Your browser doesn't support javascript.
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.586
Filtrar
1.
Int J Syst Evol Microbiol ; 70(1): 11-15, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31560297

RESUMO

Two novel strains (HT111-2T and HT170-2) of the genus Lactobacillus were isolated from Marmota himalayana faecal samples collected on the Qinghai-Tibet Plateau, PR China. The isolates were Gram-stain-positive, rod-shaped, non-spore-forming bacteria with irregular circular colonies. Phylogenetic analysis and comparison of the 16S rRNA gene sequences demonstrated that the two strains form a subcluster and are closest to Lactobacillus hamsteri JCM 6256T (97.3 %) and Lactobacillus amylolyticus DSM 11664T (97.2 %). Phylogenetic analysis of two housekeeping genes (rpoA and pheS) found that strains HT111-2T and HT170-2 had the same closest relatives as the 16S rRNA gene sequence analysis did. The G+C content of strains HT111-2T and HT170-2 were 38.8 mol%. The values of in silico DNA-DNA hybridization with known Lactobacillus species were lower than the threshold (70%). Average nucleotide identity values of strain HT111-2T with L. hamsteri JCM 6256T and L. amylolyticus DSM 11664T were 77.84 % and 76.85 %, respectively. The major fatty acids of strains HT111-2T and HT170-2 were C16 : 0, C18 : 1ω9c and C18 : 0. Results of phenotypic, chemotaxonomic and phylogenetic analyses suggest strains HT111-2T and HT170-2 represent a novel species of the genus Lactobacillus, for which the name Lactobacillus xujianguonis sp. nov. is proposed with HT111-2T (=CGMCC 1.13855T=KCTC 15803T) as the type strain.


Assuntos
Lactobacillus/classificação , Marmota/microbiologia , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Fezes/microbiologia , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Tibet
2.
J Dairy Sci ; 103(1): 379-395, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31629529

RESUMO

Dairy farmers are often challenged with the need to feed high-moisture corn (HMC) after less than 30 d of fermentation. The objective this study was to assess the effects of microbial inoculation and particle size on fermentation profile, aerobic stability, and ruminal in situ starch degradation of HMC ensiled for a short period. High-moisture corn was harvested, coarsely ground (3,798 ± 40 µm, on average) or finely ground (984 ± 42 µm, on average), then ensiled in quadruplicate vacuum pouches untreated (CON) or with the following treatments: Lactobacillus plantarum CH6072 at 5 × 104 cfu/g and Enterococcus faecium CH212 at 5 × 104 cfu/g of fresh forage (LPEF); or Lactobacillus buchneri LB1819 at 7.5 × 104 cfu/g and Lactococcus lactis O224 at 7.5 × 104 cfu/g (LBLL). Silos were allowed to ferment for 14 or 28 d. Ruminal in situ starch degradation increased when HMC was finely ground. In addition, in situ starch degradation was greater and aerobic stability increased approximately 5-fold with LBLL compared with CON and LPEF. An interaction between microbial inoculation and storage length occurred for lactic acid. At 14 d, concentrations of lactic acid were greatest in LPEF and lowest in LBLL. Lactic acid concentrations increased from 14 to 28 d with CON and LPEF, but decreased with LBLL. At 28 d, concentrations of lactic acid were lower in LBLL compared with CON and LPEF. An interaction between particle size, microbial inoculation, and storage length occurred for acetic acid and ammonia-N. At 14 and 28 d, acetic acid concentrations were greatest in finely ground LBLL followed by coarsely ground LBLL. Ammonia-N concentrations increased across all treatments from 0 to 28 d. At 14 and 28 d, concentrations of ammonia-N were greatest in finely ground LBLL and lowest in coarsely ground CON and coarsely ground LPEF. Results from this study suggest that L. buchneri LB1819 can produce acetic acid in as little as 14 d, and that by 28 d, it has the potential to improve the aerobic stability of HMC. Additionally, results indicate that L. buchneri LB1819 has the potential to improve ruminal degradation of starch by 28 d of storage. Finally, results confirm enhanced fermentation and improved ruminal starch degradation with finely ground HMC by 28 d of storage.


Assuntos
Enterococcus faecium/fisiologia , Lactobacillus/fisiologia , Silagem/análise , Amido/metabolismo , Zea mays , Ácido Acético/metabolismo , Aerobiose , Inoculantes Agrícolas , Animais , Fermentação , Lactobacillus/classificação , Tamanho da Partícula , Silagem/microbiologia , Amido/química , Zea mays/metabolismo , Zea mays/microbiologia
3.
Syst Appl Microbiol ; 42(6): 126023, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31668878

