Search results for: bacterial inactivation

Authors: M. Cynthya, V. Prabhawathi, D. Mukesh

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Food contamination occurs during post process handling. This leads to spoilage and growth of pathogenic microorganisms in the food, thereby reducing its shelf life or spreading of food borne diseases. Several methods are tried and one of which is use of antimicrobial packaging. Here, papain, a protease enzyme, is covalently immobilized with the help of glutarldehyde on polyurethane and used as a food wrap to protect food from microbial contamination. Covalent immobilization of papain was achieved at a pH of 7.4; temperature of 4°C; glutaraldehyde concentration of 0.5%; incubation time of 24 h; and 50 mg of papain. The formation of -C=N- observed in the Fourier transform infrared spectrum confirmed the immobilization of the enzyme on the polymer. Immobilized enzyme retained higher activity than the native free enzyme. The efficacy of this was studied by wrapping it over S. aureus contaminated cottage cheese (paneer) and cheese and stored at a temperature of 4°C for 7 days. The modified film reduced the bacterial contamination by eight folds when compared to the bare film. FTIR also indicates reduction in lipids, sugars and proteins in the biofilm.

Keywords: cheese, papain, polyurethane, Staphylococcus aureus

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Authors: Zanele D. Ngwenya, Mustapha Mohammed, Felix D. Dakora

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Legumes are a major source of biological nitrogen, and therefore play a crucial role in maintaining soil productivity in smallholder agriculture in southern Africa. Through their ability to fix atmospheric nitrogen in root nodules, legumes are a better option for sustainable nitrogen supply in cropping systems than chemical fertilisers. For decades, farmers have been highly receptive to the use of rhizobial inoculants as a source of nitrogen due mainly to the availability of elite rhizobial strains at a much lower compared to chemical fertilisers. To improve the efficiency of the legume-rhizobia symbiosis in African soils would require the use of highly effective rhizobia capable of nodulating a wide range of host plants. This study assessed the morphogenetic diversity, photosynthetic functioning and relative symbiotic effectiveness (RSE) of groundnut, jack bean and soybean microsymbionts in Eswatini soils as a first step to identifying superior isolates for inoculant production. According to the manufacturer's instructions, rhizobial isolates were cultured in yeast-mannitol (YM) broth until the late log phase and the bacterial genomic DNA was extracted using GenElute bacterial genomic DNA kit. The extracted DNA was subjected to enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR) and a dendrogram constructed from the band patterns to assess rhizobial diversity. To assess the N2-fixing efficiency of the authenticated rhizobia, photosynthetic rates (A), stomatal conductance (gs), and transpiration rates (E) were measured at flowering for plants inoculated with the test isolates. The plants were then harvested for nodulation assessment and measurement of plant growth as shoot biomass. The results of ERIC-PCR fingerprinting revealed the presence of high genetic diversity among the microsymbionts nodulating each of the three test legumes, with many of them showing less than 70% ERIC-PCR relatedness. The dendrogram generated from ERIC-PCR profiles grouped the groundnut isolates into 5 major clusters, while the jack bean and soybean isolates were grouped into 6 and 7 major clusters, respectively. Furthermore, the isolates also elicited variable nodule number per plant, nodule dry matter, shoot biomass and photosynthetic rates in their respective host plants under glasshouse conditions. Of the groundnut isolates tested, 38% recorded high relative symbiotic effectiveness (RSE >80), while 55% of the jack bean isolates and 93% of the soybean isolates recorded high RSE (>80) compared to the commercial Bradyrhizobium strains. About 13%, 27% and 83% of the top N₂-fixing groundnut, jack bean and soybean isolates, respectively, elicited much higher relative symbiotic efficiency (RSE) than the commercial strain, suggesting their potential for use in inoculant production after field testing. There was a tendency for both low and high N₂-fixing isolates to group together in the dendrogram from ERIC-PCR profiles, which suggests that RSE can differ significantly among closely related microsymbionts.

Keywords: genetic diversity, relative symbiotic effectiveness, inoculant, N₂-fixing

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Authors: Angelica Alvarez-Lee, Sergio Martinez-Diaz, Jose Luis Garcia-Corona, Humberto Lanz-Mendoza

