Dataset Information

Identification of thermo tolerant lactic acid bacteria isolated from silage prepared in the hot and humid climate of Southwestern Japan.

ABSTRACT: To develop high-quality silage starters adapted to hot and humid weather, 12 LAB isolates from silage produced in Kyushu and Okinawa, Japan were characterized based on their morphological features, growth curves and sugar utilization. In addition, the nucleotide sequences of the V2-V3 region of their 16S rRNA genes and the 16S-23S rRNA intergenic spacer (ITS) regions were determined. The isolates were also subjected to RAPD-PCR analysis, DNA-DNA hybridization, G+C content analysis and immuno-identification using species-specific monoclonal antibodies and SDS-PAGE profiling. Nearly all of the isolates exhibited high thermotolerance and rapid growth. Combining ITS sequence analysis, RAPD-PCR and immuno-identification enabled rapid and accurate identification of closely related LAB strains that other methods failed to appropriately differentiate; for example, L. plantarum was distinguished from L. pentosus, and L. casei was distinguished from L. rhamonsus. Using the aforementioned techniques, the isolated strains were identified as L. plantarum, L. rhamonsus, L. rapi, Pediococcus pentosaceus and P. lolii. Our findings also showed that there is greater diversity among thermophilic LABs in silage prepared in a hot and humid environment.

SUBMITTER: Doi K

PROVIDER: S-EPMC3795204 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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Identification of thermo tolerant lactic acid bacteria isolated from silage prepared in the hot and humid climate of Southwestern Japan.

Doi Katsumi K Nishizaki Yousuke Y Kimura Hidetoshi H Kitahara Maki M Fujino Yasuhiro Y Ohmomo Sadahiro S Ohshima Toshihisa T Ogata Seiya S

SpringerPlus 20130925

To develop high-quality silage starters adapted to hot and humid weather, 12 LAB isolates from silage produced in Kyushu and Okinawa, Japan were characterized based on their morphological features, growth curves and sugar utilization. In addition, the nucleotide sequences of the V2-V3 region of their 16S rRNA genes and the 16S-23S rRNA intergenic spacer (ITS) regions were determined. The isolates were also subjected to RAPD-PCR analysis, DNA-DNA hybridization, G+C content analysis and immuno-ide ...[more]

PMID: 24130959

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Project description:To investigate community synergy of lactic acid bacteria (LAB) and cleaner fermentation of oat silage, oat silages were prepared with or without (control) commercial LAB inoculants LI1 (containing Lactiplantibacillus plantarum, Lentilactobacillus buchneri, Lacticaseibacillus paracasei, and Pediococcus acidilactici) and LI2 (containing Lactiplantibacillus plantarum and Lentilactobacillus buchneri). The microbial community, LAB synergy, and cleaner fermentation were analyzed at 1, 3, 6, 15, 35, and 90 days of ensiling. The LAB inoculant improved fermentation quality, with significantly (P < 0.05) lower pH, ammonia nitrogen content, and gas production and higher lactic acid and acetic acid contents than those of the control. Enterobacteriaceae was the main bacterial community in early stage of fermentation, which utilizes sugar to produce CO2 gas, causing dry matter (DM) and energy loss. As fermentation progressed, the microbial diversity decreased, and the microbial community shifted from Gram-negative to Gram-positive bacteria. The inoculation of multispecies LAB displayed community synergy; Pediococcus acidilactici formed a dominant community in the early stage of fermentation, which produced an acid and anaerobic environment for the subsequent growth of Lentilactobacillus and Lacticaseibacillus species, thus forming a LAB-dominated microbial community. The predicted functional profile indicated that the silage inoculated with LI1 enhanced the carbohydrate metabolism pathway but inhibited the amino acid metabolism pathway, which played a role in promoting faster lactic acid production, reducing the decomposition of protein to ammonia nitrogen, and improving the fermentation quality of silage. Therefore, oat silage can be processed to high-quality and cleaner fermented feed by using an LAB inoculant, and LI1 showed better efficiency than LI2. IMPORTANCE Oat natural silage is rich in Enterobacteriaceae, increasing gas production and fermentation loss. Lactic acid bacteria interact synergistically to form a dominant community during ensiling. Pediococci grow vigorously in the early stage of fermentation and create an anaerobic environment. Lactobacilli inhibit the harmful microorganisms and result in cleaner fermentation of oat silage.

