Search results for: pathogenic Escherichia coli

Authors: T. S. Moe, T. A. Khaing

Abstract:

Chitosan is a derivative of chitin, a compound usually isolated from the shells of some crustaceans such as crab, lobster and shrimp. It has biocompatible, biodegradable, and antimicrobial properties. To use these properties of chitosan in biomedical fields, chitosan films (1%, 2%, 3% and 4%) were prepared by using l% lactic acid as solvent. The effects of chitosan films on tensile strength, elongation at break, degree of swelling, thickness, morphology, allergic and irritation reactions and antibacterial property were evaluated. Staphylococcus aureus and Escherichia coli were used as tested microorganisms. In vivo wound healing activities of chitosan films were investigated using mice model. As results, Chitosan films have similar appearance and good swelling properties and 4% chitosan film showed the better swelling activity and the greatest elongation ratio than the other chitosan films. They also showed their good activity of wound healing in mice model. Moreover, the results showed that the films did not produce any unwilling symptoms (allergy or irritation). In conclusion, it is evident that the chitosan film has the potentiality to use as wound healing biofilms in the biomedical fields.

Keywords: Chitosan, wound healing, antibacterial activity.

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Authors: N. Behidj, K. Benyounes, T. Dahmane, A. Allem

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The aim of this study was to investigate the antimicrobial activity of flavonoids isolated from the aerial part of a medicinal plant which is Thymus inodorusby the middle agar diffusion method on following microorganisms. We have Staphylococcus aureus, Escherichia coli, Pseudomonas fluorescens, AspergillusNiger, Aspergillus fumigatus and Candida albicans. During this study, flavonoids extracted by stripping with steam are performed. The yields of flavonoids is 7.242% for the aqueous extract and 28.86% for butanol extract, 29.875% for the extract of ethyl acetate and 22.9% for the extract of di - ethyl. The evaluation of the antibacterial effect shows that the diameter of the zone of inhibition varies from one microorganism to another. The operation values obtained show that the bacterial strain P fluoresces, and 3 yeasts and molds; A. Niger, A. fumigatus and C. albicansare the most resistant. But it is noted that, S. aureus is shown more sensitive to crude extracts, the stock solution and the various dilutions. Finally for the minimum inhibitory concentration is estimated only with the crude extract of Thymus inodorus flavonoid.Indeed, these extracts inhibit the growth of Gram + bacteria at a concentration varying between 0.5% and 1%. While for bacteria to Gram -, it is limited to a concentration of 0.5%.

Keywords: Antimicrobial activity, flavonoids, strains, Thymus inodorus.

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Authors: Kusumawadee Thancharoen, Rungrat Nontawong, Thanawat Junsom

Abstract:

Pathogenic bacterial flora was isolated from giant freshwater prawns, Macrobrachium rosenbergii. Infected shrimp samples were collected from BuaBan Aquafarm in Kalasin Province, Thailand, between June and September 2018. Bacterial species were isolated by serial dilution and plated on Thiosulfate Citrate Bile Salt Sucrose (TCBS) agar medium. A total 89 colonies were isolated and identified using the API 20E biochemical tests. Results showed the presence of genera Aeromonas, Citrobacter, Chromobacterium, Providencia, Pseudomonas, Stenotrophomonas and Vibrio. Maximum number of species was recorded in Pseudomonas (50.57%) with minimum observed in Chromobacterium and Providencia (1.12%).

Keywords: Biochemical test, giant freshwater prawn, isolation, salt tolerance, shrimp diseases.

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Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures.

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Authors: Ferial M. Abu-Salem, Hayam M. Ibrahim

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Plants are rich sources of bioactive compounds. In this study the photochemical screening of hexane, ethanolic and aqueous extracts of roots and latex of jojoba (Simmondsia chinensis) plant revealed the presence of saponins, tannins, alkaloids, steroids and glycosides. Ethanolic extract was found to be richer in these metabolites than hexane, aqueous extracts and latex. The extracts and latex displayed effective antimicrobial activity against Salmonella typhimurium, Bacillus cereus, Clostridium perfringens, Staphylococcus aureus, Escherichia coli, Candida albicans and Aspergillus flavus. The increase in volume of the extracts and latex caused more activity, as shown by zones of inhibition. Candida albicans growth was inhibited only by hexane extract. Jojoba latex was not effective against Candida albicans at 0.1 and 0.5 ml extracts concentration but showed 5mm zone of inhibition at (1.0 ml). Lower volume (0.1ml) of latex encouraged Aspergillus flavus growth, while at (1.00 ml) reduced its mycelial growth. Thus, jojoba root extracts and latex can be of potential natural antimicrobial agents.

Keywords: Antimicrobial activity, Jojoba (Simmondsia chinensis), latex, photochemical, root Extracts.

