Search results for: 2-DCB degrading strains

Authors: Recep Kesli, Merih Simsek, Cengiz Demir, Onur Turkyilmaz

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Objective: Goal of this study was to determine the antibiotic resistance of Staphylococcus aureus (S. aureus) and Methicillin resistant staphylococcus aureus (MRSA) strains isolated at Medical Microbiology Laboratory of ANS Application and Research Hospital, Afyon Kocatepe University, Turkey. Methods: S. aureus strains isolated between October 2012 and September 2016, from various clinical specimens were evaluated retrospectively. S. aureus strains were identified by both the conventional methods and automated identification system -VITEK 2 (bio-Mérieux, Marcy l’etoile, France), and Meticillin resistance was verified using oxacillin disk with disk-diffusion method. Antibiotic resistance testing was performed by Kirby-Bauer disc diffusion method according to CLSI criteria, and intermediate susceptible strains were considered as resistant. Results: Seven hundred S.aureus strains which were isolated from various clinical specimens were included in this study. These strains were mostly isolated from blood culture, tissue, wounds and bronchial aspiration. All of 306 (43,7%) were oxacillin resistant. While all the S.aureus strains were found to be susceptible to vancomycin, teicoplanin, daptomycin and linezolid, 38 (9.6 %), 77 (19.5 %), 116 (29.4 %), 152 (38.6 %) and 28 (7.1 %) were found to be resistant aganist to clindamycin, erythromycin, gentamicin, tetracycline and sulfamethoxazole/trimethoprim, retrospectively. Conclusions: Comparing to the Methicillin sensitive staphylococcus aureus (MSSA) strains, increased resistance rates of, trimethoprim-sulfamethoxazole, clindamycin, erythromycin, gentamicin, and tetracycline were observed among the MRSA strains. In this study, the most effective antibiotic on the total of strains was found to be trimethoprim-sulfamethoxazole, the least effective antibiotic on the total of strains was found to be tetracycline.

Keywords: antibiotic resistance, MRSA, Staphylococcus aureus, VITEK 2

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Authors: Cheng-Chih Tsai, Yu-Hsuan Liu, Cheng-Ying Ho, Chun-Chin Huang

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The aim of this study evaluated the in vitro and in vivo antimicrobial activity of selected lactic acid bacteria (LAB) against Uropathogenic Escherichia coli (UPEC) for prevention and amelioration of UTIs. We screened LAB strains with antimicrobial effects on UPEC using a well-diffusion assay, bacterial adherence to the uroepithelium cell line SV-HUC-1 (BCRC 60358), and a coculture inhibition assay. The results showed that the 7 LAB strains (Lactobacillus paracasei, L. salivarius, two Pediococcus pentosaceus strains, two L. plantarum strains, and L. crispatus) and the fermented probiotic products produced by these multi-LAB strains exhibited potent zones of inhibition against UPEC. Moreover, the LAB strains and probiotic products adhered strongly to the uroepithelium SV-HUC-1 cell line. The growth of UPEC strains was also markedly inhibited after co-culture with the LAB strains and probiotic products in human urine. In addition, the enhanced levels of IL-6, IL-8 and lactic acid dehydrogenase were significantly decreased by treatments with the LAB strains and probiotic products in UPEC-induced SV-HUC-1 cells. Furthermore, oral administration of probiotic products reduced the number of viable UPEC in the urine of UPEC-challenged BALB/c mice. Taken together, this study demonstrates that probiotic supplementation may be useful as an adjuvant therapy for the treatment of bacterial-induced urinary tract infections.

Keywords: lactic acid bacterium, SV-HUC-1 uroepithelium, urinary tract infection, uropathogenic Escherichia coli, BALB/c mice

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Authors: Omaima A. Sharaf, Tarek A. Moussa, Said M. Badr El-Din, H. Moawad

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Background: Polycyclic aromatic hydrocarbons (PAHs) pose great environmental and human health concerns for their widespread occurrence, persistence, and carcinogenic properties. PAHs releases due to anthropogenic activities to the wider environment have led to higher concentrations of these contaminants than would be expected from natural processes alone. This may result in a wide range of environmental problems that can accumulate in agricultural ecosystems, which threatened to become a negative impact on sustainable agricultural development. Thus, this study aimed to evaluate the physico-chemical, and microbial properties of the compost leachate (CL) to assess its role as nutrient and microbial source (biostimulation/bioaugmentation) for developing a cost-effective bioremediation technology for PAHs contaminated sites. Material and Methods: PAHs-degrading bacteria were isolated from CL that was collected from a composting site located in central Scotland, UK. Isolation was carried out by enrichment using phenanthrene (PHR), pyrene (PYR) and benzo(a)pyrene (BaP) as the sole source of carbon and energy. The isolates were characterized using a variety of phenotypic and molecular properties. Six different isolates were identified based on the difference in morphological and biochemical tests. The efficiency of these isolates in PAHs utilization was assessed. Further analysis was performed to define taxonomical status and phylogenic relation between the most potent PAHs-utilizing bacterial strains and other standard strains, using molecular approach by partial 16S rDNA gene sequence analysis. Results indicated that the 16S rDNA sequence analysis confirmed the results of biochemical identification, as both of biochemical and molecular identification of the isolates assigned them to Bacillus licheniformis, Pseudomonas aeruginosa, Alcaligenes faecalis, Serratia marcescens, Enterobacter cloacae and Providenicia which were identified as the prominent PAHs-utilizers isolated from CL. Conclusion: This study indicates that the CL samples contain a diverse population of PAHs-degrading bacteria and the use of CL may have a potential for bioremediation of PAHs contaminated sites.

