Hydrogen sulfide effects on the survival of lactobacilli with emphasis on the development of inflammatory bowel diseases

The gut microbiota is a complex component of humans that depends on diet, host genome, and lifestyle. The background:The study purpose is to find relations between nutrition, intestinal lactic acid bacteria (LAB) from various environments (human, animal intestine, and yogurt) and sulfate-reducing microbial communities in the large intestine; to compare kinetic growth parameters of LAB; and to determine their sensitivity to different concentration of hydrogen sulfide produced by intestinal sulfate-reducing bacteria. Methods: Microbiological (isolation and identification), biochemical (electrophoresis), molecular biology methods (DNA isolation and PCR analysis), and statistical processing (average and standard error calculations) of the results were used. The results: The toxicity of hydrogen sulfide produced by sulfate-reducing bacteria, the survival of lactic acid bacteria, and minimal inhibitory concentrations (MIC) were determined. The measured hydrogen sulfide sensitivity values were the same for L. paracasei and L. reuteri (MIC > 1.1 mM). In addition, L. plantarum and L. fermentum showed also a similar sensitivity (MIC > 0.45 mM) but significantly (p 0.45 mM). L. paracasei and L. reuteri are more sensitive to hydrogen sulfide than L. fermentum and L. plantarum. L. pentosus was sensitive to the extremely low concentration of H2S (MIC > 0.15 mM). Conclusions: The Lactobacillus species were significantly sensitive to hydrogen sulfide, which is a final metabolite of intestinal sulfate-reducing bacteria. The results are definitely helpful for a better understanding of complicated interaction among intestinal microbiota and nutrition. © 2019 by the authors. Licensee MDPI, Basel, Switzerland.Grant Agency of the Masaryk University [MUNI/A/0902/2018

The sulfate-reducing microbial communities and meta-analysis of their occurrence during diseases of small-large intestine axis

Sulfate-reducing bacteria (SRB) are often isolated from animals and people with ulcerative colitis and can be involved in the IBD development in the gut-intestine axis. The background of the research consisted of obtaining mixed cultures of SRB communities from healthy mice and mice with colitis, finding variation in the distribution of their morphology, to determine pH and temperature range tolerance and their possible production of hydrogen sulfide in the small-large intestinal environment. The methods: Microscopic techniques, biochemical, microbiological, and biophysical methods, and statistical processing of the results were used. The results: Variation in the distribution of sulfate-reducing microbial communities were detected. Mixed cultures from mice with ulcerative colitis had 1.39 times higher production of H2S in comparison with samples from healthy mice. The species of Desulfovibrio genus play an important role in diseases of the small-large intestine axis. Meta-analysis was also used for the observation about an SRB occurrence in healthy and not healthy individuals and the same as their metabolic processes. Conclusions: This finding is important for its possible correlation with inflammation of the intestine, where the present of SRB in high concentration plays a major part. It can be a good possible indicator of the occurrence of IBD.Grant Agency of the Masaryk University [MUNI/A/0902/2018

NADH and NADPH peroxidases as antioxidant defense mechanisms in intestinal sulfate-reducing bacteria

Animal and human feces typically include intestinal sulfate-reducing bacteria (SRB). Hydrogen sulfide and acetate are the end products of their dissimilatory sulfate reduction and may create a synergistic effect. Here, we report NADH and NADPH peroxidase activities from intestinal SRB Desulfomicrobium orale and Desulfovibrio piger . We sought to compare enzymatic activities under the influence of various temperature and pH regimes, as well as to carry out kinetic analyses of enzymatic reaction rates, maximum amounts of the reaction product, reaction times, maximum rates of the enzyme reactions, and Michaelis constants in cell-free extracts of intestinal SRB, D. piger Vib-7, and D. orale Rod-9, collected from exponential and stationary growth phases. The optimal temperature (35 °C) and pH (7.0) for both enzyme’s activity were determined. The difference in trends of Michaelis constants ( K m ) during exponential and stationary phases are noticeable between D. piger Vib-7 and D. orale Rod-9; D. orale Rod-9 showed much higher K m (the exception is NADH peroxidase of D. piger Vib-7: 1.42 ± 0.11 mM) during the both monitored phases. Studies of the NADH and NADPH peroxidases—as putative antioxidant defense systems of intestinal SRB and detailed data on the kinetic properties of this enzyme, as expressed by the decomposition of hydrogen peroxide—could be important for clarifying evolutionary mechanisms of antioxidant defense systems, their etiological role in the process of dissimilatory sulfate reduction, and their possible role in the development of bowel diseases

ATPase Activity of the Subcellular Fractions of Colorectal Cancer Samples under the Action of Nicotinic Acid Adenine Dinucleotide Phosphate

