Search results for: fungal pathogens

Authors: W. Lakhdari, B. Doumendji-Mitich, A. Dahliz, S. Doumendji, Y. Bouchikh, R. M'lik, H. Hammi, A. Soud

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This work was done in order to fight against Brachytrupes megacephalus, a major pest in the Algerian oasis and promote one aspect of biological control against it. He wears a hand on the isolation and identification of indigenous fungi on imagos of this insect harvested in the station of INRAA Touggourt and secondly, the study of the pathogenicity of these strains fungal on this orthoptère adults. The results obtained showed the presence of six different species of entomopathogenic fungi, it is: Aspergillus flavus, Fusarium sp, Beauveria bassiana, Penicillium sp, Metharizium anisopliae and Aspergillus Niger. The pathogenicity test using fungi Beauveria bassiana strains and Metharizium anisopliae. On adult of B. megacephalus highlights the effectiveness of these strains of predatory adults, with a mortality rate approaching 100% after 11 days.

Keywords: biological control, brachytrupes megacephalus, entomopathogenic fungi, Southeastern Algeria

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Authors: Blessing Atim Aderibigbe

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A series of amine/ester-linked hybrid compounds containing pharmacophores, such as ursolic acid, oleanolic acid, ferrocene and bisphosphonates, were synthesized in an attempt to develop potent antibacterial and anticancer agents. Their structures were analyzed and confirmed using Nuclear Magnetic Resonance, Fourier Transform Infrared Spectroscopy, and mass spectroscopy. All the synthesized hybrid compounds were evaluated for their antibacterial activities against eleven selected bacterial strains using a serial dilution method. Some of the compounds displayed significant antibacterial activity against most of the bacterial and fungal strains. In addition, the in vitro cytotoxicity of these compounds was also performed against selected cancer cell lines. Some of the compounds were also found to be more active than their parent compounds, revealing the efficacy of designing hybrid molecules using plant-based bioactive agents.

Keywords: ursolic acid, hybrid drugs, oleanolic acid, bisphosphonates

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Authors: Amal Raqib Shameran, Ghanim Aboud Al-Mola

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Respiratory syncytial virus (hRSV) is the major pathogens of respiratory tract infections (RTI) among infants and children in the world. They are classified in family Paramyxoviridae and sub-family Pneumovirinae. The current work aimed to detect the role of RSV in the lower respiratory tract infection (LRTI) in Hilla, Iraq. The samples were collected from 50 children who were admitted to hospital suffering from lower respiratory tract infections (LRTI). 50 nasal and pharyngeal swabs were taken from patients at the period from January 2010 till April 2011, hospitalized in Hilla Maternity and Children Hospital. The results showed that the proportion of children infected with hRSV accounted for 24% 12/50 with lower respiratory tract infections (LRTI) when they tested by polymerase chain reaction (RT-PCR).

Keywords: respiratory syncytial virus, respiratory tract infections, infants, polymerase chain reaction (PCR)

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Authors: Muhammad Asif Qayyoum, Bilal Saeed Khan

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Household poultry, including chickens, turkeys, ducks and geese, are affected by parasitic lice and mites. The dermanyssid mites (Acari: Dermanyssidae) are the most important parasites of poultry because they act as vectors of many pathogens of poultry and workers. Mesostigmatic mite fauna was poorly identified from Pakistan, only one species was reported before in 1971. Surveys were conducted in 2014 - 2015 to investigate the mite fauna from poultry cages in seven locations in Punjab Province, Turkey. A total of seventy-one samples were collected on cardboard and by direct litter collection. Mites were collected directly from the cardboard and 100 g samples of the litter were processed through a Berlese funnel. The collected mites were prepared for identification by using Hoyer’s medium. A total of twenty-two species belonging to the Dermanyssidae, Parasitidae, Cheyletidae, Laelapidae, Macrochelidae and Uropodidae were identified with two new species.

Keywords: poultry cages, Acari, mesostigmatic mites, Pakistan

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Authors: Payam Rasoulnia, Seyyed Mohammad Mousavi