RESUMO

Microbiota analysis of blown pack spoiled salami revealed five distinguishable Lactobacillus isolates we could not assign to a known species. Two of the isolates (TMW 1.2172T and TMW 1.1920) are rod-shaped, whilst three isolates (TMW 1.2098T, TMW 1.2118 and TMW 1.2188) appear coccus shaped or as short rods. All isolates are Gram-stain positive, facultative anaerobic, catalase and oxidase negative, non-motile and non-sporulating. Phylogenetic analysis of the 16S rRNA, dnaK, pheS and rpoA gene sequences revealed two distinct lineages within the genus Lactobacillus (L.). The isolates are members of the Lactobacillus alimentarius group with Lactobacillus ginsenosidimutans DSM 24154T (99.4% 16S similarity), Lactobacillus versmoldensis DSM 14857T (97.9%) and Lactobacillus furfuricola DSM 27174T (97.7%) as phylogenetic closest related species and L. alimentarius DSM 20249T (97.7%) and Lactobacillus paralimentarius DSM 13961T (97.5%) as closest relatives, respectively. Average Nucleotide Identity (ANI) and digital DNA-DNA hybridization (dDDH) values between the isolates and their close related type strains are lower than 80% and 25%, respectively. For both designated type strains, the peptidoglycan type is A4α l-Lys-d-Asp and the major fatty acids are C16:0, C18:1ω9c and summed feature 7. Based on phylogenetic, phenotypic and chemotaxonomic analysis we demonstrated that the investigated isolates belong to two novel Lactobacillus species for which we propose the names Lactobacillus salsicarnum with the type strain TMW 1.2098T=DSM 109451T=LMG 31401Tand Lactobacillus halodurans with the type strain TMW 1.2172T=DSM 109452T=LMG 31402T.


Assuntos
/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Produtos da Carne/microbiologia , Filogenia , Animais , Composição de Bases , Ácidos Graxos/análise , Genes Bacterianos/genética , Genoma Bacteriano/genética , Lactobacillus/química , Lactobacillus/genética , Lactobacillus/ultraestrutura , Hibridização de Ácido Nucleico , Peptidoglicano/análise , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Especificidade da Espécie , Suínos
4.
World J Microbiol Biotechnol ; 35(10): 161, 2019 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-31608422

RESUMO

Lactobacilli are considered as the most important microorganisms in regulating immune system and maintaining vaginal health. The uses and benefits of Lactobacilli as probiotics, particularly the regulation of immune system, are dependent on the strain used and a comprehensive understanding of their effects on the host. Several factors have been identified in Lactobacilli that influence the immune response, such as exopolysaccharides and proteins. The current study was designed to investigate the serum immunoreactivity of healthy women against common vaginal Lactobacilli immunoreactive proteins. Three common vaginal Lactobacillus strains (L. crispatus L1, L. gasseri L9, and L. fermentum L2) were compared for immune response. The ELISA results showed that the levels of total immunoglobulin (Ig-total) antibody for L. crispatus L1, L. fermentum L2, and L. gasseri L9 were 47%, 45% and 29%, respectively. Regarding the lower prevalence of L. fermentum L2 in comparison with the other two strains, the approximately equal levels of Ig-total compared to L. crispatus L1 and more than L. gasseri L9 indicate that L. fermentum L2 has the greater antigenicity ability. Accordingly, the immunoreactive proteins of L. fermentum L2 were identified using MALDI-TOF-MS detected by SDS-PAGE and Western blotting. These proteins included 30s ribosomal protein S4 and 50s ribosomal protein L5. Antigenic epitopes on the 3D structure of these proteins was also predicted using bioinformatics analysis. The presence of antibody in serum of healthy pre-menopausal women indicates that Lactobacilli (normal flora) proteins can stimulate host immune response. Purification and further studies of the proteins may allow their potential use as an adjuvant to improve the efficacy of vaccines.


Assuntos
Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Proteômica/métodos , Proteínas Ribossômicas/imunologia , Vagina/imunologia , Vagina/microbiologia , Adulto , Feminino , Humanos , Lactobacillus/classificação , Pessoa de Meia-Idade , Modelos Moleculares , Probióticos , Proteínas Ribossômicas/química , Proteínas Ribossômicas/isolamento & purificação , Adulto Jovem
5.
Curr Microbiol ; 76(12): 1407-1416, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31555856

RESUMO

The study provides phenotypic and molecular analyses of the antibiotic resistance in 20 Lactobacillus strains including 11 strains newly isolated from fermented plant material. According to the results of disc diffusion method, 90% of tested lactobacilli demonstrated sensitivity to clindamycin and 95% of strains were susceptible to tetracycline, erythromycin, and rifampicin. Ampicillin and chloramphenicol were found to inhibit all bacteria used in this study. The vast majority of tested strains revealed phenotypic resistance to vancomycin, ciprofloxacin, and aminoglycosides. Most of Lactobacillus strains showed high minimum inhibitory concentrations (MICs) of cefotaxime, ceftriaxone, and cefazolin and therefore were considered resistant to cephalosporins. All the strains exhibited multidrug resistance. The occurrence of resistance genes was associated with phenotypic resistance, with the exception of phenotypically susceptible strains that contained genes for tetracycline (tetK, tetL) and erythromycin (ermB, mefA) resistance. The vanX gene for vancomycin resistance was among the most frequently identified among the lactobacilli (75% of strains), but the occurrence of the parC gene for ciprofloxacin resistance was sporadic (20% of strains). Our results mainly evidence the intrinsic nature of the resistance to aminoglycosides in lactobacilli, though genes for enzymatic modification of streptomycin aadA and aadE were found in 20% of tested strains. The occurrence of extended spectrum beta-lactamases (ESBL) was unknown in Lactobacillus, but our results revealed the blaTEM gene in 80% of strains, whereas blaSHV and blaOXA-1 genes were less frequent (20% and 15% of strains, respectively).


Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana , Lactobacillus/efeitos dos fármacos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Laticínios/microbiologia , Eritromicina/farmacologia , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/metabolismo , Testes de Sensibilidade Microbiana , Tetraciclina/farmacologia , Vancomicina/farmacologia
6.
J Dairy Sci ; 102(10): 8745-8755, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31400900

RESUMO

Mongolian traditionally fermented vrum is known for its functional characteristics, and indigenous microbial flora plays a critical role in its natural fermentation. However, studies of traditionally fermented vrum are still rare. In this study, we investigated the artisanal production of traditionally fermented vrum from Inner Mongolia. In general, its physicochemical composition was characterized by 34.5 ± 8% moisture, 44.9 ± 12.1% fat, 10.6 ± 3.2% protein, and 210 ± 102°T. The total lactic acid bacteria and yeast counts ranged from 50 to 2.8 × 108 cfu/g and from 0 to 1.1 × 106 cfu/g, respectively. We studied bacterial and fungal community structures in 9 fermented vrum; we identified 5 bacterial phyla represented by 11 genera (an average relative abundance >1%) and 8 species (>1%), and 3 fungal phyla represented by 8 genera (>1%) and 8 species (>1%). Relative abundance values showed that Lactococcus and Lactobacillus were the most common bacterial genera, and Dipodascus was the predominant fungal genus. This scientific investigation of the nutritional components, microbial counts, and community profiles in Mongolian traditionally fermented vrum could help to develop future functional biomaterials and probiotics.


Assuntos
Produtos Fermentados do Leite/microbiologia , Microbiota , Animais , Bactérias/classificação , Bactérias/isolamento & purificação , Reatores Biológicos , China , Fermentação , Fungos/classificação , Fungos/isolamento & purificação , Lactobacillales , Lactobacillus/classificação , Lactobacillus/isolamento & purificação , Lactococcus , Tipagem Molecular , Probióticos
7.
Int J Syst Evol Microbiol ; 69(10): 3178-3190, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31339481

RESUMO

Fourteen Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle and yogurt. The strains were characterised using a polyphasic taxonomic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, determination of average nucleotide identity, fatty acid methyl ester analysis and an analysis of phenotypic features. The data demonstrated that the fourteen strains represented nine novel species belonging to the genus Lactobacillus, strains 54-2T, 54-5T, 33-7T, 116-2T, 184-8T, 204-8T, 8-1(1)T, 256-3T and M1575T were designated as the type strains. Strain 54-2T was phylogenetically related to the type strains of Lactobacillus composti and Lactobacillus floricola, having 96.5 and 91.6 % 16S rRNA gene sequence similarities, less than 74.6 % pheS gene sequence similarities, less than 81.6 % rpoA gene sequence similarities and less than 72.5 % ANI values. Strain 54-5T was phylogenetically related to the type strains of Lactobacillus dextrinicus and Lactobacillus concavus, exhibiting 99.1 and 97.3 % 16S rRNA gene sequence similarities, less than 83.1 % pheS gene sequence similarities, less than 93.1 % rpoA gene sequence similarities and less than 79.9 % ANI values. Strains 33-7T, 116-2T, 184-8T, 204-8T, 8-1(1)T, 256-3T and M1575T were phylogenetically related to the type strains of Lactobacillus tucceti, Lactobacillus nodensis, Lactobacillus insicii, Lactobacillus allii, Lactobacillus metriopterae, Lactobacillus terrae, Lactobacillus versmoldensis and Lactobacillus furfuricola, sharing 95.6-100 % 16S rRNA gene sequence similarities, less than 91.6 % pheS gene sequence similarities, less than 98.2 % rpoA gene sequence similarities and less than 89.4 % ANI values. Based upon the data of polyphasic characterisation obtained in the present study, nine novel species, Lactobacillus yilanensis sp. nov., Lactobacillus bayanensis sp. nov., Lactobacillus keshanensis sp. nov., Lactobacillus kedongensis sp. nov., Lactobacillus baiquanensis sp. nov., Lactobacillus jidongensis sp. nov., Lactobacillus hulinensis sp. nov., Lactobacillus mishanensis sp. nov. and Lactobacillus zhongbaensis sp. nov., are proposed and the type strains are 54-2T (=NCIMB 15154T=CCM 8896T=KCTC 21120T=LMG 31058T), 54-5T (=NCIMB 15151T=CCM 8894T), 33-7T (=NCIMB 15153T=CCM 8936T=KCTC 21118T=LMG 31166T), 116-2T (=NCIMB 15158T=CCM 8899T=KCTC 21124T=LMG 31051T), 184-8T (=NCIMB 15152T=CCM 8895T=KCTC 21131T=LMG 31050T), 204-8T (=NCIMB 15159T=CCM 8900T=KCTC 21133T=LMG 31054T), 8-1(1)T (=NCIMB 15156T=CCM 8898T=KCTC 21115T=LMG 31047T), 256-3T (=NCIMB 15160T=CCM 8901T=LMG 31048T) and M1575T (=NCIMB 15149T=CCM 8892T=LMG 31045T), respectively.


