Search results for: bacterial 16S rRNA

Authors: S. Susanti, V. P. Bintoro, A. Setiadi, S. I. Santoso, D. R. Febriandi

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Meat is a foodstuff of animal origin. It is perishable because a suitable medium for bacterial growth. That is why meat can be a potential hazard to humans. Several ways have been done to inhibit bacterial population in an effort to prolong the meat shelf-life. However, aberration sometimes happens in the practices of meat preservation, for example by using chemical material that possessed strong antibacterial activity like formaldehyde. For health reason, utilization of formaldehyde as a food preservative was forbidden because of DNA damage resulting cancer and birth defects. Therefore, it is important to seek a natural food preservative that is not harmful to the body. This study aims to reveal the potency of cashew apple as natural food preservative by measuring its antibacterial activity and marinating effect on the bacterial growth of beef and chicken meat. Antibacterial activity was measured by The Kirby-Bauer method while bacterial growth was determined by total plate count method. The results showed that inhibition zone of 10-30% cashew apple extract significantly wider compared to 0% extract on the medium of E. coli, S. aureus, S. typii, and Bacillus sp. Furthermore, beef marinated with 20-30% cashew apple extract and chicken meat marinated with 5-15% extract significantly less in the total number of bacteria compared to 0% extract. It can be concluded that marinating with 5-30% cashew apple extract can effectively inhibit the bacterial growth of beef and chicken meat. Moreover, the concentration of extracts to inhibit bacterial populations in chicken meat was reached at the lower level compared to beef. Thus, cashew apple is potential as a natural food preservative.

Keywords: bacterial growth, cashew apple, marinating, meat

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Authors: Tereza Branyšová, Martina Kračmarová, Kateřina Demnerová, Michal Ďurovič, Hana Stiborová

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Microbial deterioration threatens all objects of cultural heritage, including audio-visual materials. Fungi are commonly known to be the main factor in audio-visual material deterioration. However, although being neglected, bacteria also play a significant role. In addition to microbial contamination of materials, it is also essential to analyse air as a possible contamination source. This work aims to identify bacterial species in the archives of the Czech Republic that occur on audio-visual materials as well as in the air in the archives. For sampling purposes, the smears from the materials were taken by sterile polyurethane sponges, and the air was collected using a MAS-100 aeroscope. Metagenomic DNA from all collected samples was immediately isolated and stored at -20 °C. DNA library for the 16S rRNA gene was prepared using two-step PCR and specific primers and the concentration step was included due to meagre yields of the DNA. After that, the samples were sent to the University of Fairbanks, Alaska, for Illumina MiSeq sequencing. Subsequently, the analysis of the sequences was conducted in R software. The obtained sequences were assigned to the corresponding bacterial species using the DADA2 package. The impact of air contamination and the impact of different photosensitive layers that audio-visual materials were made of, such as gelatine, albumen, and collodion, were evaluated. As a next step, we will take a deeper focus on air contamination. We will select an appropriate culture-dependent approach along with a culture-independent approach to observe a metabolically active species in the air. Acknowledgment: This project is supported by grant no. DG18P02OVV062 of the Ministry of Culture of the Czech Republic.

Keywords: cultural heritage, Illumina MiSeq, metagenomics, microbial identification

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Authors: Kulvinder Bajwa, Narsi R. Bishnoi

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Finding a sustainable alternative for today’s petrochemical industry is a major challenge facing by researchers, scientists, chemical engineers, and society at the global level. Microorganisms are considered to be sustainable feedstock for 3rd generation biofuel production. In this study, we have investigated the potential of a native bacterial strain isolated from a petrol contaminated site for the production of biodiesel. The bacterium was identified to be Rhodococcus opacus by biochemical test and 16S rRNA. Compositional analysis of bacterial biomass has been carried out by Fourier transform infrared spectroscopy (FTIR) in order to confirm lipid profile. Lipid and biomass were optimized by combination with Box Behnken design (BBD) of response surface methodology. The factors selected for the optimization of growth condition were glucose, yeast extract, and ammonium nitrate concentration. The experimental model developed through RSM in terms of effective operational factors (BBD) was found to be suitable to describe the lipid and biomass production, which indicated higher lipid and biomass with a minimum concentration of ammonium nitrate, yeast extract, and quite higher dose of glucose supplementation. Optimum results of the experiments were found to be 2.88 gL⁻¹ biomass and lipid content 38.75% at glucose 20 gL⁻¹, ammonium nitrate 0.5 gL⁻¹ and yeast extract 1.25 gL⁻¹. Furthermore, GCMS study revealed that Rhodococcus opacus has favorable fatty acid profile for biodiesel production.

Keywords: biofuel, Oleaginious bacteria, Rhodococcus opacus, FTIR, BBD, free fatty acids

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Authors: S. S. Singh, A. Bhattacharya, S. Bhattacharya

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The eukaryotic exosome complex plays a pivotal role in RNA biogenesis, maturation, surveillance and differential expression of various RNAs in response to varying environmental signals. The exosome is composed of evolutionary conserved nine core subunits and the associated exonucleases Rrp6 and Rrp44. Rrp6p is crucial for the processing of rRNAs, other non-coding RNAs, regulation of polyA tail length and termination of transcription. Rrp6p, a 3’-5’ exonuclease is required for degradation of 5’-external transcribed spacer (ETS) released from the rRNA precursors during the early steps of pre-rRNA processing. In the parasitic protist Entamoeba histolytica in response to growth stress, there occurs the accumulation of unprocessed pre-rRNA and 5’ ETS sub fragment. To understand the processes leading to this accumulation, we looked for Rrp6 and the exosome subunits in E. histolytica, by in silico approaches. Of the nine core exosomal subunits, seven had high percentage of sequence similarity with the yeast and human. The EhRrp6 homolog contained exoribonuclease and HRDC domains like yeast but its N- terminus lacked the PMC2NT domain. EhRrp6 complemented the temperature sensitive phenotype of yeast rrp6Δ cells suggesting conservation of biological activity. We showed 3’-5’ exoribonuclease activity of EhRrp6p with in vitro-synthesized appropriate RNAs substrates. Like the yeast enzyme, EhRrp6p degraded unstructured RNA, but could degrade the stem-loops slowly. Furthermore, immunolocalization revealed that EhRrp6 was nuclear-localized in normal cells but was diminished from nucleus during serum starvation, which could explain the accumulation of 5’ETS during stress. Our study shows functional conservation of EhRrp6p in E.histolytica, an early-branching eukaryote, and will help to understand the evolution of exosomal components and their regulatory function.

Keywords: entamoeba histolytica, exosome complex, rRNA processing, Rrp6

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Authors: Azila Adnan, Giridhar R. Nair, Mark C. Lay, Janis E. Swan

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The effect of aeration on bacterial cellulose (BC) production by Gluconacetobacter xylinus DSM46604 was studied in 5-L bioreactor. Four aeration rates were applied (0.3, 0.6, 1.0 and 1.5 vvm) in the fermentation media at constant agitation rate, 150 rpm. One vvm enhanced BC concentration (4.4 g/L) and productivity (0.44 g/L/day) while greater agitation rate (1.5 vvm) decreased BC concentration (4.0 g/L) and productivity (0.40 g/L/day).

Keywords: aeration, bacterial cellulose, fermentation, gluconacetobacter xylinus

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Authors: Alok Kumar, Hari Ram, Lebin Thomas, Ved Pal Singh

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Organic solvent tolerant amylases and proteases of microbial origin are in great demand for their application in transglycosylation of water-insoluble flavanoids and in peptide synthesizing reaction in organic media. Most of the amylases and proteases are unstable in presence of organic solvent. In the present work two different bacterial strains M-11 and VP-07 were isolated from the soil sample of a dairy farm in Delhi, India, for the efficient production of extracellular amylase and protease through their screening on starch agar (SA) and skimmed milk agar (SMA) plates, respectively. Both the strains (M-11 and VP-07) were identified based on morphological, biochemical and 16S rRNA gene sequencing methods. After analysis through Ez-Taxon software, the strains M-11 and VP-07 were found to have maximum pairwise similarity of 98.63% and 100% with Bacillus subtilis subsp. inaquosorum BGSC 3A28 and Bacillus anthracis ATCC 14578 and were therefore identified as Bacillus sp. UKS1 and Bacillus sp. UKS2, respectively. Time course study of enzyme activity and bacterial growth has shown that both strains exhibited typical sigmoid growth behavior and maximum production of amylase (180 U/ml) and protease (78 U/ml) by these strains (UKS1 and UKS2) was commenced during stationary phase of growth at 24 and 20 h, respectively. Thereafter, both amylase and protease were tested for their tolerance towards organic solvents and were found to be active as well stable in p-xylene (130% and 115%), chloroform (110% and 112%), isooctane (119% and 107%), benzene (121% and 104%), n-hexane (116% and 103%) and toluene (112% and 101%, respectively). Owing to such properties, these enzymes can be exploited for their potential application in industries for organic synthesis.

Keywords: amylase, enzyme activity, industrial applications, organic solvent tolerant, protease

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Authors: Khaled Khleifat

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This study detects the bacterial species that responsible for UTI in both diabetic patients and non-diabetic patients, Jordan. 116 urine samples were investigated in order to determine UTI-causing bacteria. These samples distributed unequally between diabetic male (12) and diabetic female (25) and also non-diabetic male (13) and non-diabetic female (66). The results represent that E.coli is responsible for UTI in both diabetic and non-diabetic patients (15.5% and 29.3% respectively) with large proportion (44.8%). This study showed that not all bacterial species that isolated from the non-diabetic sample could be isolated from diabetic samples. E. coli (15.5%), P. aeruginosa (4.3%), K. pneumonia (1.7%), P. mirabilis (2.6%), S. marcescens (0.9%), S. aureus (1.7%), S. pyogenes (1.7%), E. faecalis (0.9%), S. epidermidis (1.7%) and S. saprophyticus (0.9%). But E. aerogenes, E. cloacae, C. freundii, A. baumannii and B. subtilis are five bacterial species that can’t isolate from all diabetic samples. This study shows that for the treatment of UTI in both diabetic and non-diabetic patients, Chloramphenicol (30 μg), Ciprofloxacin (5 μg) and Vancomycin (30 μg) are more favorable than other antibiotics. In the same time, Cephalothin (30μg) is not recommended.

Keywords: urinary tract infections, diabetes mellitus, bacterial species, infections

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Authors: Tessa E. Reid, Vanessa N. Kavamura, Maider Abadie, Adriana Torres-Ballesteros, Mark Pawlett, Ian M. Clark, Jim Harris, Tim Mauchline

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The deleterious effect of chemical fertilizer on rhizobacterial diversity has been well documented using 16S rRNA gene amplicon sequencing and predictive metagenomics. Biofertilization is a cost-effective and sustainable alternative; improving strategies depends on isolating beneficial soil microorganisms. Although culturing is widespread in biofertilization, it is unknown whether the composition of cultured isolates closely mirrors native beneficial rhizobacterial populations. This study aimed to determine the relative abundance of culturable plant growth-promoting rhizobacteria (PGPR) isolates within total soil DNA and how potential PGPR populations respond to chemical fertilization in a commercial wheat variety. It was hypothesized that PGPR will be reduced in fertilized relative to unfertilized wheat. Triticum aestivum cv. Cadenza seeds were sown in a nutrient depleted agricultural soil in pots treated with and without nitrogen-phosphorous-potassium (NPK) fertilizer. Rhizosphere and rhizoplane samples were collected at flowering stage (10 weeks) and analyzed by culture-independent (amplicon sequence variance (ASV) analysis of total rhizobacterial DNA) and -dependent (isolation using growth media) techniques. Rhizosphere- and rhizoplane-derived microbiota culture collections were tested for plant growth-promoting traits using functional bioassays. In general, fertilizer addition decreased the proportion of nutrient-solubilizing bacteria (nitrate, phosphate, potassium, iron and, zinc) isolated from rhizocompartments in wheat, whereas salt tolerant bacteria were not affected. A PGPR database was created from isolate 16S rRNA gene sequences and searched against total soil DNA, revealing that 1.52% of total community ASVs were identified as culturable PGPR isolates. Bioassays identified a higher proportion of PGPR in non-fertilized samples (rhizosphere (49%) and rhizoplane (91%)) compared to fertilized samples (rhizosphere (21%) and rhizoplane (19%)) which constituted approximately 1.95% and 1.25% in non-fertilized and fertilized total community DNA, respectively. The analyses of 16S rRNA genes and deduced functional profiles provide an in-depth understanding of the responses of bacterial communities to fertilizer; this study suggests that rhizobacteria, which potentially benefit plants by mobilizing insoluble nutrients in soil, are reduced by chemical fertilizer addition. This knowledge will benefit the development of more targeted biofertilization strategies.

Keywords: bacteria, fertilizer, microbiome, rhizoplane, rhizosphere

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Authors: Anish Shah, Steve A. Wakelin, Derrick Moot, Aurélie Laugraud, Hayley J. Ridgway

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A mixed pasture system of ryegrass-clover is used in New Zealand, where clovers are generally inoculated with commercially available strains of rhizobia. The community of rhizobia living in the soil and the way in which they interact with the plant are affected by different biotic and abiotic factors. In general, bacterial richness and diversity in soil varies by soil pH. pH also affects cell physiology and acts as a master variable that controls the wider soil physiochemical conditions such as P availability, Al release and micronutrient availability. As such, pH can have both primary and secondary effects on soil biology and processes. The aim of this work was to investigate the effect of soil pH on the genetic diversity and metabolic profile of Rhizobium leguminosarum strains nodulating clover. Soils were collected from 12 farms across New Zealand which had a pH(water) range of between 4.9 and 7.5, with four acidic (pH 4.9 – 5.5), four ‘neutral’ (5.8 – 6.1) and four alkaline (6.5 – 7.5) soils. Bacteria were recovered from nodules of Trifolium repens (white clover) and T. subterraneum (subterranean clover) grown in the soils. The strains were cultured and screened against a range of pH-amended media to demonstrate whether they were adapted to pH levels similar to their native soils. The strains which showed high relative growth at a given pH (~20% of those isolated) were selected for metabolic and taxonomic profiling. The Omnilog (Biolog Inc., Hayward, CA) phenotype array was used to perform assays on carbon (C) utilisation for selected strains. DNA was extracted from the strains which had differing C utilisation profiles and PCR products for both forward and reverse primers were sequenced for the following genes: 16S rRNA, recA, nodC, nodD and nifH (symbiotic).

Keywords: bacterial diversity, clover, metabolic and taxonomic profiling, pH adaptation, rhizobia

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Authors: Amel Bouderhem, Aminata Ould El Hadj Khelil, Amina N. Djrarbaoui, Aroussi Aroussi

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The present work aims to find the influence of the nature of the soil on the effectiveness of the biodegradation of hydrocarbons by a mixture of bacterial strains hydrocarbonoclastes. Processes of bioaugmentation and biostimulation trial are applied to samples of soils polluted voluntarily by the crude oil. For the evaluation of the biodegradation of hydrocarbons, the bacterial load, the pH and organic carbon total are followed in the different experimental batches. He bacterial load of the sandy soil varies among the witnesses of 45,2 .108 CFU/ml at the beginning of the experimentation to 214,07.108 CFU/ml at the end of the experiment. Of the soil silty-clay varies between 103,31 .108 CFU/ml and 614,86.108 CFU/ml . It was found a strong increase in the bacterial biomass during the processing of all samples. This increase is more important in the samples of sand bioaugmente or biomass increased from 63.16 .108 CFU/ml to 309.68 .108 CFU/ml than in soil samples silty clay- bioaugmente whose content in bacteria evolved of 73,01 .108 CFU/ml to 631.80 . 108CFU/ml

Keywords: pollution, hydrocarbons, bioremediation, bacteria hydrocarbonoclastes, ground, texture

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Authors: Reem Alajmi, Hind Al Harbi, Tahany Ayaad, Zainab Al Musawi

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Hard ticks Hyalomma spp. (family: Ixodidae) are obligate ectoparasite in their all life stages on some domestic animals mainly camels and cattle. Ticks may lead to many economic and public health problems because of their blood feeding behavior. Also, they act as vectors for many bacterial, viral and protozoan agents which may cause serious diseases such as tick-born encephalitis, Rocky-mountain spotted fever, Q-fever and Lyme disease which can affect human and/or animals. In the present study, molecular identification of ticks that attack camels in Riyadh region, Saudi Arabia based on the partial sequence of mitochondrial 16s rRNA gene was applied. Also, the present study aims to detect natural infections of collected camel ticks with Rickessia spp. and Borelia spp. using PCR/hybridization of Citrate synthase encoding gene present in bacterial cells. Hard ticks infesting camels were collected from different camels located in a farm in Riyadh region, Saudi Arabia. Results of the present study showed that the collected specimens belong to two species: Hyalomma dromedari represent 99% of the identified specimens and Hyalomma marginatum which account for 1 % of identified ticks. The molecular identification was made through blasting the obtained sequence of this study with sequences already present and identified in GeneBank. All obtained sequences of H. dromedarii specimens showed 97-100% identity with the same gene sequence of the same species (Accession # L34306.1) which was used as a reference. Meanwhile, no intraspecific variations of H. marginatum mesured because only one specimen was collected. Results also had shown that the intraspecific variability between individuals of H. dromedarii obtained in 92 % of samples ranging from 0.2- 6.6%, while the remaining 7 % of the total samples of H. dromedarii showed about 10.3 % individual differences. However, the interspecific variability between H. dromedarii and H. marginatum was approximately 18.3 %. On the other hand, by using the technique of PCR/hybridization, we could detect natural infection of camel ticks with Rickettsia spp. and Borrelia spp. Results revealed the natural presence of both bacteria in collected ticks. Rickettsial spp. infection present in 29% of collected ticks, while 35% of collected specimen were infected with Borrelia spp. The valuable results obtained from the present study are a new record for the molecular identification of camel ticks in Riyadh, Saudi Arabia and their natural infection with both Rickettsia spp. and Borrelia spp. These results may help scientists to provide a good and direct control strategy of ticks in order to protect one of the most important economic animals which are camels. Also results of this project spotlight on the disease that might be transmitted by ticks to put out a direct protective plan to prevent spreading of these dangerous agents. Further molecular studies are needed to confirm the results of the present study by using other mitochondrial and nuclear genes for tick identification.

Keywords: Camel ticks, Rickessia spp. , Borelia spp. , mitochondrial 16s rRNA gene

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Authors: Li Cheng, Zhang Yilei, Cohen Yehuda

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Two motility mechanisms were introduced into iDynoMiCs software, which adopts an individual-based modeling method. Based on the new capabilities, along with the pressure motility developed before, influence of bacterial motility on biofilm formation was studied. Simulation results were evaluated both qualitatively through 3D structure inspections and quantitatively by parameter characterizations. It was showed that twitching motility increased the biofilm surface irregularity probably due to movement of cells towards higher nutrient concentration location whereas free motility, on the other hand, could make biofilms flatter and smoother relatively. Pressure motility showed no significant influence in this study.

Keywords: iDynoMics, biofilm structure, bacterial motility, motility mechanisms

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Authors: Mohammad Mokhtari, Gires Usup, Zaidi Che Cob

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Bacteria communities as mediators of the biogeochemical process are the main component of the mangrove ecosystems. Crab burrows by increasing oxic-anoxic interfaces and facilitating the flux rate between sediment and tidal water affect biogeochemical properties of sediments. The effect of fiddler crab burrows on the density and diversity of bacteria were investigated to elucidate the effect of burrow on bacterial distribution. Samples collected from the burrow walls of three species of fiddler crabs including Uca paradussumieri, Uca rosea, and Uca forcipata. Sediment properties including grain size, temperature, Redox potential, pH, chlorophyll, water and organic content were measured from the burrow walls to assess the correlation between environmental variables and bacterial communities. Bacteria were enumerated with epifluorescence microscopy after staining with SYBR green. Bacterial DNA extracted from sediment samples and the community profiles of bacteria were determined with Terminal Restriction Fragment Length Polymorphism (T-RFLP). High endemism was observed among bacterial communities. Among the 152 observed OTU’s, 22 were found only in crab burrows. The highest bacterial density and diversity were recorded in burrow wall. The results of ANOSIM indicated a significant difference between the bacterial communities from the three species of fiddler crab burrows. Only 3% of explained bacteria variability in the constrained ordination model of CCA was contributed to depth, while much of the bacteria’s variability was attributed to coarse sand, pH, and chlorophyll content. Our findings suggest that crab burrows by affecting sediment properties such as redox potential, pH, water, and chlorophyll content induce significant effects on the bacterial communities.

Keywords: bioturbation, canonical corresponding analysis, fiddler crab, microbial ecology

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Authors: M.Ayazi, N. Golshan Ebrahimi

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The development of antibacterial surface has devoted extensive researches and important field due to the growing antimicrobial resistance strains. The superhydrophobic surface has raised attention because of reducing bacteria adhesion in the absence of antibiotic agents. Evaluating the current development antibacterial surface has to be investigating to consider the potential of applying superhydrophobic surface to reduce bacterial adhesion or role of patterned surfaces on it. In this study, we present different samples with bio-inspired hierarchical and microstructures to consider their ability in reducing bacterial adhesion. The structures have inspired from rice-like pattern and lotus-leaf that developed on the polydimethylsiloxane (PDMS) and polypropylene (PP). The results of the attachment behaviors have considered on two bacteria strains of gram-negative Escherichia coli (E. coli) bacteria and gram-positive Staphylococcus aureus (S. aureus). The reduction of bacteria adhesion on these roughness surfaces demonstrated the effectiveness of rinsing ability on removing bacterial cells on structured plastic surfaces. Results have also offered the important role of bacterial species, material chemistry and hierarchical structure to prevent bacterial adhesion.

Keywords: hierarchical structure, self-cleaning, lotus-effect, bactericidal

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Authors: Waleed S. Alwaneen

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In many countries, cow dung is used as farm manure and for biogas production. Several bacterial strains associated with cow dung such as Campylobacter, Salmonella sp. and Escherichia coli cause serious human diseases. The objective of the present study was to investigate the use of insect larvae including fruit beetle, waxworms and tiger worms to improve the breakdown of agricultural wastes and reduce their pathogen loads. Fresh cow faeces were collected from a cattle farm and distributed into plastic boxes (100 g/box). Each box was provided with 10 larvae of fruit beetle, Waxworms and Tiger worms, respectively. There were 3 replicates in each treatment including the control. Bacteria were isolated weekly from both control and cow faeces to which larvae were added to determine the bacterial populations. Results revealed that the bacterial load was higher in the cow faeces treated with fruit beetles than in the control, while the bacterial load was lower in the cow faeces treated with waxworms and tiger worms than in the control. The activities of the fruit beetle larvae led to the cow faeces being liquefied which provided a more conducive growing media for bacteria. Therefore, higher bacterial load in the cow faeces treated with fruit beetle might be attributed to the liquefaction of cow faeces.

Keywords: fruit beetle, waxworms, tiger worms, waste conditioning, composting

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Authors: Shilpi, Indu Gaur, Neha Bhadauria, Susmita Goswami, Prabir K. Paul

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Cultivation and consumption of organically grown fruits and vegetables have increased by several folds. However, the presence of Human Enteric Pathogens on the surface of organically grown vegetables causing Gastro-intestinal diseases, are most likely due to contaminated water and fecal matter of farm animals. Human Enteric Pathogens are adapted to colonize the human gut, and also colonize plant surface. Microbes on plant surface communicate with each other to establish quorum sensing. The cross talk study is important because the enteric pathogens on phylloplane have been reported to mask the beneficial resident bacteria of plant. In the present study, HEPs and bacterial colonizers were identified using 16s rRNA sequencing. Microbial colonization patterns after interaction between Human Enteric Pathogens and natural bacterial residents on tomato phylloplane was studied. Tomato plants raised under aseptic conditions were inoculated with a mixture of Serratia fonticola and Klebsiella pneumoniae. The molecules involved in cross-talk between Human Enteric Pathogens and regular bacterial colonizers were isolated and identified using molecular techniques and HPLC. The colonization pattern was studied by leaf imprint method after 48 hours of incubation. The associated protein-protein interaction in the host cytoplasm was studied by use of crosslinkers. From treated leaves the crosstalk molecules and interaction proteins were separated on 1D SDS-PAGE and analyzed by MALDI-TOF-TOF analysis. The study is critical in understanding the molecular aspects of HEP’s adaption to phylloplane. The study revealed human enteric pathogens aggressively interact among themselves and resident bacteria. HEPs induced establishment of a signaling cascade through protein-protein interaction in the host cytoplasm. The study revealed that the adaptation of Human Enteric Pathogens on phylloplane of Solanum lycopersicum involves the establishment of complex molecular interaction between the microbe and the host including microbe-microbe interaction leading to an establishment of quorum sensing. The outcome will help in minimizing the HEP load on fresh farm produce, thereby curtailing incidences of food-borne diseases.

Keywords: crosslinkers, human enteric pathogens (HEPs), phylloplane, quorum sensing

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Authors: Mian Adnan Kakakhel, NIshita Narwal, Majid Rasta, Shi Xiaotao

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The widespread use of nanoparticles means that they are significantly increasing in the aquatic ecosystem, where they are likely to pose threat to aquatic organism. In particular, the influence of nanoparticles exposure combined with varying water velocities on fish behavior remain poorly understood. Emerging evidences suggested a probable correlation between fish swimming behavior and gut bacterial dysbiosis. Therefore, the current study aimed to investigate the effects of nanomaterials in different water velocities on fish gut bacterial composition, which in results change in fish swimming behavior. The obtained findings showed that the contamination of nanoparticles was reduced as the velocity increased. However, the synergetic effects of nanoparticles and water velocity significantly (p < 0.05) decreased the bacterial composition, which plays a critical role in fish development, metabolism, digestion, enzymes production, and energy production such as Bacteroidetes and Firmicutes. This group of bacterial also support fish in swimming behavior by providing them a significant energy during movement. The obtained findings of this study suggested that the presence of nanoparticles in different water velocities have had a significant correlation with fish gut bacterial dysbiosis, as results the gut dysbiosis had been linked to the change in fish behavior. The study provides an important insight into the mechanisms by which the nanoparticles possibly affect the fish behavior.

Keywords: water velocities, fish behavior, gut bacteria, secondary metabolites, regulation

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Authors: Mubashir Hussain, Mu Lv, Xiaohan Dong, Zhiyang Li, Bin Liu, Nongyue He

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Detection and classification of microbes have a vast range of applications in biomedical engineering especially in detection, characterization, and quantification of bacterial contaminants. For identification of pathogens, different techniques are emerging in the field of biomedical engineering. Latest technology uses light scattering, capable of identifying different pathogens without any need for biochemical processing. Bacterial Pathogens Identification System (BPIS) which uses a laser beam, passes through the sample and light scatters off. An assembly of photodetectors surrounded by the sample at different angles to detect the scattering of light. The algorithm of the system consists of two parts: (a) Library files, and (b) Comparator. Library files contain data of known species of bacterial microbes in the form of binned plots, while comparator compares data of unknown sample with library files. Using collected data of unknown bacterial species, highest voltage values stored in the form of peaks and arranged in 3D histograms to find the frequency of occurrence. Resulting data compared with library files of known bacterial species. If sample data matching with any library file of known bacterial species, sample identified as a matched microbe. An experiment performed to identify three different bacteria particles: Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli. By applying algorithm using library files of given samples, results were compromising. This system is potentially applicable to several biomedical areas, especially those related to cell morphology.

Keywords: microbial identification, laser scattering, peak identification, binned plots classification

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Authors: Zakirul Islam, Yugo Kumokawa, Quoc Thinh Tran, Motoki Kubo

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Clubroot is a disease of cruciferous plant caused by soil born pathogen Plasmodiophora brassicae and can significantly limit the production through rapid spreading. The present study was designed to investigate the effect of cultivation practices (chemical and organic soils) on clubroot disease development in Brassica rapa. Disease index and root bacterial composition were investigated for both chemical and organic soils. The bacterial biomass and diversity in organic soil were higher than those in chemical soil. Disease severity was distinct for two different cultivation methods. The number of endophytic bacteria decreased in the infected root for both soils. The increased number of endophytic bacterial number led to reduce the proliferation of pathogen spore inside the root and thus reduced the disease severity in organic plants.

Keywords: clubroot disease, bacterial biomass, root infection, disease index, chemical cultivation, organic cultivation

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Authors: Sarah Almuhayya, Andrew Mcbain, Gavin Humphreys

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Background. The microbiome of the upper respiratory tract (URT) has received less research attention than other body sites. This study aims to investigate the microbial ecology of the human URT with a focus on the antagonism between the corynebacteria and staphylococci. Methods. Mucosal swabs were collected from the anterior nares and nasal turbinates of 20 healthy adult subjects. Genomic DNA amplification targeting the (V4) of the 16Sr RNA gene was conducted and analyzed using QIIME. Nasal swab isolates were cultured and identified using near full-length sequencing of the 16S rRNA gene. Isolates identified as corynebacteria or staphylococci were typed using (rep-PCR). Antagonism was determined using an agar-based inhibition assay. Results. Four major bacterial phyla (Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria) were identified from all volunteers. The typing of cultured staphylococci and corynebacteria suggested that intra-individual strain diversity was limited. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between staphylococci and corynebacteria. Despite the apparent antagonism between these genera, it was limited when investigated on agar. Of 1000 pairwise interactions, observable zones of inhibition were only reported between a single strain of C.pseudodiphtheriticum and S.aureus. Imaging under EM revealed this effect to be bactericidal with clear lytic effects on staphylococcal cell morphology. Conclusion. Nasal microbiota is complex, but culturable staphylococci and corynebacteria were limited in terms of clone type. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between these genera suggesting an antagonism or competition between these taxonomic groups.

Keywords: nasal, microbiota, S.aureus, microbioal interaction

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Authors: Ben Amar Cheba, Taha Ibrahim Zaghloul, Mohamad Hisham El-Massry, Ahmad Rafik El-Mahdy

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Seventy two bacterial strains with ability to degrade chitin were isolated during a screening program. One of the most potent isolates (strain R2) was identified as Bacillus sp. using conventional methods as well as 16S rRNA technique and submitted in the Gen Bank sequence database as Bacillus sp. R2 with a given accession number DQ 923161. This strain was able to produce high levels of extracellular chitinase. The chitinase of Bacillus sp. R2 hydrolyzed several chitinous substrates preferentially and showed a maximum activity toward the β chitin such as Calmar pen and squid bone chitins with the folds 1.47 and 1.23 respectively. The enzyme also exhibited a substrate binding capacity of more than 70% for squid chitin, shrimp shell colloidal chitin, chitosan and prawn shell chitin. The chitinase showed a moderate antifungal activity against many phytopathogenic fungi such as Aspergillus niger, A. flavus, Penicillium degitatum and Fusarium calmorum.This strain could be a suitable candidate for chitinase production on an industrial scale for using as promising antifungal biopestecide.

Keywords: antifungal activity, Bacillus sp. R2, chitinase, substrate specificity

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Authors: David Pinzauti, Simon De Jaegher, Maria D'Aguano, Manuele Biazzo

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The COVID-19 pandemic has left an indelible mark on global health, leading to a pressing need for understanding the intricate interactions between the virus and the human microbiome. This study focuses on characterizing the nasal microbiota of patients affected by COVID-19, with a specific emphasis on the comparison with unaffected individuals, to shed light on the crucial role of the microbiome in the development of this viral disease. To achieve this objective, Nanopore technology was employed to analyze the bacterial 16s rRNA full-length gene present in nasal swabs collected in Malta between January 2021 and August 2022. A comprehensive dataset consisting of 268 samples (126 SARS-negative samples and 142 SARS-positive samples) was subjected to a comparative analysis using an in-house, custom pipeline. The findings from this study revealed that individuals affected by COVID-19 possess a nasal microbiota that is significantly less diverse, as evidenced by lower α diversity, and is characterized by distinct microbial communities compared to unaffected individuals. The beta diversity analyses were carried out at different taxonomic resolutions. At the phylum level, Bacteroidota was found to be more prevalent in SARS-negative samples, suggesting a potential decrease during the course of viral infection. At the species level, the identification of several specific biomarkers further underscores the critical role of the nasal microbiota in COVID-19 pathogenesis. Notably, species such as Finegoldia magna, Moraxella catarrhalis, and others exhibited relative abundance in SARS-positive samples, potentially serving as significant indicators of the disease. This study presents valuable insights into the relationship between COVID-19 and the nasal microbiota. The identification of distinct microbial communities and potential biomarkers associated with the disease offers promising avenues for further research and therapeutic interventions aimed at enhancing public health outcomes in the context of COVID-19.

Keywords: COVID-19, nasal microbiota, nanopore technology, 16s rRNA gene, biomarkers

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Authors: Omneya M. Helmy, Mona T. Kashef

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Aminoglycosides are used in treating a wide range of infections caused by both Gram-negative and Gram positive bacteria. The presence of 16S rRNA methyl transferases (16S-RMTase) is among the newly discovered resistance mechanisms that confer high resistance to clinically useful aminoglycosides. Cephalosporins are the most commonly used antimicrobials in Egypt; therefore, this study was conducted to determine the isolation frequency of 16S rRNA methyl transferases among third generation cephalosporin-resistant clinical isolates in Egypt. One hundred and twenty three cephalosporin resistant Gram-negative clinical isolates were screened for aminoglycoside resistance by the Kirby Bauer disk diffusion method and tested for possible production of 16S-RMTase. PCR testing and sequencing were used to confirm the presence of 16S-RMTase and the associated antimicrobial resistance determinants, as well as the genetic region surrounding the armA gene. Out of 123 isolates, 66 (53.66%) were resistant to at least one aminoglycoside antibiotic. Only one Escherichia coli isolate (E9ECMO) which was totally resistant to all tested aminoglycosides, was confirmed to have the armA gene in association with blaTEM-1, blaCTX-M-15, blaCTX-M-14 and aac(6)-Ib genes. The armA gene was found to be carried on a large A/C plasmid. Genetic mapping of the armA surrounding region revealed, for the first time, the association of armA with aac(6)-Ib on the same transposon. In Conclusion, the isolation frequency of 16S-RMTase was low among the tested cephalosporin-resistant clinical samples. However, a novel composite transposon has been detected conferring high-level aminoglycosides resistance.

Keywords: aminoglcosides, armA gene, β lactmases, 16S rRNA methyl transferases

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Authors: Sanam Pudasaini, A. T. K. Perera, Ahmed Syed Shaheer Uddin, Sum Huan Ng, Chun Yang

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Development of high-efficiency and environment friendly bacterial inactivation methods is of great importance for preventing waterborne diseases which are one of the leading causes of death in the world. Traditional bacterial inactivation methods (e.g., ultraviolet radiation and chlorination) have several limitations such as longer treatment time, formation of toxic byproducts, bacterial regrowth, etc. Recently, an electroporation-based inactivation method was introduced as a substitute. Here, an electroporation-based continuous flow microfluidic device equipped with an array of micropillars is developed, and the device achieved high bacterial inactivation performance ( > 99.9%) within a short exposure time ( < 1 s). More than 99.9% reduction of Escherichia coli bacteria was obtained for the flow rate of 1 mL/hr, and no regrowth of bacteria was observed. Images from scanning electron microscope confirmed the formation of electroporation-induced nano-pore within the cell membrane. Through numerical simulation, it has been shown that sufficiently large electric field strength (3 kV/cm), required for bacterial electroporation, were generated using PDMS micropillars for an applied voltage of 300 V. Further, in this method of inactivation, there is no involvement of chemicals and the formation of harmful by-products is also minimum.

Keywords: electroporation, high-efficiency, inactivation, microfluidics, micropillar

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Authors: Ramachandran Manivannan, B. Sakthi Swaroop, Selvam Noyel Victoria

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Microbial corrosion of brass gauze by the aerobic film forming bacteria Arthrobacter sulfurous in neutral media was investigated using gravimetric studies. Maximum weight loss of 166.98 mg was observed for a period of 28 days of exposure to the bacterial medium as against the weight loss of 13.69 mg for control. The optical density studies for the bacterial culture was found to show attainment of stationary phase in 48 h. Scanning electron microscopy analysis of the samples shows the presence of pitting corrosion. The energy dispersive X-ray analysis of the samples showed increased oxygen and phosphorus content in the sample due to bacterial activity.

Keywords: Arthrobacter sulfureus, biocorrosion, brass, neutral medium

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Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

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Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

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Authors: Desouky Abd El Haleem, Sahar Zaki

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This work was conducted to develop a one-step multiplex PCR system for rapid, sensitive, and specific detection of three different bacterial pathogens, Escherichia coli, Pseudomonas aeruginosa, and Salmonella spp, directly in wastewater without prior isolation on selective media. As a molecular confirmatory test after isolation of the pathogens by classical microbiological methods, PCR-RFLP of their amplified 16S rDNA genes was performed. It was observed that the developed protocols have significance impact in the ability to detect sensitively, rapidly and specifically the three pathogens directly in water within short-time, represents a considerable advancement over more time-consuming and less-sensitive methods for identification and characterization of these kinds of pathogens.

Keywords: multiplex PCR, bacterial pathogens, Escherichia coli, Pseudomonas aeruginosa, Salmonella spp.

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Authors: Fatima Huda Hallak

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The current study aimed to evaluate the activity of bacterial insecticide Vertinic against the second star larvae of Trogoderma granarium (Everts) by four treatments: A, B, C, D, at seven concentrations: 0.001, 0.01, 0.1,1,10,100,1000 PPM. The mortality rate of larvae was 100% at concentrations 10 and 100 in treatments A and B after 24 hours and after 48 hours in treatment D at 1 PPM. The efficiency of treatment A was greater as compared to treatment B at all concentrations and all exposure times. The efficiency of treatment D was greater as compared to treatment C; for example, at 0.001, 0.01, 0.1, 1 PPM, after 120 hours, the Mortality rate of larve was 6.76, 13.33, 43.33, 100% in treatment D, which it was 0.00, 0.00, 23.33, 96.67%, respectively in the treatment C.

Keywords: bacterial insecticide, trogoderma granarium (everts), fourth star larvae, vertimic

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Authors: N. Khomarloo, F. Mousazadegan, M. Latifi, N. Hemmatinejad

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Recently, the utilization of reusable surgical gowns in order to decrease costs, environmental protection and enhance surgeon’s comfort is considered. One of the concerns in applying this kind of medical protective clothing is reduction of their resistance to bacterial penetration especially in wet state, after repeated laundering and sterilizing process. The purpose of this study is to investigate the effect of the laundering and sterilizing process on the reusable surgical gown’s resistance against bacterial wet penetration. To this end, penetration of Staphylococcus aureus bacteria in wet state after 70 washing and sterilizing cycles was evaluated on the two single-layer and three-layer reusable gowns. The outcomes reveal that up to 20 laundering and sterilizing cycles, protective property of samples improves due to fabric shrinkage, after that because of the fabric’s construction opening, the bacterial penetration increase. However, the three-layer gown presents higher protective performance comparing to the single-layer one.

Keywords: laundry, porosity, reusable surgical gown, sterilization, wet bacterial penetration

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Authors: Rifa’ah Mahmudah Bulu’

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MRSA is an important nosocomial pathogen in the world. Therefore, the prevention and effort to control MRSA is still very important to conduct. One of the preventions of MRSA, which have been reported by several studies, is Cefriaxone and Ethanol Extract of Ant Plant. This research is an experimental test to determine the potency of MRSA’s anti-bacterial with Cefriaxone (30 μg) and Ethanol Extract of Ant Plant (13 mg/ml) based on inhibition zone on LAB (Lempeng Agar Biasa). The size of inhibition zone that is formed on Cefriaxone is adjusted with CSLI criteria, which ≥ 21 mm of inhibition zone is called sensitive; ≤13 mm is called resistance and between 14-20 mm is called intermediate. This research is conducted three times. Comparative test between Cefriaxone and Ethanol Extract of Ant Plant is analyzed by Maan Whitney’s statistic method. The Result of Cefriaxone anti-bacterial potency shows the variety of inhibition zone. Cefriaxone forms approximately 16,5-20 mm with average 18,22mm of inhibition zone that make Cefriaxone’s criteria to MRSA’s inhibition is intermediate. Anti-bacterial potency of Ethanol Extract of Ant Plant is about 0,5-2 mm with average 1,17 mm of inhibition zone that prove MRSA is sensitive to Ant Plant. The conclusion of this research shows that Cefriaxone is intermediate to MRSA’s inhibition, while MRSA is sensitive to Ethanol Extract of Ant Plant, which at the end; it creates different potency of anti-bacterial between Cefriaxone and Ethanol Extract of Ant Plant.

Keywords: MRSA, cefriaxone, ant plant, CSLI, mann whitney

Procedia PDF Downloads 347