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Heavy fuel oil firing power plants produce huge amounts of ashes as solid wastes, which seriously need to be managed and processed. Recycling precious metals of V and Ni from these oil-fired ashes which are considered as secondary sources of metals recovery, not only has a great economic importance for use in industry, but also it is noteworthy from the environmental point of view. Vanadium is an important metal that is mainly used in the steel industry because of its physical properties of hardness, tensile strength, and fatigue resistance. It is also utilized in oxidation catalysts, titanium–aluminum alloys and vanadium redox batteries. In the present study bioleaching of vanadium and nickel from an oil-fired ash sample was conducted using Aspergillus niger fungus. The experiments were carried out using spent-medium bioleaching method in both Erlenmeyer flasks and also bubble column bioreactor, in order to compare them together. In spent-medium bioleaching the solid waste is not in direct contact with the fungus and consequently the fungal growth is not retarded and maximum organic acids are produced. In this method the metals are leached through biogenic produced organic acids present in the medium. In shake flask experiments the fungus was cultured for 15 days, where the maximum production of organic acids was observed, while in bubble column bioreactor experiments a 7 days fermentation period was applied. The amount of produced organic acids were measured using high performance liquid chromatography (HPLC) and the results showed that depending on the fermentation period and the scale of experiments, the fungus has different major lixiviants. In flask tests, citric acid was the main produced organic acid by the fungus and the other organic acids including gluconic, oxalic, and malic were excreted in much lower concentrations, while in the bioreactor oxalic acid was the main lixiviant and it was produced considerably. In Erlenmeyer flasks during 15 days fermentation of Aspergillus niger, 8080 ppm citric acid and 1170 ppm oxalic acid was produced, while in bubble column bioreactor over 7 days of fungal growth, 17185 ppm oxalic acid and 1040 ppm citric acid was secreted. The leaching tests using the spent-media obtained from both of fermentation experiments, were performed at the same conditions of leaching duration of 7 days, leaching temperature of 60 °C and pulp density up to 3% (w/v). The results revealed that in Erlenmeyer flask experiments 97% of V and 50% of Ni were extracted while using spent medium produced in bubble column bioreactor, V and Ni recoveries were achieved to 100% and 33%, respectively. These recovery yields indicate that in both scales almost total vanadium can be recovered, while nickel recovery was lower. With help of the bioreactor spent-medium nickel recovery yield was lower than that of obtained from the flask experiments, which it could be due to precipitation of some values of Ni in presence of high levels of oxalic acid existing in its spent medium.

Keywords: Aspergillus niger, bubble column bioreactor, oil-fired ash, spent-medium bioleaching

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Authors: Adferotimi Banso

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A laboratory investigation was conducted to determine the effect of Justica flava and Aframonium melegueta on the growth of Aspergillus niger, Rhizopus stolonifer and Fusarium species in sweet orange and mango juices. Aqueous extract (3%v/v) of Justica flava and Aframonium melegueta reduced the growth of the fungi, a combination of 2% (v/v) each of Justica flava and Aframonium melegueta extracts reduced the growth better. Partial purification of aqueous extracts of Justica flava and Aframonium melegueta showed that ethyl acetate fraction of the extracts exhibited the highest level of inhibition of growth of the test fungi compared with diethyl ether and n-hexane fractions. The results suggest that extracts of Justica flava and Aframonium melegueta may be important substitutes for conventional chemical preservatives in the processing of fruit juices.

Keywords: aqueous, fraction, mango, orange, purification, sweet

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Authors: Nahid H. Hajrah, Jamal S. M. Sabir, Neil Hall

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The jasmonic pathway is ubiquitous in plants and is crucial to plant development. It Is involved in fertility, ripening, and sex determination as well as in response to environmental stresses such as herbivory, pathogen drought or temperature shock. Essentially the jasmonic pathway acts to shut down growth in order to induce defence pathways. These pathways include the production of secondary metabolites which have evolved to defend against herbivores and pathogens but are of increasing interest due to their roll in medicine and biotechnology. Here we describe the transcriptional response of Rhazia stricta (a poisonous shrub widely used in traditional medicine) to jasmonic acid, in order to better characterize the genes involved in secondary metabolite production and its response to stress. We observe coordinated upregulation of flavonoid biosynthesis pathway leading to flavonols, flavones and anthocyanins but no similar coordination of the monoterpene indole alkaloid pathway.

Keywords: medicinal plants, Rhazia stricta, jasmonic acid, transcriptional analysis

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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: Z. Yousefniayejahr, S. Bolognini, A. Bonini, C. Campobasso, N. Poma, F. Vivaldi, M. Di Luca, A. Tavanti, F. Di Francesco

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Pathogenic bacteria represent a threat to healthcare systems and the food industry because their rapid detection remains challenging. Electrochemical biosensors are gaining prominence as a novel technology for the detection of pathogens due to intrinsic features such as low cost, rapid response time, and portability, which make them a valuable alternative to traditional methodologies. These sensors use biorecognition elements that are crucial for the identification of specific bacteria. In this context, bacteriophages are promising tools for their inherent high selectivity towards bacterial hosts, which is of fundamental importance when detecting bacterial pathogens in complex biological samples. In this study, we present the development of a low-cost and portable sensor based on the Zeno phage for the rapid detection of Staphylococcus aureus. Screen-printed gold electrodes functionalized with the Zeno phage were used, and electrochemical impedance spectroscopy was applied to evaluate the change of the charge transfer resistance (Rct) as a result of the interaction with S. aureus MRSA ATCC 43300. The phage-based biosensor showed a linear range from 101 to 104 CFU/mL with a 20-minute response time and a limit of detection (LOD) of 1.2 CFU/mL under physiological conditions. The biosensor’s ability to recognize various strains of staphylococci was also successfully demonstrated in the presence of clinical isolates collected from different geographic areas. Assays using S. epidermidis were also carried out to verify the species-specificity of the phage sensor. We only observed a remarkable change of the Rct in the presence of the target S. aureus bacteria, while no substantial binding to S. epidermidis occurred. This confirmed that the Zeno phage sensor only targets S. aureus species within the genus Staphylococcus. In addition, the biosensor's specificity with respect to other bacterial species, including gram-positive bacteria like Enterococcus faecium and the gram-negative bacterium Pseudomonas aeruginosa, was evaluated, and a non-significant impedimetric signal was observed. Notably, the biosensor successfully identified S. aureus bacterial cells in a complex matrix such as a nasal swab, opening the possibility of its use in a real-case scenario. We diluted different concentrations of S. aureus from 108 to 100 CFU/mL with a ratio of 1:10 in the nasal swap matrices collected from healthy donors. Three different sensors were applied to measure various concentrations of bacteria. Our sensor indicated high selectivity to detect S. aureus in biological matrices compared to time-consuming traditional methods, such as enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and radioimmunoassay (RIA), etc. With the aim to study the possibility to use this biosensor to address the challenge associated to pathogen detection, ongoing research is focused on the assessment of the biosensor’s analytical performances in different biological samples and the discovery of new phage bioreceptors.