Assuntos
/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Filogenia , Iogurte/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
8.
Int J Syst Evol Microbiol ; 69(10): 3237-3247, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31361212

RESUMO

Seven novel lactic acid bacterial strains, isolated from traditional Chinese pickle, were characterized using a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, determination of DNA G+C content, determination of average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH), fatty acid methyl ester (FAME) analysis and an analysis of phenotypic features. Strains 382-1T, 116-1AT, 381-7T, 203-3T, 218-3T and 398-2T were phylogenetically related to the type strains of Lactobacillus plantarumsubsp. plantarum, Lactobacillus plantarumsubsp. argentoratensis, Lactobacillus pentosus, Lactobacillus paraplantarum, Lactobacillus fabifermentans, Lactobacillus herbarum, Lactobacillus mudanjiangensis, Lactobacillus xiangfangensis, Lactobacillus plajomi and Lactobacillus modestisalitolerans, having 97.1-99.9 % 16S rRNA gene sequence similarities, less than 89.9 % pheS gene sequence similarities, less than 98.0 % rpoA gene sequence similarities, less than 91.2 % ANI values and less than 43.3 % isDDH values. Strain 778-3T was phylogenetically related to the type strains of Lactobacillus hokkaidonensis, Lactobacillus wasatchensis, Lactobacillus oligofermentans, Lactobacillus nenjiangensis, Lactobacillus vaccinostercus and Lactobacillus suebicus, exhibiting 97.0-99.4 % 16S rRNA gene sequence similarities, 78.2-82.1 % pheS gene sequence similarities, 80.0-91.5 % rpoA gene sequence similarities, less than 78.6 % ANI values and less than 22.9 % isDDH values. Based upon the data of polyphasic characterization obtained in the present study, seven novel species, Lactobacillus pingfangensis sp. nov., Lactobacillus daoliensis sp. nov., Lactobacillus nangangensis sp. nov., Lactobacillus daowaiensis sp. nov., Lactobacillus dongliensis sp. nov., Lactobacillus songbeiensis sp. nov. and Lactobacillus kaifaensis sp. nov., are proposed and the type strains are 382-1T (=NCIMB 15187T=CCM 8935T=LMG 31176T), 116-1AT (=NCIMB 15181T=CCM 8934T=LMG 31171T), 381-7T (=NCIMB 15186T=CCM 8930T), 203-3T (=NCIMB 15183T=CCM 8933T=LMG 31172T), 218-3T (=NCIMB 15184T=CCM 8932T=LMG 31173T), 398-2T (=NCIMB 15189T=CCM 8931T=LMG 31174T) and 778-3T (=NCIMB 15191T=CCM 8929T=LMG 31177T), respectively.


Assuntos
/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
9.
Genes (Basel) ; 10(7)2019 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-31336987

RESUMO

Lactobacillus is a fairly diverse genus of bacteria with more than 260 species and subspecies. Many profiling methods have been developed to carry out phylogenetic analysis of this complex and diverse genus, but limitations remain since there is still a lack of comprehensive and accurate analytical method to profile this genus at species level. To overcome these limitations, a Lactobacillus-specific primer set was developed targeting a hypervariable region in the groEL gene-a single-copy gene that has undergone rapid mutation and evolution. The results showed that this methodology could accurately perform taxonomic identification of Lactobacillus down to the species level. Its detection limit was as low as 104 colony-forming units (cfu)/mL for Lactobacillus species. The assessment of detection specificity using the Lactobacillus groEL profiling method found that Lactobacillus, Pediococcus, Weissella, and Leuconostoc genus could be distinguished, but non-Lactobacillus Genus Complex could not be detected. The groEL gene sequencing and Miseq high-throughput approach were adopted to estimate the richness and diversity of Lactobacillus species in different ecosystems. The method was tested using kurut (fermented yak milk) samples and fecal samples of human, rat, and mouse. The results indicated that Lactobacillus mucosae was the predominant gut Lactobacillus species among Chinese, and L. johnsonii accounted for the majority of lactobacilli in rat and mouse gut. Meanwhile, L. delbrueckii subsp. bulgaricus had the highest relative abundance of Lactobacillus in kurut. Thus, this groEL gene profiling method is expected to promote the application of Lactobacillus for industrial production and therapeutic purpose.


Assuntos
Proteínas de Bactérias/genética , Chaperonina 60/genética , Lactobacillus/classificação , Animais , Bovinos , Produtos Fermentados do Leite/microbiologia , Primers do DNA , Fezes/microbiologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Intestinos/microbiologia , Lactobacillus/genética , Camundongos , Tipagem Molecular , Filogenia , Ratos
10.
Microb Pathog ; 134: 103599, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31212037

RESUMO

Vaginal microbiota contributes in maintaining and protecting the urogenital niche from infections and their sequelae. Despite extensive research, microbiome studies have often ignored asymptomatic bacterial vaginosis (BV) and vulvovaginal candidiasis (VVC). The present study aimed to explore the cultivable vaginal bacterial and mycological communities in women asymptomatic for BV and VVC using multiplex PCR and species-specific PCR. Vaginal swabs collected from 199 participants asymptomatic for urogenital infections, scored by Nugent criteria indicated 73.9% had normal microbiota, 11.6% intermediate and 14.5% BV. The most frequent Lactobacillus species in normal women were L. iners (69.4%), L. crispatus (24.5%), L. reuteri (20.4%). Women with BV colonized L. iners (62.1%); L. rhamnosus (41.4%); L. salivarius (13.7%) and L. reuteri (7.2%). Furthermore, L. crispatus was associated with normal microbiota, whereas L. iners was a frequent member of normal and dysbiotic microbiota. Lactobacillus abundance and species richness reduced in asymptomatic BV. Also L. crispatus, L. fermentum, L. acidophilus and L. delbruckii were absent in these women. L. iners significantly co-existed with other Lactobacillus species, indicating its failure in independently maintaining the healthy vaginal niche. Of 30.4% women detected with Candida, 72.1% constituted non-albicans Candida. Predominance of C. albicans increased from 18.4% in healthy to 60% in women with asymptomatic BV; whereas distribution of BV related bacteria did not vary across the groups. Heterogeneous population of lactobacilli in 80.8% of normal women calls attention towards cumulative effects of these species in safeguarding the vaginal microenvironment. Since the microbiota of asymptomatic BV was different from healthy, screening and management could be encouraged to avoid further complications of infections.