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World aquaculture is always looking for improvements to achieve productions with high yields avoiding the infection by pathogenic agents. The best way to achieve this is to know the biological model to create alternative treatments that could be applied in the hatcheries, which results in greater economic gains and improvements in human public health. In the last decade, immunomodulation in shrimp culture with probiotics, organic acids and different carbon sources has gained great interest, mainly in larval and juvenile stages. Immune priming is associated with a strong protective effect against a later pathogen challenge. This work provides another perspective about immunostimulation from spawning until hatching. The stimulation happens during development embryos and generates resistance to infection by pathogenic bacteria. Massive spawnings of white shrimp L. vannamei were obtained and placed in experimental units with 700 mL of sterile seawater at 30 °C, salinity of 28 ppm and continuous aeration at a density of 8 embryos.mL⁻¹. The immunostimulating effect of three death strains of non-pathogenic bacterial (Escherichia coli, Staphylococcus aureus and Bacillus subtilis) and a pathogenic strain for white shrimp (Vibrio parahaemolyticus) was evaluated. The strains killed by heat were adjusted to O.D. 0.5, at A 600 nm, and directly added to the seawater of each unit at a ratio of 1/100 (v/v). A control group of embryos without inoculum of dead bacteria was kept under the same physicochemical conditions as the rest of the treatments throughout the experiment and used as reference. The duration of the stimulus was 12 hours, then, the larvae that hatched were collected, counted and transferred to a new experimental unit (same physicochemical conditions but at a salinity of 28 ppm) to carry out a challenge of infection against the pathogen V. parahaemolyticus, adding directly to seawater an amount 1/100 (v/v) of the live strain adjusted to an OD 0.5; at A 600 nm. Subsequently, 24 hrs after infection, nauplii survival was evaluated. The results of this work shows that, after 24 hrs, the hatching rates of immunostimulated shrimp embryos with the dead strains of B. subtillis and V. parahaemolyticus are significantly higher compared to the rest of the treatments and the control. Furthermore, survival of L. vanammei after a challenge of infection of 24 hrs against the live strain of V. parahaemolyticus is greater (P < 0.05) in the larvae immunostimulated during the embryonic development with the dead strains B. subtillis and V. parahaemolyticus, followed by those that were treated with E. coli. In summary superficial antigens can stimulate the development cells to promote hatching and can have normal development in agreeing with the optical observations, plus exist a differential response effect between each treatment post-infection. This research provides evidence of the immunostimulant effect of death pathogenic and non-pathogenic bacterial strains in the rate of hatching and oversight of shrimp L. vannamei during embryonic and larval development. This research continues evaluating the effect of these death strains on the expression of genes related to the defense priming in larvae of L. vannamei that come from massive spawning in hatcheries before and after the infection challenge against V. parahaemolyticus.

Keywords: immunostimulation, L. vannamei, hatching, survival

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Authors: Ali Bagherpour, Masoud Soltanialvar

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Campylobacter species are common bacterial pathogens associated with human gastroenteritis which are generally transmitted through foods of animal origin. This study was carried out to determine the prevalence of Campylobacter species in poultry meat and by products in the city of Dezful in Iran. Since April 2012 to July 2013, a total of 400 samples including meat (n = 100), liver (n = 100), gizzard (n = 100), and poultry heart (n = 100), were randomly collected from Dezful industrial poultry abattoir and were experimented in order to investigate presence of Campylobacter species. According to culture test, 251 samples out of 400 samples under study (69%) were contaminated with Campylobacter species. The highest prevalence of Campylobacter species was observed in poultry's liver (78.3%) and then in gizzard (75.8%), heart (65%) and meat (56.7%). The most common isolated Campylobacter were C. jejuni (90.9%) and the rest were C. coli (9.1%). There was a significant difference (P < 0.05) in the prevalence of Campylobacter species between the meat samples taken in the summer (86.7%). The results of this study indicate the importance of edible offal of poultries as the potential source of Campylobacter infections.

Keywords: Campylobacter jejuni, Campylobacter coli, poultry, meat, products

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Authors: Aminu Muhammad, Mustapha Ya’u, Hasnah Mohd Sirat

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An investigation of the chemical constituents into the stem bark of Ficus platyphylla (Moraceae) has resulted in the isolation of hordenine, epicatechin, lupeol, lupeol acetate and α-amyrin acetate. Their structures were determined using spectroscopic data as well as comparison with literature data. The antibacterial assay has been tested against Gram positive and Gram negative bacteria, while the tyrosinase inhibition assay was examined using L-Dopa as a substrate of mushroom tyrosinase enzyme. hordenine, epicatechin, lupeol, lupeol acetate and α-amyrin acetate showed minimum inhibition concentration (MIC) values in the range of 225-900 µg/mL against the bacterial strains. Lupeol, lupeol acetate and α-amyrin acetate showed significant antityrosinase activity against mushroom tyrosinase enzyme with percent inhibition of 67.7%, 66.2% and 62.2%, respectively.

Keywords: antibacterial, antityrosinase, chemical constituents, Ficus platyphylla

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Authors: K. Premakumar, D. S. Rushani, H. N. Hettiarachchi