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Authors: Waleed S. Alwaneen

Abstract:

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: B. Bina, M.H. Mehdinejad, Gunnel Dalhammer, Guna RajaraoM. Nikaeen, H. Movahedian Attar

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Coagulation of water involves the use of coagulating agents to bring the suspended matter in the raw water together for settling and the filtration stage. Present study is aimed to examine the effects of aluminum sulfate as coagulant in conjunction with Moringa Oleifera Coagulant Protein as coagulant aid on turbidity, hardness, and bacteria in turbid water. A conventional jar test apparatus was employed for the tests. The best removal was observed at a pH of 7 to 7.5 for all turbidities. Turbidity removal efficiency was resulted between % 80 to % 99 by Moringa Oleifera Coagulant Protein as coagulant aid. Dosage of coagulant and coagulant aid decreased with increasing turbidity. In addition, Moringa Oleifera Coagulant Protein significantly has reduced the required dosage of primary coagulant. Residual Al+3 in treated water were less than 0.2 mg/l and meets the environmental protection agency guidelines. The results showed that turbidity reduction of % 85.9- % 98 paralleled by a primary Escherichia coli reduction of 1-3 log units (99.2 – 99.97%) was obtained within the first 1 to 2 h of treatment. In conclusions, Moringa Oleifera Coagulant Protein as coagulant aid can be used for drinking water treatment without the risk of organic or nutrient release. We demonstrated that optimal design method is an efficient approach for optimization of coagulation-flocculation process and appropriate for raw water treatment.

Keywords: MOCP, Coagulant aid, turbidity removal, E.coliremoval, water, treatment

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Authors: Douglas N. Simões, Michele Pittol, Vanda F. Ribeiro, Daiane Tomacheski, Ruth M. C. Santana

Abstract:

The increasing demand of thermoplastic elastomers is related to the wide range of applications, such as automotive, footwear, wire and cable industries, adhesives and medical devices, cell phones, sporting goods, toys and others. These materials are susceptible to microbial attack. Moisture and organic matter present in some areas (such as shower area and sink), provide favorable conditions for microbial proliferation, which contributes to the spread of diseases and reduces the product life cycle. Compounds based on SEBS copolymers, poly(styrene-b-(ethylene-co-butylene)-b-styrene, are a class of thermoplastic elastomers (TPE), fully recyclable and largely used in domestic appliances like bath mats and tooth brushes (soft touch). Zinc oxide and zinc ions loaded in personal and home care products have become common in the last years due to its biocidal effect. In that sense, the aim of this study was to evaluate the effect of zinc as antimicrobial agent in compounds based on SEBS/polypropylene/oil/ calcite for use as refrigerator seals (gaskets), bath mats and sink squeegee. Two zinc oxides from different suppliers (ZnO-Pe and ZnO-WR) and one masterbatch of zinc ions (M-Zn-ion) were used in proportions of 0%, 1%, 3% and 5%. The compounds were prepared using a co-rotating double screw extruder (L/D ratio of 40/1 and 16 mm screw diameter). The extrusion parameters were kept constant for all materials. Tests specimens were prepared using the injection molding machine. A compound with no antimicrobial additive (standard) was also tested. Compounds were characterized by physical (density), mechanical (hardness and tensile properties) and rheological properties (melt flow rate - MFR). The Japan Industrial Standard (JIS) Z 2801:2010 was applied to evaluate antibacterial properties against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli). The Brazilian Association of Technical Standards (ABNT) NBR 15275:2014 were used to evaluate antifungal properties against Aspergillus niger (A. niger), Aureobasidium pullulans (A. pullulans), Candida albicans (C. albicans), and Penicillium chrysogenum (P. chrysogenum). The microbiological assay showed a reduction over 42% in E. coli and over 49% in S. aureus population. The tests with fungi showed inconclusive results because the sample without zinc also demonstrated an inhibition of fungal development when tested against A. pullulans, C. albicans and P. chrysogenum. In addition, the zinc loaded samples showed worse results than the standard sample when tested against A. niger. The zinc addition did not show significant variation in mechanical properties. However, the density values increased with the rise in ZnO additives concentration, and had a little decrease in M-Zn-ion samples. Also, there were differences in the MFR results in all compounds compared to the standard.

Keywords: Antimicrobial, home device, SEBS, zinc.

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Authors: Ali Mohamadi Sani

Abstract:

The aim of this study was to determine the antimicrobial effect of Helichrysum arenarium L. essential oil in "in-vitro" condition on the growth of seven microbial species including Bacillus subtilis, Escherichia coli, Staphylococcus aureus, Saccharomyces cereviciae, Candida albicans, Aspergillus flavus and Aspergillus parasiticus using micro-dilution method. The minimum inhibitory concentration (MIC) and minimum bactericidal or fungicidal concentration (MBC, MFC) were determined for the essential oil at ten concentrations. Finally, the sensitivity of tested microbes to essential oil of H. arenarium was investigated. Results showed that Bacillus subtilis (MIC=781.25 and MBC=6250 µg/ml) was more resistance than two other bacterial species. Among the tested yeasts, Saccharomyces cereviciae (MIC=97.65 and MFC=781.25 µg/ml) was more sensitive than Candida albicans while among the fungal species, growth of Aspergillus parasiticus inhibited at lower concentration of oil than the Aspergillus flavus. The extracted essential oil exhibited the same MIC value in the liquid medium against all fungal strains (48.82 µg/ml), while different activity against A. flavus and A. parasiticus was observed in this medium with MFC values of 6250 and 390.625µg/ml, respectively. The results of the present study indicated that Helichrysum arenarium L essential oil had significant (P<0.05) antimicrobial activity; therefore, it can be used as a natural preservation to increase the shelf life of food products.

 

Keywords: Helichrysum arenarium, Antimicrobial agent, Essential oil, MIC.

