scholarly journals Biocontrol Potential of Endophytic Actinobacteria against Fusarium solani, the Causal Agent of Sudden Decline Syndrome on Date Palm in the UAE

2021 ◽  
Vol 8 (1) ◽  
pp. 8
Author(s):  
Aisha A. Alblooshi ◽  
Gouthaman P. Purayil ◽  
Esam Eldin Saeed ◽  
Gaber A. Ramadan ◽  
Saeed Tariq ◽  
...  
Keyword(s):  
Date Palm ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Suitable Habitat ◽  
Cell Wall Degrading Enzymes ◽  
Root Cortex ◽  
Antifungal Metabolites ◽  
Rrna Gene Sequencing ◽  
Root Tissues

Thirty-one endophytic streptomycete and non-streptomycete actinobacteria were isolated from healthy date palm root tissues. In vitro screening revealed that the antifungal action of isolate #16 was associated with the production of cell-wall degrading enzymes, whereas with diffusible antifungal metabolites in isolate #28, albeit their production of volatile antifungal compounds. According to the 16S rRNA gene sequencing, isolates #16 and #28 were identified as Streptomyces polychromogenes UAE2 (Sp; GenBank Accession #: OK560620) and Streptomyces coeruleoprunus UAE1 (Sc; OK560621), respectively. The two antagonists were recovered from root tissues until 12 weeks after inoculation, efficiently colonized root cortex and xylem vessels, indicating that the date palm roots are a suitable habitat for these endophytic isolates. At the end of the greenhouse experiments, the development of sudden decline syndrome (SDS) was markedly suppressed by 53% with the application of Sp and 86% with Sc, confirming their potential in disease management. Results showed that the estimated disease severity indices in diseased seedlings were significantly (p < 0.05) reduced from 4.75 (scale of 5) to 2.25 or 0.67 by either Sp or Sc, respectively. In addition, conidial numbers of the pathogen significantly (p < 0.05) dropped by 38% and 76% with Sp and Sc, respectively, compared to infected seedlings with F. solani (control). Thus, the suppression of disease symptoms was superior in seedlings pre-inoculated with S. coeruleoprunus, indicating that the diffusible antifungal metabolites were responsible for F. solani retardation in these plants. This is the first report of actinobacteria naturally existing in date palm tissues acting as microbial antagonists against SDS on date palm.

scholarly journals The growth response of rice (Oryza sativa L. var. FARO 44) in vitro after inoculation with bacterial isolates from a typical ferruginous ultisol

2021 ◽  
Vol 45 (1) ◽  
Author(s):  
Musa Saheed Ibrahim ◽  
Beckley Ikhajiagbe
Keyword(s):  
16S Rrna ◽  
Bacillus Cereus ◽  
Proteus Mirabilis ◽  
Growth Response ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Rice Seedlings ◽  
Rrna Gene Sequencing

Abstract Background Rice forms a significant portion of food consumed in most household worldwide. Rice production has been hampered by soil factors such as ferruginousity which has limited phosphorus availability; an important mineral component for the growth and yield of rice. The presence of phosphate-solubilizing bacteria (PSB) in soils has been reported to enhance phosphate availability. In view of this, the present study employed three bacteria species (BCAC2, EMBF2 and BCAF1) that were previously isolated and proved P solubilization capacities as inocula to investigate the growth response of rice germinants in an in vitro setup. The bacteria isolates were first identified using 16S rRNA gene sequencing and then applied as inoculum. The inolula were prepared in three concentrations (10, 7.5 and 5.0 ml) following McFarland standard. Viable rice (var. FARO 44) seeds were sown in petri dishes and then inoculated with the three inocula at the different concentrations. The setup was studied for 28 days. Results 16S rRNA gene sequencing identified the isolates as: isolate BCAC2= Bacillus cereus strain GGBSU-1, isolate BCAF1= Proteus mirabilis strain TL14-1 and isolate EMBF2= Klebsiella variicola strain AUH-KAM-9. Significant improvement in rice germination, morphology, physiology and biomass parameters in the bacteria-inoculated setups was observed compared to the control. Germination percentage after 4 days was 100 % in the inoculated rice germinants compared to 65% in the control (NiS). Similarly, inoculation with the test isolates enhanced water-use efficiency by over 40%. The rice seedlings inoculated with Bacillus cereus strain GGBSU-1 (BiS) showed no signs of chlorosis and necrosis throughout the study period as against those inoculated with Proteus mirabilis strain TL14-1 (PiS) and Klebsiella variicola strain AUH-KAM-9 (KiS). Significant increase in chlorophyll-a, chlorophyll-b and alpha amylase was observed in the rice seedlings inoculated with BiS as against the NiS. Conclusion Inoculating rice seeds with Bacillus cereus strain GGBSU-1, Proteus mirabilis strain TL14-1 and Klebsiella variicola strain AUH-KAM-9 in an in vitro media significantly improved growth parameters of the test plant. Bacillus cereus strain GGBSU-1 showed higher efficiency due to a more improved growth properties observed.

