An overview of real-time quantitative PCR: Principles and formats for environmental microbiology studies

© Global Science Publications. A fluorescence-based real-time quantitative polymerase chain reaction (qPCR) is a powerful and commonly used molecular technique for quantifying the rRNA or DNA of targeted organisms in environmental samples. qPCR assays are easy to perform, making them capable of high throughput and can combine high sensitivity with reliable specificity. qPCR analysis is the combination of the traditional endpoint PCR attached with fluorescents to record the accumulation of the amplicons in real time during each cycle of the PCR. Detection of amplicons during the early exponential phase enables quantification of the gene numbers because they are proportional to the starting template. This review is focussed on currently used qPCR platforms, the chemistries involved in real-time PCR systems mainly applied for the environmental microbiology studies. The various factors affecting quantification of environmental microbial communities using qPCR have also been discussed

Applications of real-time quantitative PCR for environmental microbiology studies

© Global Science Publications. The real-time quantitative polymerase chain reaction (qPCR) is a robust and sensitive molecular technique to quantify phylogenetic and key functional genes of microbial populations in various natural and engineered environmental ecosystems under varying physical and chemical environmental conditions. The generated quantitative qPCR data can be used to relate variation in abundance of specific microbial groups, species, family or functional gene of interest and levels of gene expression in comparison with the change in environmental factors and or system processes. In this review, several applications of qPCR for the understanding of environmental microorganisms have been presented. The main qPCR applications reviewed are: quantification of uncultured environmental bacteria, detection of pathogens in environmental samples, the effect of antibiotics on anaerobic digester microbial community and understanding mechanism of nitrification and ammonia metabolism in engineered environmental ecosystems

Emerging contaminants of high concern for the environment: Current trends and future research

Wastewater is contaminated water that must be treated before it may be transferred into other rivers and lakes in order to prevent further groundwater pollution. Over the last decade, research has been conducted on a wide variety of contaminants, but the emerging contaminants are those caused primarily by micropollutants, endocrine disruptors (EDs), pesticides, pharmaceuticals, hormones, and toxins, as well as industrially-related synthetic dyes and dye-containing hazardous pollutants. Most emerging pollutants did not have established guidelines, but even at low concentrations they could have harmful effects on humans and aquatic organisms. In order to combat the above ecological threats, huge efforts have been done with a view to boosting the effectiveness of remediation procedures or developing new techniques for the detection, quantification and efficiency of the samples. The increase of interest in biotechnology and environmental engineering gives an opportunity for the development of more innovative ways to water treatment remediation. The purpose of this article is to provide an overview of emerging sources of contaminants, detection technologies, and treatment strategies. The goal of this review is to evaluate adsorption as a method for treating emerging pollutants, as well as sophisticated and cost-effective approaches for treating emerging contaminants

Challenges and perspectives on innovative technologies for biofuel production and sustainable environmental management

Specifically, human activities, such as those in industry and transportation, have resulted in an increase in the demand for fossil fuels, resulting in severe environmental problems.Throughout this article, we discuss the potential and challenges associated with the production of biofuels from a variety of feedstocks and advances in processing technologies utilizing a range of feedstocks. Based on the conclusion of the study, we conclude that bioenergy is a green alternative to be used for diverse energy needs, once the appropriate conversion processes are applied. The production of biofuels and their use in industries and transportation have significantly reduced the use of fossil fuels. The literature review concluded that producing biofuels from energy crops and microalgae was the most efficient and attractive method. The purpose of this review is to explain all aspects of biofuels and their sustainability criteria. With a particular focus on the role of nanotechnology in biofuel production, this article discusses the most recent advances in biofuel production. A number of emerging techniques have been investigated for improving process quality, including integrated techniques, less energy-intensive distillation strategies, and the use of microorganisms in engineering. A challenging aspect of biofuel production on a large scale remains; therefore, a novel technology must be developed in order to enhance biofuel production in order to meet the challenges and meet future energy needs

Knowledge about osteoporosis among healthy women attending a tertiary care hospital

INTRODUCTION: To determinate the knowledge on osteoporosis-risk factors and disease in three age groups of Pakistani women.METHODS: In this exploratory cross-sectional study, an osteoporosis knowledge assessment questionnaire (OKAT) was used to collect data and it was delivered through a face-to-face interview. Questions were asked about symptoms of osteoporosis, knowledge of risk factors, preventive factors and treatment. A convenience sample (n =320) comprising of three groups of healthy women aged 25-35 years, 36-45 years, and over 45 years was taken. The scoring range was 0 to 20. Among-group comparisions of means were analyzed by two-way ANOVA. To determine the overall influence of osteoporosis-risk factors, the multivariate analysis was used.RESULTS: The knowledge on osteoporosis in younger women was very poor compared to relatively older females. However, women belonging to higher socioeconomic status and better education had slightly more knowledge about osteoporosis compared to those with a low education level, regardless of age.CONCLUSION: The majority of women had modest knowledge on osteoporosis. Younger women were at increased risk for low bone mass and premature osteoporosis