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The production and consumption of preserved foods have gained much importance due to globalization, and they provide a health benefit apart from the basic nutritional functions. Therefore, a study was conducted to develop a jam incorporating carrot, banana, and lime. Considering the findings of several preliminary studies, five formulations of the jam were prepared by blending different percentages of carrot and banana including control (where the only carrot was added). The freshly prepared formulations were subjected to physicochemical and sensory analysis.Physico-Chemical parameters such as pH, TSS, titrable acidity, ascorbic acid content, total sugar and non-reducing sugar and organoleptic qualities such as colour, aroma, taste, spread ability and overall acceptability and microbial analysis (total plate count) were analyzed after formulations. Physico-Chemical Analysis of the freshly prepared Carrot –Banana Blend jam showed increasing trend in titrable acidity (from 0.8 to 0.96, as % of citric acid), TSS (from 70.05 to 67.5 0Brix), ascorbic acid content (from 0.83 to 11.465 mg/100ml), reducing sugar (from 15.64 to 20.553%) with increase in carrot pulp from 50 to 100%. pH, total sugar, and non-reducing sugar were also reduced when carrot concentration is increased. Five points hedonic scale was used to evaluate the organoleptic characters. According to Duncan's Multiple Range Test, the mean scores for all the assessed sensory characters varied significantly (p<0.05) in the freshly made carrot-banana blend jam formulations. Based on the physicochemical and sensory analysis, the most preferred carrot: banana combinations of 50:50, 100:0 and 80:20 (T1, T2, and T5) were selected for storage studies.The formulations were stored at 300 °C room temperature and 70-75% of RH for 12 weeks. The physicochemical characteristics were measured at two weeks interval during storage. The decreasing trends in pH and ascorbic acid and an increasing trend in TSS, titrable acidity, total sugar, reducing sugar and non-reducing sugar were noted with advancement of storage periods of 12 weeks. The results of the chemical analysis showed that there were significance differences (p<0.05) between the tested formulations. Sensory evaluation was done for carrot –banana blends jam after a period of 12 weeks through a panel of 16 semi-trained panelists. The sensory analysis showed that there were significant differences (p<0.05) for organoleptic characters between carrot-banana blend jam formulations. The highest overall acceptability was observed in formulation with 80% carrot and 20% banana pulp. Microbiological Analysis was carried out on the day of preparation, 1 month, 2 months and 3 months after preparation. No bacterial growth was observed in the freshly made carrot -banana blend jam. There were no counts of yeast and moulds and coliforms in all treatments after the heat treatments and during the storage period. Only the bacterial counts (Total Plate Counts) were observed after three months of storage below the critical level, and all formulations were microbiologically safe for consumption. Based on the results of physio-chemical characteristics, sensory attributes, and microbial test, the carrot –banana blend jam with 80% carrot and 20% banana (T2) was selected as best formulation and could be stored up to 12 weeks without any significant changes in the quality characteristics.

Keywords: formulations, physicochemical parameters, microbiological analysis, sensory evaluation

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Authors: Abdullah M. Almeshal, Mohammad R. Alenezi, Muhammad Moaz

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In this paper, we discuss the performance of applying hybrid spiral dynamic bacterial chemotaxis (HSDBC) optimisation algorithm on an intelligent controller for a differential drive robot. A unicycle class of differential drive robot is utilised to serve as a basis application to evaluate the performance of the HSDBC algorithm. A hybrid fuzzy logic controller is developed and implemented for the unicycle robot to follow a predefined trajectory. Trajectories of various frictional profiles and levels were simulated to evaluate the performance of the robot at different operating conditions. Controller gains and scaling factors were optimised using HSDBC and the performance is evaluated in comparison to previously adopted optimisation algorithms. The HSDBC has proven its feasibility in achieving a faster convergence toward the optimal gains and resulted in a superior performance.

Keywords: differential drive robot, hybrid fuzzy controller, optimization, path tracking, unicycle robot

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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: Mari Jaakkola, Jasmiina Haverinen, Tiina Tolonen, Vesa Virtanen

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These process monitoring of biogas plant gives valuable information of the function of the process and help to maintain a stable process. The costs of basic monitoring are often much lower than the costs associated with re-establishing a biologically destabilised plant. Reactor acidification through reactor overload is one of the most common reasons for process deterioration in anaerobic digesters. This occurs because of a build-up of volatile fatty acids (VFAs) produced by acidogenic and acetogenic bacteria. VFAs cause pH values to decrease, and result in toxic conditions in the reactor. Ammonia ensures an adequate supply of nitrogen as a nutrient substance for anaerobic biomass and increases system's buffer capacity, counteracting acidification lead by VFA production. However, elevated ammonia concentration is detrimental to the process due to its toxic effect. VFAs are considered the most reliable analytes for process monitoring. To obtain accurate results, sample storage and transportation need to be carefully controlled. This may be a challenge for off-line laboratory analyses especially when the plant is located far away from the laboratory. The aim of this study was to investigate the correlation between fatty acids, ammonium, and bacteria in the anaerobic digestion samples obtained from an industrial biogas factory. The stability of the analytes was studied comparing the results of the on-site analyses performed in the factory site to the results of the samples stored at room temperature and -18°C (up to 30 days) after sampling. Samples were collected in the biogas plant consisting of three separate mesofilic AD reactors (4000 m³ each) where the main feedstock was swine slurry together with a complex mixture of agricultural plant and animal wastes. Individual VFAs, ammonium, and nutrients (K, Ca, Mg) were studied by capillary electrophoresis (CE). Longer chain fatty acids (oleic, hexadecanoic, and stearic acids) and bacterial profiles were studied by GC-MSD (Gas Chromatography-Mass Selective Detector) and 16S rDNA, respectively. On-site monitoring of the analytes was performed by CE. The main VFA in all samples was acetic acid. However, in one reactor sample elevated levels of several individual VFAs and long chain fatty acids were detected. Also bacterial profile of this sample differed from the profiles of other samples. Acetic acid decomposed fast when the sample was stored in a room temperature. All analytes were stable when stored in a freezer. Ammonium was stable even at a room temperature for the whole testing period. One reactor sample had higher concentration of VFAs and long chain fatty acids than other samples. CE was utilized successfully in the on-site analysis of separate VFAs and NH₄ in the biogas production site. Samples should be analysed in the sampling day if stored in RT or freezed for longer storage time. Fermentation reject can be stored (and transported) at ambient temperature at least for one month without loss of NH₄. This gives flexibility to the logistic solutions when reject is used as a fertilizer.

