Search results for: fungal pathogen

Authors: Juan Jose Filgueira, Daniela Londono-Serna, Liliana Maria Hoyos

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Carnation (Dianthus caryophyllus) is one of the most important products of exportation in the floriculture industry worldwide. Fusariosis is the disease that causes the highest losses on farms, in particular the one produced by Fusarium oxysporum f.sp. dianthi, called vascular wilt. Gene identification and metabolic routes of the genes that participate in the building of the plant response to Fusarium are some of the current targets in the carnation breeding industry. The techniques for the identifying of resistant genes in the plants, is the analysis of the transcriptome obtained during the host-pathogen interaction. In this work, we report the cell transcriptome of different varieties of carnation that present differential response from Fusarium oxysporum f.sp. dianthi attack. The cells of the different hybrids produced in the outbreeding program were cultured in vitro and elicited with the parasite in a dual culture. The isolation and purification of mRNA was achieved by using affinity chromatography Oligo dT columns and the transcriptomes were obtained by using Illumina NGS techniques. A total of 85,669 unigenes were detected in all the transcriptomes analyzed and 31,000 annotations were found in databases, which correspond to 36.2%. The library construction of genic expression techniques used, allowed to recognize the variation in the expression of genes such as Germin-like protein, Glycosyl hydrolase family and Cinnamate 4-hydroxylase. These have been reported in this study for the first time as part of the response mechanism to the presence of Fusarium oxysporum.

Keywords: Carnation, Fusarium, vascular wilt, transcriptome

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Authors: Jessica P. Fiorotti, Maria C. Freitas, Caio J. B. Coutinho-Rodrigues, Mariana G. Camargo, Emily S. Mesquita, Amanda R. C. Corval, Ricardo O. B. Bitencourt, Allan F. Marciano, Diva D. Spadacci-Morena, Patricia S. Golo, Isabele C. Angelo, Vania R. E. P. Bittencourt

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The bovine tick, Rhipicephalus microplus, is an arthropod of great importance in veterinary medicine leading to anemia, weight loss, animals' leather depreciation and also acting as a vector of many pathogens. In this way, the parasitism causes a loss of 3.24 billion dollars per year in Brazil. Knowingly, entomopathogenic fungi act as natural controller of some arthropods, acting mainly by active penetration through the cuticle. However, it can also act on the hemolymph and through the production of mycotoxins. Hemocytes are responsible for the cellular immune response and participate in the processes of phagocytosis, nodulation and encapsulation and may undergo changes when challenged by pathogens. The aim of the present study was to evaluate changes in R. microplus hemocytes after inoculation of Metarhizium robertsii using transmission electron microscopy. The isolate ARSEF 2575 and 200 engorged R. microplus females were used. The groups were divided into control, in which the females were inoculated with 5 μL of sterile distilled water solution and 0.1% Tween 80, and a group inoculated with 5 μL of fungal suspension at the concentration of 10⁷ conidia mL⁻¹. The experiment was performed in duplicate and each group contained 50 females. Twenty-four hours after fungal inoculation, hemolymph was collected through the cuticle dorsal surface perforation of the tick females. After collection, the hemolymph samples were centrifuged at 500 x g for 3 minutes at 4 °C, the plasma was discarded and the hemocyte pellet was resuspended in 50 μl PBS. The suspension material was fixed in 2% glutaraldehyde in Millonig buffer for three hours. After fixation, the material was centrifuged at 500 x g for 3 minutes, the supernatant was discarded and the cells were resuspended in a wash solution. Subsequently, the cells were post-fixed with 1% osmium tetroxide in phosphate buffer for one hour at room temperature and dehydrated in increasing concentrations of ethanol, and then embedded in Epon resin. The ultrathin sections were examined under the LEO EM 906E transmission electron microscopy at 80kV. The ultrastructural results revealed that.in control group, the cells were considered intact, in which the granulocytes were observed with granules of different electrodensities, intact mitochondria and cytoplasm without vacuolization. In addition, granulocytes showed plasma membrane projections similar to pseudopodia. Plasmatocytes presented as irregularly shaped cells, with the eccentric nucleus, agranular cytoplasm and some cells presented pseudopodia. Nevertheless, in the group exposed to the fungus, most of the cells presented in degeneration. The granulocytes found had fewer granules in the cytoplasm and more vacuoles. Plasmatocytes, after treatment, presented many vacuoles also in the cytoplasm and the lysosomes presented great amount of electrodense material in their interior. Thus, the results suggest that the fungus has a depressant action in the immune system of the tick, not only by the cell degranulation, but also suggesting that this leads to morphological changes in the hemocytes and may even trigger processes such as phagocytosis.

Keywords: bovine tick, cellular defense, entomopathogenic fungi, immune response

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Authors: Sadaf Ilyas, Saba Riaz

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The extensive use of antibiotics has led to increases emergence of antibiotic-resistant organisms. Pseudomonas is a notorious opportunistic pathogen involoved in nosocomial infections and exhibit innate resistance to many antibiotics. The present study was conducted to assess the prevalence, levels of antimicrobial susceptibility and resistance mechanisms of Pseudomonas. A total of thirty clinical strains of Pseudomonas were isolated from different clinical sites of infection. All clinical specimens were collected from Chughtais Lahore Lab. Jail road, during 8-07-2010 to 11-01-2011. Biochemical characterization was done using routine biochemical tests. Antimicrobial susceptibility was determined by Kirby-Baeur method. The plasmids were isolated from all the strains and digested with restriction enzyme PstI and EcoRI. Transfer of Multi-resistance plasmid was checked via transformation and conjugation to confirm the plasmid mediated resistance to antibiotics. The prevalence of Pseudomonas in clinical specimens was found out to be 14% of all bacterial infections. IPM has shown to be the most effective drug against Pseudomonas followed by CES, PTB and meropenem, wheareas most of the Pseudomonas strains have developed significant resistance against Penicillins and some Cephalasporins. Antibiotic resistance determinants were carried by plasmids, as they conferred resistance to transformed K1 strains. The isolates readily undergo conjugation, transferring the resistant genes to other strains, illustrating the high rates of cross infection and nosocomial infection in the immunocompromised patients.