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Authors: Disciglio G., Gatta G., Libutti A., Tarantino A., Frabboni L., Tarantino E.

Abstract:

Results of a field study carried out at Trinitapoli (Puglia region, southern Italy) on the irrigation of an artichoke crop with three types of water (secondary-treated wastewater, SW; tertiary-treated wastewater, TW; and freshwater, FW) are reported. Physical, chemical and microbiological analyses were performed on the irrigation water, and on soil and yield samples.

The levels of most of the chemical parameters, such as electrical conductivity, total suspended solids, Na⁺, Ca²⁺, Mg⁺², K⁺, sodium adsorption ratio, chemical oxygen demand, biological oxygen demand over 5 days, NO₃ –N, total N, CO₃², HCO₃, phenols and chlorides of the applied irrigation water were significantly higher in SW compared to GW and TW. No differences were found for Mg²⁺, PO₄-P, K⁺ only between SW and TW. Although the chemical parameters of the three irrigation water sources were different, few effects on the soil were observed. Even though monitoring of Escherichia coli showed high SW levels, which were above the limits allowed under Italian law (DM 152/2006), contamination of the soil and the marketable yield were never observed. Moreover, no Salmonella spp. were detected in these irrigation waters; consequently, they were absent in the plants. Finally, the data on the quantitative-qualitative parameters of the artichoke yield with the various treatments show no significant differences between the three irrigation water sources. Therefore, if adequately treated, municipal wastewater can be used for irrigation and represents a sound alternative to conventional water resources.

Keywords: Artichoke, soil chemical characteristics, fecal indicators, treated municipal wastewater, water recycling.

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Authors: P. Rahnemoon, M. Sarabi Jamab, M. Javanmard Dakheli, A. Bostan

Abstract:

In recent years, tendency to use of natural antimicrobial agents in food industry has increased. Pomegranate peels containing phenolic compounds and anti-microbial agents, are counted as valuable source for extraction of these compounds. In this study, the extraction of pomegranate peel extract was carried out at different ethanol/water ratios (40:60, 60:40, and 80:20), temperatures (25, 40, and 55 ˚C), and time durations (20, 24, and 28 h). The extraction yield, phenolic compounds, flavonoids, and anthocyanins were measured. ‎Antimicrobial activity of pomegranate peel extracts were determined against some food-borne ‎microorganisms such as Salmonella enteritidis, Escherichia coli, Listeria monocytogenes, ‎‎Staphylococcus aureus, Aspergillus niger, and Saccharomyces cerevisiae by agar diffusion and MIC methods. Results showed that at ethanol/water ratio 60:40, 25 ˚C and 24 h maximum amount of phenolic compounds ‎‏(‎‏‎349.518‎‏ ‏mg gallic acid‏/‏g dried extract), ‎flavonoids (250.124 mg rutin‏/‏g dried extract), anthocyanins (252.047 ‎‏‏mg ‎cyanidin‏‎3‎‏glucoside‏/‏‎100 g dried extract), and the strongest antimicrobial activity were obtained. ‎All extracts’ antimicrobial activities were demonstrated against every tested ‎‎microorganisms‏.‎‏ Staphylococcus aureus showed the highest sensitivity among the tested ‎‎‎microorganisms.

Keywords: Antimicrobial agents, phenolic compounds, pomegranate peel, solvent extraction.

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Authors: Laidson P. Gomes, Cristina T. Andrade, Eduardo M. Del Aguila, Cameron Alexander, Vânia M. F. Paschoalin

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Chitosan is a natural polysaccharide prepared by the N-deacetylation of chitin. In this study, the physicochemical and antibacterial properties of chitosan nanoparticles, produced by ultrasound irradiation, were evaluated. The physicochemical properties of the nanoparticles were determined by dynamic light scattering and zeta potential analysis. Chitosan nanoparticles inhibited the growth of E. coli. The minimum inhibitory concentration (MIC) values were lower than 0.5 mg/mL, and the minimum bactericidal concentration (MBC) values were similar or higher than MIC values. Confocal laser scanning micrographs (CLSM) were used to observe the interaction between E. coli suspensions mixed with FITC-labeled chitosan polymers and nanoparticles.

Keywords: Chitosan nanoparticles, dynamic light scattering, zeta potential, confocal microscopy, antibacterial activity.

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Authors: N. Khazaei, M. Jouki, A. Kalbasi, H. Tavakolipour, S. Rajabifar, F. Motamedi. Sedeh, A. Jouki

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In this research saffron samples were prepared from farms and sampling was done in four states contain : sampling from fresh saffron of petal with forceps , sampling from fresh saffron of petal by hands, sampling from dried sample by warm air in shadow, sampling from dried sample which dried by dryer. Samples collected and kept in sterile tubes and containers and carried to laboratory and maintained until experiment. Microbial experiments were performed to determine microbial load such as total count, Staphylococcus aureus, coli form, E.coli, mold and yeast. Results showed that in picking and drying stages the contamination amount increases in saffron samples. There was a significant difference between the microbial load of picked up saffron by forceps and by hands, and also between dried saffron by warm air in shadow and by dryer.