Keywords: electrochemical impedance spectroscopy, bacteriophage, biosensor, Staphylococcus aureus

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Authors: Abdelaziz Wided, Khiat Nawel, Khiat Inssaf

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The present study was initiated to: Determine burned forest-inhabiting fungi in Zouagha, Terri Beinène, Mila and study the antagonistic activity of Trichoderma sp against Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp. 18 fungal strains were isolated from Soil samples taken from the forest Zouagha (Burned) in the region Mila representing 6 genera: Trichoderma sp et Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp, Rhizopus sp. The tests of dual culture method on culture medium (PDA) against Trichoderma sp et Fusarium sp, Penicillium sp, Rhizoctonia sp, Alternaria sp revealed that: Trichoderma sp could reduce l mycelium grouth of Fusarium sp23.13%, Penicillium sp33.13%, Rhizoctoniasp33.75 %and Alternaria sp 38.31% in comparaison with the witness after 6 days at room temperature. The strains of Fusarium sp ,Penicillium sp, Rhizoctonia sp et Alternaria sp showed differences sensibility to the antagoniste.

Keywords: isolation, identification, molds, burned soil of zouagha, antagonism, trichoderma sp

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Authors: S. Kumar, R. Sultana

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This study was conducted to evaluate the effect of Aspergillus species on acridid populations which are major agricultural pests of rice, sugarcane, wheat, maize and fodder crops in Pakistan. Three and replicates i.e. Aspergillus flavus, A. fumigatus and A. niger, excluding the control, were held under laboratory conditions. It was observed that consumption faecal production of acridids was significantly reduced after the pathogenic application of Aspergillus. In the control replicate, the mortality ratio for stage (N₄-N₆) was maximum on day 2^nd i.e. [F_10.7 = 18.33, P < 0.05] followed by [F_4.20 = 07.85, P < 0.05] and [F_3.77 = 06.11, P < 0.05] on 4^th and 3^rd day, respectively. Similarly, it was a minimum i.e. [F_0.48 = 84.65, P < 0.05] on the 1^st day. It was also noted that faecal production of Acridid nymphs was not significantly affected when treated with conidial concentration in H₂O formulation; however, it was significantly reduced after the contamination with conidial concentration in oil. The high morality of acridids after contamination of Aspergillus supports their use as bio-control agent for reducing pest population. The present study recommends that exploration and screening must be conducted to provide additional pathogens for evaluation as potential biological control against grasshoppers and locusts.

Keywords: acridid, agriculture, formulation, grasshoppers

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Authors: Mary Ghobrial, Sahar Wefky

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Marine macroalgae have proven to be rich source of bioactive compounds with biomedical potential, not only for human but also for veterinary medicine. Emergence of microbial disease in aquaculture industries implies serious loses. Usage of commercial antibiotics for fish disease treatment produces undesirable side effects. Marine organisms are a rich source of structurally novel biologically active metabolites. Competition for space and nutrients led to the evolution of antimicrobial defense strategies in the aquatic environment. The interest in marine organisms as a potential and promising source of pharmaceutical agents has increased in the last years. Many bioactive and pharmacologically active substances have been isolated from microalgae. Compounds with antibacterial, antifungal and antiviral activities have been also detected in green, brown and red algae. Selected species of marine benthic algae belonging to the Phaeophyta and Rhodophyta, collected from different coastal areas of Alexandria (Egypt), were investigated for their antibacterial and antifungal, activities. Macroalgae samples were collected during low tide from the Alexandria Mediterranean coast. Samples were air dried under shade at room temperature. The dry algae were ground, using electric mixer grinder. They were soaked in 10 ml of each of the solvents acetone, ethanol, methanol and hexane. Antimicrobial activity was evaluated using well-cut diffusion technique In vitro screening of organic solvent extracts from the marine macroalgae Laurencia pinnatifida, Pterocladia capillaceae, Stepopodium zonale, Halopteris scoparia and Sargassum hystrix, showed specific activity in inhibiting the growth of five virulent strains of bacteria pathogenic to fish Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio anguillarum, V. tandara, Escherichia coli and two fungi Aspergillus flavus and A. niger. Results showed that, acetone and ethanol extracts of all test macroalgae exhibited antibacterial activity, while acetone extract of the brown Sargassum hystrix displayed the highest antifungal activity. The extracts of seaweeds inhibited bacteria more strongly than fungi and species of the Rhodophyta showed the greatest activity against the bacteria rather than fungi tested. The gas liquid chromatography coupled with mass spectrometry detection technique allows good qualitative and quantitative analysis of the fractionated extracts with high sensitivity to the smaller amounts of components. Results indicated that, the main common component in the acetone extracts of L. pinnatifida and P. capillacea is 4-hydroxy-4-methyl2-pentanone representing 64.38 and 58.60%. Thus, the extracts derived from the red macroalgae were more efficient than those obtained from the brown macroalgae in combating bacterial pathogens rather than pathogenic fungi. The most preferred species over all was the red Laurencia pinnatifida. In conclusion, the present study provides the potential of red and brown macroalgae extracts for development of anti-pathogenic agents for use in fish aquaculture.