Keywords: pseudomonas, antibiotics, drug resistance, horizontal gene transfer

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Authors: Popy Bora, L. C. Bora, A. Bhattacharya, Sehnaz S. Ahmed

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Microbial bioagents, viz., Pseudomonas fluorescens, Bacillus subtilis, and Trichoderma viride were assessed for their ability to suppress grey blight caused by Pestalotiopsis theae, a major disease of tea crop in Assam. The expression of defense-related phytochemicals due to the application of these bioagents was also evaluated. The individual bioagents, as well as their combinations, were screened for their bioefficacy against P. theae in vitro using nutrient agar (NA) as basal medium. The treatment comprising a combination of the three bioagents, P. fluorescens, B. subtilis, and T. viride showed significantly the highest inhibition against the pathogen. Bioformulation of effective bioagent combinations was further evaluated under field condition, where significantly highest reduction of grey blight (90.30%), as well as the highest increase in the green leaf yield (10.52q/ha), was recorded due to application of the bioformulation containing the three bioagents. The application of the three bioformulation also recorded an enhanced level of caffeine (4.15%) and polyphenols (22.87%). A significant increase in the enzymatic activity of phenylalanine ammonia-lyase, peroxidase and polyphenol oxidase were recorded in the plants treated with the microbial bioformulation of the three bioagents. The present investigation indicates the role of microbial agents in suppressing disease, inducing plant defense response, as well as improving the quality of tea.

Keywords: enzymatic activity, grey blight, microbial bioagents, Pestalotiopsis theae, phytochemicals, plant defense, tea

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Authors: Kavita Jaidiya, Anju Chahar, Chitra Jaidiya

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The present study was conducted on 200 quarters from 50 apparently healthy cows. Samples are subjected to California Mastitis Test (CMT), cultural examination, and mPCR. Milk samples were also subjected to changes in composition Viz. fat, protein, and lactose. The prevalence of subclinical mastitis based on culture examination was 30(60/200), 36 (72/200), and 40 percent (93/200) based on CMT, culture examination, and mPCR on a quarterly basis. The prevalence of subclinical mastitis on animal basis was 40 (20/50), 46 (23/50), and 52 percent (26/50) based on CMT, Culture examination, and mPCR. The highest prevalence was observed in IVth parity on a quarterly basis and in Vth parity on cow basis. On culture examination, Staphylococcus aureus was the most prevalent organism (50.56%), followed by Streptococcus dysaglactiae (11.33%), E. coli (7.8 %), Staphylococcus agalactiae (13.48 %), Staphylococcus epidermidis (2.2 %), Streptococcus hyicus (6.94%), Streptococcus uberis (5.16%), Klebsiella pneumonia (6.74%). On isolation by bacterial mPCR, Staphylococcus spp. (42%) was the major pathogen. Organisms isolated in mixed infections are Streptococcus spp., Klebsiella pneumonia, E.coli and Pseudomonas aeruginous. The average mean value of fat, protein, and lactose content in subclinically affected milk samples were 3.40 ± 0.101, 3.009 ± 0.033, and 4.48 ± 0.03, and the mean value of fat, protein, and lactose content in normal milk were 4.13 ± 0.035, 3.39 ± 0.021, and 5.10 ± 0.016. The mean blood level of reduced glutathione in subclinical mastitis (30.44 ± 1.87 ng/ml) was lower than healthy cows (47.98 ± 4.04ng/ml). The concentration of malondialdehyde (10.026 ± 0.21mmol/L) in subclinical mastitis was significantly higher as compared to healthy group cows (2.19 ± 0.23mmol/L).

Keywords: cow, subclinical mastitis, mPCR, California Mastitis test

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Authors: Jyoti Pandey, Rajiv Kumar Bhatt

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In the absence of proper sanitation, people suffered from high levels of infectious diseases leading to high incidences of morbidity and mortality. This directly affected the ability of a country to maintain an efficient economy and implied great personal suffering among infected individuals and their families. This paper aims to estimate the catastrophic expenditure of households in terms of direct and indirect losses which a person has to face due to the illness of WASH diseases; the severity of the scenario is answered by finding out the impoverishment effect. We used the primary data survey for the objective outlined. Descriptive and analytical research types are used. The survey is done with the questionnaire formulated precisely, taking care of the inclusion of all the variables and probable outcomes. A total of 300 households is covered under this study. In order to pursue the objectives outlined, multistage random sampling of households is used. In this study, the cost of illness approach is followed for accessing economic impact. The study brought out the attention that a significant portion of the total consumption expenditure is going lost for the treatment of water and sanitation related diseases. The infectious and water vector-borne disease can be checked by providing sufficient required sanitation facility, and that 2.02% loss in income can be gained if the mechanisms of the pathogen is checked.

Keywords: water, sanitation, impoverishment, catastrophic expenditure

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Authors: Veenu Bala, Yashpal S. Chhonker, Bhavana Kushwaha, Rabi S. Bhatta, Gopal Gupta, Vishnu L. Sharma

