scholarly journals Determination of important enzymes and antimicrobial resistances of gram-positive haloalkaliphilic bacteria isolated from Salda Lake

2021 ◽  
Vol 38 (3) ◽  
pp. 375-382
Author(s):  
Pınar Çağlayan

As an extreme environment, soda lakes harbor various haloalkaliphilic microorganisms. Salda Lake is one of the natural soda lake (pH˃9) in Turkey. Haloalkaliphiles are unique microorganisms in their ability to live in high alkaline and high saline conditions, and play an important role in biodegradation and bioremediation of hydrocarbons. Hence, the aims of this study were to isolate haloalkaliphilic bacteria from water sample of Salda Lake, to identify these isolates by both conventional and molecular methods, to screen their industrially important enzymes, and to investigate their antimicrobial resistance profiles. Six isolates were identified as Bacillus horneckiae, Bacillus subtilis, Bacillus paramycoides, Bacillus pumilus, Staphylococcus epidermidis, Bacillus haynesii according to 16S rRNA gene sequencing analysis. The industrially important enzymes (amylase, cellulase, pullulanase, lipase, urease, protease, caseinase, oxidase, catalase) were produced by haloalkaliphilic isolates. These enzymes maybe used in alkaline and saline industrial processes. Although Bacillus subtilis was susceptible to all antibiotics, other isolates showed resistance to at least one antibiotic. The resistance against antibiotics were found as ampicillin/sulbactam 83%, amoxycillin/clavulanic acid 83%, ampicillin 67%, mupirocin 67%, chloramphenicol 50%, tetracycline 50%, imipenem 50%, meropenem 50%, cefadroxil 17%. These bacteria may have develope resistance to antibiotics that entering their natural environment in different ways.

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Takayuki Matsuoka ◽  
Takuya Shimizu ◽  
Tadanori Minagawa ◽  
Wakiko Hiranuma ◽  
Miki Takeda ◽  
...  

Abstract Background Bacteroides dorei is an anaerobic gram-negative bacterium first described in 2006. Because of the high similarity in mass spectra between B. dorei and Bacteroides vulgatus, discriminating between these species is arduous in clinical practice. In recent decades, 16S rRNA gene sequencing has been a complementary method for distinguishing taxonomically close bacteria, including B. dorei and B. vulgatus, at the genus and species levels. Consequently, B. dorei has been shown to contribute to some diseases, including type 1 autoimmune diabetes mellitus and atherosclerotic diseases. However, there are no reports on invasive infectious diseases caused by B. dorei. This report describes the first case of direct invasion and colonisation of human tissue by B. dorei, thus providing a warning regarding the previously proposed application of B. dorei as a live biotherapeutic for atherosclerotic diseases. Case presentation A 78-year-old Japanese man complained of intermittent chest/back pain and was diagnosed with a mycotic thoracic aortic aneurysm by enhanced computed tomography on admission. Despite strict blood pressure control and empirical antibiotic therapy, the patient’s condition worsened. To prevent aneurysmal rupture and eliminate infectious foci, the patient underwent surgical treatment. The resected specimen was subjected to tissue culture and 16S rRNA gene sequencing analysis to identify pathogenic bacteria. A few days after the surgery, culture and sequencing results revealed that the pathogen was B. dorei/B. vulgatus and B. dorei, respectively. The patient was successfully treated with appropriate antibacterial therapy and after improvement, was transferred to another hospital for rehabilitation on postoperative day 34. There was no recurrence of infection or aneurysm after the patient transfer. Conclusions This report describes the first case of invasive infectious disease caused by B. dorei, casting a shadow over its utilisation as a probiotic for atherosclerotic diseases.


Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1519
Author(s):  
Meinan Chang ◽  
Fengtao Ma ◽  
Jingya Wei ◽  
Junhao Liu ◽  
Xuemei Nan ◽  
...  

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.


2017 ◽  
Vol 2017 (1) ◽  
pp. 2256-2267
Author(s):  
Ruta Suresh Deshpande ◽  
Devi Sundaravadivelu ◽  
Pablo Campo ◽  
Jorge W. SantoDomingo ◽  
Robyn N. Conmy

Abstract 2017-271 In recent years, diluted bitumen (or dilbit) has become an important source of hydrocarbon-based fuel. While information on the degradation of crude oils has been well researched, dilbit degradation has been studied at a much lesser extent. The objective of this study was to compare biodegradation of dilbit with a conventional crude oil (CCO) under various conditions. Two different microcosm experiments were set up, one containing a mixed culture acclimated to dilbit (Kalamazoo River Enrichment, KRC) and the other having a mixed culture enriched on soil contaminated with hydrocarbons (Anderson Ferry Enrichment, AFC). The microcosms were run for 60 d at 25 °C and for 72 days at 5 °C in flasks containing sterile Bushnell Hass broth and naturally dispersed oil. Each flask was inoculated with the KRC and AFC mixed cultures, and rotated on an orbital shaker (200 rpm) at the above stated temperatures. On each sampling day, triplicates were sacrificed to determine the residual hydrocarbon concentration. Additionally, some samples were used to determine the bacterial composition using 16S rRNA gene sequencing analysis. Hydrocarbon analysis (alkanes and PAHs) was performed by gas chromatography/mass spectrometry (GC/MS/MS). Higher degradation rates were achieved at 25 °C as compared to 5 °C. All the enrichments metabolized CCO as well dilbit, but the nature and extent of the degradation was distinct. KRC meso culture was the most effective among all, as it completely removed alkanes and most of the PAHs. AFC enrichment performed differently at the two temperatures; an acclimation period (8 d) was observed at 5 °C while there was no lag at 25 °C. KRC cryo culture as well as AFC culture at both temperatures degraded alkanes completely while they were not able to metabolize heavier fractions of the oil (C2–4 homologues of 3- and 4-ring compounds). All cultures showed the presence of diverse oil degrading bacteria and the differences in their compositions affected the biodegradation. Although dilbit was biodegraded, for all the treatments except AFC at 5 °C, the rate of degradation and the extent of degradation was greater for CCO owing to the higher concentrations of lighter hydrocarbons.


