The effect of bile salts on survival and morphology of a potential probiotic strain Lactobacillus acidophilus M92

Bile tolerance is an important criterion in the selection of microbial strains for probiotic use. The survival and morphological changes of a potential probiotic strain, Lactobacillus acidophilus M92, in the presence of bile salts were examined. Lactobacillus acidophilus M92 has shown a satisfactory degree of tolerance against oxgall and individual bile salts tested, especially to taurocholate. The higher resistance of L. acidophilus M92 against taurine-conjugated bile salts relative to deconjugated and glycine-conjugated bile salts was attributed to its reaction to the stronger acidity of the former. Furthermore, bile salt hydrolase (BSH) was active when L. acidophilus M92 was grown in the presence of sodium taurocholate. The rate of BSH activity was highest at the exponential growth phase. It was hypothesised that BSH activity may be important for the bile salt resistance of this strain. The colonial and cellular morphology may also be a valuable parameter in the selection of bile salt-resistant Lactobacillus strains for probiotic use. Smooth (S) and rough (R) colonies, appeared in the original L. acidophilus M92 bacterial culture and demonstrated a different degree of bile tolerance. Rough colonies were more sensitive to bile salts than smooth ones. The R colony cells assumed a round form, probably induced by gaps in the cell wall caused by the cytotoxicity of glycodeoxycholate. This revised version was published online in July 2006 with corrections to the Cover Date.     

Growth Characteristics, Bile Sensitivity, and Freeze Damage in Colonial Variants of Lactobacillus acidophilus

Rough (R) and smooth (S) colonial variants were isolated from a heterogeneous culture of Lactobacillus acidophilus RL8K. R and S types were stable upon repeated transfer on agar, but revertant colonies did appear after broth transfers. When propagated in commercial MRS broth, R and S cultures showed similar growth characteristics, and both cell types were insensitive to freezing and frozen storage at −20°C. Alternatively, during growth in scratch MRS broth, R cultures shifted to a reduced rate of growth during the late logarithmic phase. R cells grown under these conditions were susceptible to death by freezing and injury at −20°C. Microscopically, R cells were observed as long gram-positive rods with small nonstainable blebs protruding from the cell wall. In bile sensitivity studies of R and S cells plated on MRS agar plus oxgall, the S culture was resistant to 1% bile, whereas the R culture was sensitive to 0.6% bile. Differences in the bile resistance and freeze damage of R and S cells suggest that colonial and cellular morphologies are important considerations for the selection of Lactobacillus strains as dietary adjuncts and for the development of growth conditions for preparing frozen concentrated cultures from either cell type.     

Selection of a potential probiotic Lactobacillus strain and subsequent in vivo studies

The probiotic potential of a Lactobacillus strain, isolated from pig faeces, was assessed as a probiotic in piglets. The strain was examined for resistance to pH 2.0, 0.5% oxgall and antibiotics, and antimicrobial activities against enteric pathogenic bacteria. The probiotic strain, L. reuteri BSA131, was administered through the feed to 25 1-month-old Landrace piglets. The piglets were divided into five groups of five piglets each and fed with different diets for 28 days. The daily consumption of L. reuteriBSA131 was assigned into two groups by the concentration of 10⁶ or 10⁸ freeze-dried bacteria. Fecal samples were collected before, during, and after consumption. Lactobacilli and enterobacteria cell counts were determined in the fecal samples. The liveweight gains and feed consumption of the piglets were recorded daily. This study showed that strain BSA 131 enhanced liveweight gains and feed conversion rates in piglets. It also showed a significant increase in lactobacilli cell counts and decreases in enterobacterial numbers in the fecal samples. Strain BSA 131 was considered to be a potential probiotic for piglets, especially after weaning.     

Characteristics of Lactobacillus strains contained in pharmaceuticals]

The aim of the study was to characterize lactic acid bacteria (LAB) which are components of drugs administered orally in cases of intestinal disturbances, or antibiotic--related diarrhea. Biochemical properties, growth behavior, bile tolerance, and survival at low pH of six LAB strains (four strains L. rhamnosus and two L. acidophilus) were studied. The survival at low pH was determined in MRS broth (Difco) acidified to pH 1; 2; 3; and 4. Bile tolerance was tested on MRS broth with 0.3% oxgall (Difco). Between tested strains differences in ability to grow at low pH and survival in bile were observed. Only 0.01% inoculum of all examined strains survived at pH 1. Differences between strains in survival at low pH (pH 2 and pH 3) and tolerance of bile were observed. However, after 2 h incubation at pH 4, 100% of strains stayed alive. Examined strains demonstrated good 3% bile tolerance. All strains met the criteria for probiotic strains: ability to survive at pH 3 and in the presence of bile.     