Keywords: anaerobic digestion, capillary electrophoresis, ammonium, bacteria

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Authors: Lekha Logu

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The coordination behavior of the newly synthesized Schiff base ligands, 4-bromo-2-((p-tolyl imino) methyl) phenol obtained by condensing para-toluidine with 5-bromo salicylaldehyde and N-(3,4-dichloro benzylidene)-4-methylbenzenamine obtained by condensing Para-toluidine with 3,4-dichloro benzaldehyde in ethanolic medium has been explored in this current study. The synthesized Schiff’s base ligands were complexed with lanthanide nitrate salts yielding [LnL(NO3)2(H2O)2]NO3, (Ln=Pr, Sm). Elemental analysis, conductance measurement, and spectral techniques like Nuclear Magnetic Resonance (NMR), Ultraviolet-visible (UV-Vis) and Fourier Transform Infrared (FTIR) have been used to characterize Schiff’s base ligands and their lanthanide metal complexes. An attempt has been made on these complexes for their antimicrobial activity against the gram-positive and gram-negative bacterial species like Escherichia coli, Staphylococcus aureus, Bacillus subtilis, Klebsiella pneumonia and fungal species like Canadida and Aspergillus.

Keywords: lanthanide complexes, Schiff's base, antimicrobial assay, synthesis, characterization

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

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

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

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Authors: Ayodele Odularu, Peter Ajibade, Albert Bolhuis

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This study aimed at assessing the antibacterial activities of four zinc (II) complexes. Zinc (II) complexes of nitrogen, oxygen and sulfur coordination modes were synthesized using direct substitution reaction. The characterization techniques involved physicochemical properties (molar conductivity) and spectroscopic techniques. The molar conductivity gave the non-electrolytic nature of zinc (II) complexes. The spectral studies of zinc (II) complexes were done using electronic spectra (UV-Vis) and Fourier Transform Infra-red Spectroscopy (FT-IR). Spectral data from the spectroscopic studies confirmed the coordination of the mixed ligands with zinc (II) ion. The antibacterial activities of zinc(II) complexes of were all in supportive of Overtone’s concept and Tweedy’s theory of chelation for bacterial strains of S. aureus MRSA252 and E coli MC4100 because the zones of inhibition were greater than the corresponding ligands. In summary, all zinc (II) complexes of ZEPY, ZE1PH, ZE1PY and ZE135PY all have potentials for antibacterial activities.

Keywords: antibacterial activities, spectral studies, syntheses, zinc(II) complexes

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Authors: Andrey A. Tyuftin, David Clarke, Malco C. Cruz-Romero, Declan Bolton, Seamus Fanning, Shashi K. Pankaj, Carmen Bueno-Ferrer, Patrick J. Cullen, Joe P. Kerry

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Prolonging of shelf-life is essential in order to address issues such as; supplier demands across continents, economical profit, customer satisfaction, and reduction of food wastage. Smart packaging solutions presented in the form of naturally occurred antimicrobially-active packaging may be a solution to these and other issues. Gelatin film forming solution with adding of natural sourced antimicrobials is a promising tool for the active smart packaging. The objective of this study was to coat conventional plastic hydrophobic packaging material with hydrophilic antimicrobial active beef gelatin coating and conduct shelf life trials on beef sub-primal cuts. Minimal inhibition concentration (MIC) of Caprylic acid sodium salt (SO) and commercially available Auranta FV (AFV) (bitter oranges extract with mixture of nutritive organic acids) were found of 1 and 1.5 % respectively against bacterial strains Bacillus cereus, Pseudomonas fluorescens, Escherichia coli, Staphylococcus aureus and aerobic and anaerobic beef microflora. Therefore SO or AFV were incorporated in beef gelatin film forming solution in concentration of two times of MIC which was coated on a conventional plastic LDPE/PA film on the inner cold plasma treated polyethylene surface. Beef samples were vacuum packed in this material and stored under chilling conditions, sampled at weekly intervals during 42 days shelf life study. No significant differences (p < 0.05) in the cook loss was observed among the different treatments compared to control samples until the day 29. Only for AFV coated beef sample it was 3% higher (37.3%) than the control (34.4 %) on the day 36. It was found antimicrobial films did not protect beef against discoloration. SO containing packages significantly (p < 0.05) reduced Total viable bacterial counts (TVC) compared to the control and AFV samples until the day 35. No significant reduction in TVC was observed between SO and AFV films on the day 42 but a significant difference was observed compared to control samples with a 1.40 log of bacteria reduction on the day 42. AFV films significantly (p < 0.05) reduced TVC compared to control samples from the day 14 until the day 42. Control samples reached the set value of 7 log CFU/g on day 27 of testing, AFV films did not reach this set limit until day 35 and SO films until day 42 of testing. The antimicrobial AFV and SO coated films significantly prolonged the shelf-life of beef steaks by 33 or 55% (on 7 and 14 days respectively) compared to control film samples. It is concluded antimicrobial coated films were successfully developed by coating the inner polyethylene layer of conventional LDPE/PA laminated films after plasma surface treatment. The results indicated that the use of antimicrobial active packaging coated with SO or AFV increased significantly (p < 0.05) the shelf life of the beef sub-primal. Overall, AFV or SO containing gelatin coatings have the potential of being used as effective antimicrobials for active packaging applications for muscle-based food products.