scholarly journals Live Bacillus subtilis natto Promotes Rumen Fermentation by Modulating Rumen Microbiota In Vitro

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1519
Author(s):  
Meinan Chang ◽  
Fengtao Ma ◽  
Jingya Wei ◽  
Junhao Liu ◽  
Xuemei Nan ◽  
...  
Keyword(s):  
Bacillus Subtilis ◽  
16S Rrna ◽  
Rumen Fluid ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rumen Fermentation ◽  
Rrna Gene ◽  
Bacillus Subtilis Natto ◽  
Rrna Gene Sequencing

Previous studies have shown that Bacillus subtilis natto affects rumen fermentation and rumen microbial community structure, which are limited to detect a few microbial abundances using traditional methods. However, the regulation of B. subtilis natto on rumen microorganisms and the mechanisms of microbiota that affect rumen fermentation is still unclear. This study explored the effects of live and autoclaved B. subtilis natto on ruminal microbial composition and diversity in vitro using 16S rRNA gene sequencing and the underlying mechanisms. Rumen fluid was collected, allocated to thirty-six bottles, and divided into three treatments: CTR, blank control group without B. subtilis natto; LBS, CTR with 109 cfu of live B. subtilis natto; and ABS, CTR with 109 cfu of autoclaved B. subtilis natto. The rumen fluid was collected after 0, 6, 12, and 24 h of fermentation, and pH, ammonia nitrogen (NH3-N), microbial protein (MCP), and volatile fatty acids (VFAs) were determined. The diversity and composition of rumen microbiota were assessed by 16S rRNA gene sequencing. The results revealed LBS affected the concentrations of NH3-N, MCP, and VFAs (p < 0.05), especially after 12 h, which might be attributed to changes in 18 genera. Whereas ABS only enhanced pH and NH3-N concentration compared with the CTR group (p < 0.05), which might be associated with changes in six genera. Supplementation with live B. subtilis natto improved ruminal NH3-N and propionate concentrations, indicating that live bacteria were better than autoclaved ones. This study advances our understanding of B. subtilis natto in promoting ruminal fermentation, providing a new perspective for the precise utilization of B. subtilis natto in dairy rations.

scholarly journals Findings from the microbiota of tooth apical periodontitis and the search for pathogen forgranulomatous inflammation

2021 ◽  
Author(s):  
Yuanyuan Wang ◽  
Hao Xu ◽  
Minghui Wei ◽  
Yuhong Wang ◽  
Wenzhe Wang ◽  
...  
Keyword(s):  
Granulomatous Inflammation ◽  
16S Rrna Gene Sequencing ◽  
Apical Periodontitis ◽  
Rrna Gene ◽  
Normal Microbiota ◽  
Rrna Gene Sequencing ◽  
Neisseria Subflava ◽  
Foot Pad

Abstract BackgroundOrofacial granulomatosis (OFG) is a granulomatous inflammation (GI) disease in maxillofacial region, the underlying cause of it remains unknown. Our previous study demonstrated that tooth apical periodontitis (AP) plays a significant role in the pathogenesis of OFG, we aimed here to characterize the AP bacterial signatures of OFG patients, and identify bacteria that may be important pathogens capable of inducing OFG.ResultsThe composition of AP microbiota in OFG cases and common AP controls was compared using 16S rRNA gene sequencing, the results showed a specific AP microbiota signature in OFG patients, characterized by domination of phyla Firmicutes and Proteobacteria , notably members of Streptococcus, Lactobacillus and Neisseria. To assess the pathogenicity of the potential pathogens in OFG, we isolated and successfully in vitro cultured Streptococcus, Lactobacillus casei, Neisseria subflava, Veillonella parvula and Actinomyces from OFG patients, and injected the clinical isolates into mice respectively. Ultimately, foot pad injection with N. subflava elicited granulomatous inflammation, and the virulence of N. subflava was verified based on Koch’s postulates.ConclusionsOur findings confirmed the role of bacteria in OFG, and first suggested that the component of the host normal microbiota, N. subflava is likely a pathogen for GI.