Keywords: saffron; contaminations; preparation method

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Authors: Min-Ah Woo, Min-Cheol Lim, Hyun-Joo Chang, Sung-Wook Choi

Abstract:

We developed a paper-based colorimetric sensor utilizing magnetic nanoparticles conjugated with aptamers (MNP-Apts) against E. coli O157:H7. The MNP-Apts were applied to a test sample solution containing the target cells, and the solution was simply dropped onto PVDF (polyvinylidene difluoride) membrane. The membrane moves the sample radially to form the sample spots of different compounds as concentric rings, thus the MNP-Apts on the membrane enabled specific recognition of the target cells through a color ring generation by MNP-promoted colorimetric reaction of TMB (3,3',5,5'-tetramethylbenzidine) and H₂O₂. This method could be applied to rapidly and visually detect various bacterial pathogens in less than 1 h without cell culturing.

Keywords: Aptamer, colorimetric sensor, E. coli O157:H7, magnetic nanoparticle, polyvinylidene difluoride.

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Authors: María D. Groppa, Andrea Trentini, Myriam Zawoznik, Roxana Bigi, Carlos Nadra, Patricia L. Marconi

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In the present work, a remediation bioprocess based on the use of a local isolate of the microalgae Chlorella vulgaris immobilized in alginate beads is proposed. This process was shown to be effective for the reduction of several chemical and microbial contaminants present in Cildáñez stream, a water course that is part of the Matanza-Riachuelo Basin (Buenos Aires, Argentina). The bioprocess, involving the culture of the microalga in autotrophic conditions in a stirred-tank bioreactor supplied with a marine propeller for 6 days, allowed a significant reduction of Escherichia coli and total coliform numbers (over 95%), as well as of ammoniacal nitrogen (96%), nitrates (86%), nitrites (98%), and total phosphorus (53%) contents. Pb content was also significantly diminished after the bioprocess (95%). Standardized cytotoxicity tests using Allium cepa seeds and Cildáñez water pre- and post-remediation were also performed. Germination rate and mitotic index of onion seeds imbibed in Cildáñez water subjected to the bioprocess was similar to that observed in seeds imbibed in distilled water and significantly superior to that registered when untreated Cildáñez water was used for imbibition. Our results demonstrate the potential of this simple and cost-effective technology to remove urban-water contaminants, offering as an additional advantage the possibility of an easy biomass recovery, which may become a source of alternative energy.

Keywords: Bioreactor, bioremediation, Chlorella vulgaris, Matanza-Riachuelo basin, microalgae.

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Authors: Ime R. Udotong, Samuel I. Eduok, Joseph P. Essien, Basil N. Ita

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Sediment and mangrove root samples from Iko River Estuary, Nigeria were analyzed for microbial and polycyclic aromatic hydrocarbon (PAH) content. The total heterotrophic bacterial (THB) count ranged from 1.1x107 to 5.1 x107 cfu/g, total fungal (TF) count ranged from 1.0x106 to 2.7x106 cfu/g, total coliform (TC) count ranged from 2.0x104 to 8.0x104cfu/g while hydrocarbon utilizing bacterial (HUB) count ranged from 1.0x 105 to 5.0 x 105cfu/g. There was a range of positive correlation (r = 0.72 to 0.93) between THB count and total HUB count, respectively. The organisms were Staphylococcus aureus, Bacillus cereus, Flavobacterium breve, Pseudomonas aeruginosa, Erwinia amylovora, Escherichia coli, Enterobacter sp, Desulfovibrio sp, Acinetobacter iwoffii, Chromobacterium violaceum, Micrococcus sedentarius, Corynebacterium sp, and Pseudomonas putrefaciens. The PAH were Naphthalene, 2-Methylnaphthalene, Acenapthylene, Acenaphthene, Fluorene, Phenanthene, Anthracene, Fluoranthene, Pyrene, Benzo(a)anthracene, Chrysene, Benzo(b)fluoranthene, Benzo(k)fluoranthene, Benzo(a)pyrene, Dibenzo(a,h)anthracene, Benzo(g,h,l)perylene ,Indeno(1,2,3-d)pyrene with individual PAH concentrations that ranged from 0.20mg/kg to 1.02mg/kg, 0.20mg/kg to 1.07mg/kg and 0.2mg/kg to 4.43mg/kg in the benthic sediment, epipellic sediment and mangrove roots, respectively. Total PAH ranged from 6.30 to 9.93mg/kg, 6.30 to 9.13mg/kg and 9.66 to 16.68mg/kg in the benthic sediment, epipellic sediment and mangrove roots, respectively. The high concentrations in the mangrove roots are indicative of bioaccumulation of the pollutant in the plant tissue. The microorganisms are of ecological significance and the detectable quantities of polycyclic aromatic hydrocarbon could be partitioned and accumulated in tissues of infaunal and epifaunal organisms in the study area.

Keywords: Hydrocarbonoclastic bacteria, Iko River estuary, Mangrove, Polycyclic aromatic hydrocarbon.