Keywords: active packaging, antimicrobials, edible coatings, food packaging, gelatin films, meat science

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Authors: Asmiaty Sahur, Ambo Ala, Baharuddin Patanjengi, Elkawakib Syam'un

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This study aimed to isolate and characterize the indigenous Rhizosphere bacteria producing Gibberellin Acid as plant growth isolated from local soybean of three different areas in South Sulawesi, Indonesia. Several soil samples of soybean plants were collected from the Rhizosphere of local soybeans in three different areas of South Sulawesi such as Soppeng, Bone and Takalar. There were 56 isolates of bacteria were isolated and grouped into gram-positive bacteria and gram negative bacteria .There are 35 isolates produce a thick slime or slimy when cultured on media Natrium Broth and the remaining of those produced spores. The results showed that of potential bacterial isolated produced Gibberellin Acid in high concentration. The best isolate of Rhizosphere bacteria for the production of Gibberellin Acid is with concentration 2%. There are 4 isolates that had higher concentration are AKB 19 (4.67 mg/ml) followed by RKS 17 (3.80 mg/ml), RKS 25 (3.70 mg / ml) and RKS 24 (3.29 mg/ml) respectively.

Keywords: rhizosphere, bacteria, gibberellin acid, soybeans

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Authors: M. Khallaf, R. Khalil, H. Ghetas

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In the present study, V. harveyi and V. alginolyticus were isolated from cultured seabass and seabream at Damietta Governorate, Egypt, during summer season. Isolates were biochemically and molecularly identified using primers for Vhh and Collagenase genes. The most prominent clinical observations of diseased fish were exophthalmia, abdominal distension, and multifocal cutaneous hemorrhagic ulceration on the dorsal musculature and caudal peduncle. Physicochemical characteristics of water samples indicated that the unionized ammonia, nitrate, and hydrogen sulphate concentrations were higher than the acceptable limits. Heavy metals concentrations in water samples exhibited higher concentrations than the permissible levels for fish culture, which was considered as chemical stressors that increase the prevalence of these bacterial diseases. Immune parameters were lower in diseased seabass and seabream than apparently healthy fish. Lesions of different fish organs were identified histopathologically.

Keywords: seabass, seabream, Vibrio alginolyticus, Vibrio harveyi

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Authors: Julius Lenaerts, Ahmed Elsaadawy, Mohammed Bashir

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Background Sedated and paralyzed patients have an impaired blink reflex leading to ophthalmic complications such as conjunctivitis, epithelial defects, bacterial keratitis, and more. These are entirely preventable complications through regular eye care. Methods Patients at level 3 or above (intubated/paralyzed) care in the Intensive Care Unit (ICU) were reviewed between February and April. Data was pulled from Metavision and adherence was compared to Royal College of Ophthalmology (RCOphth) recommendations[4]. Using a multi-pronged approach through posters, individual teaching sessions and faculty teaching, we aimed to educate staff about eye care in the ICU. Patients were reaudited in the period July to August. Results Out of 40 patients, only 23% were assessed for eye care needs on admission compared to 77% after teaching; eye care was only delivered 59% of the time it was due, compared to 61%; 2.5% of patients had eyedrops prescribed compared to 41%. This shows an overall increase in meeting RCOphth standards. Key messages Eye care is an overlooked aspect of patient care in the ICU, associated with avoidable ocular complications. Healthcare staff need further rigorous education on the provision and importance of eye care to reduce avoidable complications.

Keywords: ICU, eye care, risk, QIP

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Authors: Taru Singh, Shukla Das, V. G. Ramachandran

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Objectives: To develop SYBR green based real-time PCR assay to detect carbapenemases (NDM, IMP) genes in E. coli. Methods: A total of 40 E. coli from stool samples were tested. Six were previously characterized as resistant to carbapenems and documented by PCR. The remaining 34 isolates previously tested susceptible to carbapenems and were negative for these genes. Bacterial RNA was extracted using manual method. The real-time PCR was performed using the Light Cycler III 480 instrument (Roche) and specific primers for each carbapenemase target were used. Results: Each one of the two carbapenemase gene tested presented a different melting curve after PCR amplification. The melting temperature (Tm) analysis of the amplicons identified was as follows: blaIMP type (Tm 82.18°C), blaNDM-1 (Tm 78.8°C). No amplification was detected among the negative samples. The results showed 100% concordance with the genotypes previously identified. Conclusions: The new assay was able to detect the presence of two different carbapenemase gene type by real-time PCR.

Keywords: resistance, b-lactamases, E. coli, real-time PCR

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Authors: Kusno Kamil, Andi Asni, Sungkono

