Microaerophilic Conditions Research Articles

Vplyv prídavku baktérií mliečneho kvasenia na mikrobiotu a výskyt mykotoxínov v ražných silážach

In the rye silages (control and with additive) microbiota isolation and identification (total count of bacteria, coliform bacteria, enterococci, lactic acid bacteria and microscopic filamentous fungi) and occurrence of mycotoxins were evaluated. Total count of bacteria on Plate count agar at 30 °C for 48-72 hours, coliform bacteria on McConkey agar at 37 °C for 24-48 hours, enterococci on Enterococcus selective agar at 37 °C for 48-72 hours, lactic acid bacteria on De Man, Rogosa and Sharpe agar, Mayeux, Sandine and Elliker and All Purpose TWEEN® agar 37 °C for 48-72 hours in microaerophilic condition and microscopic filamentous fungi on Malt extract agar at 25 °C for 7 days were evaluated and identified with mass spectrometry. The most isolated family from both types of silage was Lactobacillaceae. The most isolated species from the control group was Lentilactobacillus buchneri (22%) and lowest number of isolated species was Mucor spp. The most isolated species from silage with additive was Lactobacillus gasseri (16%). The ELISA method (enzyme-linked immunosorbent assay) with using reader at wavelength 650 nm for detection and quantification of mycotoxins (total aflatoxins, total ochratoxins, total fumonisins, deoxynivalenol, zearalenone and T-2/HT-2 toxin) was used. The rye silages of control and with additive were characterized by occurrence of all analyzed mycotoxins, whereas deoxynivalenol was mycotoxin with the highest concentration. The rye silages with additive had significantly lower content of total aflatoxins compared to the control, while significantly higher concentration of total ochratoxins, deoxynivalenol, zearalenone and T-2/HT-2 toxin was determined.

Biodegradation of Reactive Red 198 by textile effluent adapted microbial strains.

A sustainable technology to eliminate the persistent reactive dyes from the textile effluents discharged indiscriminately in the environment is highly desirous given the explosive growth of textile industries. The present study investigated the potential of two different bacterial strains, Bacillus cereus SKB12 and Enterobacter hormaechei SKB16 isolated from the dye house effluent sludge in the biotransformation of Reactive Red 198 (RR 198). Process variables such as temperature, pH, shaking conditions and contact time were optimized for the successful decolourization of RR 198. Maximum decolourization of 80% and 85% of RR 198 was achieved at pH 6 and 7, and 40°C in microaerophilic conditions on treatment with B. cereus and E. hormaechei, respectively. High-Performance Liquid Chromatography (HPLC), and Gas Chromatography-Mass Spectrometry (GC-MS) analyses conducted further affirmed that the decolourization of RR 198 was rather due to biodegradation than biosorption through shift in wavenumbers, retention time variations and the appearance of lesser molecular weight peaks. Degradative pathway for RR 198 predicted based on the enzyme assay data and dye degraded metabolite peaks acquired through GC-MS analysis highlighted the significance of azoreductase and laccase in the degradation of RR 198 into smaller non-toxic compounds. In addition, toxicity assessment through zootoxicological and phytotoxicological experiments using brine shrimp and Vigna radiata validated the detoxified status of the metabolites thus proving the promising potentials of the bacterial strains in the remediation of azo dyes.

A Polyclonal Antibody Raised against the Burkholderia cenocepacia OmpA-like Protein BCAL2645 Impairs the Bacterium Adhesion and Invasion of Human Epithelial Cells In Vitro.