Keywords: bacteria, fungi, extracts, solvents

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Authors: Sabrina Azzi, Michal Iglewski, Véronique Nabelsi

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Diagnosis error problem is frequent and one of the most important safety problems today. One of the main objectives of our work is to propose an ontological representation that takes into account the diagnostic criteria in order to improve the diagnostic. We choose pneumonia disease since it is one of the frequent diseases affected by diagnosis errors and have harmful effects on patients. To achieve our aim, we use a semi-automated method to integrate diverse knowledge sources that include publically available pneumonia disease guidelines from international repositories, biomedical ontologies and electronic health records. We follow the principles of the Open Biomedical Ontologies (OBO) Foundry. The resulting ontology covers symptoms and signs, all the types of pneumonia, antecedents, pathogens, and diagnostic testing. The first evaluation results show that most of the terms are covered by the ontology. This work is still in progress and represents a first and major step toward a development of a diagnosis decision support system for pneumonia.

Keywords: Clinical decision support system, Diagnostic errors, Ontology, Pneumonia

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Authors: Okolie Pius Ifeanyi, Emerenini Emilymary Chima

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Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits using Molecular Nested PCR analysis and the efficacy of cell free supernatants from Lactic Acid Bacteria (LAB) isolated from fresh fruits for in vitro control of some tomato pathogens. Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples. The inhibitory potentials of supernatant obtained from LAB isolates of fruits origin that were molecularly characterized were investigated against some tomato phytopathogens using agar-well method with the view to develop biological agents for some tomato disease causing organisms. Methodology: Gram positive, catalase negative strains of LAB were isolated from fresh fruits on Man Rogosa and Sharpe agar (Lab M) using streaking method. Isolates obtained were molecularly characterized by means of genomic DNA extraction kit (Norgen Biotek, Canada) method. Standard methods were used for Nested Polymerase Chain Reaction (PCR) amplification targeting the 16S rRNA gene using universal 16S rRNA gene and LAB specific primers, agarose gel electrophoresis, purification and sequencing of generated Nested PCR products (Macrogen Inc., USA). The partial sequences obtained were identified by blasting in the non-redundant nucleotide database of National Center for Biotechnology Information (NCBI). The antimicrobial activities of characterized LAB against some tomato phytopathogenic bacteria which include (Xanthomonas campestries, Erwinia caratovora, and Pseudomonas syringae) were obtained by using the agar well diffusion method. Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%). The cell free supernatants of LAB from fresh fruits origin (Weissella cibaria, Weissella confusa, Leuconostoc paramensenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Lactobacillus pentosus) can inhibits these bacteria by creating clear zones of inhibition around the wells containing cell free supernatants of the above mentioned strains of lactic acid bacteria. Conclusion: This study shows that potentially LAB can be quickly characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer. Tomato disease causing organisms can be most likely biologically controlled by using extracts from LAB. This finding will reduce the potential hazard from the use of chemical herbicides on plant.

Keywords: nested pcr, molecular characterization, 16s rRNA gene, lactic acid bacteria

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Authors: Reenu Yadav, Vidhi Guha, Udit N. Soni, Jay Ram Patel

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Chewing gums are mobile novel drug delivery systems, with a potential for administering drugs either for local action or for systemic absorption via the buccal route. An antimicrobial chewing gum delivery system of the methanolic extracts of Beatea monosperma (barks and twigs), Cordia obliqua (leaves and seeds) and Cuminun cyminum (seeds) against periodontal diseases caused by some oral pathogens, was designed and characterized on various parameters.The results of the study support the traditional application of the plants and suggest, plant extracts possess compounds with antimicrobial properties that can be used as potential antimicrobial agents and gums can be a good carrier of herbal extracts. Developed formulation will cure/protect from various periodontal diseases. Further development and evaluations chewing gums including the isolated compounds on the commercial scale and their clinical and toxicological studies are the future challenges.