Assuntos
Candidíase Vulvovaginal/microbiologia , Microbiota/fisiologia , Vagina/microbiologia , Vaginose Bacteriana/microbiologia , Adolescente , Adulto , Biodiversidade , Candida/classificação , Candida/genética , Candida/crescimento & desenvolvimento , Candida/isolamento & purificação , Candidíase Vulvovaginal/genética , DNA Bacteriano , DNA Fúngico , Disbiose , Feminino , Humanos , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/isolamento & purificação , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , RNA Ribossômico 16S , Especificidade da Espécie , Vaginose Bacteriana/genética , Adulto Jovem
11.
Int J Syst Evol Microbiol ; 69(9): 2807-2814, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31241447

RESUMO

A Gram-stain-positive bacterial strain, 395-6.2T, was isolated from traditional pickle in Heilongjiang Province, PR China. The bacterium was characterised by a polyphasic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, fatty acid methyl ester (FAME) analysis, determination of DNA G+C content, average nucleotide identity (ANI) analysis, in silico DNA-DNA hybridisation (isDDH) and an analysis of phenotypic features. Analysis of the 16S rRNA gene sequence showed that strain 395-6.2T was phylogenetically related to Lactobacillus farciminis, Lactobacillus formosensis, Lactobacillus futsaii, Lactobacillus crustorum, Lactobacillus nuruki, Lactobacillus heilongjiangensis, Lactobacillus musae, Lactobacillus nantensis, Lactobacillus mindensis, Lactobacillus kimchiensis, Lactobacillus zhachilii, Lactobacillus alimentarius, Lactobacillus bobalius, Lactobacillus kimchii and Lactobacillus paralimentarius. Strain 395-6.2T exhibited 95.7-99.4 % 16S rRNA gene sequence similarities, 85.0-94.0 % pheS gene sequence similarities, 94.2-98.0 % rpoA gene sequence similarities to type strains of phylogenetically related species. ANI and isDDH values between strain 395-6.2T and type strains of phylogenetically related species were 77.9-87.1 % and 22.5-33.5 %, respectively. Based upon the data obtained in the present study, a novel species, Lactobacillus huachuanensis sp. nov., is proposed and the type strain is 395-6.2T (=CCM 8927T=NCIMB 15188T=LMG 31179T).


Assuntos
/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
12.
Artigo em Inglês | MEDLINE | ID: mdl-31245300

RESUMO

To investigate the parameters associated with post-treatment recurrence of bacterial vaginosis (BV), clinical factors and vaginal microbiota were examined and analyzed for BV patients who received standard metronidazole therapy. The variables associated with BV recurrence included clinical factors of past BV history, use of intravaginal device, and D7 Nugent score as well as many microbial genera, with Lactobacillus, Enterococcus, Ureaplasma, and Aerococcus being the top contributors. Co-occurrence network analysis showed that whereas overwhelming majority of interbacterial interactions were positive, negative interactions were present and connected mostly to Lactobacillus, Enterococcus, and to a less extent Ureaplasma, suggesting the importance of interbacterial antagonism for treatment outcome. The patients who were cured and recurrent also exhibited clear differences in the species composition of Lactobacillus: although L. iners remained the dominant species at all time points, L. crispatus, L. gasseri, and L. jensenii displayed apparent differences in relative abundance between the cure and recurrent groups. Based on these results, we developed a 5-component panel comprising Enterococcus, L. crispatus, Ureaplasma, Aerococcus, and L. jensenii for predicting recurrence using D7 data and showed that it generated the specificity, sensitivity, and AUC values of 0.80, 0.66, and 0.73 for the discovery cohort and 0.80, 0.67, and 0.69 for the validation cohort. Our findings highlighted key microbial components for BV recurrence and suggested that they could be used to monitor the treatment outcome.


Assuntos
Bactérias/classificação , Vagina/microbiologia , Vaginose Bacteriana/diagnóstico , Vaginose Bacteriana/microbiologia , Adolescente , Adulto , Biodiversidade , Estudos de Coortes , DNA Bacteriano , Feminino , Humanos , Lactobacillus/classificação , Metronidazol/uso terapêutico , Microbiota/genética , Pessoa de Meia-Idade , RNA Ribossômico 16S , Recidiva , Resultado do Tratamento , Vaginose Bacteriana/tratamento farmacológico , Adulto Jovem
13.
Anaerobe ; 59: 192-200, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31207299