Identification of <Emphasis Type="Italic">Lactobacillus</Emphasis> strains from breast-fed infant and investigation of their cholesterol-reducing effects

Hypercholesterolemia has been reported to be the main cause of cardiovascular diseases and the leading cause of death. Therefore, decreasing serum cholesterol level is very important for preventing the cardiovascular diseases. It has been supposed that probiotics in human gastrointestinal tract have the ability to decrease serum cholesterol level by reducing the absorption of cholesterol from the intestinal tract and the bile salt deconjugation. In this study, 28 strains of Lactobacillus spp., isolated from breast-fed infant’s feces, were identified and investigated for their bile salt deconjugation ability. The deconjugation ability of the strains was determined by the release of cholic acid resulting from the deconjugation of conjugated bile salts. Research results showed that four of the strains had bile salt deconjugation ability. The strains with deconjugation ability have been identified in species level by using biochemical test, and molecular techniques, API 50CHL test and 16S rRNA gene sequence analysis respectively. LP1, E3, and E9 strains with deconjugation activity were identified as Lactobacillus rhamnosus and GD2 strain as Lactobacillus plantarum. Even if oxgall decreases the viability of bacteria, the highest amount of cholesterol precipitation (42%) was performed by GD2 strain in the presence of 0.3% (w/v) bile. This study demonstrated that the identified Lactobacillus strains had an excellent ability to survive at low pH, a high bile deconjugation ability, and hypocholesterolemic effect in in vitro conditions.     

Selection of Lactobacillus acidophilus strains for use in “acidophilus products”

Recent studies by DNA-DNA hybridization revealed that strains now designated as L. acidophilus, can be divided into several groups and only one group should be classified as L. acidophilus. We studied several phenotypic characteristics in representative strains from the six DNA-homology groups of L. acidophilus. No group specific pattern was observed among the strains for fermentation of eight carbohydrates, growth at 15 and 45°C, resistance to 0.2% oxgall, lysis by lysozyme or sensitivity to 17 antibiotics. However, some differences among groups were observed in -galactosidase (-gal) activity and surface layer (s-layer) protein. Strains in B1 do not have a s-layer or -gal while B2 strains also lack a s-layer but do possess -gal. All strains in groups A1, A2, A3 and A4, capable of growing in lactose, have -gal activity and also have a s-layer composed of protein subunits of different molecular weights (MW). Strains in A1 homology group have a s-layer with 46 Kd protein subunits while strains in other A groups have s-layer protein subunits that varied in MW within each group. On the basis of these two traits several isolates of unknown homology groups have been tentatively placed in A1, B1 or B2 groups. L. acidophilus from A1 group showed strain variation in -gal specific activity and rate of acid production and growth. For use in dietary adjuncts, L. acidophilus strains should be selected for these three and other desirable traits. They should be maintained and grown in media containing lactose.     

In Vitro Evaluation of the Probiotic and Safety Properties of Bacteriocinogenic and Non-Bacteriocinogenic Lactic Acid Bacteria from the Intestines of Nile Tilapia and Common Carp for Their Use as Probiotics in Aquaculture

In this study, seven bacteriocinogenic and non-bacteriocinogenic LAB strains previously isolated from the intestines of Nile tilapia and common carp and that showed potent antibacterial activity against host-derived and non-host-derived fish pathogens were assayed for their probiotic and safety properties so as to select promising candidates for in vivo application as probiotic in aquaculture. All the strains were investigated for acid and bile tolerances, transit tolerance in simulated gastrointestinal conditions, for cell surface characteristics including hydrophobicity, co-aggregation and auto-aggregation, and for bile salt hydrolase activity. Moreover, haemolytic, gelatinase and biogenic amine-producing abilities were investigated for safety assessment. The strains were found to be tolerant at low pH (two strains at pH 2.0 and all the strains at pH 3.0). All of them could also survive in the presence of bile salts (0.3% oxgall) and in simulated gastric and intestinal juices conditions. Besides, three of them were found to harbour the gtf gene involved in pH and bile salt survival. The strains also showed remarkable cell surface characteristics, and 57.14% exhibited the ability to deconjugate bile salts. When assayed for their safety properties, the strains prove to be free from haemolytic activity, gelatinase activity and they could neither produce biogenic amines nor harbour the hdc gene. They did not also show antibiotic resistance, thus confirming to be safe for application as probiotics. Among them, Lactobacillus brevis 1BT and Lactobacillus plantarum 1KMT exhibited the best probiotic potentials, making them the most promising candidates.     

