Rumen methanogens are major sources of anthropogenic methane emissions, and these archaea are targets in strategies aimed at reducing methane emissions. Here we show that the poorly characterised Thermoplasmata archaea in bovine rumen are methylotrophic methanogens and that they are reduced upon dietary supplementation with rapeseed oil in lactating cows. In a metatranscriptomic survey, Thermoplasmata 16S rRNA and methylcoenzyme M reductase (mcr) transcripts decreased concomitantly with mRNAs of enzymes involved in methanogenesis from methylamines that were among the most abundant archaeal transcripts, indicating that these Thermoplasmata degrade methylamines. Their methylotrophic methanogenic lifestyle was corroborated by in vitro incubations, showing enhanced growth of these organisms upon methylamine supplementation paralleled by elevated methane production. The Thermoplasmata have a high potential as target in future strategies to mitigate methane emissions from ruminant livestock. Our findings and the findings of others also indicate a wider distribution of methanogens than previously anticipated.

Dietary doses of 2,500 ppm ZnO-Zn reduced bacterial activity (ATP accumulation) in digesta from the gastrointestinal tracts of newly weaned piglets compared to that in animals receiving 100 ppm ZnO-Zn. The amounts of lactic acid bacteria (MRS counts) and lactobacilli (Rogosa counts) were reduced, whereas coliforms (MacConkey counts) and enterococci (Slanetz counts, red colonies) were more numerous in animals receiving the high ZnO dose. Based on 16S rRNA gene sequencing, the colonies on MRS were dominated by three phylotypes, tentatively identified as Lactobacillus amylovorus (OTU171), Lactobacillus reuteri (OTU173), and Streptococcus alactolyticus (OTU180). The colonies on Rogosa plates were dominated by the two Lactobacillus phylotypes only. Terminal restriction fragment length polymorphism analysis supported the observations of three phylotypes of lactic acid bacteria dominating in piglets receiving the low ZnO dose and of coliforms and enterococci dominating in piglets receiving the high ZnO dose. Dietary doses of 175 ppm CuSO 4 -Cu also reduced MRS and Rogosa counts of stomach contents, but for these animals, the numbers of coliforms were reduced in the cecum and the colon. The influence of ZnO on the gastrointestinal microbiota resembles the working mechanism suggested for some growth-promoting antibiotics, namely, the suppression of grampositive commensals rather than potentially pathogenic gram-negative organisms. Reduced fermentation of digestible nutrients in the proximal part of the gastrointestinal tract may render more energy available for the host animal and contribute to the growth-promoting effect of high dietary ZnO doses. Dietary CuSO 4 inhibited the coliforms and thus potential pathogens as well, but overall the observed effect of CuSO 4 was limited compared to that of ZnO.

The effect of feeding dry feed (DF), nonfermented liquid feed (NFLF), and fermented liquid feed (FLF) to growing pigs on aspects of gastrointestinal ecology and on performance was investigated. Nonfermented liquid feed was prepared by mixing feed and water at a ratio of 1:2.5 immediately before feeding. Fermented liquid feed was prepared by mixing feed and water in the same ratio as NFLF, and stored in a tank at 20 degrees C for 4 d, after which half the volume was removed twice daily at each feeding and replaced with the same volume of feed and water mixture. A total of 60 pigs (initial BW of 30.7 kg) from 20 litters was used. Twenty pigs, housed individually, were allotted to each of the diets and fed restrictively. Five pigs from each diet were sacrificed at an average BW of 112 kg and digesta from the gastrointestinal tract (GI-tract) was obtained to examine variables describing some aspects of the gastrointestinal ecology. Fermented liquid feed contained high levels of lactic acid bacteria (9.4 log cfu/g) and lactic acid (approximately 169 mmol/kg), low levels of enterobacteria (<3.2 log cfu/g), and had a low pH (4.4). Nonfermented liquid feed contained 7.2 log cfu/g of lactic acid bacteria, and 6.2 log cfu/g of enterobacteria, which indicated that fermentation had started in the feed. The pigs fed FLF had the lowest levels of enterobacteria along the GI-tract (<3.2 to 5.0 log cfu/g), and those fed NFLF the highest levels (5.7 to 6.6 log cfu/g; P < or = 0.02). Fermented liquid feed caused a decrease in gastric pH from 4.4 and 4.6 for DF and NLF, to 4.0 (P = 0.003), and increased numerically the gastric concentration of lactic acid (P = 0.17) from 50 to 60 mmol/kg in the DF and NFLF treatments to 113 mmol/kg in the FLF treatment. The animals fed NFLF showed the highest weight gain (995 g/d) and feed intake (2.14 kg/d), and those fed FLF the lowest values (weight gain, 931 g/d; feed intake, 1.96 kg/d; P = 0.003 for weight gain, and P < 0.001 for feed intake). The results from the present study indicate that feeding FLF as prepared here may be a valid feeding strategy to decrease the levels of enterobacteria in the GI-tract of growing pigs, whereas feeding liquid feed that has started to ferment (high levels of enterobacteria and high pH as with NFLF) increases the presence of these undesirable bacteria. Nonetheless, higher daily feed intake and body weight gain are obtained when feeding NFLF compared with feeding FLF or DF.

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The effect of feeding a coarsely ground meal (COARSE) and a finely ground pelleted diet with 1.8% (as-fed basis) added formic acid (ACID) was compared with feeding a standard finely ground pelleted diet (STD) on the gastrointestinal ecology of growing pigs at different intervals after feeding. One hundred five castrated male growing-finishing pigs (initial BW 27 kg) were used. At a BW of 63 kg, 60 pigs were killed 0.5, 2.5, 4.5, 6.5, and 8.5 h after feeding, and samples from the gastrointestinal tract (GIT) were obtained. The remaining 45 pigs were kept on the experimental diets to a BW of 99 kg. Feeding the three diets resulted in a similar pattern of gastric pH with time, (i.e., highest pH values 0.5 h after feeding and decreasing values at the following sampling times, to reach a value of 2.12 at 8.5 h after feeding). The pH of the gastric digesta of pigs fed the ACID diet was below 4 at all sampling times, whereas the digesta from the other two dietary groups had values above pH 4 at the first sampling times. Feeding the ACID diet decreased the counts of total anaerobes in the proximal GIT (P < or = 0.007), and of lactic acid bacteria (P < or = 0.001), enterobacteria (P < or = 0.02), and yeasts (P < or = 0.01) along the GIT compared with feeding the other two diets. Feeding the COARSE diet stimulated the growth of total anaerobes and lactic acid bacteria in the stomach and distal small intestine increased the microbial diversity mainly in the stomach (P = 0.001), compared with feeding the other two diets (P < or = 0.09), and decreased the number of enterobacteria in the cecum compared with the STD diet (P = 0.03), with the same tendency in the mid-colon (P = 0.07). The concentration of lactic acid in the stomach was highest in the pigs fed the COARSE diet compared with the other two groups (P < 0.05). The concentration of formic acid was highest in the stomach and all segments of the small intestine of the pigs fed the ACID diet compared with those fed the STD and COARSE diets (P < 0.05). The results from this study suggest that feeding a coarsely ground diet and a finely ground diet with added formic acid affect the gastrointestinal ecology of pigs mainly by changing the environment in the proximal GIT. The presence of organic acids in the proximal GIT is a crucial factor contributing to the decrease in the number of enterobacteria along the GIT. The time after feeding at which samples are taken to measure characteristics describing the gastrointestinal ecology affects the results from the stomach and small intestine.