Respiratory infections by bacteria of the Burkholderia cepacia complex (Bcc) remain a life threat to cystic fibrosis (CF) patients, due to the faster lung function decline and the absence of effective eradication strategies. Immunotherapies are regarded as an attractive alternative to control and reduce the damages caused by these infections. In this work, we report the cloning and functional characterization of the OmpA-like BCAL2645 protein, previously identified and found to be immunoreactive against sera from CF patients with a record of Bcc infections. The BCAL2645 protein is shown to play a role in biofilm formation, adherence to mucins and invasion of human lung epithelial cells. The expression of the BCAL2645 protein was found to be increased in culture medium, mimicking the lungs of CF patients and microaerophilic conditions characteristic of the CF lung. Moreover, a polyclonal antibody raised against BCAL2645 was found to inhibit, by about 75 and 85%, the ability of B. cenocepacia K56-2 to bind and invade in vitro CFBE41o- human bronchial epithelial cells. These results highlight the potential of anti-BCAL2645 antibodies for the development of passive immunization therapies to protect CF patients against Bcc infections.

An in vitro assay of the effect of lysine oxidation end-product, α-aminoadipic acid, on the redox status and gene expression in probiotic Lactobacillus reuteri PL503

This study was designed to gain information about the underlying mechanisms of the effects of a food-occurring free oxidized amino acid, α-aminoadipic acid (AAA), on the probiotic Lactobacillus reuteri PL503. This bacterium was incubated in colonic-simulated conditions (37 °C for 24 h in microaerophilic conditions) and exposed to three food-compatible AAA concentrations, namely, 1 mM, 5 mM, and 10 mM. A control group with no AAA exposure was also considered. Each of the four experimental conditions was replicated three times and samplings were collected at 12, 16, 20, and 24 h. The downregulation of the uspA gene by AAA (0.5-fold decrease as compared to control) suggests that AAA is identified as a potential chemical threat. The dhaT gene, implicated in the antioxidant defense, was found to be upregulated in bacteria treated with 1 and 5 mM AAA (up to twofold increase, as compared to control), which suggest the ability of the oxidized amino acid to impair the redox status of the bacterium. In fact, AAA caused an increased production of reactive oxygen species (ROS) and the accretion of post-translational changes (protein carbonylation) in L. reuteri (up to 13 nmol allysine/mg protein vs 1.8 nmol allysine/mg protein in control). These results suggest that probiotic bacteria identify oxidized amino acids as harmful species and activate mechanisms that may protect themselves and the host against their noxious effects.

Biofilm Formation Ability of Arcobacter-like and Campylobacter Strains under Different Conditions and on Food Processing Materials.

Campylobacter jejuni is the most frequent cause of bacterial gastrointestinal food-borne infection worldwide. The transmission of Campylobacter and Arcobacter-like species is often made possible by their ability to adhere to various abiotic surfaces. This study is focused on monitoring the biofilm ability of 69 strains of Campylobacter spp. and lesser described species of the Arcobacteraceae family isolated from food, water, and clinical samples within the Czech Republic. Biofilm formation was monitored and evaluated under an aerobic/microaerophilic atmosphere after cultivation for 24 or 72 h depending on the surface material. An overall higher adhesion ability was observed in arcobacters. A chi-squared test showed no association between the origin of the strains and biofilm activity (p > 0.05). Arcobacter-like species are able to form biofilms under microaerophilic and aerobic conditions; however, they prefer microaerophilic environments. Biofilm formation has already been demonstrated at refrigerator temperatures (5 °C). Arcobacters also showed higher biofilm formation ability at the temperature of 30 °C. This is in contrast to Campylobacter jejuni NP 2896, which showed higher biofilm formation ability at temperatures of 5–30 °C. Overall, the results demonstrated the biofilm formation ability of many strains, which poses a considerable risk to the food industry, medical practice, and human health.

Bioprocess performance, transformation pathway, and bacterial community dynamics in an immobilized cell bioreactor treating fludioxonil-contaminated wastewater under microaerophilic conditions