scholarly journals Personalized Bioconversion of Panax Notoginseng Saponins Mediated by Gut Microbiota Between Chinese-diet and Western-diet Healthy Subjects

2021 ◽  
Author(s):  
Li Wang ◽  
Man-Yun Chen ◽  
Li Shao ◽  
Wei Zhang ◽  
Xiang-Ping Li ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Lactobacillus Rhamnosus ◽  
16S Rrna Gene Sequencing ◽  
Panax Notoginseng ◽  
Rrna Gene ◽  
Bifidobacterium Adolescentis ◽  
Correlative Analysis ◽  
Rrna Gene Sequencing ◽  
The Relationship

Abstract Background: Panax notoginseng saponins (PNS) as the main effective substances from P. notoginseng with low bioavailability could be bio-converted by human gut microbiota. In our previous study, PNS metabolic variations mediated by gut microbiota have been observed between high fat, high protein (HF-HP)-diet and low fat, plant fiber-rich (LF-PF)-diet subjects. In this study, we aimed to correspondingly characterize the relationship between distinct gut microbiota profiles and PNS metabolites. Methods: Gut microbiota were collected from HF-HP and LF-PF healthy adults, respectively and profiled by 16S rRNA gene sequencing. PNS were incubated with gut microbiota in vitro. A LC-MS/MS method was developed to quantify the five main metabolites yields including ginsenoside F1 (GF1), ginsenoside Rh2 (GRh2), ginsenoside compound K (GC-K), protopanaxatriol (PPT) and protopanaxadiol (PPD). The selected microbial species, Bifidobacterium adolescentis and Lactobacillus rhamnosus, were employed to metabolize PNS for the corresponding metabolites.Results: The five main metabolites were significantly different between the two diet groups. Compared with HF-HP group, the microbial genus Blautia, Bifidobacterium, Clostridium, Corynebacterium, Dorea, Enhydrobacter, Lactobacillus, Roseburia, Ruminococcus, SMB53, Streptococcus, Treponema and Weissella were enriched in LF-PF group, while Phascolarctobacterium and Oscillospira were relatively decreased. Furthermore, Spearman’s correlative analysis revealed gut microbiota enriched in LF-PF and HF-HP groups were positively and negatively associated with PNS metabolites yields, respectively. Conclusions: Our data showed gut microbiota diversity led to the personalized bioconversion of PNS.

scholarly journals Analysis of Vaginal and Endometrial Microbiota Communities in Infertile Women With a History of Repeated Implantation Failure

2020 ◽  
Author(s):  
Takuhiko Ichiyama ◽  
Keiji Kuroda ◽  
Yoko Nagai ◽  
Daichi Urushiyama ◽  
Motoharu Ono ◽  
...  
Keyword(s):  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Implantation Failure ◽  
Repeated Implantation Failure ◽  
Infertile Women ◽  
Healthy Women ◽  
History Of ◽  
Rrna Gene Sequencing ◽  
Clinical Trials Registry

Abstract Background: Repeated implantation failure (RIF) is estimated to occur in 15%–20% of infertile women undergoing in vitro fertilization-embryo transfer (IVF-ET). Molecular identification recently confirmed that the uterine microbiota may have implications for reproductive and obstetrical outcomes. One hundred forty-five women who had been diagnosed with RIF were enrolled in the study. Twenty-one healthy women were also enrolled as controls. We investigated their vaginal and endometrial microbiotas using 16S rRNA gene sequencing and compared the microbiota profiles in the patients with RIF and controls.Results: The endometrial microbiotas had higher α-diversities than did the vaginal microbiotas (p<0.001 in both patients with RIF and healthy women). The microbiota profiles showed that vaginal and endometrial samples in patients with RIF had significantly higher levels of 5 and 14 bacterial genera, respectively, than those in healthy women. These genera included Atopobium, Gardnerella, Prevotella and Megasphaera. Vaginal Lactobacillus rates in patients with RIF were significantly lower at 76.4 ± 38.9% compared with those of the controls at 91.8 ± 22.7% (p=0.018), but endometrial Lactobacillus rates did not significantly differ between the RIF patients and controls (56.2 ± 36.4% and 58.8 ± 37.0%, respectively, p=0.79) Conclusions: Impaired microbiota communities containing specific bacteria in both the endometrium and vagina were associated with implantation failure. The Lactobacillus rate in the vagina, but not the endometrium, may be a biomarker for RIF.Trial registration: UMIN Clinical Trials Registry, UMIN000031731, Registered 15 March 2018; https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000036121

scholarly journals Diversity of Nematode Microbial Antagonists from Algeria Shows Occurrence of Nematotoxic Trichoderma spp.