RESUMO

Lactobacillus sp. are well-known colonizers of human mucosa and frequently used as probiotics. Accurate species identification is crucial both for fundamental studies and biotechnology applications; however, it has been thus far challenging. The aim of this work was to develop a one-step multiplex-PCR assay for detection of ten Lactobacillus species (L. jensenii, L. fermentum, L. acidophilus, L. crispatus, L. reuteri, L. iners, L. casei, L. gasseri, L. plantarum, L. rhamnosus) directly in complex bacterial genomic DNA. A multiplex-PCR assay was optimized based on Box-Behnken experimental design, which showed to be efficient for optimization of all crucial reaction components. Nineteen Lactobacillus strains, including six collection strains and thirteen human isolates were used in order to verify the specificity and sensitivity of the assay. In addition, a set of PCR adjuvants was introduced to remove non-specific amplifications and enhance reaction yield. Among them, Triton™ X-100, Tween® 20, BSA, and dithiothreitol showed beneficial effects when compared with other adjuvants. The application of the developed method to samples that resulted from the mixing of DNA from the ten strains, resulted in amplicons of the expected sizes (from about 100 to 1000 bp). The detection limit was 1.25 ng/µl for all species with the exception of L. gasseri (0.31 ng/µl). In order to confirm the method applicability on human samples, ten vaginal fluids were enrolled in this study showing that the method can be successfully used on these biological materials. The proposed multiplex-PCR assay was shown to be selective, sensitive and efficient for detection of ten Lactobacillus species directly in human vaginal samples. This method provides a cost-effective and accessible methodology applicable to the detection of Lactobacillus species to different environments. At the same time, this approach represents a considerable improvement over other PCR-based approaches for identification of these species.


Assuntos
Lactobacillus/classificação , Lactobacillus/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Vagina/microbiologia , Análise Custo-Benefício , Feminino , Humanos , Lactobacillus/genética , Sensibilidade e Especificidade
14.
Arch Microbiol ; 201(9): 1163-1171, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31172250

RESUMO

Organic acid production including lactate and acetate is an economically attractive technology that has gained momentum worldwide over the past years. These series of action need to be performed by an esoteric and complex microbial community, in which different members have distinct roles in the establishment of a collective organization. In this study, we analyzed the bioma from bioreactors with various pH conditions of 4.0, 5.0 and 6.0 (R1, R2 and R3), respectively, involved in acidogenic digestion for stable production of various organic acids by means of high-throughput Illumina sequencing, disclosing thousands of genes and extracting more than 53 microbial genomes. At pH 5.0, the hydrolysis reaction was enhanced and thus the lactic acid fermentation was stably improved to 45.96 mm/L and acetic acid to 73.77 mm/L. R2 was found with the most suitable pH condition for stable organic acids production as Lactobacilli and Bifidobacteria were the major members. Both the members have the key roles in heterofermentation and produce higher transcripts of key encoding enzymes involved in the dominant heterofermentation pathways.


Assuntos
Acetatos/metabolismo , Bifidobacterium/genética , Reatores Biológicos/microbiologia , Ácido Láctico/metabolismo , Lactobacillus/genética , Microbiota/genética , Bifidobacterium/classificação , Bifidobacterium/metabolismo , Metabolismo dos Carboidratos , Fermentação/genética , Sequenciamento de Nucleotídeos em Larga Escala , Concentração de Íons de Hidrogênio , Hidrólise , Lactobacillus/classificação , Lactobacillus/metabolismo , Metagenômica
15.
Microbes Environ ; 34(2): 206-214, 2019 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-31167991

RESUMO

Lactic acid bacteria (LAB) exert beneficial health effects by regulating immune responses. Brassica rapa L., known as Nozawana, is commonly consumed as a lactic acid-fermented food called nozawana-zuke. Few studies have investigated changes in the bacterial community and cytokine production activities during the fermentation of B. rapa L. In order to obtain more detail information, we herein conducted a study on fresh B. rapa L. fermented for 28 d. An amplicon analysis of the 16S rRNA gene revealed that Lactobacillales predominated during fermentation, and the microbiota became less diverse on day 7 or later. Fermented B. rapa L. promoted the production of interferon (IFN)-γ and interleukin (IL)-10 by mouse spleen cells more than non-fermented vegetables. Lactobacillus curvatus was the predominant species during fermentation, followed by L. plantarum and L. brevis. L. sakei was occasionally detected. A correlation analysis showed that IFN-γ concentrations positively correlated with the numbers of L. curvatus and L. plantarum, while those of IL-10 correlated with the numbers of L. sakei in addition to these 2 species. Significantly higher levels of IFN-γ and IL-10 were induced by fermented B. rapa L. when isolated Lactobacillus strains were added as starter cultures. These results suggest that the Lactobacillus species present in fermented B. rapa L. are beneficial for manufacturing vegetables with immunomodulatory effects.


Assuntos
Brassica rapa/imunologia , Brassica rapa/microbiologia , Microbiologia de Alimentos , Fatores Imunológicos/farmacologia , Microbiota/fisiologia , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Bactérias/metabolismo , Biodiversidade , Meios de Cultivo Condicionados/farmacologia , Citocinas/metabolismo , DNA Bacteriano/genética , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Masculino , Camundongos Endogâmicos C57BL , RNA Ribossômico 16S/genética , Especificidade da Espécie , Baço/efeitos dos fármacos , Baço/metabolismo
16.
Int J Syst Evol Microbiol ; 69(8): 2340-2353, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31162009