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Authors: Ikram Mohamed Eltayeb Elsiddig, Abdel Khalig Muddather, Hiba Abdel Rahman Ali, Saad Mohamed Hussein Ayoub

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Anogeissus leiocarpus (Combretaceae) is well known for its medicinal uses in African traditional medicine, for treating many human diseases mainly skin diseases and infections. Mycetoma disease is a fungal and/ or bacterial skininfection, mainly cause by Madurella mycetomatis fungus. This study was carried out in vitro to investigate the antifungal activity of Anogeissus leiocarpus leaf extracts against the isolated pathogenic Madurella mycetomatis, by using the NCCLS modified method compared to Ketoconazole standard drug, and MTT assay. The bioactive fraction was subjected to chemical analysis implementing different chromatographic analytical methods (TLC, HPLC, and LC-MS/MS). The results showed significance antifungal activity of A. leiocarpus leaf extracts against the isolated pathogenic M. mycetomatis, compared to negative and positive controls. The chloroform fraction showed the highest antifungal activity. The chromatographic analysis of the chloroform fraction with the highest activity showed the presence of important bioactive compounds such as ellagic and flavellagic acids derivatives, flavonoids and stilbenoid, which are well known for their antifungal activity.

Keywords: Anogeissus leiocarpus, crude extracts and fractions of Anogeissus leiocarpus, in vitro susceptibility of Madurella mycetomatis, Madurella mycetomatis.

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Authors: Sang-Ho Baik

Abstract:

D-erythro-cyclohexylserine (D chiral unnatural β-hydroxy amino acid expected for the synthesis of drug for AIDS treatment. To develop a continuous bioconversion system with whole cell biocatalyst of D-threonine aldolase (D genes for the D-erythro-CHS production, D-threonine aldolase gene was amplified from Ensifer arboris 100383 by direct PCR amplication using two degenerated oligonucleotide primers designed based on genomic sequence of Shinorhizobium meliloti Sequence analysis of the cloned DNA fragment revealed one open-reading frame of 1059 bp and 386 amino acids. This putative D-TA gene was cloned into NdeI and EcoRI (pEnsi His-tag sequence or BamHI (pEnsi-DTA[2]) sequence of the pET21(a) vector. The expression level of the cloned gene was extremely overexpressed by E. coli BL21(DE3) transformed with pEnsi-DTA[1] compared to E. coli BL21(DE3) transformed with pEnsi-DTA[2]. When the cells expressing the wild used for D-TA enzyme activity, 12 mM glycine was successfully detected in HPLC analysis. Moreover, the whole cells harbouring the recombinant D-TA was able to synthesize D-erythro of 0.6 mg/ml in a batch reaction.

Keywords: About four key words or phrases in alphabetical order, separated by commas.

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Authors: Alaa Hussein Wadi

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The principal objective of a water treatment plant is to produce water that satisfies a set of drinking water quality standards at a reasonable price to the consumers. The gravel-bed flocculator provide a simple and inexpensive design for flocculation in small water treatment plants (less than 5000 m3/day capacity). The packed bed of gravel provides ideal conditions for the formation of compact settleable flocs because of continuous recontact provided by the sinuous flow of water through the interstices formed by the gravel. The field data which were obtained from the operation of the water supply treatment unit cover the physical, chemical and biological water qualities of the raw and settled water as obtained by the operation of the treatment unit. The experiments were carried out with the aim of assessing the efficiency of the gravel filter in removing the turbidity, pathogenic bacteria, from the raw water. The water treatment plant, which was constructed for the treatment of river water, was in principle a rapid sand filter. The results show that the average value of the turbidity level of the settled water was 4.83 NTU with a standard deviation of turbidity 2.893 NTU. This indicated that the removal efficiency of the sedimentation tank (gravel filter) was about 67.8 %. for pH values fluctuated between 7.75 and 8.15, indicating the alkaline nature of the raw water of the river Shatt Al-Hilla, as expected. Raw water pH is depressed slightly following alum coagulation. The pH of the settled water ranged from 7.75 to a maximum of 8.05. The bacteriological tests which were carried out on the water samples were: total coliform test, E-coli test, and the plate count test. In each test the procedure used was as outlined in the Standard Methods for the Examination of Water and Wastewater (APHA, AWWA, and WPCF, 1985). The gravel filter exhibit a low performance in removing bacterial load. The percentage bacterial removal, which is maximum for total plate count (19%) and minimum for total coliform (16.82%).

Keywords: Gravel bed flocculator, turbidity, total coliform.

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Authors: Maribel G. Guzmán, Jean Dille, Stephan Godet

Abstract:

Silver nanoparticles were prepared by chemical reduction method. Silver nitrate was taken as the metal precursor and hydrazine hydrate as a reducing agent. The formation of the silver nanoparticles was monitored using UV-Vis absorption spectroscopy. The UV-Vis spectroscopy revealed the formation of silver nanopart├¡cles by exhibing the typical surface plasmon absorption maxima at 418-420 nm from the UV–Vis spectrum. Comparison of theoretical (Mie light scattering theory) and experimental results showed that diameter of silver nanoparticles in colloidal solution is about 60 nm. We have used energy-dispersive spectroscopy (EDX), X-ray diffraction (XRD), transmission electron microscopy (TEM) and, UV–Vis spectroscopy to characterize the nanoparticles obtained. The energy-dispersive spectroscopy (EDX) of the nanoparticles dispersion confirmed the presence of elemental silver signal no peaks of other impurity were detected. The average size and morphology of silver nanoparticles were determined by transmission electron microscopy (TEM). TEM photographs indicate that the nanopowders consist of well dispersed agglomerates of grains with a narrow size distribution (40 and 60 nm), whereas the radius of the individual particles are between 10 and 20 nm. The synthesized nanoparticles have been structurally characterized by X-ray diffraction and transmission high-energy electron diffraction (HEED). The peaks in the XRD pattern are in good agreement with the standard values of the face-centered-cubic form of metallic silver (ICCD-JCPDS card no. 4-0787) and no peaks of other impurity crystalline phases were detected. Additionally, the antibacterial activity of the nanopart├¡culas dispersion was measured by Kirby-Bauer method. The nanoparticles of silver showed high antimicrobial and bactericidal activity against gram positive bacteria such as Escherichia Coli, Pseudimonas aureginosa and staphylococcus aureus which is a highly methicillin resistant strain.