Keywords: periodontal diseases, herbal chewing gum, herbal extracts, novel drug delivery systems

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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: Sahira N. Muslim, Alaa N. Mohammed, Saba Saadoon Khazaal, Batool Kadham Salman, Israa M. S. AL-Kadmy, Sraa N. Muslim, Ahmed S. Dwaish, Sawsan Mohammed Kareem, Sarah N. Aziz, Ruaa A. Jasim

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Tannase has wide applications in food, beverage, brewing, cosmetics and chemical industries and one of the major applications of tannase is the production of gallic acid. Gallic acid is used for manufacturing of trimethoprim. In the present study, a local fungal strain of Penicillium expansum A₄ isolated from spoilt apple samples gave the highest production level of tannase. Tannase was partially purified with a recovery yield of 92.52% and 6.32 fold of purification by precipitation using ammonium sulfate at 50% saturation. Tannase led to increased antimicrobial activity of ceftazidime against Pseudomonas aeruginosa and S. aureus and had a synergism effect at low concentrations of ceftazidime, and thus, tannase may be a useful adjuvant agent for the treatment of many bacterial infections in combination with ceftazidime.

Keywords: ceftazidime, Penicillium expansum, tannase, antimicrobial activity

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Authors: Jamal Elzwai, Kofi Aidoo, Alan Candlish

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Production of OTA was detected after 24hr by Aspergillus ochraceus isolate whereas at 36hr for A. carbonarius isolate and Penicillium verrucosum IMI 285522 and 60hr for A. ochraceus CBS 588.68. Highest OTA level was produced by A. carbonarius isolate followed by A. ochraceus CBS 588.68, Penicillium verrucosum IMI 285522 and finally A. ochraceus isolate. Glucosamine content of barley sample before fermentation was found to be negligible and remained almost constant during the incubation time. Glucosamine content started to increase at 12 hours after incubation with A. ochraceus isolate, A. carbonarius isolate and A. ochraceus CBS 588.68, and after 12 hours with P. verrucosum IMI 285522. Highest glucosamine content, as a result of increase in fungal biomass, was produced by A. ochraceus CBS 588.68 followed by A. ochraceus isolate, A. carbonarius isolate, and finally by P. verrucosum IMI 285522. It appears that there is a correlation between OTA synthesis and glucosamine content with A. ochraceus isolate, A. carbonarius isolate and A. ochraceus CBS 588.68 but not with P. verrucosum IMI 285522.

Keywords: chitin, barley, Ochratoxin A, Aspergiluus ochraceus, A. carbonarius, Penicillium verrucosum

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Authors: Hye-Hyun Lee, Hyo-Kon Chun, Kyung-Hee Kim Kim, Mi-Hee Kim, Saet-Byul Chu, Sang-Jong Lee, Seung-Hyeop Lee, Seung-Won Yi

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Some chemicals such as malachite green, methylene blue, and copper sulfate had been used frequently as disinfectants controlling fungal infection in aquaculture. However, their carcinogenicity, mutagenicity and teratogenicity were reported in mammals. After their accumulation in food fish and its consumers was confirmed, concerns about public health has resulted in enhanced monitoring and increased demand for eco-friendly treatments. Therefore, this study aimed to evaluate safety to fish and efficacy of sulfur solution processed by effective microorganisms (EM-PSS) against Saprolegnia parasitica, for use of a potential aquatic fungicidal disinfectant. The natural sulfur purchased from Kawah Ijen volcano, East Java, Indonesia was processed by the liquid mixture consisting of following twelve effective microorganisms (Rapha-el®; Lbiotech, Jeonnam, Korea), Lactobacillus parafarraginis, L. paracasei, L. harbinensis, L. buchneri, L. perolens, L. rhamnosus, L. vaccinostercus, Acetobacter lovaniensis, A. peroxydans, Pichia fermentans, Candida ethanolica, Saccharomycopsis schoenii isolated from fermentation process of oriental medicinal herbs including green tea, privet, and puer tea. The material was applied to in vitro antifungal activity test for Saprolegnia parasitica using agar dilution method. In addition, an acute toxicity test was performed on carp (Cyprinus carpio), eel (Anguilla japonica), and mud fish (Misgurnus mizolepis) for 96 hours. After three species of fish (n=15) were accustomed to experimental water environment for three days, the EM-PSS was added to each tank as final concentrations to be 0 to 500 ppm. The fish were taken into necropsy, and the histological sections of the gill, liver, and spleen were counter-stained with hematoxylin and eosin (H-E). And hence, no observed effect concentration (NOEC) of the solution was used for taking a medicinal bath for mudfish infected by Saprolegnia parasitica in practice. The result of in vitro antifungal activity test showed the growth inhibition of the fungus at 100 ppm, which and the lower concentrations occurred no fatal case in any fish species tested until the end of the examination. The 125 ppm of the solution, however, resulted in 13.3 %, 13.3 %, and 6.3 % of mortality in carp, eel, and mudfish, respectively. But both 250 and 500 ppm of the solution leaded lethality to all population of each fish species within 24 hours. Besides, H-E staining also showed no specific evidence for toxicity in fish at lesser than 100 ppm of EM-PSS. On the other hand, as a result of field application of the solution, no growth of fungal mycelium was found in fish bodies from gross observation 5 days post treatment. In conclusion, 100ppm of EM-PSS resulted in inhibition and treatment of Saprolegnia parasitica infection. In addition, the use of EM-PSS lower than 100 ppm is safe for fish. Therefore, EM-PSS could be used as aquatic fungicide, and also may be possible to be a potential eco-friendly disinfectant in aquaculture.