Keywords: polycyclic aromatic hydrocarbons, physico-chemical analyses, compost leachate, microbial and biochemical analyses, phylogenic relations, 16S rDNA sequence analysis

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Authors: Piña-Ronces Laura Gabriela, Reyes-Escogido María de Lourdes

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Exist a large variability in Exopolysaccharides (EPS) produced by LAB depending on carbon source, they have multiple applications in food industry mainly, but they have become important for the health. In this study, we identified EPS-producing strains belonging to the BAL group; they were previously isolated from humans. After that, we extracted and evaluated the antioxidant activity of EPS produced by all strains. Antioxidant activity was determined by DPPH method using ascorbic acid as standard for both comparison and quantification. 31 strains (51.66 %) produced EPS at concentrations between 451 and 1.561 mg/l, 16 of EPS extracted showed antioxidant effect superior to ascorbic acid at the same concentrations. EPS-producing strains were L. plantarum, L. sp and L. fermentum corresponding to Lactobacillus genus and, E. faecium, E. durans, and E. hirae of Enterococcus genus. Antioxidant activity showed by EPS from 3 strains of L. plantarum and 3 strains of E. faecium was different into specie, while the antioxidant activity determined for EPS obtained from the other strains did not show difference at specie level, but was superior to ascorbic acid. EPS produced by L. plantarum and E. hirae had the best activity, it could be considerate for selection them as a possible new alternative for therapy or treatment of diseases related whit oxidative stress. Further studies about biological functions of EPS have to be conducted for new applications in health.

Keywords: oxidative stress, lactic acid bacteria, exopolysaccharides, antioxidant activity

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Authors: Alise Senberga, Laila Dubova, Liene Strauta, Ina Alsina, Ieva Erdberga

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Legume symbiotic relationship with nitrogen fixating bacteria Rhizobim leguminosarum is an important factor used to improve the productivity of legumes, due to the fact that rhizobia can supply plant with the necessary amount of nitrogen. R. leguminosarum strains have shown different activity in fixing nitrogen. Depending on the chosen R. leguminosarum strain, host plant biochemical content can be altered. In this study we focused particularly on the changes in protein content in beans (using two different varieties) and peas (five different varieties) due to the use of several different R. leguminosarum strains (four strains for both beans and peas). Overall, the protein content increase was observed after seed inoculation with R. leguminosarum. Strain and plant cultivar interaction specification was observed. The effect of R. leguminosarum inoculation on the content of protein was dependent on the R. leguminosarum strain used. Plant cultivar also appeared to have a decisive role in protein content formation with the help of R. leguminosaru.

Keywords: legumes, protein content, rhizobia strains, soil

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Authors: Ikram Kamal, Mohamed Blaghen

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Biosurfactants are amphiphilic biological compounds consisting of hydrophobic and hydrophilic domains produced extracellularly or as part of the cell membrane by a variety of yeast, bacteria and filamentous fungi. Biosurfactant applications in the environmental industries are promising due to their biodegradability, low toxicity, and effectiveness in enhancing biodegradation and solubilization of low solubility compounds. Currently, the main application is for enhancement of oil recovery and hydrocarbon bioremediation due to their biodegradability and low critical micelle concentration (CMC). The use of biosurfactants has also been proposed for various industrial applications, such as in food additives, cosmetics, detergent formulations and in combinations with enzymes for wastewater treatment. In this study, we have investigated the potential of bacterial strains: Mannheimia haemolytica, Burkholderia cepacia and Serratia ficaria were collected aseptically from the lagoon Marchika (water and soil) in Nador, Morocco; for the production of biosurfactants. This study also aimed to optimize the biosurfactant production process by changing the variables that influence the type and amount of biosurfactant produced by these microorganisms such as: carbon sources and also other physical and chemical parameters such as temperature and pH. Emulsification index, methylene blue test, and thin layer chromatography (TLC) revealed the ability of strains used in this study to produce compounds that could emulsify gasoline. In addition, a GC/MS was used to separate and identify different biosurfactants purified.

Keywords: biosurfactants, Mannheimia haemolytica, biodegradability, Burkholderia cepacia, Serratia ficaria

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Authors: Ashraf Y. Z. Khalifa, Mohammed A. Almalki

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Medicago sativa (Alfalfa) is an important forage crop legume worldwide including Saudia Arabia due to its high nutritive value. Soil bacteria exist in root or root-nodules of Medicago sativa in either symbiotic relationships or in associations. The aim of the present study was to isolate and characterize endophytic bacteria that live in association with non-nodulated roots of Medicago sativa growing in Al-Ahsaa region, Saudia Arabia. Several bacterial strains were isolated from sterilized roots of Medicago sativa. Strains were characterized using 16S rRNA gene sequences, phylogenetic relationships analysis, morphological and biochemical characteristics. The strains utilized 50% (10 out of 20) of the different chemical substrates contained in the API20E strip. In general, many strains had the ability to ferment/oxidise all the carbohydrate tested except for rhamnose and the polyol carbohydrate, inositol. Comparative sequence analysis of the 16S rDNA gene indicated that the strains were closely related to the genus Bacillus. Furthermore, the growth parameters of Vigna sinensis were enhanced upon single-inoculation of the isolated strains, compared to the uninoculated control plants. The results highlighted that the root-nodules of Medicago sativa harbor non-nodulating bacterial strains that could have significant agricultural applications.