Keywords: Silver nanoparticles, surface plasmon, UV-Vis absorption spectrum, chemicals reduction.

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Authors: Suttijit Sriwatcharakul

Abstract:

The bioactivity studies from the weed ethanolic crude extracts from leaf, stem, pod and root of wild spider flower; Cleoma viscosa Linn. were analyzed for the growth inhibition of 6 bacterial species; Salmonella typhimurium TISTR 5562, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus TISTR 1466, Streptococcus epidermidis ATCC 1228, Escherichia coli DMST 4212 and Bacillus subtilis ATCC 6633 with initial concentration crude extract of 50 mg/ml. The agar well diffusion results found that the extracts inhibit only gram positive bacteria species; S. aureus, S. epidermidis and B. subtilis. The minimum inhibition concentration study with gram positive strains revealed that leaf crude extract give the best result of the lowest concentration compared with other plant parts to inhibit the growth of S. aureus, S. epidermidis and B. subtilis at 0.78, 0.39 and lower than 0.39 mg/ml, respectively. The determination of total phenolic compounds in the crude extracts exhibited the highest phenolic content was 10.41 mg GAE/g dry weight in leaf crude extract. Analyzed the efficacy of free radical scavenging by using DPPH radical scavenging assay with all crude extracts showed value of IC50 of leaf, stem, pod and root crude extracts were 8.32, 12.26, 21.62 and 35.99 mg/ml, respectively. Studied cytotoxicity of crude extracts on human breast adenocarcinoma cell line by MTT assay found that pod extract had the most cytotoxicity CC50 value, 32.41 µg/ml. Antioxidant activity and cytotoxicity of crude extracts exhibited that the more increase of extract concentration, the more activities indicated. According to the bioactivities results, the leaf crude extract of Cleoma viscosa Linn. is the most interesting plant part for further work to search the beneficial of this weed.

Keywords: Antimicrobial, antioxidant activity, Cleoma viscosa Linn., cytotoxicity test, total phenolic compound.

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Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le

Abstract:

Since the marine environmental conditions are extremely different from the other ones, marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411 strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411 that are effective against wide range of pathogens: both grampositive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the future.

Keywords: Marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis.

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Authors: Madiha El Awamie, Catherine Rees

Abstract:

Natural preservatives have been used as alternatives to traditional chemical preservatives; however, a limited number have been commercially developed and many remain to be investigated as sources of safer and effective antimicrobials. In this study, we have been investigating the antimicrobial activity of an extract of Glycyrrhiza glabra (liquorice) that was provided as a waste material from the production of liquorice flavourings for the food industry, and to investigate if this retained the expected antimicrobial activity so it could be used as a natural preservative. Antibacterial activity of liquorice extract was screened for evidence of growth inhibition against eight species of Gram-negative and Gram-positive bacteria, including Listeria monocytogenes, Listeria innocua, Staphylococcus aureus, Enterococcus faecalis and Bacillus subtilis. The Gram-negative bacteria tested include Pseudomonas aeruginosa, Escherichia coli and Salmonella typhimurium but none of these were affected by the extract. In contrast, for all of the Gram-positive bacteria tested, growth was inhibited as monitored using optical density. However parallel studies using viable count indicated that the cells were not killed meaning that the extract was bacteriostatic rather than bacteriocidal. The Minimum Inhibitory Concentration [MIC] and Minimum Bactericidal Concentration [MBC] of the extract was also determined and a concentration of 50 µg ml^-1 was found to have a strong bacteriostatic effect on Gram-positive bacteria. Microscopic analysis indicated that there were changes in cell shape suggesting the cell wall was affected. In addition, the use of a reporter strain of Listeria transformed with the bioluminescence genes luxABCDE indicated that cell energy levels were reduced when treated with either 12.5 or 50 µg ml^-1 of the extract, with the reduction in light output being proportional to the concentration of the extract used. Together these results suggest that the extract is inhibiting the growth of Gram-positive bacteria only by damaging the cell wall and/or membrane.

Keywords: Antibacterial activity, bioluminescence, Glycyrrhiza glabra, natural preservative.