Mode of action of acidocin D20079, a bacteriocin produced by the potential probiotic strain, <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> DSM 20079

Lactobacillus acidophilus DSM 20079 is the producer of a novel bacteriocin termed acidocin D20079. In this paper, mode of action using three various concentrations of acidocin D20079 (2,048, 128 and 11.3 AU/ml) was determined against an indicator strain L. delbrueckii subsp. lactis DSM 20076. These concentrations all led to marked decreases in both the number of viable cells and in optical density, indicating that the activity of the acidocin D20079 was bactericidal with concomitant cell lysis. Moreover, the probiotic potential of L. acidophilus DSM 20079 was analyzed for its ability to survive and retain viability at conditions (acid and bile concentrations) mimicking the gastrointestinal (GI) tract, under which it survived exposure to pH 2.0 with a 1.2 log cycle reduction in viability and where 45% of the original population survived in a medium containing 0.3% bile for 3 h.     

北疆传统发酵生奶酪中乳酸菌的耐受性及益生特性测定

【背景】北疆乳制品中生奶酪作为发酵乳制品,其中有多种微生物的参与,是优质食品级微生物的宝贵来源,然而其中蕴含的丰富的微生物资源也面临着流失。【目的】研究已筛选出的41株乳酸菌的生长曲线变化规律,并对其在低pH环境的耐酸特性及益生特性进行分析。【方法】通过人工模拟胃肠液和含0.3%、0.5%、1.0%浓度牛胆盐溶液培养,对从生奶酪中分离鉴定的41株乳酸菌菌株进行人工胃肠液、牛胆盐耐受性试验。【结果】41株乳酸菌中有6株乳酸菌(编号为QM-5、QM-27、UM-12、UM-18、NM-11和NM-14)均能在pH值分别为2.0、2.5、3.0、3.5、4.0时生长较好,在pH 2.0的环境下也保持一定程度的生长。菌株存活率大于50%,乳酸菌含量为108 CFU/mL以上。从生奶酪分离的乳酸菌有极强的耐酸、耐胆盐特性,可以预测在胃肠道环境生存。41株菌对大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)和枯草芽孢杆菌(Bacillus subtilis)均有抑制作用。【结论】通过比较耐酸性、胃肠道模拟、胆盐耐受性和抑菌特性等实验结果,初...     

Selection of Lactobacillus sp. PSC101 that produces active dietary enzymes such as amylase, lipase, phytase and protease in pigs

This study was carried out to screen lactic acid bacteria that produce active dietary enzymes, such as amylase, lipase, phytase, and protease, using a two-step process in pigs. We isolated a total of 210 and 132 strains of bacteria, grown under aerobic and anaerobic conditions, respectively, in Man Rogosa Sharpe agar containing 0.13% bile after treatment of intestinal samples at pH 3 for 30 min. From fecal samples, a total of 134 aerobic and 111 anaerobic strains were isolated in the same manner. In the second screening test, we selected four strains that produced four dietary enzymes from isolates obtained in the first screening test. Each strain was characterized as lactobacilli based on the following criteria: rod shape, negative for catalase, Gram positive, and lack of acute oral toxicity in mice. Of these four strains, we finally selected Lactobacillus spp. PSC101, which was resistant to pH 3 for 8 h and grew in the presence of 1% bile. In summary, Lactobacillus sp. PSC101 may be a strong probiotic candidate in swine due to its resistance to both acid and bile, its production of dietary enzymes that promote animal growth, and its non-toxic nature in mice.     

