Application of fluorescence in situ hybridization (FISH) to the analysis of sulfate reducing bacterial community in an oily bench scale reactor

Advances in the field of genomics and meta-genomics have led to rapid and accurate strategies for the monitoring of microbial biodiversity and have revealed its potential for biotechnological applications. In this study, fluorescent in situ hybridization (FISH) as a culture-independent molecular approach using specific CY3-labelled oligonucleotide probes was used to study the dynamics of the sulfate reduction bacterial community (SRB) of the activated sludge from an oily wastewater treatment system. The relative abundance of members of the dominant bacteria in the oily water reactor was determined by FISH for 16S rRNA using EUB338 probes, for detecting general eubacteria, and SRB385 for targeting SRBs and major species of delta-proteobacteria sulfate reducers. The percentage of cells hybridizing with probe EUB338 for the dominant bacteria decreased from 25.85 to 6.25%, while with the SRB385 probe for SRB bacteria, it increased from 7.21 to 10.20% of total cells during the reactor process. These data show that SRB bacteria dominated the active microbial community in the system. It is interesting that delta-proteobacterial SRBs occupied a high percentage and took place in an oily biological system under aerobic conditions.Keywords: Sulfate reducing bacteria, fluorescence in situ hybridization, 16S rRNA oligonucleotide probes, microbial community, dynamic

Environmental Biotechnology: Current Advances, New Knowledge Gaps, and Emerging Issues

Theterm“environmental biotechnology” encapsulates a wide and dynamic spectrum of topics including bioremediation, biofiltration, wastewater treatment, biodegradation, waste management, and biofuel production. All these disciplines are underpinned by complex interacting microbial communities; hence exciting research has been undertaken and increasingly driven by cutting-edge ecogenomic techniques. However, understanding of microbial diversity, population, and structure for sustainable environmental management remains largely a “black box.” Thus exciting findings identified new knowledge gaps and, inevitably, engendered novel research questions with inventive methodologies

Variations in genetic and chemical constituents of Ziziphus spina-christi L. populations grown at various altitudinal zonation up to 2227m height

AbstractAltitudinal gradient-defined specific environmental conditions could lead to genetics and chemical variations among individuals of the same species. By using RAPD, ISSR, GC–MS and HPLC analysis, the genetic and chemical diversity of Ziziphus spina-christi plants at various altitudinal gradient namely; Abha (2227.86m), Dala Valley (1424m), Rakhma Valley (1000m), Raheb Valley (505m) and Al-Marbh (147m) were estimated. RAPD markers revealed that the highest similarity value (40.22%) was between Raheb Valley and Al-Marbh while the lowest similarity (10.08%) was between Abha and Raheb Valley. Based on ISSR markers the highest similarity value (61.54%) was also between Raheb Valley and Al-Marbh, while the lowest similarity (26.84%) was between Abha and Rakhma Valley. GC–MS results showed the presence of various phytochemical constituents in each population. The dendrogram based on chemical compounds separated the Z. spina-christi grown at the highest elevations (Abha) from the populations in lower elevations. HPLC analysis showed that the leaves of Z. spina-christi plant contain considerable amount of vitamins including B1, B12, B2 and folic acid. In conclusion, there is a close relation between altitudinal gradients, genetic diversity and chemical constituents of the leaves of Z. spina-christi plants

Isolation of Thermoalkalophilic-?-amylase Producing Bacteria and Optimization of Potato Waste Water Medium for Enhancement of ?-amylase Production