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Authors: Abu Salim Mustafa

Abstract:

Water plays a major role in maintaining life on earth, but it can also serve as a matrix for pathogenic organisms, posing substantial health threats to humans. Although, outbreaks of diseases attributable to drinking water may not be common in industrialized countries, they still occur and can lead to serious acute, chronic, or sometimes fatal health consequences. The analysis of drinking water samples from different regions of Kuwait was performed in this study for bacterial and viral contaminations. Drinking tap water samples were collected from 15 different locations of the six Kuwait governorates. All samples were analyzed by confocal microscopy for the presence of bacteria. The samples were cultured in vitro to detect cultivable organisms. DNA was isolated from the cultured organisms and the identity of the bacteria was determined by sequencing the bacterial 16S rRNA genes, followed by BLAST analysis in the database of NCBI, USA. RNA was extracted from water samples and analyzed by real-time PCR for the detection of viruses with potential health risks, i.e. Astrovirus, Enterovirus, Norovirus, Rotavirus, and Hepatitis A. Confocal microscopy showed the presence of bacteria in some water samples. The 16S rRNA gene sequencing of culture grown organisms, followed by BLAST analysis, identified the presence of several non-pathogenic bacterial species. However, one sample had Acinetobacter baumannii, which often causes opportunistic infections in immunocompromised people, but none of the studied viruses could be detected in the drinking water samples analyzed. The results indicate that drinking water samples analyzed from various locations in Kuwait are relatively safe for drinking and do not contain many harmful pathogens.

Keywords: Drinking water, 16S rRNA, microbial diversity, viruses, Kuwait.

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Authors: Santosh Kumar, Anand Prakash, Kanak Sinha, Anita K Verma

Abstract:

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Keywords: Anti-cancer, Anti-microbial, EAC mouse cell, Tissue culture, Vernonia divergens.

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Authors: Simona Dostalova, Ana Maria Jimenez Jimenez, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

Abstract:

Various nanomaterials can be used as a drug delivery vehicles in nanomedicine, called nanocarriers. They can either be organic or inorganic, synthetic or natural-based. Although synthetic nanocarriers are easier to produce, they can often be toxic for the organism and thus not suitable for use in treatment. From naturalbased nanocarriers, the most commonly used are protein cages or viral capsids. In this work, virus bacteriophage λ was used for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2% maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000×g and 4°C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 μg·mL-1). Free drug molecules were removed by filtration. The encapsulation was verified using the absorbance for doxorubicin and atomic absorption spectrometry for platinum cytostatics. The amount of encapsulated drug linearly increased with the increasing concentration of applied drug with the determination coefficient R2=0.989 for doxorubicin; R2=0.967 for cisplatin; R2=0.989 for carboplatin and R2=0.996 for oxaliplatin. The overall encapsulation efficiency was calculated as 50% for doxorubicin; 8% for cisplatin; 6% for carboplatin and 10% for oxaliplatin.

Keywords: Bacteriophage λ, doxorubicin, platinum cytostatics, protein-based nanocarrier, viral capsid.

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Authors: S. Angalaparameswari, T.S. Mohamed Saleem, M. Alagusundaram, S. Ramkanth, V.S. Thiruvengadarajan, K. Gnanaprakash, C. Madhusudhana Chetty, G. Pratheesh

Abstract:

The present research was designed to investigate the anti-microbial activity of aristolochic acid from the root of Aristolochia bracteata. From the methanolic & ethyl extract extracts of Aristolochia bracteata aristolochic acid I was isolated and conformed through IR, NMR & MS. The percentage purity of aristolochic acid I was determined by UV & HPLC method. Antibacterial activity of extracts of Aristolochia bracteata and the isolated compound was determined by disc diffusion method. The results reveled that the isolated aristolochic acid from methanolic extract was more pure than the compound from ethyl acetate extract. The various extracts (500μg/disc) of Aristolochia bracteata showed moderate antibacterial activity with the average zone of inhibition of 7-18 mm by disc diffusion method. Among the extracts, ethyl acetate & methanol extracts were shown good anti-microbial activity and the growth of E.coli (18 mm) was strongly inhibited. Microbial assay of isolated compound (Aristolochic acid I) from ethyl acetate & methanol extracts were shown good antimicrobial activity and the zone of inhibition of both at higher concentration 50 μg/ml was similar with the standard aristolochic acid. It may be concluded that the isolated compound of aristolochic acid I has good anti-bacterial activity.

Keywords: Aristolochic acid I, Anti-microbial activity, Aristolochia bracteata, Bacillus subtilis, E.coli

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Authors: Eleftheria Tzamali, Panayiota Poirazi, Ioannis G. Tollis, Martin Reczko

Abstract:

Stable bacterial polymorphism on a single limiting resource may appear if between the evolved strains metabolic interactions take place that allow the exchange of essential nutrients [8]. Towards an attempt to predict the possible outcome of longrunning evolution experiments, a network based on the metabolic capabilities of homogeneous populations of every single gene knockout strain (nodes) of the bacterium E. coli is reconstructed. Potential metabolic interactions (edges) are allowed only between strains of different metabolic capabilities. Bacterial communities are determined by finding cliques in this network. Growth of the emerged hypothetical bacterial communities is simulated by extending the metabolic flux balance analysis model of Varma et al [2] to embody heterogeneous cell population growth in a mutual environment. Results from aerobic growth on 10 different carbon sources are presented. The upper bounds of the diversity that can emerge from single-cloned populations of E. coli such as the number of strains that appears to metabolically differ from most strains (highly connected nodes), the maximum clique size as well as the number of all the possible communities are determined. Certain single gene deletions are identified to consistently participate in our hypothetical bacterial communities under most environmental conditions implying a pattern of growth-condition- invariant strains with similar metabolic effects. Moreover, evaluation of all the hypothetical bacterial communities under growth on pyruvate reveals heterogeneous populations that can exhibit superior growth performance when compared to the performance of the homogeneous wild-type population.