Keywords: antifungal activity, effective microorganism, toxicity, saprolegnia, processed sulfur solution

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Authors: Aitor Barbero-López, Soumaya Chibily, Gerhard Scheepers, Thomas Grahn, Martti Venäläinen, Antti Haapala

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Wood decay is addressed continuously within the wood industry through use and development of wood preservatives. The increasing awareness on the negative effects of many chemicals towards the environment is causing political restrictions in their use and creating more urgent need for research on green alternatives. This paper discusses some of the possible natural extracts for wood preserving applications and compares the analytical methods available for testing their behavior and efficiency against decay fungi. The results indicate that natural extracts have interesting chemical constituents that delay fungal growth but vary in efficiency depending on the chemical concentration and substrate used. Results also suggest that presence and redistribution of preservatives in wood during exposure trials can be assessed by spectral imaging methods although standardized methods are not available. This study concludes that, in addition to the many standard methods available, there is a need to develop new faster methods for screening potential preservative formulation while maintaining the comparability and relevance of results.

Keywords: analytics, methods, preservatives, wood decay

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Authors: Laura Cochrane

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The United States Centers for Disease Control and Prevention has established three categories of biological agents based on their ease of spread and the severity of the disease they cause. Category A biological agents are the highest priority because of their high degree of morbidity and mortality, ease of dissemination, the potential to cause social disruption and panic, special requirements for public health preparedness, and past use as a biological weapon. Despite the threat of Category A biological agents, opportunities for medical intervention exist. This work summarizes public information, consolidated and reviewed across the situational usefulness and disease awareness to offer discussion to three specific Category A agents: anthrax (Bacillus anthracis), botulism (Clostridium botulinum toxin), and smallpox (variola major), and provides an overview on the management of medical countermeasures available to treat these three (3) different types of pathogens. The medical countermeasures are discussed in the setting of pre-exposure prophylaxis, post-exposure prophylaxis, and therapeutic treatments to provide a framework for requirements in public health preparedness.

Keywords: anthrax, botulism, smallpox, medical countermeasures

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Authors: Fei Jia, Xingjian Bai, Xiaowei Zhang, Wenjie Yan, Ruitong Dai, Xingmin Li, Jozef Kokini

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Pseudomonas aeruginosa is a Gram-negative, aerobic, opportunistic human pathogen that is present in the soil, water, and food. This microbe has been recognized as a representative food-borne spoilage bacterium that can lead to many types of infections. Considering the casualties and property loss caused by P. aeruginosa, the development of a rapid and reliable technique for the detection of P. aeruginosa is crucial. The whole-cell aptasensor, an emerging biosensor using aptamer as a capture probe to bind to the whole cell, for food-borne pathogens detection has attracted much attention due to its convenience and high sensitivity. Here, a low-field magnetic resonance imaging (LF-MRI) aptasensor for the rapid detection of P. aeruginosa was developed. The basic detection principle of the magnetic relaxation switch (MRSw) nanosensor lies on the ‘T₂-shortening’ effect of magnetic nanoparticles in NMR measurements. Briefly speaking, the transverse relaxation time (T₂) of neighboring water protons get shortened when magnetic nanoparticles are clustered due to the cross-linking upon the recognition and binding of biological targets, or simply when the concentration of the magnetic nanoparticles increased. Such shortening is related to both the state change (aggregation or dissociation) and the concentration change of magnetic nanoparticles and can be detected using NMR relaxometry or MRI scanners. In this work, two different sizes of magnetic nanoparticles, which are 10 nm (MN₁₀) and 400 nm (MN₄₀₀) in diameter, were first immobilized with anti- P. aeruginosa aptamer through 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) chemistry separately, to capture and enrich the P. aeruginosa cells. When incubating with the target, a ‘sandwich’ (MN₁₀-bacteria-MN₄₀₀) complex are formed driven by the bonding of MN400 with P. aeruginosa through aptamer recognition, as well as the conjugate aggregation of MN₁₀ on the surface of P. aeruginosa. Due to the different magnetic performance of the MN₁₀ and MN₄₀₀ in the magnetic field caused by their different saturation magnetization, the MN₁₀-bacteria-MN₄₀₀ complex, as well as the unreacted MN₄₀₀ in the solution, can be quickly removed by magnetic separation, and as a result, only unreacted MN₁₀ remain in the solution. The remaining MN₁₀, which are superparamagnetic and stable in low field magnetic field, work as a signal readout for T₂ measurement. Under the optimum condition, the LF-MRI platform provides both image analysis and quantitative detection of P. aeruginosa, with the detection limit as low as 100 cfu/mL. The feasibility and specificity of the aptasensor are demonstrated in detecting real food samples and validated by using plate counting methods. Only two steps and less than 2 hours needed for the detection procedure, this robust aptasensor can detect P. aeruginosa with a wide linear range from 3.1 ×10² cfu/mL to 3.1 ×10⁷ cfu/mL, which is superior to conventional plate counting method and other molecular biology testing assay. Moreover, the aptasensor has a potential to detect other bacteria or toxins by changing suitable aptamers. Considering the excellent accuracy, feasibility, and practicality, the whole-cell aptasensor provides a promising platform for a quick, direct and accurate determination of food-borne pathogens at cell-level.