In tumor cells with defects in apoptosis, autophagy allows prolonged survival. Autophagy leads to an accumulation of damaged mitochondria by autophagosomes. An acidic environment is maintained in compartments of cells, such as autophagosomes, late endosomes, and lysosomes; these organelles belong to the “acid store” of the cells. Nicotinic acid adenine dinucleotide phosphate (NAADP) may affect the release of Ca(2+) from these organelles and affect the activity of Ca(2+) ATPases and other ion transport proteins. Recently, a growing amount of evidence has shown that the variations in the expression of calcium channels or pumps are associated with the occurrence, disease-presentation, and the prognosis of colorectal cancer. We hypothesized that activity of ATPases in cancer tissue is higher because of intensive energy metabolism of tumor cells. The aim of our study was to ascertain the effect of NAADP on ATPase activity on tissue samples of colorectal cancer patients’ and healthy individuals. We tested the effect of NAADP on the activity of Na(+)/K(+) ATPase; Ca(2+) ATPase of endoplasmic reticulum (EPR) and plasma membrane (PM) and basal ATPase activity. Patients’ colon mucus cancer samples were obtained during endoscopy from cancer and healthy areas (control) of colorectal mucosa of the same patients. Results. The mean activity of Na(+)/K(+) pump in samples of colorectal cancer patients (n = 5) was 4.66 ± 1.20 μmol P(i)/mg of protein per hour, while in control samples from healthy tissues of the same patient (n = 5) this value was 3.88 ± 2.03 μmol P(i)/mg of protein per hour. The activity of Ca(2+) ATPase PM in control samples was 6.42 ± 0.63 μmol P(i)/mg of protein per hour and in cancer −8.50 ± 1.40 μmol Pi/mg of protein per hour (n = 5 pts). The mean activity of Ca(2+) ATPase of EPR in control samples was 7.59 ± 1.21 μmol P(i)/mg versus 7.76 ± 0.24 μmol Pi/mg in cancer (n = 5 pts). Basal ATPase activity was 3.19 ± 0.87 in control samples versus 4.79 ± 1.86 μmol Pi/mg in cancer (n = 5 pts). In cancer samples, NAADP reduced the activity of Na(+)/K(+) ATPase by 9-times (p < 0.01) and the activity of Ca(2+) ATPase EPR about 2-times (p < 0.05). NAADP caused a tendency to decrease the activity of Ca(2+) ATPase of PM, but increased basal ATPase activity by 2-fold vs. the mean of this index in cancer samples without the addition of NAADP. In control samples NAADP caused only a tendency to decrease the activities of Na(+)/K(+) ATPase and Ca(2+) ATPase EPR, but statistically decreased the activity of Ca(2+) ATPase of PM (p < 0.05). In addition, NAADP caused a strong increase in basal ATPase activity in control samples (p < 0.01). Conclusions: We found that the activity of Na(+)/K(+) pump, Ca(2+) ATPase of PM and basal ATPase activity in cancer tissues had a strong tendency to be higher than in the controls. NAADP caused a decrease in the activities of Na(+)/K(+) ATPase and Ca(2+) ATPase EPR in cancer samples and increased basal ATPase activity. In control samples, NAADP decreased Ca(2+) ATPase of PM and increased basal ATPase activity. These data confirmed different roles of NAADP-sensitive “acidic store” (autophagosomes, late endosomes, and lysosomes) in control and cancer tissue, which hypothetically may be connected with autophagy role in cancer development. The effect of NAADP on decreasing the activity of Na(+)/K(+) pump in cancer samples was the most pronounced, both numerically and statistically. Our data shows promising possibilities for the modulation of ion-transport through the membrane of cancer cells by influence on the “acidic store” (autophagosomes, late endosomes and lysosomes) as a new approach to the treatment of colorectal cancer

Molecular Physiology of Anaerobic Phototrophic Purple and Green Sulfur Bacteria

There are two main types of bacterial photosynthesis: oxygenic (cyanobacteria) and anoxygenic (sulfur and non-sulfur phototrophs). Molecular mechanisms of photosynthesis in the phototrophic microorganisms can differ and depend on their location and pigments in the cells. This paper describes bacteria capable of molecular oxidizing hydrogen sulfide, specifically the families Chromatiaceae and Chlorobiaceae, also known as purple and green sulfur bacteria in the process of anoxygenic photosynthesis. Further, it analyzes certain important physiological processes, especially those which are characteristic for these bacterial families. Primarily, the molecular metabolism of sulfur, which oxidizes hydrogen sulfide to elementary molecular sulfur, as well as photosynthetic processes taking place inside of cells are presented. Particular attention is paid to the description of the molecular structure of the photosynthetic apparatus in these two families of phototrophs. Moreover, some of their molecular biotechnological perspectives are discussed

Anaerobic Fermentation of Silage from the Above-Ground Biomass of Jerusalem Artichoke (Helianthus tuberosus L.) and Maize (Zea mayse L.) as a New and Promising Input Raw Material for Biogas Production