RESUMO

Thirty Gram-stain-positive bacterial strains were isolated from Chinese traditional pickle. The strains were characterized using a polyphasic taxonomic approach, including 16S rRNA gene sequence analysis, pheS gene sequence analysis, rpoA gene sequence analysis, fatty acid methyl ester analysis, determination of DNA G+C content, determination of average nucleotide identity (ANI), in silico DNA-DNA hybridization (isDDH), determination of average amino acid identity (AAI) and an analysis of phenotypic features. The data demonstrated that the 30 strains represented 11 novel species belonging to the genus Lactobacillus, strains 159-4T, 47-3T, 257-1T, 187-3T, 220-4T, 151-2BT, 137-3T, 244-4T, 218-10T, 218-6T and 112-3T were designated as the type strains. Strains 159-4T and 47-3T were related to the type strains of Lactobacillus porcinae, Lactobacillus manihotivorans, Lactobacillus nasuensis, Lactobacillus camelliae, Lactobacillus pantheris, Lactobacillus thailandensis, Lactobacillus sharpeae and Lactobacillus songhuajiangensis, having 92.0-98.9 % 16S rRNA gene sequence similarities, 58.1-84.7 % pheS gene sequence similarities and 61.4-90.3 % rpoA gene sequence similarities. Strains 257-1T and 187-3T were related to the type strains of Lactobacillus coryniformis, Lactobacillus iwatensis, Lactobacillus backii, Lactobacillus rennini and Lactobacillus bifermentans, having 93.5-99.3 % 16S rRNA gene sequence similarities, 67.7-81.8 % pheS gene sequence similarities and 77.0-96.2 % rpoA gene sequence similarities. Strains 220-4T, 151-2BT, 137-3T, 244-4T, 218-10T, 218-6T and 112-3T were closely related to the type strains of Lactobacillus paucivorans, Lactobacillus brevis, Lactobacillus hammesii, Lactobacillus senmaizukei, Lactobacillus parabrevis, Lactobacillus yonginensis, Lactobacillus koreensis and Lactobacillus cerevisiae, having 95.6-99.9 % 16S rRNA gene sequence similarities, less than 93.9 % pheS gene sequence similarities and 87.0-99.7 % rpoA gene sequence similarities. ANI, isDDH and AAI values between strains 159-4T, 47-3T, 257-1T, 187-3T, 220-4T, 151-2BT, 137-3T, 244-4T, 218-10T, 218-6T, 112-3T and type strains of phylogenetically related species were less than 92.7, 48.4 and 96.6 %, respectively, confirming that they represent 11 novel species within the genus Lactobacillus. Based upon the data of polyphasic characterization obtained in the present study, eleven novel species, Lactobacillusjixianensis sp. nov., Lactobacillusbaoqingensis sp. nov., Lactobacillusjiayinensis sp. nov., Lactobacilluszhaoyuanensis sp. nov., Lactobacilluslindianensis sp. nov., Lactobacillushuananensis sp. nov., Lactobacillustangyuanensis sp. nov., Lactobacillusfuyuanensis sp. nov., Lactobacillustongjiangensis sp. nov., Lactobacillusfujinensis sp. nov. and Lactobacillusmulengensis sp. nov., are proposed and the type strains are 159-4T (=NCIMB 15175T=CCM 8911T), 47-3T (=NCIMB 15165T=CCM 8903T=LMG 31064T), 257-1T (=NCIMB 15166T=CCM 8904T=LMG 31065T), 187-3T (=NCIMB 15172T =CCM 8910T), 220-4T (=NCIMB 15163T =CCM 8902T=KCTC 21136T), 151-2BT (=NCIMB 15164T=CCM 8913T=KCTC 21129T=LMG 31063T), 137-3T (=NCIMB 15170T=CCM 8907T=KCTC 21125T=LMG 31053T), 244-4T (=NCIMB 15168T=CCM 8906T=KCTC 21137T=LMG 31052T), 218-10T (=NCIMB 15167T=CCM 8905T=KCTC 21135T =LMG 31055T), 218-6T (=NCIMB 15171T=CCM 8908T=KCTC 21134T =LMG 31067T) and 112-3T (=NCIMB 15174T=CCM 8909T=KCTC 21123T=LMG 31049T), respectively.


Assuntos
/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
17.
Nutrients ; 11(6)2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-31159409

RESUMO

Lactobacilli are among the most common microorganisms found in kefir; a traditional fermented milk beverage produced locally in many locations around the world. Kefir has been associated with a wide range of purported health benefits; such as antimicrobial activity; cholesterol metabolism; immunomodulation; anti-oxidative effects; anti-diabetic effects; anti-allergenic effects; and tumor suppression. This review critically examines and assesses these claimed benefits and mechanisms with regard to particular Lactobacillus species and/or strains that have been derived from kefir; as well as detailing further potential avenues for experimentation.


Assuntos
Kefir/microbiologia , Lactobacillus/fisiologia , Probióticos , Humanos , Lactobacillus/classificação , Lactobacillus/genética , Especificidade da Espécie
18.
Int J Syst Evol Microbiol ; 69(7): 2147-2152, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31120413

RESUMO

A Gram-stain-positive bacterial strain, ZW163T, isolated from suancai, a traditional pickle of Heilongjiang province in China, was characterized by a polyphasic approach, including phenotypic characterization, sequence analyses of the 16S rRNA gene and two housekeeping genes (pheS and rpoA), whole genome sequencing and cellular fatty acid composition. Strain ZW163T was phylogenetically related to Lactobacillus sharpeae, Lactobacillus songhuajiangensis, Lactobacillus pantheris and Lactobacillus thailandensis, having 93.8-98.7 % 16S rRNA gene sequence similarities, 76.5-87.1 % pheS gene sequence similarities and 74.3-95.7 % rpoA gene sequence similarities to the type strains of L. sharpeae, L. songhuajiangensis, L. pantheris and L. thailandensis, respectively. Average nucleotide identity and in silico DNA-DNA hybridization values between strain ZW163T and the type strains of L. sharpeae, L. songhuajiangensis, L. pantheris and L. thailandensis were less than 77.7 % and 23.6 % respectively. Based upon the data obtained in the present study, a novel species, Lactobacillushulanensis sp. nov., is proposed and the type strain is ZW163T (=NCIMB 15193T=CCM 8926T=CCTCC AB 2019015T).