蒙古族儿童源益生特性双歧杆菌的筛选及鉴定

【目的】双歧杆菌在人和动物胃肠道中发挥着重要的生理作用,然而双歧杆菌能否耐受胃酸、肠液及胆汁酸是影响活菌制剂益生效果的关键因素,本研究旨在从蒙古族儿童粪便中分离筛选出具有良好益生特性的双岐杆菌。【方法】本文采用双岐杆菌选择性培养基对样品进行分离纯化,并对菌株进行生理生化鉴定,以耐受人工胃肠液、耐受牛胆盐为手段对各菌株的益生特性进行评价,并且对B. animalisV9进行了16S rDNA分子生物学鉴定。【结果】本文从12位健康蒙古族儿童粪便中分离得到11株双歧杆菌,经传统生理生化试验鉴定为5株B. adolescentis：A1、H3、G4、A8、V10;3株B. longum：C6、C7、D11;1株B. pseudocatenlatum：B2;1株B. bifidum：G5;1株B. animalis：V9。B. animalis V9具有较强的耐酸性,在pH2.0的人工胃液中厌氧培养3h后存活率为92.4%,而其它10株双歧杆菌在此条件下的存活率均小于31.25%;B. animalis V9在pH2.0的人工胃液中厌氧培养3h后接入pH8.0的人工肠液中消化8h,存活率为99.7%,并且可以耐受0.3%的牛胆盐。进一步对V9菌株进行16S rDNA分子生物学鉴定,发现与Bifidobacterium animalis subsp. lactic BB12的同源性为99%。【结论】本研究结果显示B. animalis V9来源安全,并且具有良好耐酸、耐胆盐益生特性,有望在乳制品及保健类产品中得到广泛的应用。     

Involvement of Trihydroxyconjugated Bile Salts in Cholesterol Assimilation by Bifidobacteria

To determine the conditions of cholesterol assimilation, various strains of Bifidobacterium species were cultured in the presence of cholesterol and bile salts. During culturing, Bifidobacterium breve ATCC 15700 assimilates cholesterol in the presence of oxgall at pH values lower than 6. This strain was selected to study the influence of conjugated (taurocholic acid) and deconjugated (cholic acid) bile salts on cholesterol assimilation. B. breve ATCC 15700 assimilated cholesterol (up to 51%) when cultures were undertaken in the presence of taurocholic acid, whereas less than 13% of the initial amount of cholesterol was measured in the cells in the presence of cholic acid. Cultured in the presence of six individual di- or trihydroxyconjugated bile salts, bifidobacteria strains assimilated cholesterol. This assimilation appeared to be more important in the presence of trihydroxyconjugated bile salts (tauro- and glycocholic acids). It is concluded that trihydroxyconjugated bile salts are involved in the assimilation of cholesterol by bifidobacteria. Received: 20 June 1996 / Accepted: 19 July 1996     

Safety,probiotic and technological properties of <Emphasis Type="Italic">Lactobacilli</Emphasis> isolated from unpasteurised ovine and caprine cheeses

Eleven Lactobacillus plantarum from Slovak ovine and caprine lump and stored cheeses, and from four commercial probiotic and yogurt cultures (Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus acidophilus) identified using a Maldi-TOF MS analysis were screened in vitro for selected aspects correlated with safety (antibiotic susceptibility patterns, biochemical and haemolytic activity, presence of genes responsible for biogenic amines production), functional traits (including acid, bile tolerance and antimicrobial activity), ecological roles (ability to produce biofilms), and technological applications (acidification and milk coagulation capacity) for assurance of their quality and diversity. The antibiotic susceptibility showed two L. plantarum strains, 19l5 and 18l4, with the presence of the non-wild-type ECOFFs (epidemiological cut-off) for clindamycin and/or gentamicin. All these strains expressed a high acid tolerance at pH 2.5 after a 4 h exposure (bacteria viability varied between 60% and 91%), and bile resistance at 0.3% oxgall ranged from 60% to 99% with no haemolytic activity. Three wild L. plantarum strains, 17l1, 16l4, 18l2, had no harmful metabolic activities, and formed strong biofilms that were measured by a crystal violet assay. Simultaneously, the acid cell-free culture supernatant (ACFCS) from L. plantarum 18l2 had a marked inhibitory effect on the viability of the pathogens as evaluated by flow-cytometry, and also exhibited fast acidification and milk coagulation. As a result, we conclude that L. plantarum 18l2 can be included as part of the created lactobacilli collection that is useful as a starter, or starter adjunct, in the dairy industry, due to its desirable safety and probiotic characteristics, together with rapid acidification capacity compared with other investigated strains from commercially accessible products.     