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According to UNAIDS 2013 estimate nearly 52% of all individuals living with HIV are now women of reproductive age (15–44 years). Seventy-five percent cases of HIV acquisition are through heterosexual contacts and sexually transmitted infections (STIs), attributable to unsafe sexual behaviour. Each year, an estimated 500 million people acquire atleast one of four STIs: chlamydia, gonorrhoea, syphilis and trichomoniasis. Trichomonas vaginalis (TV) is exclusively sexually transmitted in adults, accounting for 30% of STI cases and associated with pelvic inflammatory disease (PID), vaginitis and pregnancy complications in women. TV infection resulted in impaired vaginal milieu, eventually favoring HIV transmission. In the absence of an effective prophylactic HIV vaccine, prevention of new infections has become a priority. It was thought worthwhile to integrate HIV prevention and reproductive health services including unintended pregnancy protection for women as both are related with unprotected sex. Initially, nonoxynol-9 (N-9) had been proposed as a spermicidal agent with microbicidal activity but on the contrary it increased HIV susceptibility due to surfactant action. Thus, to accomplish an urgent need of novel woman controlled non-detergent microbicidal spermicides benzenepropanamine analogues have been synthesized. At first, five benzenepropanamine-dithiocarbamate hybrids have been synthesized and evaluated for their spermicidal, anti-Trichomonas and anti-fungal activities along with safety profiling to cervicovaginal cells. In order to further enhance the scope of above study benzenepropanamine was hybridized with thiourea as to introduce anti-HIV potential. The synthesized hybrid molecules were evaluated for their reverse transcriptase (RT) inhibition, spermicidal, anti-Trichomonas and antimicrobial activities as well as their safety against vaginal flora and cervical cells. simulated vaginal fluid (SVF) stability and pharmacokinetics of most potent compound versus N-9 was examined in female Newzealand (NZ) rabbits to observe its absorption into systemic circulation and subsequent exposure in blood plasma through vaginal wall. The study resulted in the most promising compound N-butyl-4-(3-oxo-3-phenylpropyl) piperazin-1-carbothioamide (29) exhibiting better activity profile than N-9 as it showed RT inhibition (72.30 %), anti-Trichomonas (MIC, 46.72 µM against MTZ susceptible and MIC, 187.68 µM against resistant strain), spermicidal (MEC, 0.01%) and antifungal activity (MIC, 3.12–50 µg/mL) against four fungal strains. The high safety against vaginal epithelium (HeLa cells) and compatibility with vaginal flora (lactobacillus), SVF stability and least vaginal absorption supported its suitability for topical vaginal application. Docking study was performed to gain an insight into the binding mode and interactions of the most promising compound, N-butyl-4-(3-oxo-3-phenylpropyl) piperazin-1-carbothioamide (29) with HIV-1 Reverse Transcriptase. The docking study has revealed that compound (29) interacted with HIV-1 RT similar to standard drug Nevirapine. It may be concluded that hybridization of benzenepropanamine and thiourea moiety resulted into novel lead with multiple activities including RT inhibition. A further lead optimization may result into effective vaginal microbicides having spermicidal, anti-Trichomonas, antifungal and anti-HIV potential altogether with enhanced safety to cervico-vaginal cells in comparison to Nonoxynol-9.

Keywords: microbicidal, nonoxynol-9, reverse transcriptase, spermicide

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Authors: Benarous Khedidja, Yousfi Mohamed

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Infections caused by Candida species manifest in a number of diseases, including candidemia, vulvovaginal candidiasis, endocarditis, and peritonitis. These Candida species have been reported to have lipolytic activity by secretion of lipolytic enzymes such as esterases, lipases and phospholipases. These Extracellular hydrolytic enzymes seem to play an important role in Candida overgrowth. Candidiasis is commonly treated with antimycotics such as clotrimazole and nystatin, which bind to a major component of the fungal cell membrane (ergosterol). This binding forms pores in the membrane that lead to death of the fungus. Due to their secondary effects, scientists have thought of another treatment basing on lipase inhibition but we haven’t found any lipase inhibitors used as candidiasis treatment. In this work, we are interested to lipases inhibitors such as alkaloids as another candidiasis treatment. In the first part, we have proceeded to optimize the alkaloid structures and protein 3D structure using Hyperchem software. Secondly, we have docked inhibitors using Genetic algorithm with GOLD software. The results have shown ten possibilities of binding inhibitor to Candida rugosa lipase (CRL) but only one possibility has been accepted depending on the weakest binding energy.

Keywords: marrubiin, candida rugosa lipase, docking, gold

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Authors: Rouag N., Khalifa M. W., Bencheikh A., Abed H.

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The behavior study of forty-two varieties and genotypes of chickpeas regarding root wilt disease induced by Fusarium oxysporum under the natural conditions of infection was conducted at the ITGC experimental station in Sétif. The infected plants of the different chickpea genotypes have shown multiple symptoms in the field caused by the local strain of Fusarium oxysporum f.sp.cecris belonging to race II of the pathogen. These symptoms ranged from lateral or partial wilting of some ramifications to total desiccation of the plant, sometimes combined with the very slow growth of symptomatic plants. The results of the search for sources of resistance to Fusarium wilt of chickpeas in the 42 genotypes tested revealed that in terms of infection rate, the presence of 7 groups and no genotype showed absolute resistance. While in terms of severity, the results revealed the presence of three homogeneous groups. The first group formed by the most resistant genotypes, in this case, Flip10-368C; Flip11-77C; Flip11-186C; Flip11-124C; Flip11-142C, Flip11-152C; Flip11-69C; Ghab 05; Flip11-159C; Flip11-90C; Flip10-357C and Flip11-37C while the second group is the FLIP genotype 10-382C which was found to be the most sensitive for the natural infection test. Thus, the genotypes of Cicer arietinum L., which have shown significant levels of resistance to Fusarium wilt, can be integrated into breeding and improvement programs.

Keywords: chickpea, Cicer arietinum, Fusarium oxysporum, genotype resistance

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Authors: Zhanara B. Suleimenova, Zhazira K. Saduyeva

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Filamentous fungi are major producers of enzymes that have important applications in the food and beverage industries. The overall objective of this research is a strain improvement technology for efficient industrial enzymes production. The new way of filamentous fungi cultivation method has been developed. Such technology prolong producers’ cultivation period up to 60 days and create the opportunity to obtain enzymes repeatedly in every 2-3 days of fungal cultivation. This method is based on immobilizing enzymes producers with solid support in submerged conditions of growth. Immobilizing has a range of advantages: Decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, ability of various enzymes simultaneous production, etc. Design of proposed technology gives the opportunity to increase the activity of immobilized cells culture filtrate comparing to free cells, growing in periodic culture conditions. Thus, proposed research focuses on new, more versatile, microorganisms capable of squeezing more end-products as well as proposed cultivation technology led to increased enzymatic productivity by several times.