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According to a report of the World Food and Agriculture Agency (FAO): the post-harvest fish losses in Indonesia reaches 30 percent from 170 trillion rupiahs of marine fisheries reserves, then the potential loss reaches 51 trillion rupiahs (end of 2016 data). This condition is caused by traditionally vulnerable fish catches damaged due to disruption of the cold chain of preservation. The physical and chemical changes in fish flesh increase rapidly, especially if exposed to the scorching heat in the middle of the sea, exacerbated by the low awareness of catch hygiene; many unclean catches which contain blood are often treated without special attention and mixed with freshly caught fish, thereby increasing the potential for faster fish spoilage. This background encourages research on traditional fisherman catch preservation methods that aim to find the best and most affordable methods and/or combinations of fish preservation methods so that they can help fishermen increase their fishing duration without worrying that their catch will be damaged, thereby reducing their economic value when returning to the beach to sell their catches. This goal is expected to be achieved through experimental methods of treatment of fresh fish catches in containers with the addition of anti-bacterial copper, liquid smoke solution, and the use of vacuum containers. The other three treatments combined the three previous treatment variables with an electrically powered cooler (temperature 0~4 ᵒC). As a control specimen, the untreated fresh fish (placed in the open air and in the refrigerator) were also prepared for comparison for 1, 3, and 6 days. To test the level of freshness of fish for each treatment, physical observations were used, which were complemented by tests for bacterial content in a trusted laboratory. The content of copper (Cu) in fish meat (which is suspected of having a negative impact on consumers) was also part of the examination on the 6th day of experimentation. The results of physical observations on the test specimens (organoleptic method) showed that preservation assisted by the use of coolers was still better for all treatment variables. The specimens, without cooling, sequentially showed that the best preservation effectiveness was the addition of copper plates, the use of vacuum containers, and then liquid smoke immersion. Especially for liquid smoke, soaking for 6 days of preservation makes the fish meat soft and easy to crumble, even though it doesn't have a bad odor. The visual observation was then complemented by the results of testing the amount of growth (or retardation) of putrefactive bacteria in each treatment of test specimens within similar observation periods. Laboratory measurements report that the minimum amount of putrefactive bacteria achieved by preservation treatment combining cooler with liquid smoke (sample A+), then cooler only (D+), copper layer inside cooler (B+), vacuum container inside cooler (C+), respectively. Other treatments in open air produced a hundred times more putrefactive bacteria. In addition, treatment of the copper layer contaminated the preserved fresh fish more than a thousand times bigger compared to the initial amount, from 0.69 to 1241.68 µg/g.

Keywords: fish, preservation, traditional, fishermen, boat

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Authors: Richard K. Jolley, Jonathan A. Coffman

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Since Enterococci has been implicated in oral disease, we hypothesized the transmission of avian Enterococci to humans via fecal-oral transmission facilitated by adherence to dental plaque. To demonstrate the capability of Enterococci to bind to a dental plaque we filtered avian excreta and incubated the filtrate on a sucrose-induced, Streptococcus mutans biofilm. The biofilm was washed several times with a detergent to remove bacteria binding non-specifically to the biofilm, DNA was isolated from the biofilm, 16S rDNA was amplified, sequenced by Ion Torrent DNA sequencing and analyzed with bioinformatics. Enterococci and other known bacterial pathogens were shown to adhere to the biofilm. Culturing the washed biofilm with Bile Esculin Azide (BEA) agar also confirmed the presence of Enterococci as verified with Sanger sequencing. The results suggest that Enteroccoci in avian excreta has the ability to adhere to human dental plaque and may be a mechanism of entry when humans encounter contaminated aerosols, water or food.

Keywords: Enterococci, avian excreta, dental plaque, NGS

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Authors: Bo Ram Choi, Ji Su Kim, Juyeon Cho, Hyukjin Lee

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Food contaminated by bacteria (E.coli), causes food poisoning, which occurs to many patients worldwide annually. We have introduced an application of rolling circle amplification (RCA) as a versatile biosensor and developed a diagnostic platform composed of capillary tube and microbeads for rapid and easy detection of Escherichia coli (E. coli). When specific mRNA of E.coli is extracted from cell lysis, rolling circle amplification (RCA) of DNA template can be achieved and can be visualized by beads aggregation in capillary tube. In contrast, if there is no bacterial pathogen in sample, no beads aggregation can be seen. This assay is possible to detect visually target gene without specific equipment. It is likely to the development of a genetic kit for point of care testing (POCT) that can detect target gene using microbeads.

Keywords: rolling circle amplification (RCA), Escherichia coli (E. coli), point of care testing (POCT), beads aggregation, capillary tube

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Authors: Maimuna Nontji

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The life cycle of rice plant has three phases of growth; they are the vegetative, reproductive and maturation phase. They greatly affect the life of dynamics metanotrof bacterial as reducer methane emissions in the rice field, both of population and on the rate of growth. The aim of this study was to analyze the population and growth rate of methanotrof isolates which has been isolated in previous studies. Isolates were taken at all the life cycle of rice plant. Population of analysis was conducted by standard plate count method and growth rate was analysed by logarithmic calculation. The results showed that each isolate varied in population and growth rate. The highest population was obtained in the isolates Gowa Methanotrof Reproductive (GMR 8) about 7.06 x 10 11 cfu / ml on 3 days of incubation and the lowest population was obtained in the Gowa Methanotrof Maturation (GMP 5) about 0.27 x 10 11 cfu / ml on 7 day of incubation. Some isolate were demonstrated in long growth rate about 5 days of incubation and another are 3 days.

Keywords: emission, methanotrof, methane, population

Procedia PDF Downloads 433

Authors: W. Mazari, K. Boucherit, Z. Boucherit-Otmani, M. N. Rahmoun, M. Benabdallah

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The hospital or any other establishment of care can be considered as an ecosystem where the patient comes into contact with a frightening microbial universe and a risk to contract infection that is referred to as nosocomial or health care-associated. In these last years, the incidence of these infections has risen sharply. Several microorganisms are the cause of these nosocomial infections and the emergence of resistance of the microbial strains against antibiotics creates a danger to public health. The search for new antimicrobial agents to overcome this problem has produced interesting compounds through chemical synthesis, which plays a very important role in the research and discovery of new drugs. It is in this framework that our study was conducted at our laboratory and it involves evaluating the antibacterial activity of thirteen 2-pyridone derivatives synthesized by two methods, the diffusion disc method and the dilution method against eight Gram negative bacterial strains. The results seem interesting especially for two products that have shown the best activities against Escherichia coli ATCC 25922 and Enterobacter cloacae ATCC 13047 with CMI of 512µg/ml.