Keywords: magnetic resonance imaging, meat spoilage, P. aeruginosa, transverse relaxation time

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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: Hattie Hope Makumbe, Bhekisisa Dlamini, Frederick Tabit

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Listeria monocytogenes is one of the most important foodborne pathogens associated with ready-to-eat (RTE) food. This study investigated the incidence of Listeria monocytogenes in 80 RTE food sold in the formal (dairy and processed meat) and informal markets (vegetable salads, beef stew, and rice) of Johannesburg, South Africa. High Enterobacteriaceae, S. aureus, and E. coli counts were obtained, which ranged from 1.9-7.5 log CFU/g. Listeria monocytogenes microbial counts in the food samples ranged from 3.5-6.0 log colony forming unit per gram except in cooked rice. The Listeria monocytogenes isolates were identified using biochemical tests and confirmed with the Biolog identification system and PCR analyses. The percentage incidence for Listeria monocytogenes in ready to eat food was 12.5%. When Minimum Inhibitory Concentrations were under consideration, all disinfectants were effective against Listeria monocytogenes strains. For antimicrobial work, rates of resistance amongst the antibiotics ranged from 17-100%. Therefore, more effective preventive control strategies for Listeria monocytogenes are needed to reduce the prevalence of the pathogen in RTE food that is sold in Johannesburg.

Keywords: Listeria monocytogenes, Listeria species, ready to eat food, sanitiser efficacy

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Authors: Gauravi Agarkar, Mahendra Rai

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Endophytic fungi from medicinal plants are important source of numerous pharmacologically important compounds. In the present investigation, the endophytic fungi were isolated from three medicinal plants; Andrographis paniculata, Rauwolfia serpentina and Tridax procumbens. Endophytic Colletotrichum sp. were identified on the basis of cultural and morphological characteristics as well as internal transcribed spacer (ITS) sequence analysis. Antibacterial and antifungal activity of the ethyl acetate and methanol extract of endophytic Colletotrichum sp. was evaluated against seven different human pathogenic bacteria and six Candida sp. The extracts were effective and showed significant activity against all the test pathogens. In case of yeast Candida, the combined effect of extracts and standard antibiotic was enhanced greatly showing synergistic activity. Further, the extracts were assayed for Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal/Fungicidal Concentration (MBC/MFC) where, MIC values were in the range of 100-250 μg/ml. These results suggest that the endophytic Colletotrichum sp. isolated from the medicinal plants are capable of producing promising antimicrobial metabolites.

Keywords: antimicrobial, colletotrichum, endophytic fungi, medicinal plants

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Authors: Sang-Myung Jung, Gwang Heum Yoon, Hoo Chul Lee, Hwa Sung Shin

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Ceramide, a component of stratum corneum at epidermis, helps to construct a rigid and dense skin barrier to prevent pathogens that cause atopic dermatitis. However, ceramide was too hydrophobic to be directly absorbed into stratum corneum and has risks of side effects by excessive treatment. To overcome the obstacles, ceramide was embedded into PLGA nanoparticles coated with chitosan. PLGA and chitosan have been known as biocompatible materials. PLGA was squeezed when faced with water and pumped ceramide out of PLGA nanoparticle. In addition, the chitosan coating layer helped initial adherence of nanoparticles to skin and regulate ceramide release until removed. This coating was degraded at weakly acid state like skin surface, finally ceramide release could be controlled. Finally, the nanoparticle was demonstrated to be non-cytotoxic and regenerate stratum corneum of atopic dermatitis model. Overall the nanoparticle is suggested as a novel and effective nanodrug to apply atopic dermatitis.

Keywords: nanoparticle, controlled release, atopic dermatitis, chitosan coating, ceramide

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Authors: Susmita Sharma, Pankaj Pathak

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Biosolids are by-products of municipal and industrial wastewater treatment process. The generation of the biosolids is increasing at an alarming rate due to the implementation of strict environmental legislation to improve the quality of discharges from wastewater treatment plant. As such, proper management and safe disposal of sewage sludge have become a worldwide topic of research. Biosolids, rich in organic matter and essential micro and macronutrients; can be used as a soil conditioner, to cut fertilizer costs and create favorable conditions for vegetation. However, it also contains pathogens and heavy metals which are undesirable as they are harmful to both humans and the environment. Therefore, for safe utilization of biosolids for land application purposes, categorization of the contaminant and pathogen is mandatory. In this context, biosolids collected from a wastewater treatment plant in Maharashtra are utilized to determine its physical, chemical and microbiological attributes. This study would ascertain, if the use of these materials from the specific site, are suitable for agriculture. Further, efforts have also been made to present the internationally acceptable legal standards and guidelines for biosolids management or application.