Plants ◽  
2020 ◽  
Vol 9 (8) ◽  
pp. 941
Author(s):  
Nawal Benttoumi ◽  
Mariantonietta Colagiero ◽  
Samira Sellami ◽  
Houda Boureghda ◽  
Abdelaziz Keddad ◽  
...  
Keyword(s):  
Pcr Amplification ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Fusarium Spp ◽  
Trichoderma Spp ◽  
Bacillus Spp ◽  
Phytoparasitic Nematodes ◽  
Meloidogyne Spp ◽  
Rrna Gene Sequencing

Fungi and bacteria associated to phytoparasitic nematodes Globodera rostochiensis and Meloidogyne spp. in Algeria were identified and characterized. Trichoderma spp. showed the highest prevalence in the cysts of G. rostochiensis. A number of isolates were identified through PCR amplification and the sequencing of the internal transcribed spacer (ITS)1-2 and Rpb2 gene regions. The most represented species were T. harzianum and T. afroharzianum. The latter and T. hirsutum were reported for the first time in Algeria. Fusarium spp., including F. oxysporum and F. solani, comprised a second group of fungi found in cysts. Taxa associated to females of Meloidogyne spp. included T. harzianum, Fusarium spp. and other hyphomycetes. To assess the efficacy of Trichoderma spp., two assays were carried out in vitro with the culture filtrates of two T. afroharzianum and T. harzianum isolates, to check their toxicity versus the second stage juveniles of M. incognita. After 24–48 h exposure, a mortality significantly higher than the control was observed for both filtrates at 1% dilutions. The TRI genes involved in the production of trichothecenes were also amplified with the PCR from some Trichoderma spp. isolates and sequenced, supporting a putative role in nematode toxicity. Bacteria isolated from the cysts of G. rostochiensis included Brucella, Rhizobium, Stenotrophomonas and Bacillus spp., identified through 16S rRNA gene sequencing. The potential of the microbial isolates identified and their mechanisms of action are discussed, as part of a sustainable nematode management strategy.

In vitro and in vivo characterization and strain safety of Lactobacillus reuteri NCIMB 30253 for probiotic applications

2012 ◽  
Vol 58 (6) ◽  
pp. 776-787 ◽  
Author(s):  
Imran Sulemankhil ◽  
Mathieu Parent ◽  
Mitchell Lawrence Jones ◽  
Zhenqian Feng ◽  
Alain Labbé ◽  
...  
Keyword(s):  
Lactobacillus Reuteri ◽  
16S Rrna Gene Sequencing ◽  
Human Consumption ◽  
Rrna Gene ◽  
Sprague Dawley ◽  
Dose Study ◽  
Clinically Significant ◽  
Rrna Gene Sequencing

Lactobacillus reuteri NCIMB 30253 was shown to have potential as a probiotic by reducing the proinflammatory chemokine interleukin-8. Moreover, this strain was evaluated, by in vitro and in vivo techniques, for its safety for human consumption. The identity of the strain was investigated by metabolic profiling and 16S rRNA gene sequencing, and in vitro safety evaluations were performed by molecular and metabolic techniques. Genetic analysis was confirmed by assessing the minimum inhibitory concentration to a panel of antibiotics, showing that the strain was susceptible to 8 antibiotics tested. The ability of the strain to produce potentially harmful by-products and antimicrobial compounds was evaluated, showing that the strain does not produce biogenic amines and does not show bacteriocin activity or reuterin production. A 28-day repeated oral dose study was conducted in normal Sprague–Dawley rats to support the in vivo strain safety. Oral administration of the strain resulted in no changes in general condition and no clinically significant changes to biochemical and haematological markers of safety relative to vehicle control treated animals. This comprehensive assessment of safety of L. reuteri NCIMB 30253 supports the safety of the strain for use as a probiotic.

scholarly journals Abundant DNase I-Sensitive Bacterial DNA in Healthy Porcine Lungs and Its Implications for the Lung Microbiome