Research of new input raw materials for biogas plants is a very actual topic. There are only a very few studies dealing with the possibility of using silage prepared from the above-ground parts of the Jerusalem artichoke (Helianthus tuberosus L.) and maize (Zea mayse L.) for methane production. This study deals with the determination of methane production and methane content in biogas during the fermentation of maize silage with dissimilar additions of the biomass of the Jerusalem artichoke (JA). Except for the effect of the JA's addition on the yield of methane, we also studied its potential influence on the inhibition of the process of anaerobic digestion and the bacterial and methanogenic archaeal composition of anaerobic digestate. There were five model silages prepared; two of them contained only maize or JAs, and the remaining three were mixtures of maize and JA silages (30%wt; 50%wt and 70%wt). The fermentation tests showed that the JA addition (from 30 to 70%wt) resulted in the production of biogas decreasing, on average, by 15%. Based on the performed metagenomic analysis, we cannot confirm an essential influence of JA biomass addition on the composition of the community of microorganisms during fermentation.O

The Possibility of Using Spent Coffee Grounds to Improve Wastewater Treatment Due to Respiration Activity of Microorganisms

Spent coffee ground (SCG) may affect wastewater treatment processes due to high coffee consumption worldwide. The impact of the main chemical compounds present in SCG on respiration activity of sewage sludge was investigated. The results showed approximately two times higher respiration in the samples where various types of SCG were present in comparison with samples without SCG. During intense microbial metabolism, statistically significant (p &lt; 0.05) decreases in caffeine, total polyphenols, and chlorogenic acid contents after processing and in filtrate was observed. The monitored compounds (caffeine, polyphenols, and chlorogenic acid) deteriorated due to their probable inclusion in microbiological metabolism. Increase in respiration activity of microorganisms in the presence of cheap waste material such as coffee grounds can help to improve wastewater treatment. The research was focused on spent coffee grounds&rsquo; impact on the respiratory activity of microorganisms in the activated sludge taken from small and large wastewater treatment plants. The impact was measured in more detail due to the inclusion of different coffee species (Robusta and Arabica) in diverse concentrations. The novelty of the study can also be seen through the literature overview, where information cannot be found about SCG influence on the respiration activity of microbial communities, and data on the possible SCG aerobic degradation or utilization by a sewage sludge bacterial consortium has also never been reported. The study has shown the possibility of improving wastewater treatment due to respiration activity of microorganisms in the presence of cheap waste material such as coffee grounds

Characterisation of actinomycetes community from the heavy metals polluted soil

The isolation of actinomycetes was performed from soil samples influenced by car-traffic. The acute toxicity of soil leaches was tested by the help of Microtox® bioassay testing system which uses freeze dried luminescent bacteria Photobacterium phosphoreum as the test organisms. The content of heavy metals in biomass of soil microorganisms and in whole soil samples was determinate. 115 strains of actinomycetes were isolated and their total numbers in soil samples were estimated. The acute toxicity of soil influenced the total numbers of actinomycetes. By the help of DNA-DNA reassociation procedure the generic diversity of bacteria was estimated. The identification and differentiation of streptomycetes from the total isolated actinomycetes was made using specific morphological criteria and the gas chromatography-fatty acid methyl ester (GC-FAME) analysis. FAME method is adequate only for differentiation of members of genus Streptomyces from other actinomycetes because of their characteristical profile of fatty acids

Microbiological characteristics of bioaerosol at the composting plant

The diversion of biodegradable waste from landfill is of key importance in developing a sustainable waste strategy for the next decade and beyond. The proliferation of waste treatment technologies such as mechanical biological treatment, anaerobic digestion and composting will be paramount in achieving this strategic goal. Composting plant is one of the end technology, which is widely used in waste processing of the biodegradable waste. These wastes originate from the maintenance of green areas in the cities and the municipalities and from the separatelly collected biodegradable waste from the citizens. There is also possible to process other biodegradable materials whose origin may be in other technologies of waste management at the composting plant. The most commonly used technology of composting is windrow system. Technological operations, which are necessary for the proper conduct of the composting process, may have negative influence on the environment in the immediate vicinity of composting plant. As pollutants we can mark particular odor and microorganisms. The largest group of microorganisms in the monitored air were psychrophilic and mesophilic bacteria and microscopic thermotolerant fungi. The amount of thermophillic actinomycetes ranged from 10 to 84.000 CFU∙m−3 (colony forming units per m3). Furthermore, it was confirmed that the maximum air contamination has been found during aeration of windrow by compost turner and during the sieving of the mature compost. For each indicator, the increase in concentrations due to the turning of compost windrow as compared to the background concentration obtained in natural environments and upwind of composting plants was determined. At a distance of 150 m from the composting plant, only low numbers of indicator organisms at a regular occurrence in the air has been found

Influence of heavy metals occurrence on respiratory activity of microorganisms in the compost

This work deals with the influence of selected heavy metals, copper, nickel, cobalt, zinc and cadmium on respiratory activity of microorganisms during the composting process. We focused on comparing the respiratory activity of microorganisms in the compost after composting process in the presence of different concentrations of above mentioned metals and the measured results were confronted with the valid Czech standard for industrial composts. The results show that the high inhibitory effect at low concentrations, about 1 mg/dm3, has cobalt, for which the limit concentration is no set by Czech standard. Other heavy metals exhibit an inhibitory effect at concentrations higher than maximum allowable concentration, which is set in Czech standard for compost class I and II, with regard to their application to agricultural land