Keywords: Medicago sativa, endophytic bacteria, Pisum sativum, Vigna sinensis

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Authors: Abdulrahman Abdulhamid Arabo, Raji Arabi Bamanga, Mujiburrahman Fadilu, Musa Abubakar, Fatima Abdullahi Shehu, Hafeez Muhammad Yakasai, Nasiru Abdullahi

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The aim of this study was to isolate and identify biosurfactant-producing and diesel alkanes degrading bacteria. For this reason, bacteria isolated from the diesel-contaminated site were screened for their potential to produce biosurfactants and degrade diesel alkanes. Primary selection of diesel degraders was carried out by using the conventional enrichment culture technique, where 12 bacterial strains were isolated based on their ability to grow on minimal media supplemented with diesel as the sole carbon source, which was followed by qualitative screening methods for potential biosurfactant production. Isolate B11 was the only candidate that showed positive signs for drop collapse, foaming, hemolytic test, oil displacement of more than 22 ± 0.05 mm, and emulsification (E24) of 14 ± 0.30%. The effect of various culture parameters (incubation time, diesel concentration, nitrogen source, pH and temperature) on the biodegradation of diesel was evaluated. The optimum incubation time was confirmed to be 120 days for isolate B11, and the optimum PH was confirmed as 8.0 for the isolate; similarly, the optimum temperature was confirmed as 35oC. In addition, diesel oil was used as the sole carbon source for the isolates. The favorable diesel concentration was 12.5 % (v/v) for the isolate. The isolate has shown degradative ability towards Tridecane (C13), dodecane, 2, 6, 10-trimethyl- (C15), Tetradecane (C14), 2,6,10-Trimethyltridecane (C16), Pentadecane (C15). It degraded between 0.27% - 9.65% of individual diesel oil alkanes. The strain has exhibited the potential of degrading diesel oil n-alkanes and was identified as Alcaligenes species strain B11 (MZ027604) using the 16S rRNA. Sequencing.

Keywords: diesel oil, biosurfactant, Alcaligenes sp, biodegradation

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Authors: Khaled Khleifat, Muayyad Abboud, Ahmad Almustafa

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The effect of metabolic inhibitor/uncoupler(s) (CCCP and NaN3) and sulfhydryl reagents (dithiothreitol, 2 mercaptoethanol glutathione) on cadmium uptake was investigated in Enterobacter aerogenes strains. They include a transformed strain bearing the Vitreoscillahemoglobin gene, vgb as well as control strains that lack this transformed gene. The vgb-harboring strains showed better uptake of cadmium than vgb-lacking strains. Under low aeration, there was 2 fold enhancement of Cd+2 uptake in vgb-harboring strains compared with 1.6-fold enhancement under high aeration. The CCCP caused 36, 40 and 58% inhibition in cadmium uptake of parental, pUC9 harboring and VHb expressing cells, respectively. Similarly, the sodium azide exerted 44, 38 and 55% inhibition in Cd+2 uptake of parental, pUC9 harboring and VHb expressing cells, respectively. Less extensive inhibition of Cd+2 uptake in the range of 11 to 39% was observed with sulfhydryl reagents.

Keywords: bacterial hemoglobin, VHb, Cd uptake, biosorption

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Authors: Fdhila Walid, Bayar Sihem, Khouidi Bochra, Maâtouk Fethi, Ben Amor Feten, Hajer Hentati, Mahdhi Abdelkarim

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The presence of resistant bacteria in the oral cavity can be the major cause of dental antibiotic prophylaxis failure. Multidrug efflux has been described for many organisms, including bacteria and fungi as part of their drugs resistance strategy. The potential use of probiotic bacteria can be considered as a new alternative in the prevention or cure of oral cavity diseases. In this study, different Bacillus strains isolated from the environment were isolated and characterized using biochemical and molecular procedures. The inhibitory activity against different pathogenic bacteria and yeast strains was tested using diffusion agar assay method. Our data revealed that the tested strains have an antimicrobial effect against the pathogenic strains such as Streptococcus mutants. The inhibitory effect was variable depending from the probiotic and pathogenic strains. The obtained result demonstrated that Bacillus can be used as a potential candidates probiotic and help in the prevention and treatment of oral infections, including dental caries, periodontal disease and halitosis. Our data, partly encourage the use of probiotic strains because they do not produce acid which can contribute to faster installation decay and these are spore-forming bacteria that can withstand the stress of the oral cavity (acids, alkalis, and salty foods).

Keywords: probiotic, pathogenic bacteria, yeast, oral cavity

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Authors: A. Cheriguene, F. Chougrani

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A total of 120 wild lactic acid bacteria were isolated from goat’s milk collected from different areas in Western Algeria. The strains were screened for production and technological properties such as acid production, aminopeptidase activity, autolytic properties, antimicrobial activity, and exopolysaccharide production. In general most tested isolates showed a good biomass separation when collected by centrifugation; as for the production of the lactic acid, results revealed that our strains are weakly acidifying; nevertheless, lactococci showed a best acidifying activity compared to lactobacilli. Aminopeptidase activity was also weak in most strains; but, it was generally higher for lactobacilli compared to lactococci. Autolytic activity was generally higher for most strains, more particularly lactobacilli. Antimicrobial activity was detected in 50% of the isolates, particularly in lactobacilli where 80% of strains tested were able to inhibit the growth of other strains. The survey of the profile of the texture, the proteolysis as well as the development of the flavor in the Domiati cheese made on the basis of our isolated strains have been led during the ripening. The sensory assessment shows that the cheese salted in milk received the best scores in relation to cheese salted after drainage. Textural characteristics, such as hardness, cohesiveness, gumminess, and chewiness decreased in the two treatments during the 60 days of ripening. Otherwise, it has been noted that adhesiveness and adhesive force increased in the cheese salted in milk.

