Repeated ruminal acidosis challenges in lactating dairy cows at high and low risk for developing acidosis: feeding, ruminating, and lying behavior.

An experiment was conducted to determine whether the susceptibility to ruminal acidosis, as defined through differences in days in milk (DIM), milk production level, and ration composition, influences cow feeding, ruminating, and lying behavior and whether these behaviors change during an acute bout of ruminal acidosis. Eight ruminally cannulated cows were assigned to 1 of 2 acidosis risk levels: low risk (LR, mid-lactation cows fed a 60:40 forage:concentrate ratio diet) or high risk (HR, early lactation cows fed a 45:55 forage:concentrate diet). As a result, diets were intentionally confounded with DIM and milk production to represent 2 different acidosis risk scenarios. Cows were exposed to an acidosis challenge in each of three 14-d periods. Each period consisted of 3 baseline days, a feed restriction day (restricting total mixed ration to 50% of ad libitum intake), an acidosis challenge day (1 h meal of 4 kg of ground barley/wheat before allocating the total mixed ration), and a recovery phase. Feeding, rumination, and standing/lying behavior were recorded for 2 baseline days, on the challenge day, and 1 and 4 d after the challenge day for each cow. Across the study, there were no differences in measures of standing, lying, or feeding behavior between the 2 groups of cows. The HR cows did, on average, spend less time ruminating (491 vs. 555 min/d) than the LR cows, resulting in a lesser percentage of observed cows ruminating across the day (44.6 vs. 48.1%). The acidosis challenge resulted in changes in behavior in all cows. Compared with the baseline, feeding time increased on the first day after the challenge (395 vs. 310 min/d), whereas lying time decreased (565 vs. 634 min/d). Rumination time decreased the first day following the challenge (436 min/d) relative to the baseline (533 min/d), but increased the following day (572 min/d). Fewer cows were observed to be ruminating at a given time on the first day following the challenge as compared with the baseline period. Despite this, on a herd level, numerous observations of the proportion of cows ruminating at any one time would need to be taken to accurately detect an acute bout of acidosis using changes in rumination behavior. Overall, these results suggest that risk of acidosis may have little overall effect on general behavior, with the exception of rumination. Furthermore, an acute bout of acidosis alters behavioral patterns of lactating dairy cows, particularly rumination behavior, and identification of these changes in behavior through repeated measurements may assist in the detection of an acidosis event within a herd.

Repeated ruminal acidosis challenges in lactating dairy cows at high and low risk for developing acidosis: feed sorting.

An experiment was conducted to determine whether the susceptibility of cows to ruminal acidosis influences feed sorting and whether feed sorting changes during a bout of ruminal acidosis. Eight ruminally cannulated cows were assigned to 1 of 2 acidosis risk levels: low risk (LR, mid-lactation cows fed a 60% forage diet) or high risk (HR, early lactation cows fed a 45% forage diet). As a result, diets were intentionally confounded with milk production to represent 2 different acidosis risk scenarios. Cows were exposed to an acidosis challenge in each of two 14-d periods. Each period consisted of 3 baseline days, a feed restriction day (restricting TMR to 50% of ad libitum intake), an acidosis challenge day (1-h meal of 4 kg of ground barley/wheat before allocating the TMR), and a recovery phase. Ruminal pH was measured continuously for the first 9 d of each period using an indwelling system. Feed and orts were sampled for 2 baseline days, on the challenge day, and 1 and 3 d after the challenge day for each cow and subjected to particle size analysis. The separator contained 3 screens (18, 9, and 1.18 mm) and a bottom pan to determine the proportion of long, medium, short, and fine particles, respectively. Sorting was calculated as the actual intake of each particle size fraction expressed as a percentage of the predicted intake of that fraction. All cows sorted against the longest and finest TMR particles and sorted for medium-length particles. Sorting was performed to a greater extent by the HR cows, and this sorting was related to low ruminal pH. Both HR and LR cows altered their sorting behavior in response to acidosis challenges. For the HR cows, severe acidosis was associated with increased sorting for the longer particles in the diet and against the shorter particles, likely to lessen the effects of the very.

Repeated ruminal acidosis challenges in lactating dairy cows at high and low risk for developing acidosis: ruminal pH.