Fludioxonil is a post-harvest fungicide contained in effluents produced by fruit packaging plants, which should be treated prior to environmental dispersal. We developed and evaluated an immobilized cell bioreactor, operating under microaerophilic conditions and gradually reduced hydraulic retention times (HRTs) from 10 to 3.9 days, for the biotreatment of fludioxonil-rich wastewater. Fludioxonil removal efficiency was consistently above 96%, even at the shortest HRT applied. A total of 12 transformation products were tentatively identified during fludioxonil degradation by using liquid chromatography coupled to quadrupole time-of-flight Mass spectrometry (LC-QTOF-MS). Fludioxonil degradation pathway was initiated by successive hydroxylation and carbonylation of the pyrrole moiety and disruption of the oxidized cyanopyrrole ring at the NH-C bond. The detection of 2,2-difluoro-2H-1,3-benzodioxole-4-carboxylic acid verified the decyanation and deamination of the molecule, whereas its conversion to the tentatively identified compound 2,3-dihydroxybenzoic acid indicated its defluorination. High-throughput amplicon sequencing revealed that HRT shortening led to reduced α-diversity, significant changes in the β-diversity, and a shift in the bacterial community composition from an initial activated sludge system typical community to a community composed of bacterial taxa like Clostridium, Oligotropha, Pseudomonas, and Terrimonas capable of performing advanced degradation and/or aerobic denitrification. Overall, the immobilized cell bioreactor operation under microaerophilic conditions, which minimizes the cost for aeration, can provide a sustainable solution for the depuration of fludioxonil-contaminated agro-industrial effluents.

Detection of clarithromycin resistance and 23SrRNA point mutations in clinical isolates of Helicobacter pylori isolates: Phenotypic and molecular methods

Background and objectivesPeptic ulcer disease, chronic gastritis, and stomach cancer are all caused by H. pylori. The most notable drug for the treatment is the antibiotic clarithromycin, which is currently the drug of choice. H. pylori clarithromycin resistance has been associated with point mutations in 23srRNA, the most prominent of which are A2143 and A2144G. In H. pylori bacteria, methylase synthesis, macrolide-inactivating enzyme activity, and active efflux have all been found to be resistance mechanisms. The goal of the study is to determine how resistant H. pylori is to clarithromycin and what the minimum inhibitory concentration is for various antimicrobials. Furthermore, gastro-endoscopy will be performed on Iraqi patients to detect the presence of A2143G and A2144G point mutations in Helicobacter pylori infections, as diagnosed from the pyloric region and other anatomical regions. MethodsOne hundred fifteen samples were collected from patients strongly suspected of H. pylori infection presented for upper gastrointestinal endoscopy at Ramadi Teaching Hospitals and Private Clinics for the period from January 2020 until February 2021. Specimens were cultured on brain heart infusion agar containing various antibiotics and were incubated at 37 °C under microaerophilic conditions. For identification of H. pylori, isolates of the biochemical tests and RT-PCR assay were applied. The Epsilometer test was used in the antibiotic susceptibility testing as dependent on the CLSI standard. The Restriction Fragment Length Polymorphism technique was used to determine point mutations. ResultsIn total, 55 (47.8%) Helicobacter pylori isolates were cultured from the 115 biopsy specimens, among which 16 (29.1%), 38 (69.1%), 20 (36.4%), and 40 (72.7%) revealed some degree of resistance to levofloxacin, clarithromycin, ciprofloxacin, and metronidazole, respectively. The frequency of A2144G and A2143 point mutations were 23 (60.5%) and 19 (50%), respectively. ConclusionsAccording to our results, Helicobacter pylori showed high resistance to clarithromycin. Our results demonstrate the requirement for antibiotic susceptibility testing and molecular methods in selecting drug regimens.

Peptoniphilus nemausensis sp. nov. A new Gram-positive anaerobic coccus isolated from human clinical samples, an emendated description of the genus Peptoniphilus and an evaluation of the taxonomic status of Peptoniphilus species with not validly published names