Keywords: lactic acid bacteria, technological properties, acidification, exopolysaccharide, bacteriocin, textural properties

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Authors: Nita Rukminasari

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The aim of this study is to select microalgae strains, which were isolated from agriculture and municipal wastewater drain, Reno, Nevada that has highest growth rate and lipid contents. The experiments in this study were carried out in two consecutive stages. The first stage is aimed at testing the survival capability of all isolated microalgae strains and determining the best candidates to grow in centrate cultivation system. The second stage was targeted at determination the highest growth rate and highest lipid content of the selected top performing algae strain when cultivated on centrate wastewater. 26 microalgae strains, which were isolated from municipal and agriculture waste water, were analyzed using Flow cytometer for FACS of lipid with BODIPY and Nile Red as a lipid dyes and they grew on 96 wells plate for 31 days to determine growth rate as a based line data for growth rate. The result showed that microalgae strains which showed a high mean of fluorescence for BODIPY and Nile Red were F3.BP.1, F3.LV.1, T1.3.1, and T1.3.3. Five microalgae strains which have high growth rate were T1.3.3, T2.4.1. F3.LV.1, T2.12.1 and T3.3.1. In conclusion, microalgae strain which showed the highest starch content was F3.LV.1. T1.3.1 had the highest mean of fluorescence for Nile Red and BODIPY. Microalgae strains were potential for biofuel feedstock such as F3.LV.1 and T1.3.1, those microalgae strains showed a positive correlation between growth rate at stationary phase, biomass and meant of fluorescence for Nile Red and BODIPY.

Keywords: agriculture and municipal wastewater, biofuel, centrate, microalgae

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Authors: M. Raissy, M. Shahrani

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The present study was done to evaluate the presence of tetracycline resistance genes in Lactococcus garvieae isolated from cultured rainbow trout, West Iran. The isolates were examined for antimicrobial resistance using disc diffusion method. Of the 49 strains tested, 19 were resistant to tetracycline (38.7%), 32 to enrofloxacin (65.3%), 21 to erythromycin (42.8%), 20 to chloramphenicol and trimetoprim-sulfamethoxazole (40.8%). The strains were then characterized for their genotypic resistance profiles. The results revealed that all 49 isolates contained at least one of the tetracycline resistance genes. Tet (A) was found in 89.4% of tetracycline resistant isolates and the frequency of other gene were as follow: tet (E) 42.1%, tet (B) 47.3%, tet (D) 15.7%, tet (L) 26.3%, tet (K) 52.6%, tet (G) 36.8%, tet (34) 21%, tet (S) 63.1%, tet (C) 57.8%, tet (M) 73.6%, tet (O) 42.1%. The results revealed high levels of antibiotic resistance in L. garvieae strains which is a potential danger for trout culture as well as for public health.

Keywords: Lactococcus garvieae, tetracycline resistance genes, rainbow trout, antimicrobial resistance

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Authors: Seung Min Kim, Lyu Jin Jun, Joon Bum Jeong

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The Myxosporea to cause emaciation disease in the olive flounder (Paralichthys olivaceus) is a pathogen to cause severe losses in the aquafarming industry in Korea. The 3,362 bp of DNA nucleotide sequences of four myxosporean strains (EM-HM-12, EM-MA-13, EM-JJ-14, and EM-MS-15) detected by PCR method from olive flounder suffering from emaciation disease in Korea during 2012-2015 were sequenced and deposited in GenBank database (GenBank accession numbers: KU377574, KT321705, KU377575 and KU377573, respectively). The homologies of DNA nucleotide sequences of four strains were compared to each other and were more than 99.7% homologous between the four strains. All of the strains were identified as Parvicapsula petunia based on the results of phylogenetic analysis. The results in this study would be useful for the research of emaciation disease in olive flounder of Korea.

Keywords: disease, emaciation, olive flounder, phylogenetic analysis

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Authors: Alouache Souhila, Messai Yamina, Torres Carmen, Bakour Rabah

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Objective: It has often been reported an association between antibiotic resistance and virulence. However, resistance to quinolones seems to be an exception, it tends instead to be associated with an attenuation of virulence, particularly in clinical strains. The purpose of this study was to evaluate the potential virulence of 28 quinolone-resistant E. coli strains recovered from water at the inflow (n=16) and outflow (n=12) of an urban wastewater treatment plant (WWTP). Methods: E. coli isolates were selected on Tergitol-7 agar supplemented with 2µg/ml of ciprofloxacin, they were screened by PCR for 11 virulence genes related to Extraintestinal pathogenic E. coli (ExPEC): papC, papG, afa/draBC, sfa/foc, kpsMTII, iutA, iroN, hlyF, ompT, iss and traT. The phylogenetic groups were determined by PCR and clonal relationship was evaluated by ERIC-PCR. Results: Genotyping by ERIC-PCR showed 7 and 12 DNA profiles among strains of wastewater (inflow) and treated water (outflow), respectively. Strains were assigned to the following phylogenetic groups: B2 (n = 1, 3.5%), D (n = 3, 10.7%), B1 (n = 10, 35.7%.) and A (n = 14, 50%). A total of 8 virulence-associated genes were detected, traT (n=19, 67.8%), iroN (n= 16, 57 .1%), hlyF (n=15, 53 .5%), ompT (n=15, 53 .5%), iss (n=14, 50%), iutA (n=9, 32.1%) , sfa/foc (n=7, 25%) and kpsMTII (n=2, 7.1%). Combination of virulence factors allowed to define 16 virulence profiles. The pathotype APEC was observed in 17.8% (D=1, B1=4) and human ExPEC in 7% (B2=1, D=1) of strains. Conclusion: The study showed that quinolone-resistant E. coli strains isolated from wastewater and treated water in WWTP harbored virulence genes with the presence of APEC and human ExPEC strains.

