Assessment of ruminal bacterial populations and protozoal generation time in cows fed different methionine sources.

Methionine supplemented as 2-hydroxy-4-(methylthio)-butanoic acid (HMB) has been suggested to alter bacterial or protozoal populations in the rumen. Our objective was to determine if source of Met would change microbial populations in the rumen and to compare those results to samples from the omasum. The ruminal and omasal samples were collected from cows fed control (no Met), dl-Met, HMB, or the isopropyl ester of HMB (HMBi; estimated 50% rumen protection) in a replicated 4 x 4 Latin square design. In one square, changes in protozoal populations were determined using microscopic counts and denaturing gradient gel electrophoresis (DGGE), whereas changes in bacterial populations were determined using DGGE and ribosomal intergenic spacer length polymorphism (RIS-LP). Neither the protozoal counts nor the DGGE banding patterns derived from protozoa were different among the dietary treatments or for ruminal vs. omasal samples. As revealed by both DGGE and RIS-LP, bacterial populations clustered by treatments in ruminal and especially in omasal samples. Using cows from both Latin squares, the flow of protozoal cells from the rumen was quantified by multiplying protozoal cell count in omasal fluid by the omasal fluid flow (using CoEDTA as a liquid flow marker) or was estimated by rumen pool size of cells multiplied by either the ruminal dilution rate of CoEDTA (after termination of CoEDTA dosing) or the passage rate of Yb-marked particles. Compared with the omasal fluid flow measurement (16.4 h), protozoal generation time was approximated much more closely using the particulate than the fluid passage rate from the rumen (generation times of 15.7 and 7.5 h, respectively). There seems to be minimal selective retention of protozoal genera in the rumen in dairy cattle fed every 2 h. Data support the validity of the omasal sampling technique under our conditions.

Effects of 2-hydroxy-4-(methylthio) butanoic acid (HMB) on microbial growth in continuous culture.

2-Hydroxy-4-(methylthio) butanoic acid (HMB) positively affects milk composition and yield, potentially through ruminal actions. Four continuous culture fermenters were used to determine the optimal concentration of HMB for digestibility of organic matter (OM), neutral detergent fiber (NDF), acid detergent fiber (ADF), and hemicellulose and synthesis of microbial N. A highly degradable mix of hay and grain was used as a basal diet to simulate a typical lactation diet. Three concentrations of HMB (0, 0.055, and 0.110%) and one concentration of dl-Met (0.097%) were infused into the fermenters according to a 4 x 4 Latin square design. Digesta samples were collected during the last 3 d of each of the four 10-d experimental periods. Digestibility of OM, hemicellulose, and NDF was largely insensitive to treatment. Digestibility of ADF showed a quadratic effect to supplementation of HMB, with 0.055% having lower digestibility than 0 or 0.110%. Total production of VFA was not influenced by HMB supplementation, but differences in concentration and production of individual VFA were seen. Isobutyrate increased linearly with increasing HMB supplementation. Propionate concentration decreased linearly with increased HMB supplementation, but propionate production showed a quadratic trend (P = 0.13). A higher concentration of acetate was detected for dl-Met compared with the highest HMB concentration. There were trends (P < 0.15) for dl-Met to decrease the production of isobutyrate and to lower the concentration of butyrate when compared with HMB. Microbial efficiency was not different among treatments. The proportion of bacterial N produced from NH3-N decreased linearly with increasing HMB, and bacteria receiving dl-Met synthesized more N from NH3-N than those receiving HMB. These data suggest that supplementation of HMB may have a sparing effect on branched chain volatile fatty acids because the fatty acids are not needed to provide carbon for synthesis of valine, isoleucine and leucine with ammonia. Comparisons of bacterial community structure in the fermenter effluent samples using PCR amplicons containing the ribosomal intergenic spacer region and its flanking partial 16S ribosomal RNA gene showed no distinct banding patterns, though treatments tended to group together. Both Met and HMB affect the rumen microbial population, but Met supplied as dl-Met does not act identically to that supplied as HMB.

Effect of whole and expanded-expelled cottonseed on milk yield and blood gossypol.

Thirty-two primiparous and 12 multiparous Holstein cows were randomly assigned at calving to treatments to determine the effects of type and amount of cottonseed product on plasma gossypol, milk production, and composition, and conjugated linoleic acid concentration in milk fat. Rations consisted of corn silage, corn grain, soybean meal, and cottonseed hulls, and contained on average 16.8% crude protein and 25.3% acid detergent fiber on a dry matter basis. On a dry matter basis, diets contained one of the following: 1) 14% whole cottonseed; 2) 14% expanded-expelled cottonseed; 3) 21% expanded-expelled cottonseed; or 4) 28% expanded-expelled cottonseed. Cows remained on treatment from 30 through 120 d in milk. Dry matter intakes were not significantly different, but intakes of crude protein, acid detergent fiber, and fat were higher for multiparous cows fed whole cottonseed. Multiparous cows fed whole cottonseed had higher yields of milk, fat-corrected milk, crude protein, fat and solids-not-fat than those fed any level of expanded-expelled cottonseed. Concentrations of milk fat, protein, and SNF were not affected by treatment. Although there were treatment differences in fat intake, there were no production differences in primiparous cows. Milk production efficiency (fat-corrected milk/dry matter intake) was not affected by treatment for either multiparous or primiparous cows. Cows fed 14% whole or 14% expanded-expelled cottonseed had similar levels of total plasma gossypol and plasma levels of the negative isomer of gossypol. Increasing the level of expanded-expelled cottonseed in the diet increased both total plasma gossypol and the negative isomer. In this experiment, multiparous but not primiparous cows fed whole cottonseed produced more milk than those fed expanded-expelled cottonseed at 14 to 28% of the diet dry matter, however, feed efficiencies were similar for all treatments.