A Gram-positive, anaerobic coccus isolated from a human surgical site infection was previously shown to belong to an unknown species of the genus Peptoniphilus initially proposed as ‘Peptoniphilus nemausus’ sp. nov., based on both 16S rRNA gene sequence identity of 97.9% with the most closely related species Peptoniphilus coxii and an individualized phylogenetic branching within the genus Peptoniphilus. A polyphasic characterization of the novel species is proposed herein. Whole genome sequence analysis showed an average nucleotide identity value of 84.75% and digital DNA-DNA hybridization value of 28.9% against P. coxii type strain. The strain displayed unique features among members of the genus Peptoniphilus, as it was able to hydrolyze aesculin, and produced acetate as the major metabolic end-product without associated production of butyrate. Growth was observed under microaerophilic conditions. From all these data, the isolate is confirmed as belonging to a new Peptoniphilus species, for which the name Peptoniphilus nemausensis sp. nov. is proposed. The type strain is 1804121828T (=LMG 31466T = CECT 9935T). A database survey using a highly polymorphic partial sequence of the 16S rRNA gene of P. nemausensis revealed P. nemausensis to be a particularly rare skin-associated species in humans. An emendated description of the Peptoniphilus genus is proposed based on a review of the characteristics of the 12 new species with validly published names since the genus description in 2001 and of P. nemausensis. Finally, the relationships between members of the genus Peptoniphilus were explored based on whole genome sequence analysis in order to clarify the taxonomic status of not yet validly published species showing that three pairs of species should be considered as synonyms: Peptoniphilus timonensis and ‘Peptoniphilus phoceensis’, Peptoniphilus lacydonensis and ‘Peptoniphilus rhinitidis’, Peptoniphilus tyrrelliae and Peptoniphilus senegalensis.

Amelioration of disulfonated Acid Red and hexavalent chromium phytotoxic effects on Triticum aestivum using bioremediating and plant growth-promoting Klebsiella pneumoniae SK1

Bacterial strain identified as Klebsiella pneumoniae SK1, based on 16S rDNA sequence study, was isolated from a textile mill discharge point at Dada Nagar industrial area in Kanpur, India in media containing 100 µg/ml each of disulfonated azo dye Acid Red 249 (AR) and hexavalent chromium [Cr(VI)]. SK1 is efficient in the concomitant decolorization and reduction of 100 µg/ml toxic AR and Cr(VI) respectively in 48 h under microaerophilic conditions. The efficacy of AR decolorization was found to be preeminent in static conditions, 37°C, 2-4% salinity and 7-9 pH range. 99% decolorization for initial AR concentration of 100 µg/ml in 48 h, and 91% for 250 µg/ml and 77% for 500 µg/ml was recorded in 72 h respectively in the presence of 100 µg/ml Cr(VI). Significant reductive changes in spectroscopic absorption spectra were observed for SK1 treated AR+Cr(VI) amended media with respect to controls. FITR spectroscopy was used to ascertain the breakage of the azo dye bond and the formation of biodegradative metabolites. Additionally, SK1 was found to be positive for indole acetic acid, ammonia, phosphate and potassium solubilization and biofilm formation. In plant bioassay, in vitro SK1 treated AR+Cr(VI) TSB media was used to treat Triticum aestivum in the soil environment. In comparison to untreated control, plants treated with bioremediation media shows increased percent germination, root and shoot length with a complete reversal of phytotoxic effects of the AR+Cr(VI) treated plants. In this study, environmental K. pneumoniae SK1 shows concomitant azo dye and Cr(VI) remediation with plant growth-promoting activity.

Genome analysis of Pseudomonas sp. OF001 and Rubrivivax sp. A210 suggests multicopper oxidases catalyze manganese oxidation required for cylindrospermopsin transformation