Keywords: E. coli, quinolone-resistance, virulence, WWTP

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Authors: Asad Ali, Asif Jamal

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The agriculture productivity losses due to microbial pathogens have been a serious issue in Pakistan and rest of the world. Present work was designed to isolate soil borne microorganisms having the antagonistic ability against notorious phytopathogens. From the initial collection of 23 bacterial isolates, two potent strains of Bacillus were screened on the basis of their comparative efficacy against devastating fungal pathogens. The strains AK-1 and AK-5 showed excellent inhibitory indexes against the majority of tested fungal strains. It was noted that both strains of Bacillus showed significant biocontrolling activity against Aspergillus flavus, Fusarium moniliforme, Colletotricum falcatum, Botrytis cinerea, Aspergillus niger, Fusarium oxysporum, Phythopthora capsici and Rhizopus oryzae. The strain AK-1 was efficient to suppress Aspergillus species and Rhizopus oryzae while AK-5 expressed significant antagonistic activity against Fusarium, Botrytis and Colletotricum species. On the basis of in vitro assay, it can be postulated that the Bacillus strains AK-1 and AK-5 can be used as bio-protective agent against various plant diseases. In addition, their applications as natural pesticides could be very helpful to prevent the adverse effects of chemical pesticides.

Keywords: bacillus species, biocontrol agent, biopesticides, phytopathogens

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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

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Authors: Shah Md. Asraful Islam, Shabina Yeasmin, Fatima Aktar Mousumi

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The experiment was conducted to evaluate the antagonistic potential of three commercially available Trichoderma strains viz., T. harzianum (armigera), T. harzianum (Ispahani), and T. viride against Colletotrichum musae isolates from three banana varieties viz., sagar, sobri, and katali. Mycelial growth rates of C. musae isolates were observed, the highest mycelial growth (11.62, 15.75, and 23.12 mm diameter) was observed by C. musae from sagor banana at 1, 2 and 3 days after inoculation, respectively. All the Trichoderma strains were capable of growth inhibition of C. musae isolates. After 4 days of duel culture, the highest mycelial growth reduction (10.33 mm diameter) was observed by the interaction between T. harzianum (armigera) with C. musae from sagor banana. Moreover, the highest growth inhibition (46.29%) was observed by the interaction between T. harzianum (armigera) with C. musae from the sobri banana. All the Trichoderma strains fully affected the viability of all the Colletotrichum isolates. Interestingly, both cultural filtrates and mycelial powders of all the Trichoderma strains showed a very nice inhibitory effect against C. musae isolates, where cultural filtrates were more potential than that of mycelial powders. So, all the tested Trichoderma strains may be used for the control of banana anthracnose disease.

Keywords: biological control, banana, anthracnose, Trichoderma, Colletotrichum

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Authors: Maria Manzoor, Iram Gul, Muhammad Arshad

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Bacterial strains were isolated from contaminated soil collected from Rawalpindi and Islamabad. The strains were investigated for lead resistance and their effect on Pb solubility and PGPR activity. Incubation experiments were carried for inoculated and unoculated soil containing different levels of Pb. Results revealed that few stains (BTM-4, BTM-11, BTM-14) were able to tolerate Pb up to 600 mg L-1, whereas five strains (BTM-3, BTM-6, BTM-10, BTM-21 and BTM-24) showed significant increase in solubility of Pb when compared to all other strains and control. The CaCl2 extractable Pb was increased by 13.6, 6.8, 4.4 and 2.4 folds compared to un-inoculated control soil at increased soil Pb concentration (500, 1000, 1500 and 200 mg kg-1, respectively). The selected bacterial strains (11) were further investigated for plant growth promotion activity through PGPR assays including. Germination and root elongation assays were also conducted under elevated metal concentration in controlled conditions to elucidate the effects of microbial strains upon plant growth and development. The results showed that all the strains tested in this study, produced significantly varying concentrations of IAA, siderophores and gibberellic acid along with ability to phosphorus solubilization index (PSI). The results of germination and root elongation assay further confirmed the beneficial role of the microbial strains in elevating metal stress through PGPR activity. Among all tested strains, BTM-10 significantly improved plant growth. 1.3 and 2.7 folds increase in root and shoot length was observed when compared to control. Which may be attributed to presence of important plant growth promoting enzymes (IAA 74.6 μg/ml; GA 19.23 μg/ml; Sidrophore units 49% and PSI 1.3 cm). The outcome of this study indicates that these Pb tolerant and solubilizing strains may have the potential for plant growth promotion under metal stress and can be used as mediator when coupled with heavy metal hyperaccumulator plants for phytoremediation of Pb contaminated soil.

Keywords: Pb resistant bacteria, Pb mobilizing bacteria, Phytoextraction of Pb, PGPR activity of bacteria

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Authors: Aimeurnadjette, Hammoudi Abd Erahmen, Bordjibaouahiba

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For many years, the pesticides used in agriculture have been responsible for environmental degradation, particularly noticeable in the areas of intensive agriculture, particularly through contamination of surface and groundwater. Our study was conducted to isolate and identify the microflora of water polluted by pesticides in an area with an agricultural vocation (Ben M'Hidi) subject to the pesticide effect for several years. Isolated fungal strains were identified based on the morphology of their vegetative and reproductive apparatus. The micromycètes were obtained; they belong mainly to the genera Aspergillus, Penicillium and Trichoderma. Furthermore, most bacterial strains characterized in this work, are that of the genus Aeromonas, Pseudomonas that are widely represented in the study of the biodegradation of pesticides.