Keywords: filamentous fungi, immobilization, industrial enzymes production, strain improvement

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Authors: Rute Faria, Digo Moraes, Amanda Santos, Dione Morais, Maria Sartori

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Seed vigor is a fundamental component for the good performance of the entire soybean production process. Seeds with mechanical damage at harvest time will be more susceptible to fungal and insect attack during storage, which will invariably reduce their vigor to the field, compromising uniformity and final stand performance. Harvesters, even the most modern ones, when not properly regulated or operated, can cause irreversible damages to the seeds, compromising even their commercialization. Therefore, the control of an efficient harvest is necessary in order to guarantee a good quality final product. In this work, the damage caused by two different harvesters (one rented, and another one) was evaluated, traveling in two speeds (4 and 8 km / h). The design was completely randomized in 2 x 2 factorial, with four replications. To evaluate the physiological quality seed germination and vigor tests were carried out over a period of six months. A multivariate analysis of Principal Components (PCA) and clustering allowed us to verify that the leased machine had better performance in the incidence of immediate damages in the seeds, but after a storage period of 6 months the vigor of these seeds reduced more than own machine evidencing that such a machine would bring more damages to the seeds.

Keywords: Glycine max (L.), cluster analysis, PCA, vigor

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Authors: M. A. Gad, A. Z. Al-Herrawy

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Microsporidia comprises various pathogenic species can infect humans by means of water. Moreover, chlorine disinfection of drinking-water has limitations against this protozoan pathogen. A total of 48 water samples were collected from two drinking water treatment plants having two different filtration systems (slow sand filter and rapid sand filter) during one year period. Samples were collected from inlet and outlet of each plant. Samples were separately filtrated through nitrocellulose membrane (142 mm, 0.45 µm), then eluted and centrifuged. The obtained pellet from each sample was subjected to DNA extraction, then, amplification using genus-specific primer for microsporidia. Each microsporidia-PCR positive sample was performed by two species specific primers for Enterocytozoon bieneusi and Encephalitozoon intestinalis. The results of the present study showed that the percentage of removal for microsporidia through different treatment processes reached its highest rate in the station using slow sand filters (100%), while the removal by rapid sand filter system was 81.8%. Statistically, the two different drinking water treatment plants (slow and rapid) had significant effect for removal of microsporidia. Molecular identification of microsporidia-PCR positive samples using two different primers for Enterocytozoon bieneusi and Encephalitozoon intestinalis showed the presence of the two pervious species in the inlet water of the two stations, while Encephalitozoon intestinalis was detected in the outlet water only. In conclusion, the appearance of virulent microsporidia in treated drinking water may cause potential health threat.

Keywords: removal, efficacy, microsporidia, drinking water treatment plants, PCR

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Authors: Laurentius J. M. Rumokoy, Jimmy Posangi, Wisje Lusia Toar, Julio Lopez Aban

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The threat of pathogen infection agents to the neonates is a major health problem to the new born life livestock. Neonate losses became an important case in the world as well as in Indonesia. This condition can be triggered by an infection with nematode in conjunction with a failure of immunoglobulin passive transfer. The study was conducted to evaluate the role of the curcuma combined with IgG colostrum on the development of parasites in the gut of CD1 mice. Animal experiments were divided in four groups (G) based on the treatment: G1 (infection only); G2 (curcuma+infection), G3 (IgG + infection) and G4 (curcuma+IgG+infection). The parameters measured were EPG (eggs per gram) and female in the intestine. The results obtained showed that the treatment has no a significant influence on the number of eggs per gram of feces in the group infected compared to the control group without receiving IgG nor curcuma. However, the EGP response tended to decrease at day 6 in G3 and G4 with a minimum number at zero eggs. This performant showed that the immunoglobulin-G and curcuma substances could slightly decreased the number of eggs in animal infected with Strongyloides. The results obtained showed also that the treatment has no significant difference (P > 0.05) on female larva in the gut of MCD1 experimental. In other side, we found that the best performance to inhibit the female quantity in the gut was the treatment with IgG and infection of parasite in G3. In this treatment, the minimum number was five female only in the gut. The results described IgG response was better than the curcuma single use in reducing the female parasite in the gut. This positive response of IgG compared to other controls group was associated with the function of colostrum antibodies.

Keywords: parasites, livestock, curcuma, colostrums

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Authors: Elizabeth Starzycka-Korbas, Michal Starzycki, Wojciech Rybinski, Mirosława Dabert

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For a few years, the populations of pathogenic fungi occurring in winter oilseed rape in Malyszyn were analyzed. Brassica napus L. in Poland and in the world is a source of energy for both the men (oil), and animals, as post-extraction middling, as well as a motor fuel (oil, biofuel) therefore studies of this type are very important. The species composition of pathogenic fungi can be an indicator of seed yield. The occurrence of oilseed rape pathogens during several years were analyzed using the sequencing method DNA ITS. The results were compared in the gene bank using the program NCBI / BLAST. In field conditions before harvest of oilseed rape presence of pathogens infesting B. napus has been assessed. For example, in 2015, 150 samples have been isolated and applied to PDA medium for the identification of belonging species. From all population has been selected mycelium of 83 isolates which were sequenced. Others (67 isolates) were pathogenic fungi of the genus Alternaria which are easily to recognize. The population of pathogenic species on oilseed rape have been identified after analyzing the DNA ITS and include: Leptosphaeria sp. 38 (L. maculans 25, L. biglobosa 13), Alternaria sp. 29, Fusarium sp. 3, Sclerotinia sclerotiorum 7, heterogeneous 6, total of 83 isolates. The genus Alternaria sp. fungi wear the largest share of B. napus pathogens in particular years. Another dangerous species for oilseed rape was Leptosphaeria sp. Populations of pathogens in each year were different. The number of pathogens occurring in the field and their composition is very important for breeders and farmers because of the possible selection of the most resistant genotypes for sowing in the next growing season.