BackgroundCylindrospermopsin is a highly persistent cyanobacterial secondary metabolite toxic to humans and other living organisms. Strain OF001 and A210 are manganese-oxidizing bacteria (MOB) able to transform cylindrospermopsin during the oxidation of Mn2+. So far, the enzymes involved in manganese oxidation in strain OF001 and A210 are unknown. Therefore, we analyze the genomes of two cylindrospermopsin-transforming MOB, Pseudomonas sp. OF001 and Rubrivivax sp. A210, to identify enzymes that could catalyze the oxidation of Mn2+. We also investigated specific metabolic features related to pollutant degradation and explored the metabolic potential of these two MOB with respect to the role they may play in biotechnological applications and/or in the environment.ResultsStrain OF001 encodes two multicopper oxidases and one haem peroxidase potentially involved in Mn2+ oxidation, with a high similarity to manganese-oxidizing enzymes described for Pseudomonas putida GB-1 (80, 83 and 42% respectively). Strain A210 encodes one multicopper oxidase potentially involved in Mn2+ oxidation, with a high similarity (59%) to the manganese-oxidizing multicopper oxidase in Leptothrix discophora SS-1. Strain OF001 and A210 have genes that might confer them the ability to remove aromatic compounds via the catechol meta- and ortho-cleavage pathway, respectively. Based on the genomic content, both strains may grow over a wide range of O2 concentrations, including microaerophilic conditions, fix nitrogen, and reduce nitrate and sulfate in an assimilatory fashion. Moreover, the strain A210 encodes genes which may convey the ability to reduce nitrate in a dissimilatory manner, and fix carbon via the Calvin cycle. Both MOB encode CRISPR-Cas systems, several predicted genomic islands, and phage proteins, which likely contribute to their genome plasticity.ConclusionsThe genomes of Pseudomonas sp. OF001 and Rubrivivax sp. A210 encode sequences with high similarity to already described MCOs which may catalyze manganese oxidation required for cylindrospermopsin transformation. Furthermore, the analysis of the general metabolism of two MOB strains may contribute to a better understanding of the niches of cylindrospermopsin-removing MOB in natural habitats and their implementation in biotechnological applications to treat water.

Role of air scouring in anaerobic/anoxic tanks providing nitrogen removal by mainstream anammox conversion in a hybrid biofilm/suspended growth full-scale WWTP in China.

A full-scale wastewater treatment plant in China experienced unintentional anammox bacterial enrichment on biofilm carriers placed in the anaerobic and anoxic zones of an anaerobic/anoxic/oxic process under ambient temperatures and without bioaugmentation. Here, we show that microaerophilic conditions resulting from air scouring needed for biofilm carrier suspension in the anaerobic/anoxic zones can support a robust nitritation/anammox process. Results from an in situ on/off air scouring test showed that air scouring strongly induced both ammonia and total inorganic nitrogen removal in the anaerobic/anoxic zones. Ammonium concentration in the anaerobic and anoxic tanks remained constant or even slightly increased when air scouring was off, indicating that air scouring made a noticeable difference in nitrogen profiles in the anaerobic/anoxic zones. Various batch tests further indicated that partial denitrification is not likely to generate nitrite for anammox bacteria. Robust nitritation, and anammox on the carriers, can occur at low dissolved oxygen conditions, as measured in the full-scale facility. The observations show that mainstream deammonification without sidestream bioaugmentation at moderate temperature is feasible and further optimization by a more dedicated design can result in improved nitrogen removal in cases when chemical oxygen demand is limited in mainstream wastewater treatment. PRACTITIONER POINTS: Microaerophilic conditions in a full-scale IFAS reactor caused mainstream anammox in moderate temperate area. Robust nitritation, and anammox on the carriers, can occur at low dissolved oxygen conditions in anaerobic/anoxic tanks with air scouring. Anammox can function well with conventional nitrification and denitrification process at mainstream conditions for stable nitrogen removal.

Microbial degradation of azo dyes by textile effluent adapted, Enterobacter hormaechei under microaerophilic condition

Landmark and sustainable eco-friendly dye treatment processes are highly desirous to ameliorate their effect on the environment. The present study investigated the azo dye degradation efficiency of adapted Enterobacter hormaechei SKB 16 from textile effluent polluted soil in optimized culture conditions. The adapted bacteria strain was identified by standard microbiological and molecular techniques. E. hormaechei was tested individually for the decolourizing of Reactive Yellow 145 (RY 145) and Reactive Red F3B (RR 180) dyes under optimized conditions of pH, temperature and dye concentration on decolourization were studied. The adapted bacteria strain exhibited maximum decolourization (98 %) of Reactive yellow 145 and Reactive red 180 in 100 ppm concentration at pH 7, temperature 37 °C after 98 h of incubation. The enzyme analyses revealed that azo reductase and laccase played major roles in the cleavage of the azo bond and desulfonation respectively of both dyes during degradation. The metabolites were further characterized by Fourier Transform Infrared Spectroscopy (FT-IR), High-Performance Liquid Chromatography (HPLC), and Gas Chromatography-Mass Spectrometry (GCMS). Thereafter, degradation was deduced based on changes of the functional group, variation in retention times and mass/charge ratio and molecular weight. This study elucidated the promising potentials of adapted SKB 16 strain in the eco-friendly removal of textile azo dyes. In addition, repeatability and sustainability are enhanced due to effective management of time which would have been spent on rigorous and extensive screening process.