Keywords: isolated, strains, polluted, pesticides

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Authors: Hieu Van Dong, Giang Thi Huong Tran, Giap Van Nguyen, Tung Duy Dao, Vuong Nghia Bui, Le Thi My Huynh, Yohei Takeda, Haruko Ogawa, Kunitoshi Imai

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Chicken anemia virus (CAV) has a ubiquitous and worldwide distribution in chicken production. Our group previously reported high seroprevalence of CAV in chickens in northern Vietnam. In the present study, 330 tissue samples collected from commercial and breeder chicken farms in eleven provinces in northern Vietnam were tested for the CAV infection. We found that 157 out of 330 (47.58%) chickens were positive with CAV genes by real-time PCR method. Nine CAV strains obtained from the different location and time were forwarded to the full-length sequence of CAV VP1 gene. Phylogenetic analysis of the Vietnamese CAV vp1 gene indicated that the CAVs circulating in northern Vietnam were divided into three distinct genotypes, II, III, and V, but not clustered with the vaccine strains. Among the three genotypes, genotype III was the major one widely spread in Vietnam, and that included three sub-genotypes, IIIa, IIIb, and IIIc. The Vietnamese CAV strains were closely related to the Chinese, Taiwanese, and USA strains. All the CAV isolates had glutamine at amino acid position 394 in the VP1 gene, suggesting that they might be highly pathogenic strains. One strain was defined to be genotype V, which had not been reported for Vietnamese CAVs. Additional studies are required to further evaluate the pathogenicity of CAV strains circulating in Vietnam.

Keywords: chicken anemia virus, genotype, genetic characteristics, Vietnam

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Authors: Paula Buldres, Jorge Toledo

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Objectives: This study aimed to characterize potential probiotic strains isolated from canine breast milk for use in dogs with enteropathies. Methodology: Six canine breast milk strains, one canine colostrum strain, and one control porcine breast milk strain were characterized. According to its functional properties of resistance to acids, different concentrations of bile salts, and pancreatin, its presumptive properties of safety and inhibitory effect on pathogens, non-cytotoxic characteristics, and adhesion to the intestine. The immunomodulatory effect of formulations with better probiotic characterization in vitro and in vivo was also analyzed. Results: Two strains characterized as potential probiotics were obtained, which corresponded to the canine strains (TUCO-16 and TUCO-17), presenting resistance to acidic pH, bile salts, and pancreatin, as well as an inhibitory effect on pathogenic Escherichia coli, Salmonella sp., and Clostridium perfringens. Strains TUCO-16 and TUCO-17 induced a significant increase in the expression of TNF-α and IL-8 in canine macrophages, respectively. Expression analyses of pattern recognition receptors in DH82 cells suggest that TUCO-16 and TUCO-17 might increase the TLR2 expression marker, and porcine strain (TUCO-4) increases the NOD2 expression marker. Based on the count obtained and the encapsulation yield, the best formulations correspond to FOS-Inulin for the TUCO-17 and TUCO-4 strains; Maltodextrin-Inulin for TUCO-16. All the strains are non-cytotoxic. The strain that showed the highest adhesion to intestinal epithelial cells was TUCO-17 with the FOS-Inulin formulation. On the other hand, the probiotics decreased the expression of pro-inflammatory markers in vivo, both in the intestine and in the spleen of mice. Conclusion: The combination of these three strains under study (TUCO-16, TUCO-17, and TUCO-4) would cover the probiotic properties in formulation and immunomodulation of all the markers under study.

Keywords: probiotics, gastrointestinal infec, dog, probiotic formulation, immunomodulatory probiotics

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Authors: Marija Ratkova Manovska, Mirko Prodanov, Dean Jankuloski, Katerina Blagoevska

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Staphylococci, which are widely found in the environment, animals, humans, and food products, include Staphylococcus aureus (S. aureus), the most significant pathogenic species in this genus. The virulence and toxicity of S. aureus are primarily attributed to the presence of specific genes responsible for producing toxins, biofilms, invasive components, and antibiotic resistance. Staphylococcal food poisoning, caused by the production of staphylococcal enterotoxins (SEs) by these strains in food, is a common occurrence. Globally, S. aureus food intoxications are typically ranked as the third or fourth most prevalent foodborne intoxications. For this study, a total of 333 milk samples and 1160 dairy product samples were analyzed between 2016 and 2020. The strains were isolated and confirmed using the ISO 6888-1:1999 "Horizontal method for enumeration of coagulase-positive staphylococci." Molecular analysis of the isolates, conducted using conventional PCR, involved detecting the 23s gene of S. aureus, the nuc gene, the mecA gene, and 11 genes responsible for producing enterotoxins (sea, seb, sec, sed, see, seg, seh, sei, ser, sej, and sep). The 23s gene was found in 93 (75.6%) out of 123 isolates of Staphylococcus spp. obtained from milk. Among the 76 isolates from dairy products, either S. aureus or the 23s gene was detected in 49 (64.5%) of them. The mecA gene was identified in three isolates from raw milk and five isolates from cheese samples. The nuc gene was present in 98.9% of S. aureus strains from milk and 97.9% from dairy products. Other Staphylococcus strains carried the nuc gene in 26.7% of milk strains and 14.8% of dairy product strains. Genes associated with SEs production were detected in 85 (69.1%) strains from milk and 38 (50%) strains from dairy products. In this study, 10 out of the 11 SEs genes were found, with no isolates carrying the see gene. The most prevalent genes detected were seg and sei, with some isolates containing up to five different SEs genes. These findings indicate the presence of enterotoxigenic staphylococci strains in the tested samples, emphasizing the importance of implementing proper sanitation and hygienic practices, utilizing safe raw materials, and ensuring adequate handling of finished products. Continued monitoring for the presence of SEs is necessary to ensure food safety and prevent intoxication.

Keywords: dairy products, milk, Staphylococci, enterotoxins, SE genes

Procedia PDF Downloads 42

Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

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A huge of dark color palm oil mill effluent (POME) cannot pass the discharge standard. It has been identified as the major contributor to the pollution load into ground water. Here, lignin-degrading yeast isolated from a termite nest was tested to treat the POME. Its lignin-degrading and decolorizing ability was determined. The result illustrated that Galactomyces sp. was successfully grown in POME. The decolorizing test demonstrated that 40% of Galactomyces sp. could reduce the color of POME (50% v/v) about 74-75% in 5 days without nutrient supplement. The result suggested that G. reessii has a potential to apply for decolorizing the dark wastewater like POME and other industrial wastewaters.