Screening of Lactobacilli derived from chicken feces and partial characterization of Lactobacillus acidophilus A12 as an animal probiotics

This study was performed to screen and select Lactobacillus strains from chicken feces for probiotic use in animals. Of these strains, strain A12 had the highest immunostimulatory effect. Therefore, strain A12 was characterized as a potential probiotic. Strain A12 was tentatively identified as Lactobacillus acidophilus A12, using the API 50 CHL kit based on a 99.9% homology. L. acidophilus A12 was highly resistant to artificial gastric juice (pH 2.5) and bile acid (oxgall). Based on results from the API ZYM kit, leucine arylamidase, crystine arylamidase, acid phosphatase, naphthol-AS-BI-phosphohydrolase, alpha-galactosidase, beta- galactosidase, alpha-glucosidase, beta-glucosidase, and N-acetyl-beta- glucosamidase were produced by strain A12. L. acidophilus A12 showed resistance to several antibiotics (nisin, gentamycin, and erythromycin). The amount of interleukin (IL)-1alpha in 20x concentrated supernatant from L. acidophilus A12 was approximately 156 pg/ml. With regard to antioxidant activity, L. acidophilus A12 supernatant showed 60.6% DPPH radical scavenging activity. These results demonstrate the potential use of L. acidophilus A12 as a health-promoting probiotics.     

Genetics of antagonistic action and drug resistance inLactobacillus acidophilus

Lactobacillus acidophilus has been recommended as a dietary adjunct because of its antagonistic action toward intestinal pathogens and anti-carcinogenic and hypocholesterolemic activities. ManyL. acidophilus strains harbour plasmids and such strains generally produce bacteriocin(s). Resistance to antibiotics has also been shown to be linked with plasmids. Gene transfer and cloning systems are being developed forL. acidophilus which should permit the rapid genetic characterization of desired species and their modification to obtain predetermined traits. Drug resistance determinants and production of antibiotic-like substances may serve as suitable markers for the study and development of these genetic systems. Recent developments in gene transfer systems have been reviewed here.     

Acid-bile, antibiotic resistance and inhibitory properties of propionibacteria isolated from Turkish traditional home-made cheeses

In this study, a total of 29 Propionibacterium spp. were isolated from traditional home-made Turkish cheese samples. As a result of the identification, isolates were identified as Propionibacterium freudenreichii subsp. freudenreichii (15 strains), Propionibacterium jensenii (12), and Propionibacterium thoenii (2). All isolates and 5 reference strains were examined for their abilities to survive at pH 2.0, 3.0, 4.0, 5.0 and in the presence of 0.06, 0.15 and 0.30% bile salts, their influence on the growth of food-borne and spoilage bacteria, as well as their sensitivity against 11 selected antibiotics. Only seven propionibacteria strains survived in both the acidic and bile salt environments. Propionibacterium spp. strains strongly inhibited growth of the Escherichia coli ATCC 11229 and Shigella sonnei Mu:57 strains (91%). All propionibacteria strains were sensitive to a majority of the antibiotics used in the investigations. Overall, dairy propionibacteria showed high antibacterial activity, resistance to pH 4.0, 5.0, high resistance to bile salts and will provide an alternative source to Lactobacillus and Bifidobacterium as probiotic culture.     

Serotyping of and hippurate hydrolysis by Campylobacter jejuni isolates from human patients,poultry and pigs in the Netherlands

Three hundred strains of Campylobacter jejuni, isolated from human patients, poultry and pigs were serotyped according to the Penner-Lauwers system and furthermore tested for hippurate hydrolysis. Serotyping showed a close relationship between human and chicken strains, whereas there was little relationship between human and pig strains. A similar conclusion was drawn from the results of the hippurate hydrolysis test: 86% of human and 94% of poultry strains were positive, whereas 91% of pig strains were negative.These data confirm the conclusions from epidemiological investigations, according to which poultry is a major source of human campylobacteriosis, while pigs are rarely so.     