2010 ◽  
Vol 4 (1) ◽  
pp. 123-131 ◽  
Author(s):  
Jens JØrgen Christensen ◽  
Brita Bruun ◽  
Ute Wolff Sönksen ◽  
Lisbeth Nielsen ◽  
Annemarie Hesselbjerg ◽  
...  

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Kaoru Haro ◽  
Midori Ogawa ◽  
Mitsumasa Saito ◽  
Koichi Kusuhara ◽  
Kazumasa Fukuda

AbstractNasopharyngeal colonization by bacteria is a prerequisite for progression to respiratory disease and an important source of horizontal spread within communities. We aimed to perform quantitative analysis of the bacterial cells and reveal the microbiota of the nasal discharge in children at the species level based on highly accurate 16S rRNA gene sequencing. This study enrolled 40 pediatric patients with rhinorrhea. The bacterial cells in the nasal discharge were counted by epifluorescence microscopic analysis. The microbiota was analyzed by using the 16S rRNA gene clone library sequencing method. We demonstrated that a high abundance (median 2.2 × 107 cells/mL) of bacteria was contained in the nasal discharge of children. Of the 40 samples, 37 (92.5%) were dominated by OTUs corresponding to Haemophilus aegyptius/influenzae, Moraxella catarrhalis/nonliquefaciens, or Streptococcus pneumoniae. These samples showed higher cell abundance and lower alpha diversity than the remaining three samples in which the other bacteria coexisted. In addition, 12 sequences with low homology to type strains were considered as previously unknown bacterial lineages. In conclusion, the nasal discharge of most young children contains a large amount of respiratory pathogens and several unknown bacteria, which could not only cause endogenous infection but also be a source of transmission to others.


2021 ◽  
Vol 9 (5) ◽  
pp. 1030
Author(s):  
Ke Liu ◽  
Siyu Chen ◽  
Jing Huang ◽  
Feihong Ren ◽  
Tingyu Yang ◽  
...  

The oral microbiota can be affected by several factors; however, little is known about the relationship between diet, ethnicity and commensal oral microbiota among school children living in close geographic proximity. In addition, the relationship between the oral and gut microbiota remains unclear. We collected saliva from 60 school children from the Tibetan, Han and Hui ethnicities for a 16S rRNA gene sequencing analysis and comparison with previously collected fecal samples. The study revealed that Bacteroidetes and Proteobacteria were the dominant phyla in the oral microbiota. The Shannon diversity was lowest in the Tibetan group. A PCA showed a substantial overlap in the distribution of the taxa, indicating a high degree of conservation among the oral microbiota across ethnic groups while the enrichment of a few specific taxa was observed across different ethnic groups. The consumption of seafood, poultry, sweets and vegetables was significantly correlated with multiple oral microbiotas. Furthermore, 123 oral genera were significantly associated with 191 gut genera. A principal coordinate analysis revealed that the oral microbiota clustered separately from the gut microbiota. This work extends the findings of previous studies comparing microbiota from human populations and provides a basis for the exploration of the interactions governing the tri-partite relationship between diet, oral microbiota and gut microbiota.


Interação ◽  
2021 ◽  
Vol 21 (2) ◽  
pp. 38-48
Author(s):  
Renato Ventresqui Oliveira ◽  
Lívio da Silva Amaral ◽  
Cristiane Aparecida Milagres ◽  
Celso Tadeu Barbosa Santos ◽  
Afonso Pelli ◽  
...  

Bacillus subtilis is a spore-forming bacterium and an important food contaminant. The aim of this study was to analyze the ability of B. subtilis spores to survive under conditions of low pH and high temperature. The package was purchased at a local supermarket, in Uberaba, Minas Gerais. A sample was collected, diluted and plated on Brain-Heart-Infusion agar (BHI). After incubation, suspected colonies of B. subtilis were transferred to BHI agar. Cell morphology, the presence of spores and Gram stain were examined, and the isolate was identified by 16S rRNA gene sequencing . The microscope evaluation indicated the presence of spores. The thermal tolerance of the spores was evaluated by the addition of 3x109spores/mL in test tubes containing peptone water. Heat treatments were carried out at 80 and 90°C at different incubation times (0, 10, 20, 30, 40, 50 and 60 min). After heating, the tubes were cooled and the number of viable spores was determined in BHI Agar. For the analysis of spore survival, D and Z values were calculated. Tolerance to acid conditions was evaluated using BHI broth with different pH values. After incubation, the bacterial concentration was determined by determining viable cell count on BHI Agar medium. The vegetative cells were transferred to the BHI broth and the pH was adjusted to different values (3, 4 or 5). Sampling were taken 8, 12 and 24 h after incubation. The samples were serially diluted in peptone water and spread in BHI Agar to determine the viable cell count . The 16S rRNA gene sequencing indicated high similarity (99.99%) with B. subtilis. D values were 17.01 min at 80°C and 13.42 min at 90°C. The Z-value was 97.13°C. B. subtilis was not able to grow at pH 3 and pH 4, but its survival was confirmed after the growth of colonies on BHI agar. At pH 5, B. subtilis grew after 24 h and the final pH changed to 7. Our results suggest that the spores of B. subtilis isolated from fruit juice-added soy beverage are tolerant to low pH and high temperature.


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