Isolation Procedure for CP E. coli from Caeca Samples under Review towards an Increased Sensitivity.

Due to the increasing reports of carbapenemase-producing Enterobacteriaceae (CPE) from livestock in recent years, the European Reference Laboratory for Antimicrobial Resistances (EURL-AR) provided a protocol for their recovery from caecum and meat samples. This procedure exhibited limitations for the detection of CPE with low carbapenem MIC values. Therefore, it was modified by a second, selective enrichment in lysogeny broth with cefotaxime (CTX 1 mg/L) and with meropenem (MEM 0.125 mg/L) at 37 °C under microaerophilic conditions. By Real-time PCR, these enrichments are pre-screened for the most common carbapenemase genes. Another adaptation was the use of in-house prepared MacConkey agar with MEM and MEM+CTX instead of commercial selective agar. According to the EURL-method, we achieved 100% sensitivity and specificity using the in-house media instead of commercial agar, which decreased the sensitivity to ~75%. Comparing the method with and without the second enrichment, no substantial influence on sensitivity and specificity was detected. Nevertheless, this enrichment has simplified the CPE-isolation regarding the accompanying microbiota and the separation of putative colonies. In conclusion, the sensitivity of the method can be increased with slight modifications.

Effect of low oxygen concentration on activation of inflammation by Helicobacter pylori

The gastrointestinal tract of the human body is characterized by a highly unique oxygenation profile, where the oxygen concentration decreases toward the lower tract, not found in other organs. The epithelial cells lining the mucosa where Helicobacter pylori resides exist in a relatively low oxygen environment with a partial pressure of oxygen (pO2) below 58 mm Hg. However, the contribution of hypoxia to H. pylori-induced host immune responses remains elusive. In this study, we investigated the inflammasome activation induced by H. pylori under hypoxic, compared with normoxic, conditions. Our results indicated that the activation of caspase-1 and the subsequent secretion of IL-1β were significantly enhanced in infected macrophages under 1% oxygen, compared with those under a normal 20% oxygen concentration. The proliferation of H. pylori under aerobic conditions was 3-fold higher than under microaerophilic conditions, and the bacterial growth was more dependent on CO2 than on oxygen. Also, we observed that hypoxia-induced cytokine production as well as HIF-1α accumulation were both decreased when murine macrophages were treated with an HIF-1α inhibitor, KC7F2. Furthermore, hypoxia enhanced the phagocytosis of H. pylori in an HIF-1α-dependent manner. IL-1β production was also affected by the HIF-1α inhibitor in a mouse infection model, suggesting the important role of HIF-1α in the host defense system during infection with H. pylori. Our findings provide new insights into the intersection of low oxygen, H. pylori, and inflammation and disclosed how H. pylori under low oxygen tension can aggravate IL-1β secretion.

Methanotrophs: Discoveries, Environmental Relevance, and a Perspective on Current and Future Applications.