Keywords: Bacterial polymorphism, clique identification, dynamic FBA, evolution, metabolic interactions.

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Authors: D. Berdous, H. Ferfera-Harrar

Abstract:

Superabsorbent polymers (SAPs) or hydrogels with three-dimensional hydrophilic network structure are high-performance water absorbent and retention materials. The in situ synthesis of metal nanoparticles within polymeric network as antibacterial agents for bio-applications is an approach that takes advantage of the existing free-space into networks, which not only acts as a template for nucleation of nanoparticles, but also provides long term stability and reduces their toxicity by delaying their oxidation and release. In this work, SAP/nanosilver nanocomposites were successfully developed by a unique green process at room temperature, which involves in situ formation of silver nanoparticles (AgNPs) within hydrogels as a template. The aim of this study is to investigate whether these AgNPs-loaded hydrogels are potential candidates for antimicrobial applications. Firstly, the superabsorbents were prepared through radical copolymerization via grafting and crosslinking of acrylamide (AAm) onto chitosan backbone (Cs) using potassium persulfate as initiator and N,N’-methylenebisacrylamide as the crosslinker. Then, they were hydrolyzed to achieve superabsorbents with ampholytic properties and uppermost swelling capacity. Lastly, the AgNPs were biosynthesized and entrapped into hydrogels through a simple, eco-friendly and cost-effective method using aqueous silver nitrate as a silver precursor and curcuma longa tuber-powder extracts as both reducing and stabilizing agent. The formed superabsorbents nanocomposites (Cs-g-PAAm)/AgNPs were characterized by X-ray Diffraction (XRD), UV-visible Spectroscopy, Attenuated Total reflectance Fourier Transform Infrared Spectroscopy (ATR-FTIR), Inductively Coupled Plasma (ICP), and Thermogravimetric Analysis (TGA). Microscopic surface structure analyzed by Transmission Electron Microscopy (TEM) has showed spherical shapes of AgNPs with size in the range of 3-15 nm. The extent of nanosilver loading was decreased by increasing Cs content into network. The silver-loaded hydrogel was thermally more stable than the unloaded dry hydrogel counterpart. The swelling equilibrium degree (Q) and centrifuge retention capacity (CRC) in deionized water were affected by both contents of Cs and the entrapped AgNPs. The nanosilver-embedded hydrogels exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus bacteria. These comprehensive results suggest that the elaborated AgNPs-loaded nanomaterials could be used to produce valuable wound dressing.

Keywords: Antibacterial activity, nanocomposites, silver nanoparticles, superabsorbent hydrogel.

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Authors: M. Naimi, M. B. Khaled

Abstract:

The aim of the present work was to test in vitro inhibition of food pathogens and spoilage bacteria by crude bacteriocins from autochthonous lactic acid bacteria. Thirty autochthonous lactic acid bacteria isolated previously, belonging to the genera: Lactobacillus, Carnobacterium, Lactococcus, Vagococcus, Streptococcus, and Pediococcus, have been screened by an agar spot test and a well diffusion assay against Gram-positive and Gram-negative harmful bacteria: Bacillus cereus, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 6538, and Pseudomonas aeruginosa under conditions means to reduce lactic acid and hydrogen peroxide effect to select bacteria with high bacteriocinogenic potential. Furthermore, crude bacteriocins semiquantification and heat sensitivity to different temperatures (80, 95, 110°C, and 121°C) were performed. Another exploratory test concerning the response of St. aureus ATCC 6538 to the presence of crude bacteriocins was realized. It has been observed by the agar spot test that fifteen candidates were active toward Gram-positive targets strains. The secondary screening demonstrated an antagonistic activity oriented only against St. aureus ATCC 6538, leading to the selection of five isolates: L_m14, L_m21, L_m23, L_m24, and L_m25 with a larger inhibition zone compared to the others. The ANOVA statistical analysis reveals a small variation of repeatability: L_m21: 0.56%, L_m23: 0%, L_m25: 1.67%, L_m14: 1.88%, L_m24: 2.14%. Conversely, slight variation was reported in terms of inhibition diameters: 9.58± 0.40, 9.83± 0.46 and 10.16± 0.24 8.5 ± 0.40 10 mm for, L_m21, L_m23, L_m25, L_m14and L_m24, indicating that the observed potential showed a heterogeneous distribution (BMS = 0.383, WMS = 0.117). The repeatability coefficient calculated displayed 7.35%. As for the bacteriocins semiquantification, the five samples exhibited production amounts about 4.16 for L_m21, L_m23, L_m25 and 2.08 AU/ml for L_m14, L_m24. Concerning the sensitivity the crude bacteriocins were fully insensitive to heat inactivation, until 121°C, they preserved the same inhibition diameter. As to, kinetic of growth , the µmax showed reductions in pathogens load for L_m21, L_m23, L_m25, L_m14, L_m24 of about 42.92%, 84.12%, 88.55%, 54.95%, 29.97% in the second trails. Inversely, this pathogen growth after five hours displayed differences of 79.45%, 12.64%, 11.82%, 87.88%, 85.66% in the second trails, compared to the control. This study showed potential inhibition to the growth of this food pathogen, suggesting the possibility to improve the hygienic food quality.

Keywords: Exploratory test, lactic acid bacteria, crude bacteriocins, spoilage, pathogens.

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