Keywords: B. napus, DNA ITS Sequencing, pathogenic fungi, population

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Authors: Mahdi Asghari Ozma

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Background and Objective:Maclurapomifera (Rafin.) Schneider, known as osage orange, is a north american native plant which has multiple applications in herbal medicine. The extract of this plant has many therapeutic effects, including antimicrobial, anti-tumor, anti-inflammation, etc., that discussed in this study. Materials and Methods: For this study, the keywords "Maclurapomifera", "osage orange, ""herbal medicine ", and "plant extract" in the databases PubMed and Google Scholar between 2002 and 2021 were searched, and 20 articles were chosen, studied and analyzed. Results: Due to the increased resistance of microbes to antibiotics, the need for antimicrobial plants is increasing. Maclurapomifera is one of the plants with antimicrobial properties that can affect all microbes, especially Gram-negative bacteria, and fungi. This plant also has anti-tumor, anti-inflammatory, anti-oxidant, anti-aging, antiviral, anti-fungal, anti-ulcerogenic, anti-diabetic, and anti-nociceptive effects, which can be used as a substance with many amazing therapeutic applications. Conclusion: These results suggest that the extract of Maclurapomifera can be used in clinical medicine as a remedial agent, which can be substituted for chemical drugs or help them in the treatment of diseases.

Keywords: maclura pomifera, osage orange, herbal medicine, plant extract

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Authors: Alisha Akya, Afsaneh salami

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Background and Objective: Pseudomonas aeruginosa is an opportunistic pathogen associated with nosocomial infections. One of the major concerns for the treatment of P. aeruginosa infections is its resistant to a variety of antibiotics. The purpose of this study was to assess the dissemination of p. aeruginosa isolates obtained from major hospitals in Kermanshah, west of Iran. Materials and Methods: Antibiotic susceptibility testing was performed using the minimal inhibitory concentrations. Mettalo-beta-lactamase was investigated using the double disk diffusion (DDST) test and PCR. Molecular typing was performed by pulsed-field gel electrophoresis (PFGE). Results: The 60 P. aeruginosa isolates, 30 (50%) were resistant to gentamicin, 38 (63/3%) to piperacilin, 42 (70%) to ceftazidime, and 45 (75%) to cefepime. Twenty-nine (48/3%) isolates were MBLs producer based on the DDST test. Five (8/3%) isolates were positive for VIM gene and 4 of them were from burn specimens. PFGE analysis among MBLs producers revealed 12 distinct genotype patterns. A pattern covering the highest number of strains was determined as the dominant clone. Conclusions: Our study showed that P. aeruginosa strains can be spread between patients in hospitals or acquired from different environmental sources. P. aeruginosa isolates were highly resistant to antibiotics and, therefore, the susceptibility of isolates to antibiotics should be tested before treatment. Given the clinical significance of MBLs producing isolates, identification of these organisms is essential in the hospitals in order to get a better therapeutic response and control of bacterial dissemination.

Keywords: clonal dissemination, mettalo-beta-lactamase, Pseudomonas aeruginosa, PFGE

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Authors: Sehrish Iftikhar, Kiran Nawaz, Ahmad A. Shahid, Waheed Anwar, Muhammad S. Haider

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Early blight caused by Alternaria solani Sorauer is one of the most serious foliage diseases of the potato (Solanum tuberosum L.). This disease causes huge crop losses and has major economic importance worldwide. The antifungal activity for three medicinal plants (Foeniculum vulgare, Syzygium aromaticum, and Eucalyptus citriodora) against Alternaria solani has been evaluated. The inhibitory potential of selected essential oils on the radial mycelial growth and germination of spore was measured in vitro at various concentrations (5%, 2.5%. 1.25%, 0.625%, and 0.312%) using agar well diffusion assay. Essential oil of E. citriodora was most effective causing 85% inhibition of mycelial growth and 88% inhibition of spore germination at 0.625% and 1.25% concentrations. Essential oil of Foeniculum vulgare also caused 80% and 82% inhibition of the above mentioned parameters but at double the concentrations 1.25% and 2.5%. While essential oil of Syzygium aromaticum was least effective in controlling the mycelial growth and spore germination with 76% and 77% inhibition at 1.25% and 2.5%. All the selected essential oils, especially E. citriodora, showed marked antimicrobial activity significant at higher concentration. These results suggest that the use of essential oils for the control of A. solani can reduce environmental risks related with commercial fungicides, lower cost for control, and the chances for resistance development. Additional studies are essential to evaluate the potential of essential oils as natural treatments for this disease.

Keywords: clove, essential oils, fennel, potato

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Authors: M. Caruffo, P. Navarrete, C. G. Feijoo, L. Sáenz

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Disease prevention through the use of vaccines has been crucial to achieve the current level of production in the salmon industry. However, vaccines have been developed based largely on inactivated bacterial formulations, using the whole pathogen. These formulations have demonstrated excellent efficacy against extracellular bacterial pathogens. However diseases with the greatest economic impacts correspond to intracellular bacterial and viral pathogens, vaccines based on these types of agents have shown a discrete effectiveness. It is for these reasons that the development of subunit vaccines based on defined antigens offers a promising solution. The main problem is that subunit vaccines offer a low immunogenicity, since they lack immunostimulatory elements, so that the development of new adjuvants platforms becomes an important challenge for this type of formulations. We evaluate the effect of a formulation based on proteoliposomes of Vibrio anguillarum administered by immersion as a new adjuvant strategy, allowing efficient stimulation of the innate immune system. Proteoliposomes physicochemical properties were evaluated in its ability to produce an inflammatory process. Using zebrafish (Danio rerio) larvae as a model species and the transgenic line (Tg(mpx: GFP)i114) allowed us to track the neutrophil migration in real time. Additionally we evaluated the gene expression of some molecular markers involved in the development of the innate immune response characterizing the adjuvant capacity of the formulation.