Assuntos
/microbiologia , Microbiologia de Alimentos , Lactobacillus/classificação , Filogenia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Genes Bacterianos , Lactobacillus/isolamento & purificação , Hibridização de Ácido Nucleico , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
19.
Food Microbiol ; 82: 218-230, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31027777

RESUMO

Structure of lactic acid bacteria biota in ivy flowers, fresh bee-collected pollen (BCP), hive-stored bee bread, and honeybee gastrointestinal tract was investigated. Although a large microbial diversity characterized flowers and fresh BCP, most of lactic acid bacteria species disappeared throughout the bee bread maturation, giving way to Lactobacillus kunkeei and Fructobacillus fructosus to dominate long stored bee bread and honeybee crop. Adaptation of lactic acid bacteria was mainly related to species-specific, and, more in deep, to strain-specific features. Bee bread preservation seemed related to bacteria metabolites, produced especially by some L. kunkeei strains, which likely gave to lactic acid bacteria the capacity to outcompete other microbial groups. A protocol to ferment BCP was successfully set up, which included the mixed inoculum of selected L. kunkeei strains and Hanseniaspora uvarum AN8Y27B, almost emulating the spontaneous fermentation of bee bread. The strict relationship between lactic acid bacteria and yeasts during bee bread maturation was highlighted. The use of the selected starters increased the digestibility and bioavailability of nutrients and bioactive compounds naturally occurring in BCP. Our biotechnological protocol ensured a product microbiologically stable and safe. Conversely, raw BCP was more exposed to the uncontrolled growth of yeasts, moulds, and other bacterial groups.


Assuntos
Abelhas/microbiologia , Microbiologia de Alimentos , Pólen/metabolismo , Pólen/microbiologia , Própole/metabolismo , Animais , Anti-Infecciosos , Fermentação , Flores/microbiologia , Trato Gastrointestinal/microbiologia , Hanseniaspora/metabolismo , Hedera , Lactobacillales/classificação , Lactobacillales/crescimento & desenvolvimento , Lactobacillales/isolamento & purificação , Lactobacillales/metabolismo , Lactobacillus/classificação , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/isolamento & purificação , Lactobacillus/metabolismo , Interações Microbianas , Microbiota , Pólen/química , Especificidade da Espécie
20.
Food Microbiol ; 82: 259-268, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31027782

RESUMO

The aim of this study is to examine the biogeography of the microbial communities associated to the Spanish-style green olive fermentations in the province of Seville (Andalucía, south-western Spain). Also, to understand how microorganisms colonize and persist in non-sterile food fermentations across a specific table olive producing area, i.e. a specific "agroecosystem". The microbial diversity, bacteria and yeast, in 30 ten-ton fermenters of three different fermentations yards (patios) along the olive fermentation was studied. A total of 951 microbial isolates were obtained which were clustered according to their RAPD profile. A total of 376 distinct genotypes were identified, belonging to 57 different microbial species, 41 bacterial and 16 yeast species. Up to 16 bacterial species had not been described before in table olives. Only the species Lactobacillus pentosus showed a ubiquitous presence in all 30 fermenters. Pediococcus parvulus, Lactobacillus collinoides/paracollinoides, Lactobacillus coryniformis, Lactobacillus plantarum, Pichia manshurica and Candida thaimueangensis were found in every patio. Cosmopolitan strains, up to 15, were shared by the three patios and belonged to the species L. pentosus (12 strains), P. parvulus (1), L. collinoides/paracollinoides (1) and P. manshurica (1). To expand our biodiversity analyses to the "regional" level, we have compared our results with those obtained from two previously studied patios of similar characteristics and in the same geographical area. PERMANOVA analysis of the microbial community composition revealed significant differences among different patios in their structure at every fermentation stage. In contrast, SIMPER analyses showed that, as fermentation progressed, the overall dissimilarities among patios were reduced. Discriminant species were identified for each fermentation stage. Among these, L. pentosus and P. parvulus were "eu-constant" species, while L. collinides/paracollinoides and Marinilactibacillus psychrotolerans group were "constant" species that could be considered microbial key taxa based on the occurrence stability index. The characteristic and, presumably, well adapted microbiota associated to the Spanish-style olive fermentations at the specific geographic area described here is a valuable natural resource which should be preserved conveniently. To our knowledge, this is the first study on the microbial biogeography of table olive fermentations, both at the species and strain levels.


Assuntos
Bactérias/isolamento & purificação , Biodiversidade , Reatores Biológicos/microbiologia , Olea/microbiologia , Leveduras/isolamento & purificação , Bactérias/classificação , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Ecossistema , Fermentação , Microbiologia de Alimentos , Variação Genética , Concentração de Íons de Hidrogênio , Lactobacillus/classificação , Lactobacillus/genética , Lactobacillus/crescimento & desenvolvimento , Lactobacillus/isolamento & purificação , Microbiota/genética , Sais , Espanha , Leveduras/classificação , Leveduras/genética , Leveduras/crescimento & desenvolvimento
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA