Search results for: bacillus bacteria study

Authors: Karima Ould Yerou, B. Meddah, A. Tir Touil

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The intestinal flora also called the intestinal microbiota, consists of different bacteria and other microorganisms which occur naturally in the gastrointestinal tract organs components. These intestinal bacteria are present in their millions and help the functioning of the body in particular allowing aid to degradation of certain molecules into absorbable substrates. They also protect against invasion of the gut by other pathogenic bacteria, that is to say which may be responsible for disease. Factors like stress, antibiotics and diet can affect the balance of intestinal flora and in case of imbalance, digestive disorders type bloating, diarrhea or vomiting may occur. Rattus norvegicus of bad weight of 100 kg, an Algerian firm West are scarified and isolation of their ileum and colon respectively two Bactrian strains Escherichia coli and Lactobacillus are then purified and identified.

Keywords: intestinal flora, Rattus norvegicus, Escherichia coli, lactobacillus, West Algerian farm

Procedia PDF Downloads 315

Authors: K. Haggag, M. Salama, H. El-Sayed

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Polyester fabrics were sanitized by exposing them to vaporized water under the influence of conventional heating or microwave irradiation. Hydrogen peroxide was added the humid sanitizing environment as a disinfectant. The said sanitization process was found to be effective towards two types of bacteria, namely Escherichia coli ATCC 2666 (G –ve) and Staphylococcus aureus ATCC 6538 (G +ve). The effect of the sanitization process on some of the inherent properties of polyester fabrics was monitored.

Keywords: polyester, fabric, sanitization, microwave, bacteria

Procedia PDF Downloads 346

Authors: Malalage Mudara Peiris

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Objectives of the study are: the biosynthesis of silver nanoparticles (AgNPs) using Escherichia coli, Acinetobacter baumannii and Staphylococcus aureus, characterization of silver nanoparticles and determination of antimicrobial activity against E. coli, P. aeruginosa, S. aureus, MRSA, and C. Albicans. Methods: E. coli (ATCC 25922), A. baumanii (clinical strain), S. aureus (clinical strain) cultured in nutrient broth medium were used for biosynthesis of AgNPs. Culture conditions (AgNO3 concentration, pH, incubation time and temperature) were optimized. Characterization of synthesized NPs was done by UV-Visible spectroscopy. The antimicrobial activity of the synthesized NPs was studied using the good diffusion assay against E. coli, S. aureus, MRSA (Methicillin-resistant Staphylococcus aureus), P. aeruginosa and C. Albicans. Results: All the selected bacteria produced silver nanoparticles at alkaline pH above 0.3 g/L AgNO3 concentration. The optimum reaction temperature was 60oC. According to the UV-Visible spectroscopy, the maximum absorbance was found to be around 420 - 430 nm indicating the presence of AgNPs. According to the good diffusion results, AgNPs produced by S. aureus resulted in the larger zone of inhibition (ZOI) against the selected pathogens, while AgNPs produced by E. coli showed comparatively smaller ZOI. In general, biosynthesized AgNPs were highly effective against gram-negative bacteria compared to gram-positive bacterial and fungal species. Conclusions: Green AgNPs produced by each bacterium show antimicrobial activity against the selected pathogens. AgNPs produced by S. aureus are the most effective NPs among tested AgNPs, while AgNPs produced by E. coli are the least effective. Further characterization of NPs is required to study the physical properties of silver NPs.

Keywords: green nanotechnology, silver nanoparticles, bacteria, antimicrobial activity

Procedia PDF Downloads 185

Authors: A. A. Dehpour, B. Eslami, S. Rezaie, S. F. Hashemian, F. Shafie, M. Kiaie

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The aims of study were investigation on chemical composition essential oil and the effect of extract of Coronilla varia on antimicrobial and cytotoxicity activity. The essential oils of Coronilla varia is obtained by hydrodistillation and analyzed by (GC/MS) for determining their chemical composition and identification of their components. Antibacterial activity of plant extract was determined by disc diffusion method. The effect of hydroalcolic extracts from Cornilla varia investigated on MCF7 cancer cell line by MTT assay. The major components were Caryophyllene Oxide (60.19%), Alphacadinol (4.13%) and Homoadantaneca Robexylic Acid (3.31%). The extracts from Coronilla varia had interesting activity against Proteus mirabilis in the concentration of 700 µg/disc and did not show any activity against Staphylococus aureus, Bacillus subtillis, Klebsiella pneumonia and Entrobacter cloacae. The positive control, Ampicillin, Chloramphenicol and Cenphalothin had shown zone of inhibition resistant all bacteria. Corohilla varia ethanol extract could inhibit the proliferation of MCF7 cell line in RPMI 1640 medium. IC50 5(mg/ml) was the optimum concentration of extract from Coronilla varia inhibition of cell line growth. The MCF7 cancer cell line and Proteus mirabilis were more sensitive to Coronilla varia ethanol extract.

Keywords: Coronilla varia, essential oil, antibacterial, anticancer, hela cell line

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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: Wilailak Siripornadulsil, Siriyanapat Tasaku, Jutamas Buahorm, Surasak Siripornadulsil

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The objectives of this study were to isolate LAB from various sources, dietary supplement, Thai traditional fermented food, and freshwater fish and to characterize their potential as probiotic cultures. Out of 1,558 isolates, 730 were identified as LAB based on isolation on MRS agar supplemented with a bromocresol purple indicator and CaCO3 and gram-positive, catalase and oxidase negative characteristics. Eight isolates showed the potential probiotic properties including tolerance to acid, bile salt and heat, proteolytic, amylolytic and lipolytic activities and oxalate-degrading capability. They all showed the antimicrobial activity against some Gram-negative and Gram-positive pathogenic bacteria. Based on 16S rDNA sequence analysis, they were identified as Enterococcus faecalis BT2 and MG30, Leconostoc mesenteroides SW64 and Pediococcus pentosaceous BD33, CF32, NP6, PS34 and SW5. The health beneficial effects and food safety will be further investigated and developed as a probiotic or protective culture used in Nile tilapia belly flap meat fermentation.

Keywords: probiotic, lactic acid bacteria, pathogen, protective culture

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Authors: Karthick Gopalan, Selvamohan Thankiah

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Biosurfactants are lipid compounds produced by microbes, which are amphipathic molecules consisting of hydrophophic and hydrophilic domains. In the present investigation, ten bacterial strains were isolated from petroleum oil contaminated sites near petrol bunk. Oil collapsing test, haemolytic activity were used as a criteria for primary isolation of biosurfactant producing bacteria. In this study, all the bacterial strains gave positive results. Among the ten strains, two were observed as good biosurfactant producers, they utilize the diesel as a sole carbon source. Optimization of biosurfactant producing bacteria isolated from petroleum oil contaminated sites was carried out using different parameters such as, temperature (20ºC, 25ºC, 30ºC, 37ºC and 45ºC), pH (5,6,7,8 & 9) and nitrogen sources (ammonium chloride, ammonium carbonate and sodium nitrate). Biosurfactants produced by bacteria were extracted, dried and quantified. As a result of optimization of parameters the suitable values for the production of more amount of biosurfactant by the isolated bacterial species was observed as 30ºC (0.543 gm/lt) in the pH 7 (0.537 gm/lt) with ammonium nitrate (0.431 gm/lt) as sole carbon source.

Keywords: isolation and identification, biosurfactant, microorganism, bioaugmentation

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Authors: Arfan Ali, Idrees Ahmad Nasir

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The critical evaluation of tolerance in tomato plants against the induced saline soil were assessed by transcript analysis of genes coding for products potentially involved in stress tolerance. A reverse transcriptase PCR experiment was performed with Hsp90-1, MT2, and GR1like protein genes using RNA isolated from different tissues of tomato plants. Four strains of Bacillus magisterium were inoculated with 100 Mm & 200 Mm concentrations of salt. Eleven treatments each ten replica pots were installed in green house experiment and the parameters taken into account were morphological (length, weight, number of leaves, leaf surface area), chemical (anthocyanin, chlorophyll-a, chlorophyll-b, carotenoids) and biological (gene expression). Results bare a response i.e. highest response of MT2 like gene was at 24 hpi and the highest levels of GR1 like protein transcript accumulation were detected at 36 hpi. The chemical and morphological parameters at diverse salt concentrations bequeath superlative response amongst strains which candidly flank on Zm7 and Zm4. Therefore, Bacillus magisterium Zm7 strains and somehow Zm4 strain can be used in saline condition to make plants tolerant. The overall performance of strains Zm7, Zm6, and Zm4 was found better for all studied traits under salt stress conditions. Significant correlations among traits root length, shoot length, number of leaves, leaf surface area, carotenoids, anthocyanin, chlorophyll-a and chlorophyll-b were found and suggested that the salt tolerance in tomato may be improved through the use of PGPR strains.

Keywords: Bacillus magisterium, gene expression glutathione reductase, metallothionein, PGPR, Rhizobacteria, saline

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Authors: Yong Geun Yun, Jong Hui kim, Sang Ho Baik

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This study was carried out to develop a fermented dandelion (Tarazacum officinale) beverage using lactic acid bacteria with cinnamoyl esterase (CE) activity isolated from human feces. Lactic acid bacteria were screened based on bacterial survival ability in dandelion extract and CE activity. Dandelion extract fermented by Lactobacillus acidophilus F-46 (LA-F46) maintained approximately 105-106 log CFU/mL over an 8 days period. After fermented dandelion beverage (FDB) with LA-46 for 8 days at 37oC the pH was decreased from pH 7.0 to 3.5. Antioxidant activity by using DPPH radical scavenging activity of the prepared FDB was significantly increased compared to that of non-fermented dandelion beverage (NFDB). Moreover, CE activity was significantly enhanced during fermentation and showed the approximately 4.3 times increased concentration of caffeic acid up to 9.91 mg/100 mL after 8 days of incubation compared to NFDB. Therefore, it concluded that dandelion can be a good source for preparing a functional beverage and fermentation by LA-F46 enhanced the food functionality with enhanced caffeic acids.

Keywords: cinnamoyl esterase, dandelion, fermented beverage, lactic acid bacteria

Procedia PDF Downloads 380

Authors: Eze Catherine Chinwe, J. D. Njoku

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Background: Palm oil mill effluent is the voluminous liquid waste that comes from the sterilization and clarification sections of the oil palm milling process. The raw effluent contains 90-95% water and includes residual oil, soil particles, and suspended solids. Palm oil mill effluent is a highly polluting material and much research has been dedicated to means of alleviating its threat to the environment. Objectives: 1. To compare Physico-chemical and microbiological analysis of soil samples from POME and non-POME sites. 2. To make recommendations on how best to handle POME in the study area. Methods: Quadrant approach was adopted for sampling POME (A) and Non POME (B) locations. Qualities were determined using standard analytical procedures. Conclusions: Results of the analysis were obtained in the following range; pH (3.940 –7.435), dissolved oxygen (DO) (1.582–6.234mg/l), biological oxygen demand (BOD) (50–5463mg/l etc. For the various locations, the population of total heterotrophic bacteria (THB) ranged from 1.36x106–2.42x106 cfu/ml, the total heterotrophic fungi (THF) ranged from 1.22–3.05 x 104 cfu/ml. The frequency of occurrence revealed the microbial isolates Pseudomonas sp., Bacillus sp., Staphylococcus, as the most frequently occurring isolates. Analysis of variance showed that there were significant differences (P<0.05) in microbial populations among locations. The discharge of industrial effluents into the soil in Nigeria invariably results in the presence of high concentrations of pollutant in the soil environment.

Keywords: effluents, mirobial composition, soil samples, isiala mbano

Procedia PDF Downloads 287

Authors: A. T. Laiche, M. L. Tlil, Benine B., S. Bechoua

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The aim of this study is to isolate and select, from camel milk from El-Oued region in Algeria, a strains of lactic acid bacteria and possessing probiotic properties ; and to evaluate their potential effect on intestinal disorders in Wistar ratsmThe results relating to the selection of probiotic strains confirms that the Thermophilic streptococcus exhibits the best probiotic activity performance, with a resistance important to different degrees of pH and to bile salts, and a remarkable antibacterial activity and resistance to antibiotics compared to the other four isolated strains. In the in vivo study, diseases are induced in rats at the level of the digestive system, it was reported that the administration of Escherichia coli and castor oil caused an intestinal disorders. The microscopic observation of the histological section of the intestine showed a damaged intestinal structure and some symptoms of its irritation, including a decrease in the height of the villi and the presence of others destroyed cells, and after treatment with Streptococcus thermophilus, the microscopic observation of the cut of the histological section of the intestine showed almost complete disappearance of the mentioned symptoms, The dosage of the hematological parameters by complete blood count (CBC) is in agreement with the results of the histological sections.

Keywords: camel milk, probiotic, pathogenic bacteria, intestinal disorders, lactic acid bacteria

Procedia PDF Downloads 135

Authors: Nesrine Lenchi, Salima Kebbouche-Gana, Laddada Belaid, Mohamed Lamine Khelfaoui, Mohamed Lamine Gana

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Saline lakes are extreme hypersaline environments that are considered five to ten times saltier than seawater (150 – 300 g L-1 salt concentration). Hypersaline regions differ from each other in terms of salt concentration, chemical composition and geographical location, which determine the nature of inhabitant microorganisms. In order to explore the diversity of moderate and extreme halophiles Bacteria in Chott Tinsilt (East of Algeria), an isolation program was performed. In the first time, water samples were collected from the saltern during pre-salt harvesting phase. Salinity, pH and temperature of the sampling site were determined in situ. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions. Isolates were obtained by plating out the samples in complex and synthetic media. In this study, seven halophiles cultures of Bacteria were isolated. Isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (oxidase, catalase, nitrate reductase and urease), and optimization of growth conditions were done. The results indicated that the salinity optima varied from 50 to 250 g L-1, whereas the optimum of temperature range from 25°C to 35°C. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. The results showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Idiomarina, Halobacillus Thalassobacillus and Planococcus genera and what may represent a new bacterial genus.

Keywords: bacteria, Chott, halophilic, 16S rRNA

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Authors: Zahid Rehman, TorOve Leiknes

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Quorum sensing (QS) is the process by which bacteria communicate with each other through small signaling molecules, such as N-acylhomoserine lactones (AHLs). Also, certain bacteria have the ability to degrade AHL molecules by a process referred to as quorum quenching (QQ); therefore, QQ can be used to control bacterial infections and biofilm formation. In this study, we aimed to identify new species of bacteria with QQ activities. To achieve this, sediments from Red Sea were collected either in the close vicinity of Sea grass or from area with no vegetation. From these samples, we isolated 72 bacterial strains and tested their ability to degrade/inactivate AHL molecules. Chromobacterium violaceum based bioassay was used in initial screening of isolates for QQ activity. The QQ activity of the positive isolates was further confirmed and quantified by employing liquid chromatography and mass spectrometry. These analyses showed that isolated bacterial strain could degrade AHL molecules with different acyl chain length and modifications. Sequencing of 16S-rRNA genes of positive isolates revealed that they belong to three different genera. Specifically, two isolates belong to genus Erythrobacter, four to Labrenzia and one isolate belongs to Bacterioplanes. Time course experiment showed that isolate belonging to genus Erythrobacter could degrade AHLs faster than other isolates. Furthermore, these isolates were tested for their ability to inhibit formation of biofilm and degradation of 3OXO-C12 AHLs produced by P. aeruginosa PAO1. Our results showed that isolate VG12 is better at controlling biofilm formation. This aligns with the ability of VG12 to cause at least 10-fold reduction in the amount of different AHLs tested.

Keywords: quorum sensing, biofilm, quorum quenching, anti-biofouling

Procedia PDF Downloads 146

Authors: S. Alqarni, J. C. McCann, A. Palladino, J. J. Loor

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Seven fistulated Holstein cows were used from 3 weeks prepartum to 4 weeks postpartum to determine the relative abundance of 7 different species of ruminal microorganisms. The prepartum diet was based on corn silage. In the postpartum, diet included ground corn, grain by-products, and alfalfa haylage. Ruminal digesta were collected at five times: -14, -7, 10, 20, and 28 days around parturition. Total DNA from ruminal digesta was isolated and real-time quantitative PCR was used to determine the relative abundance of bacterial species. Eubacterium ruminantium and Selenomonas ruminantium were not affected by time (P>0.05). Megasphaera elsdenii and Prevotella bryantii increased significantly postpartum (P<0.001). Conversely, Butyrivibrio proteoclasticus decreased gradually from -14 through 28 days (P<0.001). Fibrobacter succinogenes was affected by time being lowest at day 10 (P=0.02) while Anaerovibrio lipolytica recorded the lowest abundance at -7 d followed by an increase by 20 days postpartum (P<0.001). Overall, these results indicate that changes in diet after parturition affect the abundance of ruminal bacteria, particularly M. elsdenii (a lactate-utilizing bacteria) and P. bryantii (a starch-degrading bacteria) which increased markedly after parturition likely as a consequence of a higher concentrate intake.

Keywords: rumen bacteria, transition cows, rumen metabolism, peripartal period

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Authors: Khadijeh Abhari, Seyed Shahram Shekarforoush, Saeid Hosseinzadeh

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Probiotics have been considered as an approach to treat and prevent a wide range of inflammatory diseases. The spore forming probiotic strain Bacillus coagulans has demonstrated anti-inflammatory and immune-modulating effects in both animals and humans. The prebiotic, inulin, also potentially affects the immune system as a result of the change in the composition or fermentation profile of the gastrointestinal microbiota. An in vivo trial was conducted to evaluate the effects of probiotic B. coagulans, and inulin, either separately or in combination, on down regulate immune responses and progression of rheumatoid arthritis using induced arthritis rat model. Forty-eight male Wistar rats were randomly divided into 6 groups and fed as follow: 1) control: Normal healthy rats fed by standard diet, 2) Disease control (RA): Arthritic induced (RA) rats fed by standard diet, 3) Prebiotic (PRE): RA+ 5% w/w long chain inulin, 4) Probiotic (PRO): RA+ 109 spores/day B. coagulans by orogastric gavage, 5) Synbiotic (SYN): RA+ 5% w/w long chain inulin and 109 spores/day B. coagulans and 6) Treatment control: (INDO): RA+ 3 mg/kg/day indomethacin by orogastric gavage. Feeding with mentioned diets started on day 0 and continued to the end of study. On day 14, rats were injected with complete Freund’s adjuvant (CFA) to induce arthritis. Arthritis activity was evaluated by biochemical parameters and paw thickness. Biochemical assay for Fibrinogen (Fn), Serum Amyloid A (SAA), TNF-α and Alpha-1-acid glycoprotein (α1AGp) was performed on day 21, 28 and 35 (1, 2 and 3 weeks post RA induction). Pretreatment with PRE, PRO and SYN diets significantly inhibit SAA and Fn production in arthritic rats (P < 0.001). A significant decrease in production of pro-inflammatory cytokines, TNF-α, was seen in PRE, PRO and SYN groups (P < 0.001) which was similar to the effect of the anti-inflammatory drug Indomethacin. Further, there were no significant anti-inflammatory effects observed following different treatments using α1AGp as a RA indicator. Pretreatment with all supplied diets significantly inhibited the development of paw swelling induced by CFA (P < 0.001). Conclusion: Results of this study support that oral intake of probiotic B. coagulans and inulin are able to improve biochemical and clinical parameters of induced RA in rat.

Keywords: rheumatoid arthritis, bacillus coagulans, inulin, animal model

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Authors: Teodora Mocan, Lucian Mocan, Cornel Iancu, Flaviu A. Tabaran, Bartos Dana, Matea Cristian

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Antimicrobial resistance, which threatens the efficacy of the existing antibiotics represents a worldwide public health issue. At the current time, vancomycin is the only responsive treatment although has significant cytotoxicity, is partially effective and it is poorly retained by infected tissues. From a clinical point of view, attractive alternative approaches for treating such Meticillin-Resistant Staphylococcus Aureus (MRSA) strains would be using agents that cause physical damage to the bacteria. Modular nanopharmaceuticals systems are being designed to address all of these multifunctional capabilities for the ideal bacterial treatment, with the ability to mix and match appropriate functions. Here we present a novel method of selective laser photothermal ablation of MRSA bacteria mediated by gold nanoparticles bound to PBP antibody against PBP protein located on the MRSA surface.

Keywords: MRSA, laser, nanoparticle, antibody

Procedia PDF Downloads 247

Authors: Vita Lele, Vadims Bartkevics, Iveta Pugajeva, Paulina Zavistanaviciute, Daiva Zadeike, Grazina Juodeikiene, Elena Bartkiene

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Different combinations of appropriate technological properties showing lactic acid bacteria (Pediococcus pentosaceus VLGL183 and Enterococcus pseudoavium VLGL 234, Lactobacillus plantarum VLGL135 and Pediococcus pentosaceus VLGL183, Pediococcus pentosaceus VLGL183 and Lactobacillus brevis VLGL173, Pediococcus pentosaceus VLGL183 and Leuconostoc mesenteroides VLGL242, Pediococcus pentosaceus VLGL183 and Lactobacillus curvatus VLGL51, Lactobacillus plantarum VLGL135 and Lactobacillus curvatus VLGL51) for wheat sourdough production were used, and the influence of different sourdoughs on wheat bread quality parameters was evaluated. The highest overall acceptability (135.8 mm in 140 mm hedonic scale) of the bread produced with L. plantarum VLGL135 and P. pentosaceus VLGL183 sourdough was established. Also, bread produced with above mention sourdough, has the highest specific volume, shape coefficient, moisture content, and porosity, 3.40 ml /g; 2.59, 33.7 %, and 76.6 %, respectively. It was found, that the used sourdoughs reduce acrylamide content in bread (from 29.5 to 67.2%), just, the isolated lactic acid bacteria strains could be recommended for higher quality and safer bread production.

Keywords: acrylamide, lactic acid bacteria, quality, sourdough, wheat bread

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Authors: Munir Ashraf, Shagufta Riaz

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Personal protective equipment (PPE) and bioactive textiles are highly important for the health care of front line hospital workers, patients, and the general population to be safe from highly infectious diseases. This was even more critical in the wake of COVID-19 outbreak. Most of the medical textiles are inactive against various viruses and bacteria, hence there is a need to wash them frequently to avoid the spread of microorganisms. According to survey conducted by the world health organization, more than 500 million people get infected from hospitals, and more than 13 million died due to these hospitals’ acquired deadly diseases. The market available PPE are though effective against the penetration of pathogens and to kill bacteria but, they are not breathable and active against different viruses. Therefore, there was a great need to develop textiles that are not only effective against bacteria, fungi, and viruses but also are comfortable to the medical personnel and patients. In the present study, waterproof breathable, and biologically active textiles were developed using antiviral and antibacterial nanomaterials. These nanomaterials like TiO₂, ZnO, Cu, and Ag were immobilized at the surface of cotton fabric by using different silane coupling agents and electroless deposition that they retained their functionality even after 30 industrial laundering cycles. Afterwards, the treated fabrics were coated with a waterproof breathable film to prevent the permeation of liquid droplets, any particle or microorganisms greater than 80 nm. The developed cotton fabric was highly active against bacteria and viruses. The good durability of nanomaterials at the cotton surface after several industrial washing cycles makes this fabric an ideal candidate for bioactive textiles used in the medical field.

Keywords: antibacterial, antiviral, cotton, durable

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Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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Authors: Debamitra Chakravorty, Pratap K. Parida

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Cold-adapted proteases enhance wash performance in low-temperature laundry resulting in a reduction in energy consumption and wear of textiles and are also used in the dehairing process in leather industries. Unfortunately, the possible drawbacks of using cold-adapted proteases are their instability at higher temperatures. Therefore, proteases with broad temperature stability are required. Unfortunately, wild-type cold-adapted proteases exhibit instability at higher temperatures and thus have low shelf lives. Therefore, attempts to engineer cold-adapted proteases by protein engineering were made previously by directed evolution and random mutagenesis. The lacuna is the time, capital, and labour involved to obtain these variants are very demanding and challenging. Therefore, rational engineering for cold stability without compromising an enzyme's optimum pH and temperature for activity is the current requirement. In this work, mutations were rationally designed with the aid of high throughput computational methodology of network analysis, evolutionary conservation scores, and molecular dynamics simulations for Savinase from Bacillus lentus with the intention of rendering the mutants cold stable without affecting their temperature and pH optimum for activity. Further, an attempt was made to incorporate a mutation in the most stable mutant rationally obtained by this method to introduce oxidative stability in the mutant. Such enzymes are desired in detergents with bleaching agents. In silico analysis by performing 300 ns molecular dynamics simulations at 5 different temperatures revealed that these three mutants were found to be better in cold stability compared to the wild type Savinase from Bacillus lentus. Conclusively, this work shows that cold adaptation without losing optimum temperature and pH stability and additionally stability from oxidative damage can be rationally designed by in silico enzyme engineering. The key findings of this work were first, the in silico data of H5 (cold stable savinase) used as a control in this work, corroborated with its reported wet lab temperature stability data. Secondly, three cold stable mutants of Savinase from Bacillus lentus were rationally identified. Lastly, a mutation which will stabilize savinase against oxidative damage was additionally identified.

Keywords: cold stability, molecular dynamics simulations, protein engineering, rational design

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Authors: Anza-Vhudziki Mboyi, Ilunga Kamika, Maggy Momba

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The aim of this study was to ascertain the survival limit and capability of commonly found wastewater protozoan (Aspidisca sp, Trachelophyllum sp, and Peranema sp) and bacterial (Bacillus licheniformis, Brevibacillus laterosporus, and Pseudomonas putida) species to remove COD while exposed to commercial nanomaterials under varying pH conditions. The experimental study was carried out in modified mixed liquor media adjusted to various pH levels (pH 2, 7 and 10), and a comparative study was performed to determine the difference between the cytotoxicity effects of commercial zinc oxide (nZnO) and silver (nAg) nanomaterials (NMs) on the target wastewater microbial communities using standard methods. The selected microbial communities were exposed to lethal concentrations ranging from 0.015 g/L to 40 g/L for nZnO and from 0.015 g/L to 2 g/L for nAg for a period of 5 days of incubation at 30°C (100 r/min). Compared with the absence of NMs in wastewater mixed liquor, the relevant environmental concentration ranging between 10 µg/L and 100 µg/L, for both nZnO and nAg caused no adverse effects, but the presence of 20 g of nZnO/L and 0.65 g of nAg/L significantly inhibited microbial growth. Statistical evidence showed that nAg was significantly more toxic compared to nZnO, but there was an insignificant difference in toxicity between microbial communities and pH variations. A significant decrease in the removal of COD by microbial populations was observed in the presence of NMs with a moderate correlation of r = 0.3 to r = 0.7 at all pH levels. It was evident that there was a physical interaction between commercial NMs and target wastewater microbial communities; although not quantitatively assessed, cell morphology and cell death were observed. Such phenomena suggest the high resilience of the microbial community, but it is the accumulation of NMs that will have adverse effects on the performance in terms of COD removal.

Keywords: bacteria, biological treatment, chemical oxygen demand (COD) and nanomaterials, consortium, pH, protozoan

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Authors: Francis Gyapong, Denis Yar

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The use of cell phones has become an indispensable tool in the hospital's settings. Cell phones are used in hospitals without restrictions regardless of their unknown microbial load. However, the indiscriminate use of mobile devices, especially at health facilities, can act as a vehicle for transmitting pathogenic bacteria and other microorganisms. These potential pathogens become exogenous sources of infection for the patients and are also a potential health hazard for self and as well as family members. These are a growing problem in many health care institutions. Innovations in mobile communication have led to better patient care in diabetes, asthma, and increased in vaccine uptake via SMS. Notwithstanding, the use of cell phones can be a great potential source for nosocomial infections. Many studies reported heavy microbial contamination of cell phones among healthcare workers and communities. However, limited studies have been reported in our region on bacterial contamination on cell phones among healthcare workers. This study assessed microbial contamination of cell phones of health care workers (HCWs) at the Mampong Municipal Government Hospital (MMGH), Ghana. A cross-sectional design was used to characterize bacterial microflora on cell phones of HCWs at the MMGH. A total of thirty-five (35) swab samples of cell phones of HCWs at the Laboratory, Dental Unit, Children’s Ward, Theater and Male ward were randomly collected for laboratory examinations. A suspension of the swab samples was each streak on blood and MacConkey agar and incubated at 37℃ for 48 hours. Bacterial isolates were identified using appropriate laboratory and biochemical tests. Kirby-Bauer disc diffusion method was used to determine the antimicrobial sensitivity tests of the isolates. Data analysis was performed using SPSS version 16. All mobile phones sampled were contaminated with one or more bacterial isolates. Cell phones from the Male ward, Dental Unit, Laboratory, Theatre and Children’s ward had at least three different bacterial isolates; 85.7%, 71.4%, 57.1% and 28.6% for both Theater and Children’s ward respectively. Bacterial contaminants identified were Staphylococcus epidermidis (37%), Staphylococcus aureus (26%), E. coli (20%), Bacillus spp. (11%) and Klebsiella spp. (6 %). Except for the Children ward, E. coli was isolated at all study sites and predominant (42.9%) at the Dental Unit while Klebsiella spp. (28.6%) was only isolated at the Children’s ward. Antibiotic sensitivity testing of Staphylococcus aureus indicated that they were highly sensitive to cephalexin (89%) tetracycline (80%), gentamycin (75%), lincomycin (70%), ciprofloxacin (67%) and highly resistant to ampicillin (75%). Some of these bacteria isolated are potential pathogens and their presence on cell phones of HCWs could be transmitted to patients and their families. Hence strict hand washing before and after every contact with patient and phone be enforced to reduce the risk of nosocomial infections.

Keywords: mobile phones, bacterial contamination, patients, MMGH

Procedia PDF Downloads 75

Authors: Abdurrahim A. Elouzi, Abdurrauf M. Gusbi, Ali M. Elgerbi

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Antibiotic resistance has become a global public-health problem, thus it is imperative that new antibiotics continue to be developed. Major problems are being experienced in human medicine from antibiotic resistant bacteria. Moreover, no new chemical class of antibiotics has been introduced into medicine in the past two decades. The aim of the current study presents experimental results that evaluate the capability of bio surfactant rhamnolipid on enhancing the efficacy of hydrophobic antibiotics. Serial dilutions of azithromycin and clarithromycin were prepared. A bacterial suspension (approximately 5 X 105 CFU) from an overnight culture in MSM was inoculated into 20 ml sterile test tube each containing a serial 10-fold dilution of the test antibiotic(s) in broth with or without 200 mgL-1 rhamnolipid. The tubes were incubated for 24 h with vigorous shaking at 37°C. Antimicrobial activity in multiple antibiotic-resistant gram-negative bacteria pathogens and gram-positive bacteria were assessed using optical density technique. The results clearly demonstrated that the presence of rhamnolipid significantly improved the efficiency of both antibiotics. We hypothesized that the addition of rhamnolipid at low concentration, causes release of LPS which results in an increase in cell surface hydrophobicity. This allows increased association of cells with hydrophobic antibiotics resulting in increased cytotoxicity rates.

Keywords: hydrophobic antibiotics, biosurfactant, rhamnolipid, azithromycin, clarithromycin

Procedia PDF Downloads 493

Authors: H. K. Ratre, M. Roy, S. Roy, M. S. Parmar, V. Bhagat

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Mastitis is a common problem of dairy industries. Reduction in milk production and an irreparable damage to the udder associated with the disease are common causes of culling of dairy cows. Milk from infected animals is not suitable for drinking and for making different milk products. So, it has a major economic importance in dairy cattle. The aims of this study were to investigate the bacteriological panorama in milk from udder quarters with subclinical mastitis and to carried out for the molecular characterization of the major isolated organisms, from subclinical mastitis-affected cows in and around Durg and Rajnandgaon district of Chhattisgarh. Isolation and identification of bacteria from the milk samples of subclinical mastitis-affected cows were done by standard and routine culture procedures. A total of 78 isolates were obtained from cows and among the various bacteria isolated, Staphylococcus spp. occupied prime position with occurrence rate of 51.282%. However, other bacteria isolated includeStreptococcus spp. (20.512%), Micrococcus spp. (14.102%), E. coli (8.974%), Klebsiela spp. (2.564%), Salmonella spp. (1.282%) and Proteus spp. (1.282%). Staphylococcus spp. was isolated as the major causative agent of subclinical mastitis in the studied area. Molecular characterization of Staphylococus aureusisolates was done for genetic expression of the virulence genes like ‘nuc’ encoding thermonucleaseexoenzyme, coa and spa by PCR amplification of the respective genes in 25 Staphylococcus isolates. In the present study, 15 isolates (77.27%) out of 20 coagulase positive isolates were found to be genotypically positive for ‘nuc’ where as 20 isolates (52.63%) out of 38 CNS expressed the presence of the same virulence gene. In the present study, three Staphylococcus isolates were found to be genotypically positive for coa gene. The Amplification of the coa gene yielded two different products of 627, 710 bp. The amplification of the gene segment encoding the IgG binding region of protein A (spa) revealed a size of 220 and 253bp in twostaphylococcus isolates. The X-region binding of the spa gene produced an amplicon of 315 bp in one Staphylococcal isolates. Staphylococcus aureus was found to be major isolate (51.28%) responsible for causing subclinical mastitis in cows which also showed expression of virulence genesnuc, coa and spa.

Keywords: mastitis, bacteria, characterization, expression, gene

Procedia PDF Downloads 193

Authors: Aliasghar Golmohammadian, Sona Rostampour Yasouri

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One of the most important intestinal pathogenic strains is E. coli O₁₅₇:H₇. This pathogenic bacterium is transmitted to humans through water and food. E. coli O₁₅₇:H₇ is the main cause of Hemorrhagic colitis (HC), Hemolytic Uremic Syndrome (HUS), Thrombotic Thrombocytopenic Purpura (TTP) and in some cases death. Since E. coli O₁₅₇:H₇ can be transmitted through the consumption of different foods, including vegetables, agricultural products, and fresh dairy products, this study aims to identify and isolate E. coli O₁₅₇:H₇ from wastewater by PCR technique. One hundred twenty samples of water and wastewater were collected by Falcom Sterile from Shahrood and Neka cities. The samples were checked for colony formation after appropriate centrifugation and cultivation in the specific medium of Sorbitol MacConkey Agar (SMAC) and other diagnostic media of E. coli O₁₅₇:H₇. Also, the plates were observed macroscopically and microscopically. Then, the necessary phenotypic tests were performed on the colonies, and finally, after DNA extraction, the PCR technique was performed with specific primers related to rfbE and stx2 genes. The number of 5 samples (6%) out of all the samples examined were determined positive by PCR technique with observing the bands related to the mentioned genes on the agarose gel electrophoresis. PCR is a fast and accurate method to identify the bacteria E. coli O₁₅₇:H₇. Considering that E. coli bacteria is a resistant bacteria and survives in water and food for weeks and months, the PCR technique can provide the possibility of quick detection of contaminated water. Moreover, it helps people in the community control and prevent the transfer of bacteria to healthy and underground water and agricultural and even dairy products.

Keywords: E. coli O₁₅₇:H₇, PCR, water, wastewater

Procedia PDF Downloads 39

Authors: S. N. Al-Bahry, S. K. Al-Musharafi, I. Y. Mahmoud

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Recycled treated sewage effluents (TSE) is used for agriculture, Public park irrigation and industrial purposes. TSE was found to play a major role in the distribution of antibiotic resistant bacteria into the environment. Fecal coliform and enterococci counts were significantly higher during summer compared to winter seasons. Oman has low annual rainfall with annual average temperature varied between 15-45oC. The main source of potable water is from seawater desalination. Resistance of the isolates to 10 antibiotics (Amikacin, Ampicillin, chloramphenicol, gentamycine, minocylin, nalidixicacid, neomycin, streptomycin, Tetracycline, Tobramycin, and Trimethoprim) was tested. Both fecal coliforms and enterococci were multiple resistant to 2-10 antibiotics. However, temperature variation during summer and winter did not affect resistance of the isolates to antibiotics. The significance of this investigation may be indicator to the environmental TSE pollution.

Keywords: antibiotic resistance, bacteria, environment, sewage treated effluent

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Authors: Rashad R. Al-Hindi

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The aim of this study was to isolate and identify lactic acid bacteria and evaluate their therapeutic and food preservation importance. Ninety-three suspected lactic acid bacteria (LAB) were isolated from thirteen different raw and fermented milk of indigenous sources in the Kingdom of Saudi Arabia. The identification of forty-six selected LAB strains and genetic relatedness were performed based on 16S rDNA gene sequence comparison. The LAB counts in certain samples were higher under microaerobic than anaerobic conditions. The identified LAB belonged to genera Enterococcus (16 strains), Lactobacillus (9 strains), Weissella (10 strains), Streptococcus (8 strains) and Lactococcus (3 strains). Phylogenetic tree generated from the full-length (~1.6 kb) sequences confirmed previous findings. Utilization of shorter 16S rDNA sequences (~1.0 kb) also discriminated among strains of which V2 region was the most effective. None of the strains exhibited resistance to clinically relevant antibiotics or undesirable hemolytic activity, while they differed in other probiotic characteristics, e.g., tolerance to acidic pH, resistance to bile salt, and antibacterial activity. In conclusion, the isolates Lactobacillus casei MSJ1, Lactobacillus casei Dwan5, Lactobacillus plantarum EyLan2 and Enterococcus faecium Gail-BawZir8 are likely the best probiotic LAB and we speculate that studying the synergistic effects of bacterial combinations might result in the occurrence of more effective probiotic potential. We argue that the raw and fermented milk of animals hosted in Saudi Arabia, especially stirred yogurt (Laban) made from camel milk, are rich in LAB with promising probiotics potential.

Keywords: fermented foods, lactic acid bacteria, probiotics, Saudi Arabia

Procedia PDF Downloads 174

Authors: G. Amariei, K. Boltes, R. Rosal, P. Leton

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Wastewater treatment plants (WWTPs) are supposed to hold an important place in the reduction of emerging contaminants, but provide an environment that has potential for the development and/or spread of adaptation, as bacteria are continuously mixed with contaminants at sub-inhibitory concentrations. Reviewing the literature, there are little data available regarding the use of adapted bacteria forming activated sludge community for toxicity assessment, and only individual validations have been performed. Therefore, the aim of this work was to study the toxicity of Triclosan (TCS) and Ibuprofen (IBU), individually and in binary combination, on adapted activated sludge (AS). For this purpose a battery of biomarkers were assessed, involving oxidative stress and cytotoxicity responses: glutation-S-transferase (GST), catalase (CAT) and viable cells with FDA. In addition, we compared the toxic effects on adapted bacteria with unadapted bacteria, from a previous research. Adapted AS comes from three continuous-flow AS laboratory systems; two systems received IBU and TCS, individually; while the other received the binary combination, for 14 days. After adaptation, each bacterial culture condition was exposure to IBU, TCS and the combination, at 12 h. The concentration of IBU and TCS ranged 0.5-4mg/L and 0.012-0.1 mg/L, respectively. Batch toxicity experiments were performed using Oxygraph system (Hansatech), for determining the activity of CAT enzyme based on the quantification of oxygen production rate. Fluorimetric technique was applied as well, using a Fluoroskan Ascent Fl (Thermo) for determining the activity of GST enzyme, using monochlorobimane-GSH as substrate, and to the estimation of viable cell of the sludge, by fluorescence staining using Fluorescein Diacetate (FDA). For IBU adapted sludge, CAT activity it was increased at low concentration of IBU, TCS and mixture. However, increasing the concentration the behavior was different: while IBU tends to stabilize the CAT activity, TCS and the mixture decreased this one. GST activity was significantly increased by TCS and mixture. For IBU, no variations it was observed. For TCS adapted sludge, no significant variations on CAT activity it was observed. GST activity it was significant decreased for all contaminants. For mixture adapted sludge the behaviour of CAT activity it was similar to IBU adapted sludge. GST activity it was decreased at all concentration of IBU. While the presence of TCS and mixture, respectively, increased the GST activity. These findings were consistent with the viability cells evaluation, which clearly showed a variation of sludge viability. Our results suggest that, compared with unadapted bacteria, the adapted bacteria conditions plays a relevant role in the toxicity behaviour towards activated sludge communities.

Keywords: adapted sludge community, mixture, PPCPs, toxicity

Procedia PDF Downloads 382

Authors: K. Abdul Halim, E. L. Yong

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Palm oil industry is a major revenue earner in Malaysia, despite the growth of the industry is synonymous with a massive production of agro-industrial wastewater. Through the oil extraction processes, palm oil mill effluent (POME) contributes to the largest liquid wastes generated. Due to the high amount of organic compound, POME can cause inland water pollution if discharged untreated into the water course as well as affect the aquatic ecosystem. For more than 20 years, Malaysia adopted the conventional biological treatment known as lagoon system that apply biological treatment. Besides having difficulties in complying with the standard, a large build up area is needed and retention time is higher. Although anaerobic digester is more favorable, this process comes along with enormous volumes of sludge and methane gas, demanding attention from the mill operators. In order to reduce the sludge production, denitrifiers are to be removed first. Sulfate reducing bacteria has shown the capability to inhibit the growth of methanogens. This is expected to substantially reduce both the sludge and methane production in anaerobic digesters. In this paper, the effectiveness of sulfate reducing bacteria in minimizing sludge and methane will be examined.

Keywords: methane reduction, palm oil mill effluent, sludge minimization, sulfate reducing bacteria, sulfate reduction

Procedia PDF Downloads 412

Authors: Ergun Sakalar, Kubra Bilgic

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Faster detection and characterization of pathogens are the basis of the evoid from foodborne pathogens. Salmonella spp., Shigella spp. and Listeria monocytogenes are common foodborne bacteria that are among the most life-threatining. It is important to rapid and accurate detection of these pathogens to prevent food poisoning and outbreaks or to manage food chains. The present work promise to develop a sensitive, species specific and reliable PCR based detection system for simultaneous detection of Salmonella spp., Shigella spp. and Listeria monocytogenes. For this purpose, three genes were picked out, ompC for Salmonella spp., ipaH for Shigella spp. and hlyA for L. monocytogenes. After short pre-enrichment of milk was passed through a vacuum filter and bacterial DNA was exracted using commercially available kit GIDAGEN®(Turkey, İstanbul). Detection of amplicons was verified by examination of the melting temperature (Tm) that are 72° C, 78° C, 82° C for Salmonella spp., Shigella spp. and L. monocytogenes, respectively. The method specificity was checked against a group of bacteria strains, and also carried out sensitivity test resulting in under 10² CFU mL⁻¹ of milk for each bacteria strain. Our results show that the flourescence based triplex qPCR method can be used routinely to detect Salmonella spp., Shigella spp. and L. monocytogenes during the milk processing procedures in order to reduce cost, time of analysis and the risk of foodborne disease outbreaks.

Keywords: evagreen, food-born bacteria, pathogen detection, real-time pcr

Procedia PDF Downloads 220