.001), did not differ between the milk and the dough stages in the 2 barley crops, but increased in the wheat silage (P = 0.034). The NDF digestibility decreased between the heading and milk stages in all crops (P < 0.001), whereas it decreased in 1 barley crop (P < 0.001), increased in the other barley (P = 0.025), and was unchanged in the wheat between the milk and dough stages of maturity. Starch digestibility was less in the 2 barley crops compared with the wheat at the dough stage of maturity (P < 0.001). The feeding value of the whole-crop barley and wheat declined between the heading and milk stages of maturity, but thereafter the effect of maturity on the feeding value was minor.

Influence of vitamins A, D3 and E status on post-mortem meat quality in steers under winter housing or pasture finishing systems.

We investigated the influence of Swedish recommended vitamins A, D3 and E supplementation levels on muscle tenderness and fatty acid (FA) composition under indoor or outdoor finishing programmes. Swedish Red breed steer calves were divided into vitamin supplemented (n = 12) and non-supplemented (n = 15) groups while on pasture prior to the finishing period. This trial began at the beginning of the winter housing period during which the steers were fed a 55 : 45 dry matter barley : grass silage diet indoors. The indoor finished group was comprised of vitamin supplemented (n = 6) and non-supplemented (n = 8) steers slaughtered after about 155 days on feed. Vitamin supplemented steers were provided with 100 g mineral supplement providing 400 000 IU vitamin A, 100 000 IU D3 and 3000 IU E daily as recommended for Swedish production practices. In spring, outdoor finished vitamin supplemented (n = 6) and non-supplemented (n = 7) steers grazed semi-natural grassland for an additional 120 days before slaughter. During pasture, vitamin supplemented steers had free-choice access to a mineral supplement containing vitamins A, D3 and E. The mineral supplement for the non-supplemented steers did not contain vitamins A, D3 and E and was provided at the same amount as the vitamin supplemented steers. Shear force values were similar between vitamin supplemented and non-supplemented steers after ageing 2, 7 and 14 days within indoor and outdoor finishing programmes. The shear force values had decreased by 14 days of ageing within all programmes. The µ- and m-calpain activity did not differ between vitamin supplemented and non-supplemented steers for either the indoor or outdoor finishing programmes. The calpastatin activity was higher for the indoor, vitamin supplemented steers. Indoor finished vitamin supplemented steers had a greater proportion of C18:1c-9 and total monounsaturated fatty acids, whereas the non-supplemented steers had a greater proportion of total saturated fatty acids. We concluded that the meat quality from steers not receiving vitamin supplementation was similar to that of steers receiving vitamins A, D3 and E supplementation at Swedish recommended levels under indoor and outdoor finishing programmes.

Effects of physical form and stage of maturity at harvest of whole-crop barley silage on intake, chewing activity, diet selection and faecal particle size of dairy steers.

This study examined the effects of physical form and stage of maturity at harvest of whole-crop barley silage (WCBS) on feed intake, eating and rumination activity, diet selection and faecal particle size in dairy steers. Whole-crop barley was harvested and ensiled in round bales. Eight dairy steers (live weight (LW): 350 ± 10 kg) in a duplicated 4 × 4 Latin square design were fed WCBS harvested at heading or dough stage of maturity in long form or chopped in a 2 × 2 factorial treatment arrangement. The WCBS was supplemented with soybean meal. Daily dry matter (DM) intake increased by 7% (P < 0.05) due to chopping of dough-stage silage but was unaffected by chopping of heading-stage silage. The steers fed chopped, but not those fed long dough-stage silage, selected for starch in the WCBS (P < 0.001). The neutral detergent fibre (NDF) intake was 5% higher (P < 0.01) for heading than for dough-stage silage and was associated with lower concentration of indigestible NDF (96 v. 170 g/kg DM). Rate of intake of DM and NDF was 37% higher (P < 0.001), daily eating time was 24% shorter (P < 0.001) and daily chewing time was 8% shorter (P < 0.05) for chopped silage compared with long silage but there was no effect of maturity. Daily rumination time was not affected by treatments, whereas rumination and chewing times per kg NDF intake were 15% and 13% higher (P < 0.05), respectively, for dough-stage than for heading-stage silage. The proportion of faecal particles retained on a 1 mm screen was 30% higher (P < 0.001) due to chopping and 45% higher (P < 0.001) due to delayed harvest. Chopping the dough-stage silage reduced the proportion of grain in faeces from 97 to 43 g/kg DM (P < 0.05) indicating higher starch digestibility. In conclusion, chopping increased DM intake of WCBS when harvested at dough stage but not at heading stage of maturity.