Keywords: heterocyclic derivatives, chemical synthesis, antimicrobial activity, biotechnology

Procedia PDF Downloads 343

Authors: Kessara Seneesrisakul, Erdogan Gulari, Sumaeth Chavadej

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The complex structure of lignocellulose leads to great difficulties in converting it to fermentable sugars for the ethanol production. The major hydrolysis impediments are the crystallinity of cellulose and the lignin content. To improve the efficiency of enzymatic hydrolysis, microbial pretreatment of corncob was investigated using two bacterial strains of Bacillus subtilis A 002 and Cellulomonas sp. TISTR 784 (expected to break open the crystalline part of cellulose) and lignin-degrading fungus, Phanerochaete sordida SK7 (expected to remove lignin from lignocellulose). The microbial pretreatment was carried out with each strain under its optimum conditions. The pretreated corncob samples were further hydrolyzed to produce reducing glucose with low amounts of commercial cellulase (25 U•g-1 corncob) from Aspergillus niger. The corncob samples were determined for composition change by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscope (SEM). According to the results, the microbial pretreatment with fungus, P. sordida SK7 was the most effective for enhancing enzymatic hydrolysis, approximately, 40% improvement.

Keywords: corncob, enzymatic hydrolysis, glucose, microbial pretreatment

Procedia PDF Downloads 566

Authors: Kashif Jaeel, Bingru Huang

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Stress-induced accumulation of ethylene exacerbates drought damages in plants, and suppressing stress induction of ethylene may promote plant tolerance to heat stress. The objective of this study was to investigate the effects of endophytic bacteria (Paraburkholderia aspalathi) with 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase enzymes in suppressing ethylene production on plant tolerance to heat stress and underlying physiological mechanisms of P. aspalathi-regulation in creeping bentgrass (Agrostis stolonifera). A novel strain of P. aspalathi, ‘WSF23’, with ACC deaminase activity was used to inoculate the roots of plants (cv. ‘Penncross’) subjected to heat stress in controlled-environment chambers. Inoculation with WSF23 bacteria resulted in improved shoot and root growth during heat stress. The differential changes in metabolite regulation due to the bacterial inoculation could contribute to ACC deamination bacteria-improved heat tolerance in cool-season grass species.

Keywords: rhizobacteria, grass, heat, plant metabolism, soil bacteria

Procedia PDF Downloads 49

Authors: Somdutt Mujwar, Kamal Raj Pardasani

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Cholera pandemics are caused by facultative pathogenic Vibrio cholera bacteria persisting in the countries having warmer climatic conditions as well as the presence of large water bodies with huge amount of organic matter, it is responsible for the millions of deaths annually. Presently the available therapy for cholera is Oral Rehydration Therapy (ORT) with an antibiotic drug. Excessive utilization of life saving antibiotics drugs leads to the development of resistance by the infectious micro-organism against the antibiotic drugs resulting in loss of effectiveness of these drugs. Also, many side effects are also associated with the use of these antibiotic drugs. This riboswitch is explored as an alternative drug target for Vibrio cholera bacteria to overcome the problem of drug resistance as well as side effects associated with the antibiotics drugs. The bacterial riboswitch is virtually screened with 24407 legends to get possible drug candidates. The 10 ligands showing best binding with the riboswitch are selected to design a pharmacophore, which can be utilized to design lead molecules by using the phenomenon of bioisosterism.

Keywords: cholera, drug design, ligand, riboswitch, pharmacophore

Procedia PDF Downloads 331

Authors: F. Allag, S. Bouharati, M. Belmahdi, R. Zegadi

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The rapid expansion of deserts in recent decades as a result of human actions combined with climatic changes has highlighted the necessity to understand biological processes in arid environments. Whereas physical processes and the biology of flora and fauna have been relatively well studied in marginally used arid areas, knowledge of desert soil micro-organisms remains fragmentary. The objective of this study is to conduct a diversity analysis of bacterial communities in unvegetated arid soils. Several biological phenomena in hot deserts related to microbial populations and the potential use of micro-organisms for restoring hot desert environments. Dry land ecosystems have a highly heterogeneous distribution of resources, with greater nutrient concentrations and microbial densities occurring in vegetated than in bare soils. In this work, we found it useful to use techniques of artificial intelligence in their treatment especially artificial neural networks (ANN). The use of the ANN model, demonstrate his capability for addressing the complex problems of uncertainty data.