The primary objective of this experiment was to determine whether lactating dairy cows that are at high (HR) or low (LR) risk for experiencing ruminal acidosis, because of their diet and stage of lactation, differ in their response to an acidosis challenge. A secondary objective was to determine whether the severity of acidosis changes with repeated challenges. The experiment was a completely randomized design with 2 groups (risk scenarios, HR vs. LR) and 3 periods corresponding to 3 repeated acidosis challenges. Eight lactating ruminally cannulated cows were assigned to 1 of 2 groups: HR, early lactation cows fed a 45% forage diet, or LR, midlactation cows fed a 60% forage diet. Cows were exposed to 3 acidosis challenges, each separated by 14 d. The challenge consisted of restricting total mixed rations to 50% of ad libitum intake for 24 h, followed by a 1-h meal of 4 kg of ground barley-wheat before allocating the total mixed rations. Ruminal pH was measured continuously for 9 of the 14 d each period using an indwelling system. Subacute acidosis (SARA) was described at 2 thresholds: pH <5.8 and pH <5.5. As expected, HR cows had lower ruminal pH profiles (curves) compared with LR cows: mean pH (5.81 vs. 6.21) and nadir pH (5.13 vs. 5.53). The HR cows also experienced SARA to a greater extent than LR cows during the experiment (pH <5.8, 10.6 vs. 3.5 h/d; pH <5.5, 5.9 vs. 1.6 h/d). The pH profiles of cows in both risk categories decreased with each challenge period; mean pH was 6.13, 6.03, 5.77, and nadir pH was 5.52, 5.34, and 5.14 in periods 1, 2, and 3, respectively. The challenges caused a similar decrease in pH for cows in both risk categories, but because the HR cows had a lower baseline pH, they experienced more severe SARA with each subsequent challenge. Feed restriction the day before administering the acidosis challenge caused ruminal pH to gradually increase. On the challenge day, the entire grain allotment was consumed by all cows in period 1, six cows in period 2, and only 3 cows in period 3. The pH plummeted immediately after each grain challenge. Ruminal pH remained very low during the first day after the challenge for all cows, but LR cows began their recovery more quickly than HR cows. Regardless of risk category, with each successive challenge, the pH decrease on the challenge day was more severe: nadir pH on the challenge day was 5.19, 5.07, and 4.90 and duration of SARA (pH <5.8) was 12.2, 13.4, and 15.8 h/d in periods 1, 2, and 3. This study indicates that cows become more prone to acidosis over time even though they decrease intake of the challenge grain to avoid acidosis. The severity of each subsequent bout of acidosis increases, especially for cows fed diets low in physically effective fiber and at high acidosis risk. Therefore, a bout of acidosis that occurs due to improper feed delivery or poor diet formulation can have long-term consequences on cow health and productivity.

Nitrogen and mineral balance of lambs artificially infected with Haemonchus contortus and fed tanniferous sainfoin (Onobrychis viciifolia).

Tanniferous temperate legumes are assumed to possess anthelmintic properties, but it is unclear whether this is the direct result of condensed tannins (CT) or is mediated indirectly via an improved metabolic protein supply. A metabolism experiment was conducted to differentiate between these factors by feeding the CT plant sainfoin (19.7% CP in DM) to lambs infected with the abomasal blood-sucking nematode Hemonchus contortus. A total of 18 infected lambs were fed sainfoin either untreated or treated with polyethylene glycol, a CT-inactivating agent, or a grass-clover mixture (13.2% CP in DM) over 3 wk (n = 6). Six uninfected lambs received the grass-clover mixture as a control. In addition to indicators of infection (fecal egg count, packed-cell volume, abomasal worm burden, and serum protein), nutrient digestibility, the balance of N and selected minerals, ruminal fluid characteristics, and plasma AA levels were determined mostly in the final experimental week. The specific effects of the sainfoin CT, the extra CP with sainfoin, and the infection were statistically evaluated by contrast analysis. The sainfoin CT exerted no beneficial effects on resilience to nematode infection and exerted only minor effects on ruminal ammonia or blood urea concentrations and the excretory pattern of N. Plasma alanine, aspartate, and proline concentrations tended to be greater (P < or = 0.09) because of the sainfoin CT, whereas the other AA remained unaffected. Intake of the mineral supplement was lower (P < 0.001) for lambs fed sainfoin compared with those fed sainfoin treated with polyethylene glycol. Feeding the high-protein sainfoin instead of the grass-clover mixture increased (P < 0.001) N retention and apparent OM digestibility, whereas digestibility of NDF and ADF were decreased (P < 0.001). Feeding sainfoin also decreased (P < or = 0.04) plasma alanine, glycine, isoleucine, and total nonessential AA compared with the grass-clover mixture. Although fecal egg count, worm burden, and packed cell volume were not affected by the greater CP supply associated with sainfoin feeding, serum albumin level was increased (P = 0.008). The lack of effects of sainfoin on resilience to nematode infection might have been the result of the unexpectedly low CT content (3.6% in DM) of the material used. It cannot be excluded that longer term feeding of this batch of sainfoin might have been effective.

Effects of supplemental hay and corn silage versus full-time grazing on ruminal pH and chewing activity of dairy cows.