Digestibility of whole-crop barley and oat silages in dairy heifers.

This study evaluated the digestibility of whole-crop cereal silage (WCCS) made from oats and six-rowed barley harvested at the heading, early milk and early dough stages, and two-rowed barley harvested at the early milk and early dough stages of maturity. The eight WCCSs were fed to 32 Swedish Red heifers in a changeover design over three periods of 28 days each. The heifers were first fed ad libitum for 17 days and then at 0.95 of ad libitum for 11 days of each period. During the last 5 days all faeces and orts were collected to determine the digestibility of the silages. Only the maturity stage effect was significant for the WCCS organic matter (OM) digestibility and the average OM digestibility was higher at the heading stage (698 g/kg) than at early milk (647 g/kg) and early dough (652 g/kg) stages of maturity. For neutral detergent fibre (NDF) digestibility the crop × maturity stage effect was significant. The NDF digestibility decreased from the heading to the early milk stage for both six-rowed barley (746 to 607 g/kg) and oats (698 to 596 g/kg). There was no further significant decrease in NDF digestibility for six-rowed barley at the early dough stage (577 g/kg), but for two-rowed barley it decreased from the early milk (682 g/kg) to the early dough (573 g/kg) stage, and also for oats the NDF digestibility was lowest at the early dough stage (507 g/kg). The decrease in NDF digestibility during maturation was to a large extent compensated by an increase in starch concentration in the crops. The starch digestibility was lower for six-rowed barley at early dough stage (948 g/kg) than at early milk stage (977 g/kg), and was also lower compared with oats (979 g/kg) at early dough stage. The average crude protein (CP) digestibility was higher at the heading (646 g/kg) and the early milk (642 g/kg) stages than at the early dough stage (599 g/kg), and oats had higher average CP digestibility (650 g/kg) than six-rowed (613 g/kg) and two-rowed (624 g/kg) barley. Delaying the harvest of WCCS from the heading to the early milk and dough stage of maturity will decrease the OM digestibility; as a result there is a decreased NDF digestibility.

Administration of Pediococcus acidilactici or Saccharomyces cerevisiae boulardii modulates development of porcine mucosal immunity and reduces intestinal bacterial translocation after Escherichia coli challenge.

In this study, the influence of the probiotics, Pediococcus acidilactici (PA) and Saccharomyces cerevisiae boulardii (SCB), on intestinal immune traits and resistance to enterotoxigenic Escherichia coli (ETEC) infection was evaluated in pigs. Two weeks before farrowing, 30 sows and their future litters were allocated to the following treatments: 1) control group without antibiotic or probiotic treatment (CTRL), 2) control with antibiotic (tiamulin) added to weanling feed (ABT), or litters treated with 3) PA, 4) SCB, or 5) PA+SCB from 24 h after birth. During lactation, PA, SCB, or PA+SCB were given to piglets 3 times a week by gavage. After weaning at 21 d of age, probiotics or ABT were added to the diet. Four pigs per litter were chosen to evaluate performance and blood concentrations of folic acid and vitamin B(12). Three of these were orally challenged with an ETEC strain on d 49 to 51 and killed on d 52. Three piglets from the rest of the litter were slaughtered on d 18 and 3 others on d 24. Blood, ileum, and mesenteric lymph node (MLN) samples were taken to characterize leukocyte populations, determine IgA concentrations in ileal flushes, and evaluate bacterial translocation in MLN. No treatment effect on postweaning performance and on blood concentrations of folic acid and vitamin B(12) was observed. In the ileum, the percentage of CD4(-)CD8(+low) T cells was greater (P = 0.05) in 18-d-old nursed piglets treated with PA than in those of the CTRL and PA+SCB groups. In the MLN, the percentage of CD8(+) T cells was not affected by any of the treatments at d 18 and 24 but decreased (P = 0.006) after weaning. In the blood, CD8(+) T cells were not affected by treatments or weaning. After the ETEC challenge (d 52), bacterial translocation to MLN was reduced (P = 0.05) in pigs treated with PA, SCB, PA+SCB, or ABT compared with CTRL. No treatment effect was observed on blood leukocyte populations after ETEC challenge, although a time effect (d 42 vs. 52) indicated that blood CD4(+) and gammadelta-T lymphocytes were increased (P < 0.05) on d 52 compared with d 42, whereas CD4(-)CD8(+low) T lymphocytes and monocytes were markedly reduced (P < 0.01). Finally, the IgA concentration in ileal flushes collected on d 42 and 52 was greater in SCB and CTRL piglets than in ABT and PA piglets. In conclusion, probiotics may have the potential to modulate establishment of lymphocyte populations and IgA secretion in the gut and to reduce bacterial translocation to MLN after ETEC infection.