Keywords: adjuvants, vaccine development, zebrafish, innate immunity

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Authors: Houda Haddad, Radhia Mejri, , Alyne Rodrigues de Araujo, Amira Zairi

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Nosocomial infections represent one of the biggest health problems nowadays. Acinetobacter baumannii is known as an opportunistic pathogen in humans, affecting people with compromised immune systems, and is becoming increasingly important as a hospital-derived infection. It is known that in recent years, more and more bacteria have become multidrug-resistant (MDR), and for this reason, the development of new drugs is a priority. However, these products must not affect the human body, and therefore, cytotoxicity studies are mandatory. In this context, antimicrobial peptides with potential antibacterial proprieties could be an alternative. In this research, we describe the synthesis and the bioactivity of dermaseptins and their derivatives against Acinetobacter baumannii. The cytotoxicity of these dermaseptins was investigated on the HEp-2 cell line by the MTT cell viability assay. Thereafter, we studied morphological alterations caused by the action of one of the active peptides on the bacterial membrane using atomic force microscopy (AFM). The cytotoxicity of dermaseptins was concentration-dependent at microgram concentrations. It was observed that all tested analogs exhibit antibacterial activity with Minimum Inhibitory Concentrations (MICs) ranging from 3.125 to 12.5 μg/mL and Minimum Bactericidal Concentrations (MBCs) ranging from 6.25 to 25 μg/mL. Microscopic images obtained by AFM revealed morphological changes on the surface of treated bacteria caused by K4S4(1-16), as well as significant surface alterations. Overall, these findings demonstrate that dermaseptins might constitute new lead structures for the development of potent antibacterial agents against Acinetobacter baumannii infections.

Keywords: dermaseptin B2, dermaseptin S4, analogs, Acinetobacter baumannii, healthcare-associated infections, antibacterial activity

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Authors: H. Parameshwar Naik, Shripad Kulkarni

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Powdery mildew is a serious disease among the fungal in high humid areas with varied temperature conditions. In recent days disease becomes very severe due to uncertain weather conditions and unique character of the disease is, it produces white mycelia growth on upper and lower leaf surfaces and in severe conditions it leads to defoliation. Results of the experiment revealed that sowing of crop in the I fortnight (FN) of July recorded the minimum mean disease severity (7.96 %) followed by crop sown in II FN of July (13.19 %) as against the crop sown in II FN of August (41.44 %) and I FN of September (33.78 %) and the I fortnight of October (33.77 %). In the first date of sowing infection started at 45 DAS and progressed till 73 DAS and it was up to 14.66 Percent and in second date of sowing disease progressed up to 22.66 percent and in the third date of sowing, it was up to 59.35 percent. Afterward, the disease started earlier and progressed up to 66.15 percent and in sixth and seventh date of sowing disease progressed up to 43.15 percent and 59.85 percent respectively. Disease progress is very fast after 45 days after sowing and highest disease incidence was noticed at 73 DAS irrespective of dates of sowing. From the results of the present study, it is very clear that disease development will be very high if crop sown in between 1st fortnight of August and the 1st fortnight of September.

Keywords: cucumber, India, Karnataka, powdery mildew

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Authors: Sanjay Bari, Vinod Ugale, Kamalkishor Patil

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Over the past several years the emergence of micro-organisms resistant to nearly all the class of antimicrobial agents has become a serious public health concern. In the present research, we report the synthesis and in-vitro antimicrobial activity of a new series of novel quinazolino-thiadiazoles 3 (a-j). The synthesized compounds were confirmed by melting point, IR, 1H-NMR, 13C NMR and Mass spectroscopy. In general, the results of the in-vitro antibacterial activity are encouraging, as out of 10 compounds tested, Compound 3f and 3i with a 4-chloro phenyl and 4-nitro phenyl at C-2 of thiadiazolyl of quinazolino-thiadiazoles, displayed the excellent antibacterial and antifungal activities against all the tested microorganisms (Bacterial and Fungal strain) with MIC values of 62.5 μg/mL. It is worth to mention that the combination of two biologically active moieties quinazoline and thiadiazole profoundly influences the biological activity. While evaluating the antimicrobial activity, it was observed that compounds having electron withdrawing groups on thiazole has shown profound activity in comparison to compounds having electron releasing groups. As a result of this study, it can be concluded that halogen substituent on thiazole ring increases antimicrobial activity. Possible improvements in the antimicrobial activity can be further achieved by slight modifications in the substituent’s and/or additional structural activity investigations to have good antimicrobial activity.

Keywords: antifungal, antimicrobial, quinazolino-thiazoles, synthesis

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Authors: Mohammad A. M. Kewisha

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Xerophilic fungi , which are responsible for many cases of biodeterioration monuments, have been known as an interesting source of antimicrobial compounds. Sixty nine fungal strains, isolated from different localities and species inside Egyptian museums, were screened for antimicrobial activity against some bacterial species and unicellular fungi. The most potent antimicrobial activity was obtained by Asperigillus fumigatus which was identified by ITS4 ……. and showed activity against Staphylococcus aureus with 20 mm and C. albicans with18 mm of inhibition zone. Different parameters were optimized to enhance this activity. The culture grown under stationary conditions for 8 days at 30°C and pH 8 gave the best antimicrobial activity. Moreover, both starch and yeast extract showed the most suitable carbon and nitrogen sources, respectively. The antimicrobial compound was purified and subjected to spectroscopic characterization, which revealed that the antimicrobial compound might be 5,7 ethoxy, 4\,5\ methoxy isorhamnetin -3- O- galactoside. This study suggests that Aspergillus fumagates as a potential candidate offering a better scope for the production, purification and isolation of broad-spectrum antimicrobial compounds. These findings will facilitate the scale-up and further purification to ascertain the compounds responsible for antimicrobial activity, which can be exploited for the treatment of biodeterioration monuments and pharmaceutical applications.