2013 ◽  
Vol 79 (19) ◽  
pp. 5936-5941 ◽  
Author(s):  
Alejandro A. Pezzulo ◽  
Patrick H. Kelly ◽  
Boulos S. Nassar ◽  
Cedric J. Rutland ◽  
Nicholas D. Gansemer ◽  
...  
Keyword(s):  
Bacterial Diversity ◽  
Defense Mechanisms ◽  
16S Rrna Gene Sequencing ◽  
Dnase I ◽  
Rrna Gene ◽  
Bacterial Dna ◽  
Lung Microbiome ◽  
Rrna Gene Sequencing ◽  
Porcine Lungs

ABSTRACTHuman lungs are constantly exposed to bacteria in the environment, yet the prevailing dogma is that healthy lungs are sterile. DNA sequencing-based studies of pulmonary bacterial diversity challenge this notion. However, DNA-based microbial analysis currently fails to distinguish between DNA from live bacteria and that from bacteria that have been killed by lung immune mechanisms, potentially causing overestimation of bacterial abundance and diversity. We investigated whether bacterial DNA recovered from lungs represents live or dead bacteria in bronchoalveolar lavage (BAL) fluid and lung samples in young healthy pigs. Live bacterial DNA was DNase I resistant and became DNase I sensitive upon human antimicrobial-mediated killingin vitro. We determined live and total bacterial DNA loads in porcine BAL fluid and lung tissue by comparing DNase I-treated versus untreated samples. In contrast to the case for BAL fluid, we were unable to culture bacteria from most lung homogenates. Surprisingly, total bacterial DNA was abundant in both BAL fluid and lung homogenates. In BAL fluid, 63% was DNase I sensitive. In 6 out of 11 lung homogenates, all bacterial DNA was DNase I sensitive, suggesting a predominance of dead bacteria; in the remaining homogenates, 94% was DNase I sensitive, and bacterial diversity determined by 16S rRNA gene sequencing was similar in DNase I-treated and untreated samples. Healthy pig lungs are mostly sterile yet contain abundant DNase I-sensitive DNA from inhaled and aspirated bacteria killed by pulmonary host defense mechanisms. This approach and conceptual framework will improve analysis of the lung microbiome in disease.

scholarly journals Biodiversity of dominant cultivable endophytic bacteria inhabiting tissues of six different cultivars of maize (Zea mays L. ssp. mays) cropped under field conditions

2015 ◽  
Vol 64 (2) ◽  
pp. 163-170 ◽  
Author(s):  
KATARZYNA PISARSKA ◽  
STANISŁAW JERZY PIETR
Keyword(s):  
Endophytic Bacteria ◽  
16S Rrna Gene Sequencing ◽  
Field Conditions ◽  
Temperate Climate ◽  
Rrna Gene ◽  
Maize Cultivars ◽  
Bacillus Simplex ◽  
Rrna Gene Sequencing ◽  
Arthrobacter Nicotinovorans

Endophytic bacteria (EnB) play a crucial role in plant development. This study was an attempt to isolate and identify dominant cultivable EnB inhabiting young seedlings germinated in vitro and leaves of six maize cultivars grown under field conditions at temperate climate zone with culture-dependent approach. We isolated bacteria from field cropped maize only. Strains were identified based on 16S rRNA gene sequencing. In particular, members of Actinobacteria, Bacteroidetes, Firmicutes and α- and γ-Proteobacteria were found. Species of two genus Pseudomonas and Bacillus were dominant among them. Higher diversity of EnB was found in plants collected from Kobierzyce, where we identified 35 species from 16 genera with 22 species uniquely found at this field. On the contrary, from maize leaves collected at Smolice we identified 24 species representing 10 genera with 10 species uniquely isolated from this field. However, none of species was common for all cultivars at both locations. Among isolated EnB six species only, Pseudomonas clemancea, Pseudomonasfluorescens, Bacillus megaterium, Bacillus simplex, Arthrobacter nicotinovorans and Arthrobacter nitroguajacolicus, were found in aboveground parts of the same cultivar grown on both tested fields. The fact that the same cultivars, sown from the same lots of seeds, under field conditions on two different locations were colonized with noticeably different associations of cultivable EnB suggest that cultivar genotype is an important factor selecting endophytic bacteria from local agro-environment. To our knowledge this is first report about the significant variation of diversity of cultivable endophytic bacteria inhabiting aboveground parts of the same maize cultivars grown at different locations.

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