Escherichia coli: on-farm contamination of animals.

Escherichia coli is one of the main inhabitants of the intestinal tract of most mammalian species, including humans, and birds. Shiga toxin-producing E. coli (STEC), also called verotoxinogenic E. coli, usually do not cause disease in animals but may cause watery diarrhoea, haemorrhagic colitis, and/or haemolytic uraemic syndrome in humans. Zoonotic STEC include the O157:H7 strains and, with increasing frequency, certain non-O157 strains. The importance of non-O157 zoonotic strains is probably underestimated as they have been less well characterised and are more difficult to detect in samples than O157:H7. Another large subset of STEC strains has been isolated from animals but has not, at the present time, been associated with disease in animals or humans. Cattle and other ruminants are the most important reservoir of zoonotic STEC, which are transmitted to humans through the ingestion of foods or water contaminated with animal faeces, or through direct contact with the infected animals or their environment. The main sources of STEC infection of cattle on-farm are the drinking water, the feed, and the immediate environment of the animal. Risk factors that have been identified for infection of animals with O157 STEC include age, weaning, movement of the animals, season, feed composition, and the ability of the bacteria to persist in the environment. On-farm control of the zoonotic risk of human infection with STEC should primarily target the main source of contamination: the animal reservoir. Various strategies to reduce intestinal colonisation of cattle by zoonotic STEC have been tried with varying results, including vaccination, treatment with probiotics, such as direct-fed microbials or competitive exclusion, administration of bacteriophages, and modification of the diet.

Intake, digestibility, and composition of orchardgrass and alfalfa silages treated with cellulase, inoculant, and formic acid fed to lambs.

The objectives of this study were to determine the effect of a cellulase (from Trichoderma longibrachiatum) alone or combined with a bacterial inoculant (Lactobacillus plantarum and Pediococcus cerevisiae) or formic acid on composition, intake, and digestibility of orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.) silages. Orchardgrass and alfalfa were harvested at the early heading stage and at the early bloom stage of maturity and wilted to approximately 22 and 32% DM, respectively. Forages were then ensiled in 100-L sealed barrels for at least 60 d before they were fed to lambs. Silage treated with cellulase had lower (P < .001) pH and lower (P < .001) acetic acid and NH3 N concentrations than untreated silage of both plant species and a higher (P = .004) lactic acid concentration than the control treatment of alfalfa silage. Fermentation characteristics of cellulase-treated silages, especially of alfalfa, were further enhanced by use of inoculant. Formic acid addition increased (P < .001), reducing sugar concentration of cellulase-treated orchardgrass and alfalfa silage by 90 and 154%, respectively, and decreased (P < .001) NH3 N concentration of cellulase-treated alfalfa silage by 19%. Averaged across plant species, cellulase, combined with inoculant or formic acid, resulted in 8 and 13% greater (P = .03) DMI, respectively, than the control silage. Extensive enzymatic cell-wall degradation during ensiling decreased (P = .003) NDF intake of cellulase-treated orchardgrass silage by 25% and decreased (P = .001) cellulose intake by 23%, when averaged across plant species. Addition of formic acid increased (P = .003) NDF intake of cellulase-treated orchardgrass silage by 19%. Averaged across species, cellulase application decreased (P < .05) silage NDF digestibility by 18%. Greater sugar and lower acetic acid, NH3 N, and NDF concentrations resulted in greater DMI of cellulase-treated silage than of control silage, when cellulase was combined with formic acid or inoculant.

Enzyme, bacterial inoculant, and formic acid effects on silage composition of orchardgrass and alfalfa.