Keywords: antimicrobial activity, asperigillus fumigatus, Identification by ITS4, Staphylococcus aureus, C.albicans

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Authors: Siti Helmyati, Euis Nurdiyawati, Joko Susilo, Endri Yuliati, Siti Fadhilatun Nashriyah, Kurnia Widyastuti

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Background: Iron fortification is one of the most effective and sustainable strategies to overcome anemia. It contradictively, has negative effect on gut microbiota balance. Pathogenic bacteria required iron for their growth. The iron source have greatly affect iron absorption in the intestine. Probiotic can inhibit the growth of pathogen. Lactobacillus plantarum Dad 13, Indonesian local isolate provides many benefits for health while fructo-oligosaccharides (FOS) provides selective substrates for probiotics’ growth. Objective: To determine the effect of iron fortification (NaFeEDTA and FeSO4) on antibacterial activity of synbiotic fermented milk. Methods: The antibacterial activity test was performed using the disc diffusion method. Paper discs were soaked in three kinds of synbiotic fermented milk, which are: 1) fortified with NaFeEDTA, 2) FeSO4 and 3) control. Escherichia coli was inoculated on nutrient agar medium. The ability of inhibition was shown by the formation of clear zone around the paper disc and measured in diameter (mm). Results: Synbiotic fermented milk fortified with iron (either NaFeEDTA or FeSO4) had antibacterial activity against Escherichia coli with diameter of clear zone were 6.53 mm and 12.3 mm, respectively (p<0.05). Compared to control (10.73 mm), synbiotic fermented milk fortified with FeSO4 had similar antibacterial activity (p>0.05). Conclusions: In vitro, synbiotic fermented milk fortified with NaFeEDTA and FeSO4 had different antibacterial activity against Escherichia coli. Iron fortification compound affected the antibacterial activity of synbiotic fermented milk.

Keywords: lactobacillus plantarum Dad 13, FOS, NaFeEDTA, FeSO4, antibacterial activity

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Authors: Francisco Alvarado, Luz Ramírez

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Corneal diseases are one of the main causes of visual impairment and affect almost 4 million, and this study assesses the effects of deep anterior lamellar keratoplasty (DALK) with porcine corneal stroma and postoperative topical treatment with tacrolimus in patients with infectious keratitis. No patient was observed with clinical graft rejection. Among the cases: 2 were positive to fungal culture, 2 with Aspergillus and the other 8 cases were confirmed by bacteriological culture. Corneal diseases are one of the main causes of visual impairment and affect almost 4 million. This study assesses the effects of deep anterior lamellar keratoplasty (DALK) with porcine corneal stroma and postoperative topical treatment with tacrolimus in patients with infectious keratitis. Receiver bed diameters ranged from 7.00 to 9.00 mm. No incidents of Descemet's membrane perforation were observed during surgery. During the follow-up period, no corneal graft splitting, IOP increase, or intolerance to tacrolimus were observed. Deep anterior lamellar keratoplasty seems to be the best option to avoid xenograft rejection, and it could help new surgical techniques in humans.

Keywords: ophthalmology, cornea, corneal transplant, xenografts, surgical innovations

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Authors: T. O. Adejumo, O. S. Akinyemi

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Powders and oils of Aframomum melegueta, Garcinia kola and Piper guineense were tested as preservatives for the storage of maize grains for four weeks. The concentrations of the powders were 32.5gkg-1, 65.0gkg-1 and 97.5gkg-1 maize, while those of oils were 0.85mlkg-1, 0.50mlkg-1 and 0.75mlkg-1 maize respectively. Powders of the three botanicals at 97.5gkg-1 maize possessed insecticidal effect on Sitophilus zeamais and also inhibitory activities on Aspergillus flavus, A. fumigatus, A. niger and Fusarium verticillioides, while little effect was observed for other concentrations. Oils of the three botanicals at 0.50mlkg-1 and 0.75mlkg-1 maize showed an insecticidal effect on S. zeamais and also inhibitory activities on A. flavus, A. fumigatus, A. niger, F. verticillioides, Penicillium and Rhizopus species. Oils showed more potential as a protectant against fungal and insect pest in storage maize grains than powders. Powders and oils of A. melegueta, G. kola and P. guineense could be successfully used as biopesticides.

Keywords: aframomum melegueta, garcinia kola, maize, powder, oils, piper guineense

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Authors: Ssembatya Joseph Mary

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Introduction: Cryptococcal infection in the Central Nervous System (CNS) is frequently seen in human immunodeficiency virus (HIV) patients and others with low immunity as an opportunistic fungal infection. However, CNS cryptococcal granuloma (cryptococcoma) in immunocompetent patients is rare. We present a case of CNS cryptococcoma in an immunocompetent patient and review the literature to illustrate the diagnosis and treatment of such lesions. Case presentation: A 62-year-old, HIV-negative, immunocompetent female patient with no known chronic illness presented with 5 months history of a progressive headache associated with on and off episodic generalized tonic-clonic convulsions. She had been to several hospitals before she was referred to our center with a diagnosis of a brain tumor. Before referral and despite a negative CSF analysis result, she had received treatment for bacterial meningitis with no success. At Mbarara Regional Referral Hospital (MRRH), she had surgery with an excision biopsy which showed features consistent with cryptococcosis on histology. The patient had a successful adjuvant treatment with antifungal drugs following surgery. Conclusion: The diagnosis of a parasitic CNS infection, particularly cryptococcal infection mimicking neoplastic lesions in an immunocompetent patient, was unusual. Surgical management of such lesions from different reports has a bad outcome and management remains totally conservative.