Methane is the final product of the anaerobic decomposition of organic matter. The conversion of organic matter to methane (methanogenesis) as a mechanism for energy conservation is exclusively attributed to the archaeal domain. Methane is oxidized by methanotrophic microorganisms using oxygen or alternative terminal electron acceptors. Aerobic methanotrophic bacteria belong to the phyla Proteobacteria and Verrucomicrobia, while anaerobic methane oxidation is also mediated by more recently discovered anaerobic methanotrophs with representatives in both the bacteria and the archaea domains. The anaerobic oxidation of methane is coupled to the reduction of nitrate, nitrite, iron, manganese, sulfate, and organic electron acceptors (e.g., humic substances) as terminal electron acceptors. This review highlights the relevance of methanotrophy in natural and anthropogenically influenced ecosystems, emphasizing the environmental conditions, distribution, function, co-existence, interactions, and the availability of electron acceptors that likely play a key role in regulating their function. A systematic overview of key aspects of ecology, physiology, metabolism, and genomics is crucial to understand the contribution of methanotrophs in the mitigation of methane efflux to the atmosphere. We give significance to the processes under microaerophilic and anaerobic conditions for both aerobic and anaerobic methane oxidizers. In the context of anthropogenically influenced ecosystems, we emphasize the current and potential future applications of methanotrophs from two different angles, namely methane mitigation in wastewater treatment through the application of anaerobic methanotrophs, and the biotechnological applications of aerobic methanotrophs in resource recovery from methane waste streams. Finally, we identify knowledge gaps that may lead to opportunities to harness further the biotechnological benefits of methanotrophs in methane mitigation and for the production of valuable bioproducts enabling a bio-based and circular economy.

Isolation and enumeration of probiotic microorganisms from fermented idli batter

Probiotics are defined as nonpathogenic living microorganisms, that have beneficial effects on host health. The most important and frequently used functional food compounds are probiotics and fermented foods. Functional foods have numerous beneficial nutritional effects. Probiotics dairy foods, idli, dhokla, pickles etc are traditional Indian foods that beneficially affect the host. They improve survival and implantation of live microbial supplements. In our study, probiotics were isolated from Fermented Idli batter. There is a difference observed in the pH of fermented idli batter. It decreases from 6 to 4.3-5.3 during fermentation. Yeasts were isolated from the fermented batter on Sabouraud's agar. Serial dilutions were performed, furthermore spread plate of these dilutions upto 10-5 was done on Rogosa agar plates and incubated for 72 h in a candle jar under microaerophilic condition. Colonies of lactic acid bacteria were observed on Rogosa agar plate. The count of bacteria in the fermented batter was 108 cells/ml. This experiment shows the presence of probiotic microorganisms, both bacteria and yeasts. Lactic acid bacteria and yeasts facilitate idli batter fermentation thereby enhancing their nutritional value and development of desirable texture and flavor.

Glycosaminoglycan Metabolism by Commensal and Pathogenic Bacteria of the Urinary Tract

Urinary tract infections (UTIs) are among the most common adult bacterial infections. When a patient suffers ≥3 UTIs per year, it is defined as recurrent UTI (rUTI). Current rUTI therapies rely on antimicrobials to achieve sterility in the urinary tract (UT). However, research has shown that urine is not sterile in healthy individuals and that the urinary microbiota has not been fully characterized. A constant flux of urine containing electrolytes, osmolytes, amino acids, and carbohydrates supply nutrients to support urinary microbiota. An overlooked carbon source in the UT is the glycosaminoglycan (GAG) layer lining the urothelium. GAGs are linear heteropolysaccharides containing repeating disaccharide units composed of uronic acid (or galactose) and amino sugars. Previous studies have determined that species belonging to the genera Lactobacillus, Bifidobacterium, and Bacteroides derived from the human gut express enzymes that degrade GAGs into metabolizable disaccharides. However, the ability of the urinary microbiota and invading uropathogens to metabolize GAGs has not been assessed. To fill this knowledge gap, we screened a library of urinary bacteria clinically isolated from women with and without rUTI for the ability to degrade and utilize GAGs such as chondroitin sulfate (CS), heparan sulfate (HS), and hyaluronic acid (HA). To test this, we developed a plate-based, semi-quantitative GAG degradation and growth assay. Briefly, isolates were inoculated in modified MRS lacking carbohydrates and supplemented with CS, HS, HA, glycogen, glucose, or maltose. The resuspension was added to a microtiter plate and incubated at 35°C for 72 hours in microaerophilic conditions. Optical density at 600nm (OD600) was measured to record growth. Plates were centrifuged and the cell-free supernatant was added to 1% bovine serum albumin (BSA), which complexes with GAGs to form a white precipitate in the presence of acetic acid. To calculate percent GAG degradation, OD600 was recorded after addition of acetic acid and compared to bacteria-free controls. Using this method, we screened 16 representative isolates of four Lactobacillus species (L. crispatus, L. gasseri, L. delbrueckii, L. vaginalis), Streptococcus agalactiae, and Escherichia coli. Preliminary results show that all urinary Lactobacilli and E. coli exhibited no GAG degradation. However, all 4 Lactobacillus species were able to utilize maltose as a sole carbon source. Two S. agalactiae strains were able to degrade HA, but no other GAGs. Although urinary Lactobacilli were unable to degrade GAGs in vitro, these findings suggest that urinary Lactobacilli may rely on a bacterial consortia or host enzymes to degrade and utilize GAGs. Future work will focus on screening more common urinary microbiota species and analyzing the ability of urinary Lactobacilli to utilize GAGs in the presence of host GAG degradative enzymes and in co-culture with S. agalactiae. Upon completion, this work will define the metabolic requirements of key urinary microbiota and elucidate metabolic interactions between the urinary microbiota, host environment, and invading uropathogens.