We evaluated the effects of cellulase (from Trichoderma longibrachiatum) application rates on neutral detergent fiber (NDF) concentration and fermentation products of orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.) silages harvested with decreasing dry matter (DM) digestibility. Additionally, the impacts of inoculant (Lactobacillus plantarum and Pediococcus cerevisiae), pectinase (from Aspergillus niger), or formic acid on silage composition were studied. Forages wilted to a DM content of about 320 g/kg were ensiled in laboratory silos for 60 d. Cellulase, combined with inoculant, was applied at 2, 10, and 20 ml/kg of herbage (at least 2500 IU/ml). Cellulase at 10 ml/kg was also applied alone or in combination with pectinase and inoculant or formic acid. The NDF concentration of orchardgrass silage decreased with increasing cellulase up to 20 ml/kg, at which NDF content was decreased by 30%. The NDF concentration of alfalfa silage decreased with increasing cellulase application up to 10 ml/kg, at which NDF content was decreased by 13%. Immature plants were more responsive to cellulase treatment than mature plants. Cellulase at 2 ml/kg combined with inoculant improved fermentation characteristics of the silages but generally, there was no effect on silage fermentation by higher cellulase applications, resulting in an accumulation of sugar. The improved fermentation of orchardgrass treated with cellulase and inoculant was mostly related to the effect of inoculant, whereas cellulase alone improved fermentation characteristics of alfalfa silage and this effect was enhanced by addition of inoculant. Decreased NDF and increased sugar concentrations did not improve the in vitro DM digestibility of cellulase-treated silages.

Multiple phosphorylated forms of the Saccharomyces cerevisiae Mcm1 protein include an isoform induced in response to high salt concentrations.

The Saccharomyces cerevisiae Mcm1 protein is an essential multifunctional transcription factor which is highly homologous to human serum response factor. Mcm1 protein acts on a large number of distinctly regulated genes: haploid cell-type-specific genes, G2-cell-cycle-regulated genes, pheromone-induced genes, arginine metabolic genes, and genes important for cell wall and cell membrane function. We show here that Mcm1 protein is phosphorylated in vivo. Several (more than eight) isoforms of Mcm1 protein, resolved by isoelectric focusing, are present in vivo; two major phosphorylation sites lie in the N-terminal 17 amino acids immediately adjacent to the conserved MADS box DNA-binding domain. The implications of multiple species of Mcm1, particularly the notion that a unique Mcm1 isoform could be required for regulation of a specific set of Mcm1's target genes, are discussed. We also show here that Mcm1 plays an important role in the response to stress caused by NaCl. G. Yu, R. J. Deschenes, and J. S. Fassler (J. Biol. Chem. 270:8739-8743, 1995) showed that Mcm1 function is affected by mutations in the SLN1 gene, a signal transduction component implicated in the response to osmotic stress. We find that mcm1 mutations can confer either reduced or enhanced survival on high-salt medium; deletion of the N terminus or mutation in the primary phosphorylation site results in impaired growth on high-salt medium. Furthermore, Mcm1 protein is a target of a signal transduction system responsive to osmotic stress: a new isoform of Mcm1 is induced by NaCl or KCl; this result establishes that Mcm1 itself is regulated.

Kinetics of cell-wall digestion of orchardgrass and alfalfa silages treated with cellulase and formic acid.

The objectives of this study were to determine the effects of cellulase (from Trichoderma longibrachiatum) combined with formic acid, applied before ensiling, on the subsequent concentration and composition of the cell wall and on the extent and rate of in situ cell-wall digestion of orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.). Treated and control forages of both plant species were ensiled for at least 60 d before being ruminally digested by two fistulated cows. Analyses of NDF, ADF, and acid detergent lignin were conducted sequentially on original and digested samples. Data were fitted with a first-order, nonlinear model to estimate extents and rates of digestion of potentially digestible NDF, cellulose, and hemicellulose. The concentration of indigestible residue and the discrete lag time before digestion were also determined for the cell-wall components. After ensiling, the mean NDF concentration of treated silages was 19% lower than that of control silages; the effect was greater for orchardgrass than for alfalfa. The extent of digestion of NDF, cellulose, and hemicellulose, respectively, was 33, 37, and 27% lower for treated silages than for control silages. Treatment effects on the extent of digestion varied between plant species. Cellulose from treated orchardgrass was digested 19% more slowly than cellulose from the control silage. Indigestible residue concentrations of NDF, cellulose, and hemicellulose, respectively, were 7, 8, and 7% lower in treated silages than in control silages. Thus, extensive cell-wall degradation by cellulase during ensiling resulted in less digestible cell-wall material for ruminal digestion but greater total cell-wall degradation, including that during ensiling and ruminal incubation, especially during early digestion in the rumen.