Keywords: Cryptococcal meningitis, immunocompetent patient, Uganda, low resource setting

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Authors: Deepika Biswas

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Acinetobacter baumannii, a hospital-acquired pathogen, causes nosocomial infections including pneumonia, urinary tract infection, and secondary meningitis. Carbapenem is most effective antibiotics against it. Its increased resistance to carbapenems has been a rising global concern. Antibiotics such as carbapenem are unable to use on hospital setups to eradicate A. baumannii, hence different concentrations of disinfectants including phenol; sodium hypochlorite and ethanol are increasingly being used. The objective of the present study is to find an effective concentration of above disinfectants against carbapenem-resistant strain RS307 of A. baumannii. Growth kinetics of RS307 has been determined using UV-Vis spectrophotometer in the presence and absence of disinfectants in triplicate and its standard deviation has also been calculated which make the results more reliable. Differential growth curves were plotted, which showed the effective concentration among all the concentrations of phenol, sodium hypochlorite and ethanol. On disc diffusion assay, antimicrobial effect was observed by comparing all the concentrations of disinfectants to check its synergy with imipenem, most effective carbapenem. All the results collectively revealed that 0.5% phenol, 0.5% sodium hypochlorite, and 70% ethanol could preferably be used as disinfectant for hospital setup against the carbapenem-resistant strain of A. baumannii. SDS PAGE analysis showed differential expression in the protein profile of A. baumannii after treatment. The present study highlighted that few disinfectants even in low concentration had shown better antimicrobial activity hence may be recommended for regular use in the hospitals, which will be cost effective and less harmful.

Keywords: Acenatobacter bomunii, phenol, sodium hypoclirite, ethanol, carbapenem resistance, disinfectant

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Authors: Minhas Alam, Muhammad Hidayat Rasool, Mohsin Khurshid, Bilal Aslam

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Acinetobacter baumannii is a notorious bacterial pathogen that is an emerging nightmare in clinical settings and is mainly involved in severe nosocomial infections. However, the data related to carbapenem-resistant A. baumannii (CRAB) from veterinary settings is limited, especially in developing countries like Pakistan. To investigate the genetic diversity and molecular basis of carbapenem resistance in Acinetobacter baumannii isolates from Cattle, a total of 1960 samples were collected from cattle from Punjab, Pakistan. The isolates were analyzed by routine microbiological procedures and confirmed by polymerase chain reaction (PCR). The isolates were further screened for antimicrobial susceptibility and the presence of multiple antimicrobial-resistant determinants by PCR. Multilocus sequence typing (MLST) was performed. The results of the current study revealed that the overall prevalence of A. baumannii in cattle was 3.28% (65/1980). Among cattle 27.7% (18/65) were found CRAB strains. The CRAB isolates harbor class D β- lactamases genes, e-g, blaOXA-23 and blaOXA-51, 94.4% (17/18). CRAB isolates carry class B β- lactamases gene blaIMP, and only one isolate carries the blaNDM-1 gene. The MLST results of CRAB isolates from cattle demonstrated 5 STs and one new ST. The commonly found sequence types in CRAB isolates were ST2 (n=10, 55.5%), followed by ST642 (n=5, 27.8%) and ST600 & ST889 (n=1, 5.55%). The presence of CRAB isolates in cattle indicates an alarming situation in Punjab, Pakistan. Immediate control measures should be taken to stop the transmission of CRAB isolates within cattle, to the environment, and to clinical settings.

Keywords: acinetobacter baumannii, carbapenemases, veterinary, drug resistance

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Authors: Congping Lin

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Intracellular transport in fungi has a number of important roles in, e.g., filamentous fungal growth and cellular metabolism. Two basic mechanisms for intracellular transport are motor-driven trafficking along microtubules (MTs) and diffusion. Mathematical modelling has been actively developed to understand such intracellular transport and provide unique insight into cellular complexity. Based on live-cell imaging data in Ustilago hyphal cells, probabilistic models have been developed to study mechanism underlying spatial organization of molecular motors and organelles. In particular, anther mechanism - stochastic motility of dynein motors along MTs has been found to contribute to half of its accumulation at hyphal tip in order to support early endosome (EE) recycling. The EE trafficking not only facilitates the directed motion of peroxisomes but also enhances their diffusive motion. Considering the importance of spatial organization of early endosomes in supporting peroxisome movement, computational and experimental approaches have been combined to a whole-cell level. Results from this interdisciplinary study promise insights into requirements for other membrane trafficking systems (e.g., in neurons), but also may inform future 'synthetic biology' studies.

Keywords: intracellular transport, stochastic process, molecular motors, spatial organization

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Authors: Zonglin Yang, Tatsuya Akiyama, Kerry S. Williamson, Michael J. Franklin, Thiruvarangan Ramaraj

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Pseudomonas aeruginosa is an opportunistic pathogen that forms surface-associated microbial communities (biofilms) on artificial implant devices and on human tissue. Biofilm infections are difficult to treat with antibiotics, in part, because the bacteria in biofilms are physiologically heterogeneous. One measure of biological heterogeneity in a population of cells is to quantify the cellular concentrations of ribosomes, which can be probed with fluorescently labeled nucleic acids. The fluorescent signal intensity following fluorescence in situ hybridization (FISH) analysis correlates to the cellular level of ribosomes. The goals here are to provide computationally and statistically robust approaches to automatically quantify cellular heterogeneity in biofilms from a large library of epifluorescent microscopy FISH images. In this work, the initial steps were developed toward these goals by developing an automated biofilm detection approach for use with FISH images. The approach allows rapid identification of biofilm regions from FISH images that are counterstained with fluorescent dyes. This methodology provides advances over other computational methods, allowing subtraction of spurious signals and non-biological fluorescent substrata. This method will be a robust and user-friendly approach which will enable users to semi-automatically detect biofilm boundaries and extract intensity values from fluorescent images for quantitative analysis of biofilm heterogeneity.

Keywords: image informatics, Pseudomonas aeruginosa, biofilm, FISH, computer vision, data visualization

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