Sixty one thermoalkalophilic bacteria were isolated from soil samples in Saudi Arabia’s southern region. Isolate TA-38, obtained from the Tanomah region, showed the best performance for enzyme production and was submitted for further study. It was identified as Bacillus axarquiensis based on 16S rRNA gene sequencing studies. The feasibility of using potato waste water as a simple and cheap medium for the production of ?-amylase was evaluated compared with starch broth medium. The production of ?-amylase in the potato waste water medium was only 13.8% less than that of the starch medium. Maximum enzyme production was achieved after 48 hours of cultivation at the beginning of the stationary phase at pH 10.0 and 50 0C. The appropriate addition of starch; nitrogen; phosphate; and calcium to potato waste water significantly enhanced the production of ?-amylase. The enzyme production reached a maximum of 64.5 Uml-1 with the potato wastewater adding with 0.5 % starch; 0.4 % yeast extract; 0.04% CaCl2-2H2O and 0.05 % KH2PO4.  The optimization of the potato waste water medium led to an approximately 4.02 fold increase in the production of ?-amylase compared to starch broth medium. Data indicated that the potato waste water contained substrates which could be used by bacterial isolate for the production of ?-amylase production and the developed procedure was cost effective since it requires only a slightly addition of nutrients to the medium. Keywords: Isolation; ?-amylase; 16S rRNA; Production; Potato waste water; Thermoalkaliphilic bacteria

Enhanced biomass production of pseudomonas flourescens for waste water treatment in shake flask and semi industrial scale bioreactor

Pseudomonas fluorescens is a biological denitrification bacterium that able to transform of oxidized nitrogen compounds (nitrate and nitrite) into harmless nitrogen gas. Nitrification followed by denitrification is a reliable and economical method for removing nitrogen from wastewater. However, the process is slow and affected by limitation of organic sources. Therefore in order to increase efficiency P. fluorescens as nitrogen removal in the waste water treatment, it is recommended to study medium composition that affects the growth of P. fluorescens for industrial scale cell mass production. Moreover, little information on cell mass production is available. Therefore, the objective of this research is to develop an industrial culture medium and a cultivation strategy for the mass production of P. fluorescens in a semi-industrial scale. At beginning of research, several medium formulations were tested for their cell growth potential. The best medium yielded a cell mass of only 2.08 g L -1 in shake flask cultures and was optimized using a classical approach (changing factors one by one). Sucrose and yeast extract were selected as the best carbon and nitrogen source respectively for the new medium composition. Only 10 g L -1 for sucrose concentration and 3 g L -1 of yeast extract concentration required for cell growth and no significant increase in cell mass for both concentration above than that. Thus, the final cell mass of optimized medium from classical method achieving a maximum of 3.02 g L -1 increased up to 45.19 %. Then optimized medium was applied in bioreactor for batch cultivation by controlled and uncontrolled pH condition. It is found that controlled pH showed 39.56 % more of cell mass compared to uncontrolled pH yielded a cell mass of only 2.78 g L -1. As a result, fermentation with controlled pH gives higher potential of production of cell mass at industrial scale

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

Surgical site infection after gastrointestinal surgery in high-income, middle-income, and low-income countries: a prospective, international, multicentre cohort study

Background: Surgical site infection (SSI) is one of the most common infections associated with health care, but its importance as a global health priority is not fully understood. We quantified the burden of SSI after gastrointestinal surgery in countries in all parts of the world. Methods: This international, prospective, multicentre cohort study included consecutive patients undergoing elective or emergency gastrointestinal resection within 2-week time periods at any health-care facility in any country. Countries with participating centres were stratified into high-income, middle-income, and low-income groups according to the UN's Human Development Index (HDI). Data variables from the GlobalSurg 1 study and other studies that have been found to affect the likelihood of SSI were entered into risk adjustment models. The primary outcome measure was the 30-day SSI incidence (defined by US Centers for Disease Control and Prevention criteria for superficial and deep incisional SSI). Relationships with explanatory variables were examined using Bayesian multilevel logistic regression models. This trial is registered with ClinicalTrials.gov, number NCT02662231. Findings: Between Jan 4, 2016, and July 31, 2016, 13 265 records were submitted for analysis. 12 539 patients from 343 hospitals in 66 countries were included. 7339 (58·5%) patient were from high-HDI countries (193 hospitals in 30 countries), 3918 (31·2%) patients were from middle-HDI countries (82 hospitals in 18 countries), and 1282 (10·2%) patients were from low-HDI countries (68 hospitals in 18 countries). In total, 1538 (12·3%) patients had SSI within 30 days of surgery. The incidence of SSI varied between countries with high (691 [9·4%] of 7339 patients), middle (549 [14·0%] of 3918 patients), and low (298 [23·2%] of 1282) HDI (p < 0·001). The highest SSI incidence in each HDI group was after dirty surgery (102 [17·8%] of 574 patients in high-HDI countries; 74 [31·4%] of 236 patients in middle-HDI countries; 72 [39·8%] of 181 patients in low-HDI countries). Following risk factor adjustment, patients in low-HDI countries were at greatest risk of SSI (adjusted odds ratio 1·60, 95% credible interval 1·05–2·37; p=0·030). 132 (21·6%) of 610 patients with an SSI and a microbiology culture result had an infection that was resistant to the prophylactic antibiotic used. Resistant infections were detected in 49 (16·6%) of 295 patients in high-HDI countries, in 37 (19·8%) of 187 patients in middle-HDI countries, and in 46 (35·9%) of 128 patients in low-HDI countries (p < 0·001). Interpretation: Countries with a low HDI carry a disproportionately greater burden of SSI than countries with a middle or high HDI and might have higher rates of antibiotic resistance. In view of WHO recommendations on SSI prevention that highlight the absence of high-quality interventional research, urgent, pragmatic, randomised trials based in LMICs are needed to assess measures aiming to reduce this preventable complication