BENEFICIAL EFFECTS OF PROBIOTICS BASED THERAPY FOR IMMUNE HOMEOSTASIS

Several approaches have been used to demonstrate that signals derived from gut microbiota are critical for the development of the immune system. Probiotics are live microorganisms that have beneficial effects on host health, including extended lifespan, when they are administered or present in adequate quantities. However, the mechanisms by which probiotics stimulate host longevity remain unclear and very poorly understood. Probiotic supplements are needed to determine the effectiveness as a non-chemical approach to promote health and welfare. Our studies focus on the assessment of the molecular impact of probiotic administration involved in homeostasis and immunity. Shrimps received the recommended doses of microbial pack containing Bacillus species andRhodococcus species. The fermentation was carried at 37°C for 72 hours under microaerophilic condition. The parameters like pH, microbial load, acidity and concentration of reducing sugar had been measured.This probiotic pack when used either alone or in combination with traditional dairy starter, significantly improved the nutritional properties and the shelf life of the product. The present work will be valuable to elaborate novel functional food based on these original probiotic properties.

National and Regional US Antibiotic Resistance to Helicobacter pylori: Lessons From a Clinical Trial

National and Regional US Antibiotic Resistance to Helicobacter pylori: Lessons From a Clinical Trial

Bioremedial approach of Pseudomonas stutzeri SPM-1 for textile azo dye degradation.

The optimization of the bacterium Pseudomonas stutzeri SPM-1, obtained from textile wastewater dumping sites of Surat, Gujarat was studied for the degradation of the textile azo dye Procion Red-H3B. The strain showed significant activities of azoreductase (95%), laccase (76%) and NADH-DCIP reductase (88%) at 12, 10 and 8h of growth, respectively, indicating the evidence for reductive cleavage of the dye. The optimization was carried on phenanthrene enrichment medium followed by exposing it to variable environmental factors and nutritional sources. The complete decolourization of dye (50mg/L) happened within 20h of incubation at pH 8 and temperature 32 ± 0.2°C under microaerophilic condition. Decolourization was monitored with the shifting of absorbance peak in UV-Vis spectrophotometry and HPLC analysis. The changes in the functional groups were confirmed by the presence of new peaks in FT-IR data. GC-MS analysis helped in recognizing the degraded dye compounds thus elucidating the proposed pathway for Procion Red-H3B. The potential of bioremediation process was completed by phytotoxicity test using two plants Vigna radiata and Cicer arietinum. Our study concludes that the strain Pseudomonas stutzeri SPM-1, with its rapid decolourization efficiency holds noteworthy prospective in industrial application for textile wastewater treatment.