Weissella oryzae sp. nov., isolated from fermented rice grains.

A taxonomic study was conducted on two Gram-reaction-positive, catalase-negative, irregular short-rod-shaped or coccoid lactic acid bacteria, designated strains SG25(T) and SG23, that were isolated from grains of fermented Japanese rice (Oryza sativa L. subsp. japonica). A phylogenetic analysis based on 16S rRNA gene sequence data clearly showed that the strains belonged to the genus Weissella and were most closely related to Weissella soli LMG 20113(T) (with a sequence similarity of 96.9 % for each novel strain). The peptidoglycan of each strain contained the amino acids glutamic acid, lysine, serine and alanine in a molar ratio of 1.0 : 1.2 : 0.5 : 3.0, respectively. On the basis of the unusual phenotypic characteristics of the novel strains and the low levels of DNA-DNA relatedness recorded between each novel strain and Weissella soli JCM 12536(T), strains SG25(T) and SG23 represent a single novel species in the genus Weissella, for which the name Weissella oryzae sp. nov. is proposed. The type strain is SG25(T) ( = JCM 18191(T)  = DSM 25784(T)).

Lactobacillus oryzae sp. nov., isolated from fermented rice grain (Oryza sativa L. subsp. japonica).

The taxonomic position of three Lactobacillus-like micro-organisms (strains SG293(T), SG296 and SG310) isolated from fermented rice grain (Oryza sativa L. subsp. japonica) in Japan was investigated. These heterofermentative lactic acid bacteria were Gram-stain-positive, rod-shaped, facultatively anaerobic, non-motile, non-spore-forming and did not show catalase activity. 16S rRNA gene sequence analysis of strain SG293(T) revealed that the type strains of Lactobacillus malefermentans (98.3 %), Lactobacillus odoratitofui (96.2 %), Lactobacillus similis (96.1 %), Lactobacillus kimchicus (96.1 %), Lactobacillus paracollinoides (95.9 %) and Lactobacillus collinoides (95.7 %) were the closest neighbours. Additional phylogenetic analysis on the basis of pheS and rpoA gene sequences, as well as biochemical and physiological characteristics, indicated that these three strains were members of the genus Lactobacillus and that the novel isolates had a unique taxonomic position. The predominant cellular fatty acids were C18 : 1ω9c and C19 : 1 cyclo 9,10. Because low DNA-DNA hybridization values among the isolates and Lactobacillus malefermentans JCM 12497(T) were observed, it is proposed that these unidentified isolates be classified as a novel species of the genus Lactobacillus, Lactobacillus oryzae sp. nov. The type strain is SG293(T) (= JCM 18671(T) = DSM 26518(T)).

Effects of rice conservation methods on lactation, blood metabolites, and rumen fermentation in dairy cows.

We investigated the effect of rice grain conservation methods on feed intake, milk production, blood metabolites, and rumen fermentation in dairy cows. Raw rice grain was dried before crushing (DRY), ensiled after crushing (ENS-A), or ensiled before crushing (ENS-B). Twelve multiparous Holstein cows were used in a replicated 3 × 3 Latin square design with three dietary treatments comprising ad libitum access to one of three total mixed rations (TMRs; containing DRY, ENS-A, or ENS-B at 17% of dietary dry matter) plus a standard allowance of 2.0 kg/day of dairy concentrates. The dietary treatments did not affect the feed intake, milk yield, or milk composition. The selected blood constituents were not influenced by the rice conservation method. The ruminal lactic acid and total volatile fatty acid (VFA) concentrations and the VFA proportion in the cows were not influenced by the rice conservation method. These results demonstrate that the rice grain conservation method has little impact on lactation performance when cows are fed a TMR containing 17% treated rice grain (dry matter basis).

Effect of different methods for conserving rice grain on in situ ruminal degradation and in vivo nutrient digestion and rumen fermentation in steers.

We investigated the effects of different rice conservation techniques on in situ ruminal degradation and in vivo nutrient digestibility and rumen fermentation in steers. Raw rice grain was dried before crushing (DRY), ensiled after crushing (ENS-A), or ensiled before crushing (ENS-B). Six ruminally cannulated steers were used in a replicated 3 × 3 Latin square design with three dietary treatments: diets containing DRY, ENS-A, or ENS-B at 36% of the dietary dry matter. The in situ rapidly degradable fraction and effective ruminal degradability were higher for ensiled rice than for DRY, and higher for ENS-A than for ENS-B. The ruminal pH was lower and the lactic acid and total volatile acid concentrations were higher for the steers fed ensiled rice than those fed the DRY diet, but a treatment effect was not observed in the comparison between ENS-A and ENS-B. The whole-tract digestibility of crude protein and ether extract was improved when the rice grain was ensiled, but there were no differences in nutrient digestibility between ensiling methods. These results show that ensiling treatment can be a strategy to improve the nutrient value of rice grain, but the ensiling method has little impact on in vivo digestion.

Changes of fumonisin production in rice grain during ensiling.

We assessed fumonisin production during the ensiling of rice grain. Rice grain was harvested at the full-ripe stage and prepared as rough rice, crushed rough rice, brown rice or crushed brown rice. Each material was ensiled under six conditions: (1) no fungus, anaerobic; (2) no fungus, aerobic; (3) water added, anaerobic; (4) water and fumonisin-producing fungus added, anaerobic; (5) water and fumonisin-producing fungus added, aerobic; or (6) fumonisin-producing fungus added to autoclaved material, aerobic. After 40 days of ensilage, we analyzed the silage fermentative quality and fumonisin concentration. The fermentative quality of all materials was good in treatments (3) and (4) (pH < 4), reasonable in treatment (5) (pH = 5∼6) and unacceptable in treatments (1) and (2) (pH > 6.5). The fumonisin concentration was low in all materials in treatments (1) to (4), slightly increased in the three materials other than rough rice in treatment (5), and enormously increased in all materials in treatment (6). The results indicate that the fumonisin-producing fungus does not produce fumonisin in anaerobic conditions. It is important that an anaerobic condition be maintained during ensiling in order to reduce the fumonisin content in rice grain silage.