Keywords: decolorization, palm oil mill effluent, termite, yeast

Procedia PDF Downloads 183

Authors: Kidane workelul, Li xu, Xiaoyang Pang, Jiaping Lv

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Dairy products are exceptionally ideal media for the growth of microorganisms because of their high nutritional content. There are several ways that milk might get contaminated throughout the milking process, including how the raw milk is transported and stored, as well as how long it is kept before being processed. Psychrotrophic bacteria are among the one which can deteriorate the quality of milk mainly their heat resistance proteas and lipase enzyme. For this research purpose 8 selected strains of Psychrotrophic bacteria (Entrococcus hirae, Pseudomonas fluorescens, Pseudomonas azotoformans, Pseudomonas putida, Exiguobacterium indicum, Pseudomonas paralactice, Acinetobacter indicum, Serratia liquefacients)are chosen and try to determine their characteristics based on the research methodology protocol. Thus, the 8 selected strains are cultured, plated incubate, extracted their genomic DNA and genome DNA was amplified, the purpose of the study was to identify their Psychrotrophic properties, lipase hydrolysis positive test, their optimal incubation temperature, designed primer using the noble strain P,flourescens conserved region area in target with lipA gene, optimized primer specificity as well as sensitivity and PCR detection for lipase positive strains using the design primers. Based on the findings both the selected 8 strains isolated from stored raw milk are Psychrotrophic bacteria, 6 of the selected strains except the 2 strains are positive for lipase hydrolysis, their optimal temperature is 20 to 30 OC, the designed primer specificity is very accurate and amplifies for those strains only with lipase positive but could not amplify for the others. Thus, the result is promising and could help in detecting the Psychrotrophic bacteria producing heat resistance enzymes (lipase) at early stage before the milk is processed and this will safe production loss for the dairy industry.

Keywords: dairy industry, heat-resistant, lipA, milk, primer and psychrotrophic

Procedia PDF Downloads 22

Authors: Nacéra Benoussaid, Lehalali Meriem, Benchabane Messaoud

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Our work focuses on the production of compound antibiotic effect of volatile nature namely hydrogen cyanide and the production and identification of molecules phénazinique by some strains of fluorescent Pseudomonas spp isolated from the rhizosphere of some trees for a possible use as bio pesticides antifungal effect and/or antibiotic. We tested the production of hydrogen cyanide of 21 strains of Pseudomonas spp. fluorescent among them 19 strains (90, 47%) showed a positive cyanogenesis.The antagonism test executed in vitro showed that Pseudomonas strains have a higher anti fungal effect relative to their antibacterial effect with diameters of inhibition zones up to 3, 9 cm recorded by the strain F48 against Coleosporiumsp compared with recorded results against bacteria with a maximum inhibition of 1, 26 cm among this antagonistic strain.Three strains were selected by testing for producing phénazines namely PI9, BB9 and F20. The effect of the antimicrobial activity was performed on different culture media (GN, King B, ISP2 and PDA). The results of our study allowed us to retain the King B medium as ideal medium for the production of secondary metabolite. The produced phenazinique compounds was extracted from various organic solvents, and after the results of antibiographie against germs - targets, the extracts of ethyl acetate gave the best results compared to dichloromethane and hexane.The Analysis of these compounds of antibiotic phenazinique effect within layer chromatography (CCM) and high performance liquid chromatography( HPLC) indicate that both strains PI9 and F20 are productive of phenazine-1-carboxylic acid (PCA). The BB9 strain is suspected to be productive of another phenazinique compound.

Keywords: Pseudomonas ssp. fluorescents, antagonism in vitro, secondary metabolite, phenazines, biopesticide.

Procedia PDF Downloads 488

Authors: Mohamed M. Bumadian, D. James Gilmour

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Three bacterial species were isolated from the River Wye (Derbyshire, England) and identified using 16S rRNA gene sequencing as Serratia proteamaculans, Aeromonas hydrophila and Klebsiella pneumoniae. Respiration rates of the strains were measured in order to determine the metabolic activity under salt stress. The highest respiration rates of all three strains were found at 0.17 M and 0.5 M NaCl and then the respiration rate decreased with increasing concentrations of NaCl. In addition, the effect of increasing concentrations of NaCl on malate dehydrogenase activity was determined using cell-free extracts of the three strains. Malate dehydrogenase activity was stimulated at NaCl concentrations up to 0.5 M, and a small level of activity remained even at 3.5 M NaCl. The pH optimum of the malate dehydrogenase in cell-free extracts of all strains was higher than pH 7.5.

Keywords: fresh water, halotolerant pathogenic bacteria, 16S rRNA gene, cell-free extracts, respiration rates, malate dehydrogenase

Procedia PDF Downloads 430

Authors: Rupa Rani, Vipin Kumar

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Endosulfan, an organochlorine pesticide is of environmental concern due to its apparent persistence and toxicity. It has been reported as contaminants in soil, air, and water and is bioaccumulated and magnified in ecosystems. The combined use of microorganisms and plants has great potential for remediating soil contaminated with organic compounds such as pesticides. The objective of this study was to evaluate whether the bacterial inoculation influences plant growth promotion, endosulfan degradation in soil and endosulfan accumulation in different plant parts. Lycopersicon esculentum L. (Tomato) was grown in endosulfan spiked soil and inoculated with endosulfan tolerant bacterial strains. Endosulfan residues from different parts of plants and soil were extracted and estimated by using gas chromatograph equipped with 63Ni electron capture detector (GC-ECD). The inoculation of bacterial strains into the soil with plants showed a beneficial effect on endosulfan degradation and plant biomass production. Maximum endosulfan (90%) degradation was observed after 120 days of bacterial inoculation in the soil. Furthermore, there was significantly less endosulfan accumulation in roots and shoots of bacterial strains inoculated plants as compared to uninoculated plants. The results show the effectiveness of inoculated endosulfan tolerant bacterial strains to increase the remediation of endosulfan contaminated soil.