An in vitro study of probiotic activity exhibited by Lactobacillus acidophilus and Lactobacillus rhamnosus on oral isolates of Streptococcus mutans and Candida albicans

Background: Oral infections caused by microorganisms have led to increased risk of oral health problems like dental caries (DC). Streptococcus mutans and Candida albicans are the organisms responsible for DC. The goal of the presented study was to investigate the potential of probiotics to prevent and treat DC. An in vitro assay was developed to investigate several probiotic strains for their ability to inhibit the aforementioned oral pathogens. Methods: 40 oral isolates of Streptococcus mutans and 51 oral isolates of Candida albicans were tested for probiotic activity against Lactobacillus acidophilus and Lactobacillus rhamnosus using agar overlay interference technique as prescribed by Fleming et al. Results: The zone of inhibition shown by L. acidophilus was higher than L. rhamnosus against Streptococcus mutans and Candida albicans. Conclusions: In conclusion the two probiotic strains L. acidophilus and L. rhamnosus exhibited inhibitory activity on S. mutans and C. albicans respectively in vitro

Study of total seed storage protein in indigenous Brassica species based on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE)

Genetic diversity was studied in 234 accessions of locally collected Brassica species for total seed protein content through sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). These accessions were collected from different locations of Pakistan. After the study of these accessions on SDS-PAGE, 28 reproducible bands were used for cluster analysis and with the help of these bands, genetic diversity were estimated. Out of 28, four major bands were observed. Dendogram was constructed and the accessions were divided into two main groups comprising 11 clusters. The results obtained from these clusters showed minimum genetic diversity in these accessions on SDS-PAGE level. Due to low genetic diversity on SDS-PAGE level, it is suggested that two dimensional (2D)- electrophoresis can be used for protein study.Key words: Genetic variation, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), protein electrophoresis, cluster analysis, brassica

Bacterial community structure in anaerobic digesters of a full scale municipal wastewater treatment plant - Case study of dubai, united arab emirates

© 2019, International Centre for Sustainable Development of Energy, Water and Environment Systems SDEWES. All rights reserved. A highly complex microbial community involved in anaerobic sludge digesters plays vital roles in sludge treatment. The data on microbial ecology is important to accomplish efficient operation of the anaerobic digesters. This study is aimed at monitoring the bacterial community of three full-scale anaerobic digesters of a full-scale municipal wastewater treatment Plant in Dubai, United Arab Emirates. Fluorescent in-situ hybridization technique was applied to identify the bacterial groups and quantitative polymerase chain reaction to compare the richness of bacterial and archaeal domain. Results of the fluorescent in-situ hybridization technique analysis showed that the phylum Proteobacteria was most abundant followed by cytophage-Flavobacterium group of Bacteroides, Firmicutes and Actinobacteria. Among proteobacterial subclass Delta- and Alpha- were dominating than Gamma- and Beta-proteobacteria. The genus Desulfobacter and Desulfobacterium were the dominant groups hybridizing 70-76% of total 4\u27, 6\u27- diamidino - 2 phenylindole stained cells. The quantitative polymerase chain reaction results showed that Bacterial domain was dominating in all three digesters compared to the archaeal domain

Assessing methanogenic archaeal community in full scale anaerobic sludge digester systems in Dubai, United Arab Emirates

© 2018 Khan et al. Introduction: Anaerobic digestion for methane production comprises of an exceptionally diverse microbial consortium, a profound understanding about which is still constrained. In this study, the methanogenic archaeal communities in three full-scale anaerobic digesters of a Municipal Wastewater Treatment Plant were analyzed by Fluorescence in situ hybridization and quantitative real-time Polymerase Chain Reaction (qPCR) technique. Methods & Materials: Fluorescence in situ hybridization (FISH) was performed to detect and quantify the methanogenic Archaea in the sludge samples whereas qPCR was carried out to support the FISH analysis. Multiple probes targeting domain archaea, different orders and families of Archaea were used for the studies. Results and Discussion: In general, the aceticlastic organisms (Methanosarcinaceae & Methanosaetaceae) were more abundant than the hydrogenotrophic organisms (Methanobacteriales, Methanomicrobiales, Methanobacteriaceae & Methanococcales). Both FISH and qPCR indicated that family Methanosaetaceae was the most abundant suggesting that aceticlastic methanogenesis is probably the dominant methane production pathway in these digesters. Conclusion: Future work involving high-throughput sequencing methods and correlating archaeal communities with the main operational parameters of anaerobic digesters will help to obtain a better understanding of the dynamics of the methanogenic archaeal community in wastewater treatment plants in United Arab Emirates (UAE) which in turn would lead to improved performance of anaerobic sludge digesters