Keywords: desert soil, climatic changes, bacteria, vegetation, artificial neural networks

Procedia PDF Downloads 378

Authors: Fatemeh Moradian, Razieh Ghorbani, Poria Biparva

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Growing resistance of microorganisms to potent antibiotics has renewed a great interest towards investigating bactericidal properties of nanoparticles and their Nano composites as an alternative. The use of metal nanoparticles to combat bacterial infections is one of the most wide spread applications of nanotechnology in the field of antibacterial. Nanomaterials have unique properties compared to their bulk counterparts. In this report, we demonstrate the antimicrobial activity of zerovalent Iron(ZVI) and Ag(silver) nanoparticles against Gram-negative bacteria E.coli(DH5α) and Pseudomonas aeruginosa. At first ZVI and Ag nanoparticles were synthesized by chemical reduction method and using scanning electron microscopy (SEM) the nanoparticle size determined. Different concentrations of Ag and ZVI nanoparticles were added to bacteria on nutrient agar medium. Minimum inhibitory concentration (MIC) of Ag and Fe nanoparticles for P. aeruginosa were 5µM and 1µg as well as for E.coli were 6µM. and 10 µg, respectively. Among the two nanoparticles, ZVI showed that the greatest antimicrobial activity against E.coli and Ag nanoparticle on P.aeruginosa. Results suggested that the bactericidal effect of metal nanoparticles has been attributed to their small size as well as high surface to volume ratio and NPs could be used as an effective antibacterial material.

Keywords: bactericidal properties, MIC, nanoparticle, SEM

Procedia PDF Downloads 579

Authors: Abeer M. Algeblawi

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Fifteen isolates of Agrobacterium tumefaciens were obtained from crown gall samples collected from six locations (Tripoli, Alzahra, Ain-Zara, Alzawia, Alazezia in Libya) from Grape (Vitis vinifera L.), Pear (Pyrus communis L.), Peach (Prunus persica L.) and Alexandria in Egypt from Guava (Psidium guajava L.) trees, Artichoke (Cynara cardunculus L.) and Sugar beet (Beta vulgaris L.). Total DNA was extracted from the eight isolates as well as the identification of six isolates used into Polymerase Chain Reaction (PCR) analysis and Random Amplified Polymorphic DNA (RAPD) technique were used. High similarity (55.5%) was observed among the eight A. tumefaciens isolates (Agro1, Agro2, Agro3, Agro4, Agro5, Agro6, Agro7, and Agro8). The PCR amplification products were resulting from the use of two specific primers (virD2A-virD2C). Analysis induction six isolates of A. tumefaciens obtained from different hosts. A visible band was specific to A. tumefaciens of (220 bp, 224 bp) and 338 bp produced with total DNA extracted from bacterial cells.

Keywords: Agrobacterium tumefaciens, crown gall, identification, molecular characterization, PCR, RAPD

Procedia PDF Downloads 124

Authors: Hummera Rafique, Aamer Saeed

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Synthesis of structural analogues of various well known bioactive natural 3,4-dihydroisocoumarins viz. Scorzocreticin, Annulatomarin, Montroumarin, and Thunberginol B, have been carried out starting from 3,5-dimethoxy-4-methylphenyl acetic acid. 3,5-Dimethoxy-4-methylphenyl acetic acid was then condensed with various aryl acid chlorides (a-e) to afford the corresponding 6,8-dimethoxy-7-methyl-3-aryl isocoumarins (5a-e). The alkaline hydrolysis of isocoumarins yields keto-acids (3a-e), which were then reduced to hydroxyacids, followed by cyclodehydration with acetic anhydride furnish corresponding 3,4-dihydroisocoumarins (7a-e). Finally, demethylation of 3,4-dihydroisocoumarins was carried out to afford 6,8-dihydroxy-7-methyl-3-aryl-3,4-dihydroisocoumarins (7a-e). Antibacterial evaluation of all the synthesized compounds were carried out against ten bacterial strains, it was concluded that isocoumarins (5a-e) and 3,4-dihydroisocoumarins (7a-e) are more active against gram positive bacteria then gram negative. However, the 6,8-dihydroxy-3,4-dihydroisocoumarin derivatives (8a-e) are more active against gram negative then gram positive.

Keywords: 3, 5-Dimethoxy-4-methylhomophthalic acid, natural 3, 4-Dihydroisocoumarin analogues, antibacterial activity, isocoumarins, demethylation

Procedia PDF Downloads 390

Authors: I. Sani, A. Abdulhamid, F. Bello, Isah M. Fakai

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This research has focused on the application of green fluorescent protein (GFP) as a new technique for direct monitoring of fermentation processes involving cultured bacteria. To use GFP as a sensor for pH and oxygen, percentage ratio of red fluorescence to green (% R/G) was evaluated. Assessing the magnitude of the % R/G ratio in relation to low or high pH and oxygen concentration, the bacterial strains were cultivated under aerobic and anaerobic conditions. SCC1 strains of E. coli were grown in a 5 L laboratory fermenter, and during the fermentation, the pH and temperature were controlled at 7.0 and 370C respectively. Dissolved oxygen tension (DOT) was controlled between 15-100% by changing the agitation speed between 20-500 rpm respectively. Effect of reducing the DOT level from 100% to 15% was observed after 4.5 h fermentation. There was a growth arrest as indicated by the decrease in the OD650 at this time (4.5-5 h). The relative fluorescence (green) intensity was decreased from about 460 to 420 RFU. However, %R/G ratio was significantly increased from about 0.1% to about 0.25% when the DOT level was decreased to 15%. But when the DOT was changed to 100%, a little increase in the RF and decrease in the %R/G ratio were observed. Therefore, GFP can effectively detect and indicate any change in pH and oxygen level during fermentation processes.

Keywords: Escherichia coli SCC1, fermentation process, green fluorescent protein, red fluorescence

Procedia PDF Downloads 485