Project description:IntroductionTo prepare high-quality silage, we studied the chemical composition, silage fermentation, characterization, and identification of lactic acid bacteria (LAB) associated with the silage fermentation of native grass on the Inner Mongolian Plateau.MethodsLAB were isolated from fresh native grass and their silage, and silages were prepared using a small-scale fermentation system with 2-3 cm length in plastic bags.ResultsThe dominant species of native grasses used were Stipa baicalensis, Leymus chinensis, Cleistogenes squarrosa, Melissilus ruthenicus and Pulsatilla turczaninovii, which contained 47.83-59.43 % moisture, 55.12-67.74 % neutral detergent fiber (NDF), and 8.72-14.55 crude protein (CP), and these nutrients did not change greatly during ensiling. Good preservation with a relatively low pH (below 4.44) and high (p < 0.05) lactic acid content (>0.58) was obtained after ensiling. Based on the morphological and biochemical characteristics, these isolates were divided into 12 groups (A-L). All isolate strains were gram-positive and catalase-negative bacteria that produce lactic acid from glucose. Group A-K were cocci, while group L was rod-shaped. Group A-E formed D-lactic acid, but group H-K formed L-lactic acid, and other groups formed DL-lactic acid. Group A-E were heterofermentative, and Group F-L were homofermentative types of LAB. According to the 16S rRNA gene sequences analysis, strains were identified as genus Leuconostoc (A, B, and C), Weissellla (D, E), Pediococcus (F, G), Enterococcus (H, I, J and K), and Lactiplantibacillus (L). Enterococcus (E.) faecium (29.17%, percentage of total isolates) and Pediococcus (P.) acidilactici (18.75%) were the most frequently occurring dominant species.DiscussionThis study suggests that the native grasses contained abundant LAB species, and they can be used as good-quality silages in animal husbandry. In addition, the strains P. acidilactici and E. faecium were the most frequently isolated from native grass silages as dominant species which can be a potentially excellent inoculant for native grass silage.

Project description:Hot and humid weather exposes animals to high temperature and relative humidity that ultimately reduce their ability to disperse body heat. To avoid serious consequences of heat stress, it is imperative to understand animal physiological responses and biochemical changes during a state of altered body homeostasis across different seasons of the year. This study evaluated seasonal dynamics of physiological, oxidative, and metabolic responses of Nili-Ravi buffaloes to hot and humid climate. Twenty non-lactating multiparous buffaloes were enrolled for this 1-year study. Meteorological data were recorded twice daily to calculate temperature humidity index (THI). Physiological parameters including rectal temperature (RT), body surface temperature (BST), and respiratory rate (RR) were measured weekly. Blood samples were collected once in each season (spring, summer, autumn, and winter) to analyze biochemical and antioxidant parameters. We also measured activities of liver enzymes including alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The results revealed a significantly higher THI value (82) during summer which resulted in a significant increase in RR and BST as compared to winter. Higher oxidative stress was observed in summer owing to significantly higher malondialdehyde (MDA) content and lower levels of serum antioxidant enzymes (GPx, SOD, and CAT) as compared to other seasons. Moreover, serum cortisol was also significantly higher while adrenocorticotropic hormone (ACTH), Triiodothyronine (T3), insulin, and growth hormone contents were significantly lower in summer. Contrarily, plasma thyroxin (T4) level was higher in summer. THI showed a positive correlation with physiological responses but a negative correlation with antioxidant parameters. Our study provides practical insights on the adaptive physiology of buffaloes and has several implications regarding the alleviation of heat stress in buffaloes to enhance the efficiency of production and reproduction under tropical climate. Our study suggests the use of appropriate cooling strategies to effectively manage the non-lactating buffaloes to avoid performance losses and animal welfare issues in summer season.

Dataset Information

Identification of thermo tolerant lactic acid bacteria isolated from silage prepared in the hot and humid climate of Southwestern Japan.

Publications

Identification of thermo tolerant lactic acid bacteria isolated from silage prepared in the hot and humid climate of Southwestern Japan.

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