Pooled analysis of WHO Surgical Safety Checklist use and mortality after emergency laparotomy

Background The World Health Organization (WHO) Surgical Safety Checklist has fostered safe practice for 10 years, yet its place in emergency surgery has not been assessed on a global scale. The aim of this study was to evaluate reported checklist use in emergency settings and examine the relationship with perioperative mortality in patients who had emergency laparotomy. Methods In two multinational cohort studies, adults undergoing emergency laparotomy were compared with those having elective gastrointestinal surgery. Relationships between reported checklist use and mortality were determined using multivariable logistic regression and bootstrapped simulation. Results Of 12 296 patients included from 76 countries, 4843 underwent emergency laparotomy. After adjusting for patient and disease factors, checklist use before emergency laparotomy was more common in countries with a high Human Development Index (HDI) (2455 of 2741, 89.6 per cent) compared with that in countries with a middle (753 of 1242, 60.6 per cent; odds ratio (OR) 0.17, 95 per cent c.i. 0.14 to 0.21, P <0001) or low (363 of 860, 422 per cent; OR 008, 007 to 010, P <0.001) HDI. Checklist use was less common in elective surgery than for emergency laparotomy in high-HDI countries (risk difference -94 (95 per cent c.i. -11.9 to -6.9) per cent; P <0001), but the relationship was reversed in low-HDI countries (+121 (+7.0 to +173) per cent; P <0001). In multivariable models, checklist use was associated with a lower 30-day perioperative mortality (OR 0.60, 0.50 to 073; P <0.001). The greatest absolute benefit was seen for emergency surgery in low- and middle-HDI countries. Conclusion Checklist use in emergency laparotomy was associated with a significantly lower perioperative mortality rate. Checklist use in low-HDI countries was half that in high-HDI countries.Peer reviewe

Global economic burden of unmet surgical need for appendicitis

Background: There is a substantial gap in provision of adequate surgical care in many low-and middle-income countries. This study aimed to identify the economic burden of unmet surgical need for the common condition of appendicitis. Methods: Data on the incidence of appendicitis from 170 countries and two different approaches were used to estimate numbers of patients who do not receive surgery: as a fixed proportion of the total unmet surgical need per country (approach 1); and based on country income status (approach 2). Indirect costs with current levels of access and local quality, and those if quality were at the standards of high-income countries, were estimated. A human capital approach was applied, focusing on the economic burden resulting from premature death and absenteeism. Results: Excess mortality was 4185 per 100 000 cases of appendicitis using approach 1 and 3448 per 100 000 using approach 2. The economic burden of continuing current levels of access and local quality was US $92 492 million using approach 1 and$73 141 million using approach 2. The economic burden of not providing surgical care to the standards of high-income countries was $95 004 million using approach 1 and$75 666 million using approach 2. The largest share of these costs resulted from premature death (97.7 per cent) and lack of access (97.0 per cent) in contrast to lack of quality. Conclusion: For a comparatively non-complex emergency condition such as appendicitis, increasing access to care should be prioritized. Although improving quality of care should not be neglected, increasing provision of care at current standards could reduce societal costs substantially