Grazing young, highly digestible swards with and without supplemental hay or corn silage (5.5 kg of DM/d) offered overnight was tested for its effects on ruminal pH and chewing activity. A double 3 x 3 Latin square arrangement with 6 rumen-cannulated Brown Swiss cows (29 kg/d of milk) was applied. Herbage intake was quantified by controlled-release alkane capsules. Chewing activity was determined using an automatic microcomputer-based system for digital recording of the jaw movements. Except during milking, ruminal pH was measured continuously over 7 d by applying a device consisting of an indwelling pH electrode and a data-recording unit integrated in the cannula's cover. The grazing system had no significant effect on body weight, milk yield or composition (except milk urea), or total DM intake (13.5, 13.8, and 15.7 kg/d with full-time grazing, hay, and corn silage supplementation). No differences occurred for ruminating time per day and time per kilogram of DM intake. Full-time grazing cows spent more time eating per day (+26%) and time per kilogram of DM intake (+31%) than the other cows. Ruminal pH and time with pH <5.8 at night did not differ. Throughout the day, hay-supplemented cows had a significantly lower pH (-0.23) than full-time grazing cows, and the period of pH <5.8 was longer compared with corn-silage fed cows (77 vs. 11 min). Nocturnal supplement feeding gave no advantage over full-time grazing, and supplemental hay led to lower daytime pH.

Role of 5S RNA in assembly and function of the 50S subunit from Escherichia coli.

Total reconstitution experiments performed under various conditions revealed that 5S RNA plays an important role during the last assembly step in vitro leading to an active 50S particle. For the preceding steps this RNA species is dispensable. However, 50S RNA can be integrated efficiently during any of the assembly steps in vitro. The 47S particle, reconstituted in two steps and lacking 5S RNA, shows low but significant activity in many functional tests. High activity could be obtained by incubating this particle with 5S RNA alone, demonstrating the importance of the 5S RNA in generating an active ribosomal conformation. In particular, the activity of the peptidyltransferase (peptidyl-tRNA:aminoacyl-tRNA N-peptidyltransferase; EC 2.3.2.12) center is drastically influenced by 5S RNA. No significant factor-dependent tRNA binding to the A-site was observed with the 47S particle, in contrast to the corresponding P-site binding. The elongation factor G dependent GTPase activity was not affected by the lack of 5S RNA.

Total reconstitution of functionally active 50S ribosomal subunits from Escherichia coli.

Total reconstitution of 50S subunits from E. coli was achieved by a two-step incubation procedure. In the first step, 23S RNA, 5S RNA, and the total proteins from 50S subunits were incubated for 20 min at 40 degrees in the presence of 4 mM Mg(++) and 400 mM NH(4)Cl. In the second step, the Mg(++) concentration was raised to 20 mM and the incubation was performed for 90 min at 50 degrees . No requirement for 30S subunits or other components (e.g., polyamine) was found. The reconstituted particle has the same sedimentation coefficient as the native 50S subunit and is highly active in protein synthesis with natural (R17 RNA) and artificial [poly(U)] messengers as well as in tests for peptidyltransferase (fragment assay) and for binding of antibiotics (chloramphenicol).

Ruminal methanogenesis as influenced by individual fatty acids supplemented to complete ruminant diets.

The objective of the present study was to investigate the effects of seven different pure fatty acids on rumen fermentation using the rumen simulation technique (RUSITEC). The fatty acids were supplied to a complete ruminant diet at a proportion of 50 g x kg(-1) dietary dry matter and compared with an unsupplemented control. Methane release and methanogenic counts were suppressed by the fatty acids C12 : 0, C14 : 0 and C18 : 2 whereas C8 : 0, C10 : 0, C16 : 0 and C18 : 0 showed no corresponding effects. Apart from C12 : 0 and C18 : 2, C8 : 0 and C10 : 0 also adversely affected ciliate protozoa suggesting independence from the methane-suppressing effect of medium-chain fatty acids (MCFA). Although MCFA but not C18 : 2 reduced ruminal fibre degradation, the influence on other fermentation traits remained low. In conclusion, the supply of certain fatty acids to ruminant diets seems to have the potential to reduce methane release.

Secondary structure model for 23S ribosomal RNA.

A secondary structure model for 23S ribosomal RNA has been constructed on the basis of comparative sequence data, including the complete sequences from E. coli. Bacillus stearothermophilis, human and mouse mitochondria and several partial sequences. The model has been tested extensively with single strand-specific chemical and enzymatic probes. Long range base-paired interactions organize the molecule into six major structural domains containing over 100 individual helices in all. Regions containing the sites of interaction with several ribosomal proteins and 5S RNA have been located. Segments of the 23S RNA structure corresponding to eucaryotic 5.8S and 25 RNA have been identified, and base paired interactions in the model suggest how they are attached to 28S RNA. Functionally important regions, including possible sites of contact with 30S ribosomal subunits, the peptidyl transferase center and locations of intervening sequences in various organisms are discussed. Models for molecular 'switching' of RNA molecules based on coaxial stacking of helices are presented, including a scheme for tRNA-23S RNA interaction.