Keywords: organochlorine pesticides, endosulfan, degradation, plant-bacteria partnerships

Procedia PDF Downloads 123

Authors: Meenakshi Chaudhary, V .S. Randhawa, M. Jais, R. Dutta

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Introduction: An outbreak of Multi drug resistant S. Typhi (i.e. resistance to chloramphenicol, ampicillin, and trimethoprim-sulfamethoxazole) occurred in 1990's in India which peaked in 1992-93 and resulted in the change of drug of choice from chloramphenicol to ciprofloxacin for enteric fever. Currently an emergence of Ciprofloxacin susceptible S. Typhi isolates in the region is being reported which appears to be chromosomally mediated. Methodology: Six hundred sixty four strains were randomly selected from the time period between January 2008-December 2011 at the National Salmonella Phage Typing Centre, LHMC, New Delhi. The strains were representative of the north, central and south zones of India. All isolates were subjected to serotyping, biotyping, phage typing and then to antimicrobial susceptibility testing by CLSI disk diffusion (CLSI) technique to Ciprofloxacin, Cefotaxime, Ampicillin, Chloramphenicol, Trimethoprim-Sulfomethoxazole and Tetracycline. Subsequently MIC of the isolates was determined by E-test (AB-Biodisc). Results: More than 80% of the tested strains had intermediate susceptibility to ciprofloxacin. The E test revealed the MIC (Ciprofloxacin) of these strains to be in the range of 0.12 to 0.5 µg/ml. Sixty nine percent of ciprofloxacin intermediate susceptible strains belonged to Phage type E1 and fourteen percent of these were Vi- Negative i.e these could not be typed by the phage typing scheme of Craigie and Yen. All the strains remained susceptible to cefotaxime. Conclusion: Predominant isolation of intermediate susceptible S. Typhi strains from India would alter the recommendations of empiric treatment of enteric fever in the region. Alternative to the low cost ciprofloxacin will have to be sought or increased dosage and/or duration of ciprofloxacin will have to be recommended. The reasons for the trend of increase in percentage of intermediate susceptible S. Typhi strains are not clear but may be attributed partly to the revision of CLSI guidelines in 2013.

Keywords: salmonella typhi, decreased ciprofloxacin susceptibility, ciprofloxacin, minimum inhibitory concentration

Procedia PDF Downloads 281

Authors: C. J. Tien, C. S. Chen, S. F. Huang, Z. X. Wang

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Algae in soil ecosystems can produce organic matters and oxygen by photosynthesis. Heterocyst-forming cyanobacteria can fix nitrogen to increase soil nitrogen contents. Secretion of mucilage by some algae increases the soil water content and soil aggregation. These actions will improve soil quality and fertility, and further increase abundance and diversity of soil microorganisms. In addition, some mixotrophic and heterotrophic algae are able to degrade petroleum hydrocarbons. Therefore, the objectives of this study were to analyze the effects of algal addition on the degradation of total petroleum hydrocarbons (TPH), diversity and activity of bacteria and algae in the diesel-contaminated soil under different nutrient contents and frequency of plowing and irrigation in order to assess the potential bioremediation technique using edaphic algae. The known amount of diesel was added into the farmland soil. This diesel-contaminated soil was subject to five settings, experiment-1 with algal addition by plowing and irrigation every two weeks, experiment-2 with algal addition by plowing and irrigation every four weeks, experiment-3 with algal and nutrient addition by plowing and irrigation every two weeks, experiment-4 with algal and nutrient addition by plowing and irrigation every four weeks, and the control without algal addition. Soil samples were taken every two weeks to analyze TPH concentrations, diversity of bacteria and algae, and catabolic genes encoding functional degrading enzymes. The results show that the TPH removal rates of five settings after the two-month experimental period were in the order: experiment-2 > expermient-4 > experiment-3 > experiment-1 > control. It indicated that algal addition enhanced the degradation of TPH in the diesel-contaminated soil, but not for nutrient addition. Plowing and irrigation every four weeks resulted in more TPH removal than that every two weeks. The banding patterns of denaturing gradient gel electrophoresis (DGGE) revealed an increase in diversity of bacteria and algae after algal addition. Three petroleum hydrocarbon-degrading algae (Anabaena sp., Oscillatoria sp. and Nostoc sp.) and two added algal strains (Leptolyngbya sp. and Synechococcus sp.) were sequenced from DGGE prominent bands. The four hydrocarbon-degrading bacteria Gordonia sp., Mycobacterium sp., Rodococcus sp. and Alcanivorax sp. were abundant in the treated soils. These results suggested that growth of indigenous bacteria and algae were improved after adding edaphic algae. Real-time polymerase chain reaction results showed that relative amounts of four catabolic genes encoding catechol 2, 3-dioxygenase, toluene monooxygenase, xylene monooxygenase and phenol monooxygenase were appeared and expressed in the treated soil. The addition of algae increased the expression of these genes at the end of experiments to biodegrade petroleum hydrocarbons. This study demonstrated that edaphic algae were suitable biomaterials for bioremediating diesel-contaminated soils with plowing and irrigation every four weeks.

Keywords: catabolic gene, diesel, diversity, edaphic algae

Procedia PDF Downloads 243