Probiotic characteristics of lactic acid bacteria isolated from kimchi

Aims: The present work was aimed at identifying strains of lactic acid bacteria (LAB) from kimchi, with properties suitable for use as starter cultures in yogurt fermentation. Methods and Results: A total of 2344 LAB strains were obtained from two different sources, one group consisted of commercial LAB strains from kimchi, and the second group consisted of those strains isolated from various types of kimchi. The LAB strains from both groups were screened for resistance to biological barriers (acid and bile salts), and the four most promising strains were selected. Further analysis revealed that KFRI342 of the four selected strains displayed the greatest ability to reduce the growth of the cancer cells, SNU‐C4. The in vivo efficacy of strains in quinone reductase induction assay was evaluated, and the extent of DNA strand breakage in individual cells was investigated using the comet assay. Strain KFRI342 was identified as Lactobacillus acidophilus by 16S rRNA sequence analysis, showed protection against tumour initiation and imparted immunostimulation as well as protection against DNA damage. Conclusions: Strain KFRI342, which showed probiotic characteristics reducing cancer cell growth, could be a suitable starter culture for yogurt fermentation because of its strong acid production and high acid tolerance. Significance and Impact of the Study: This is the first report to describe a bacterium, isolated from kimchi, Lact. acidophilus KFRI342 which has the probiotic characteristics and the acid tolerance needed for its use as a starter culture in yogurt fermentation.     

Selection of Competitive Strains of Rhizobium Nodulating Phaseolus Vulgaris and Adapted to Environmental Conditions in Egypt,Using the Gus-Reporter Gene Technique

Two Rhizobium etli strains, EBRI 2 and EBRI 26, isolated from Egypt were tested for nodulation competitiveness on beans using Rhizobium tropici CIAT 899G as the competing strain. The insertion of the gus-reporter transposon mTn5ssgusA30 did not alter the nodulation or nitrogen fixation capacity of mutant strain CIAT 899G compared to the wild type. At neutral pH, R. etli strains EBRI 2 and EBRI 26 were more competitive than CIAT 899G with the bean cultivar Saxa. These two strains gave nodule occupancies of 52.1 and 61.1% competing with equal cell numbers of CIAT 899G. Nodule occupancies from these two native strains increased with the bean cultivar Giza 6 from Egypt to 66 and 67.5%. Based on these results, cultivar Giza 6 was used to select the most competitive strains under stress of salinity or alkalinity as a major problem for a large part of Egyptian soils. Under stress of salinity (0.2% NaCl or 34.2 mM NaCl), the salt-sensitive strain EBRI 2 was more competitive than the salt-resistant strain EBRI 26. Strain EBRI 2 gave 87.4% but strain EBRI 26 gave 63.7% nodule occupancy against CIAT 899G. The same trend of results was observed under stress of alkalinity (pH 8). Strain EBRI 2 occupied 83% while Strain EBRI 26 occupied 53.2%.     

<Emphasis Type="Italic">Oceanobacillus manasiensis</Emphasis> sp. nov., a moderately halophilic bacterium isolated from the salt lakes of Xinjiang,China

Three Gram reaction positive, rod-shaped, moderately motile halophilic bacterial strains, designated YD3-56^T, YD16, and YH29, were isolated from the sediments of Manasi and Aiding salt lakes in the Xinjiang region of China, respectively. The strains grew optimally at 30–37°C, pH 8–11, in the presence of 5–10% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strains were closely related to members of the genus Oceanobacillus, exhibiting 99.1–99.2% similarity to O. kapialis KCTC 13177^T, 99.2–99.3% to O. picturae KCTC 3821^T, and 94.2–96% sequence similarity to other described Oceanobacillus species. SDS-PAGE of whole cell proteins preparations demonstrated that the strains exhibited high similarity to each other, but distinguished from O. kapialis KCTC 13177^T and O. picturae KCTC 3821^T (75%). DNA-DNA hybridization revealed that the similarity between the representative strain YD3-56^T and O. kapialis KCTC 13177^T was 35.3%, and the similarity between YD3-56^T and O. picturae KCTC 3821^T was 22.3%. Chemotaxonomic analysis of the strains showed menaquinone-7 was the predominant respiratory quinine. Major cellular fatty acids were anteiso-C_15:0 and anteiso-C_17:0. The polar lipid pattern for strain YD3-56^T predominantly contained phosphatidylcholine, and trace to moderate amounts of phosphatidyl ethanolamine and hydroxy-phosphatidyl ethanolamine. The diamino acid in murein was meso-diaminopimelic acid. The DNA G+C content of the strains was 39.7–40.1 mol%. On the basis of these results, the three strains should be classified as a novel species of the genus Oceanobacillus, for which the name Oceanobacillus manasiensis sp. nov. has been proposed, with the type strain as YD3-56^T (=CGMCC 1.9105^T =NBRC 105903^T).