Keywords: biosolids, sewage, heavy metal, sustainable agriculture

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Authors: T. Büyüksırıt, H. Kuleaşan

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Natural antimicrobials are used to preserve foods that can be found in plants, animals, and microorganisms. Antimicrobial substances are natural or artificial agents that produced by microorganisms or obtained semi/total chemical synthesis are used at low concentrations to inhibit the growth of other microorganisms. Food borne pathogens and spoilage microorganisms are inactivated by the use of antagonistic microorganisms and their metabolites. Yeasts can produce toxic proteins or glycoproteins (toxins) that cause inhibition of sensitive bacteria and yeast species. Antimicrobial substance producing phenotypes belonging different yeast genus were isolated from different sources. Toxins secreted by many yeast strains inhibiting the growth of other yeast strains. These strains show antimicrobial activity, inhibiting the growth of mold and bacteria. The effect of antimicrobial agents produced by yeasts can be extremely fast, and therefore may be used in various treatment procedures. Rapid inhibition of microorganisms is possibly caused by microbial cell membrane lipopolysaccharide binding and in activation (neutralization) effect. Antimicrobial agents inhibit the target cells via different mechanisms of action.

Keywords: antimicrobial agents, yeast, toxic protein, glycoprotein

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Authors: Dipali Nagaonkar, Mahendra Rai

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Nanotechnology has experienced significant developments in engineered nanomaterials in the core-shell arrangement. Nanomaterials having nanolayers of silver and gold are of primary interest due to their wide applications in catalytical and biomedical fields. Further, mycosynthesis of nanoparticles has been proved as a sustainable synthetic approach of nanobiotechnology. In this context, we have synthesized silver and silver (core)-gold (shell) bimetallic nanoparticles using a fungal extract of Fusarium graminearum by sequential reduction. The core-shell deposition of nanoparticles was confirmed by the red shift in the surface plasmon resonance from 434 nm to 530 nm with the aid of the UV-Visible spectrophotometer. The mean particle size of Ag and Ag-Au nanoparticles was confirmed by nanoparticle tracking analysis as 37 nm and 50 nm respectively. Quite polydispersed and spherical nanoparticles are evident by TEM analysis. These mycosynthesized bimetallic nanoparticles were tested against some pathogenic bacteria and Candida sp. The antimicrobial analysis confirmed enhanced anticandidal and antibacterial potential of bimetallic nanoparticles over their monometallic counterparts.

Keywords: bimetallic nanoparticles, core-shell arrangement, mycosynthesis, sequential reduction

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Authors: Seyed Reza Aghili, Tahereh Shokohi, Shirin Sadat Hashemi Fesharaki, Mohammad Ali Boroumand, Bahar Salmanian

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Introduction: Intravenous catheter-associated fungal infection as nosocomial infection continue to be a deep problem among hospitalized patients, decreasing quality of life and adding healthcare costs. The capacity of catheters in the spread of candidemia in heart failure patients is obvious. The aim of this study was to evaluate the prevalence and genetic identification of Candida species in heart disorder patients. Material and Methods: This study was conducted in Tehran Hospital of Cardiology Center (Tehran, Iran, 2014) during 1.5 years on the patients hospitalized for at least 7 days and who had central or peripheral vein catheter. Culture of catheters, blood and skin of the location of catheter insertion were applied for detecting Candida colonies in 223 patients. Identification of Candida species was made on the basis of a combination of various phenotypic methods and confirmed by sequencing the ITS1-5.8S-ITS2 region amplified from the genomic DNA using PCR and the NCBI BLAST. Results: Of the 223 patients samples tested, we identified totally 15 Candida isolates obtained from 9 (4.04%) catheter cultures, 3 (1.35%) blood cultures and 2 (0.90%) skin cultures of the catheter insertion areas. On the base of ITS region sequencing, out of nine Candida isolates from catheter, 5(55.6%) C. albicans, 2(22.2%) C. glabrata, 1(11.1%) C. membranifiaciens and 1 (11.1%) C. tropicalis were identified. Among three Candida isolates from blood culture, C. tropicalis, C. carpophila and C. membranifiaciens were identified. Non-candida yeast isolated from one blood culture was Cryptococcus albidus. One case of C. glabrata and one case of Candida albicans were isolated from skin culture of the catheter insertion areas in patients with positive catheter culture. In these patients, ITS region of rDNA sequence showed a similarity between Candida isolated from the skin and catheter. However, the blood samples of these patients were negative for fungal growth. We report two cases of catheter-related candidemia caused by C. membranifiaciens and C. tropicalis on the base of genetic similarity of species isolated from blood and catheter which were treated successfully with intravenous fluconazole and catheter removal. In phenotypic identification methods, we could only identify C. albicans and C. tropicalis and other yeast isolates were diagnosed as Candida sp. Discussion: Although more than 200 species of Candida have been identified, only a few cause diseases in humans. There is some evidence that non-albicans infections are increasing. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition are considered to be associated with candidemia in hospitalized heart failure patients. Identifying the route of infection in candidemia is difficult. Non-albicans candida as the cause of candidemia is increasing dramatically. By using conventional method, many non-albicans isolates remain unidentified. So, using more sensitive and specific molecular genetic sequencing to clarify the aspects of epidemiology of the unknown candida species infections is essential. The positive blood and catheter cultures for candida isolates and high percentage of similarity of their ITS region of rDNA sequence in these two patients confirmed the diagnosis of intravenous catheter-associated candidemia.

Keywords: catheter-associated infections, heart failure patient, molecular genetic sequencing, ITS region of rDNA, Candidemia

Procedia PDF Downloads 315