Search results for: resistant microbial strains

Authors: G. R. Hashemi Tabar, G. R. Razmi, F. Mirshekar

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Echinococcus granulosus have ten strains from G1 to G9. Each strain is related to special intermediated host. The morphology, epidemiology, treatment and control in these strains are different. There are many morphological and molecular methods to differentiate of Echinococcus strains. However, using both methods were provided better information about identification of each strain. The aim of study was to identify Echinococcus granulosus strain of hydrated cysts in slaughtered sheep using morphological and molecular methods in Mashhad area. In the present study, the infected liver and lung with hydatid cysts were collected and transferred to laboratory. The hydatid cyst liquid was extracted and morphological characters of rostellar hook protosclocies were measured using micrometer ocular. The total length of large blade length of large hooks, total length of small and blade length of small hooks, and number of hooks per protoscolex were 23± 0.3μm, 11.7±0.5 μm, 19.3±1.1 μm,8±1.1 and 33.7±0.7 μm, respectively. In molecular section of the study, DNA each samples was extracted with MBST Kit and development of PCR using special primers (EgF, EgR) which amplify fragment of ITS1 gen. The PCR product was digested with Bsh1236I enzyme. Based on pattern of PCR-RLFP results (four band forming), G1, G2 and G3 strain of Echinococcus granulosus were obtained. Differentiation of three strains was done using sequencing analysis and G1 strain was diagnosed. The agreement between the molecular results with morphometric characters of rosetellar hook was confirmed the presence of G1 strain of Echinococcus in the slaughtered sheep of Mashhad area.

Keywords: Echinococcus granulosus, Hydatid cyst, PCR, sheep

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Authors: Mst. Tuhina-Khatun, Mohamed Hanafi Musa, Mohd Rafii Yosup, Wong Mui Yun, Md. Aktar-Uz-Zaman, Mahbod Sahebi

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Rice is a staple crop of economic importance of most Asian people, and blast is the major constraints for its higher yield. Heritability of plants traits helps plant breeders to make an appropriate selection and to assess the magnitude of genetic improvement through hybridization. Diversity of crop plants is necessary to manage the continuing genetic erosion and address the issues of genetic conservation for successfully meet the future food requirements. Therefore, an experiment was conducted to estimate heritability and to determine the diversity of 27 blast resistant upland rice genotypes based on 18 quantitative traits using randomized complete block design. Heritability value was found to vary from 38 to 93%. The lowest heritability belonged to the character total number of tillers/plant (38%). In contrast, number of filled grains/panicle, and yield/plant (g) was recorded for their highest heritability value viz. 93 and 91% correspondingly. Cluster analysis based on 18 traits grouped 27 rice genotypes into six clusters. Cluster I was the biggest, which comprised 17 genotypes, accounted for about 62.96% of total population. The multivariate analysis suggested that the genotype ‘Chokoto 14’ could be hybridized with ‘IR 5533-55-1-11’ and ‘IR 5533-PP 854-1’ for broadening the gene pool of blast resistant upland rice germplasms for yield and other favorable characters.

Keywords: blast resistant, diversity analysis, heritability, upland rice

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Authors: Eusèbe Gnonlonfoun, Xavier Framboisier, Michel Fick, Emmanuel Rondags

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Pathogenic fungi represent a generic problem for cereals, including barley, as they can produce a number of thermostable toxic metabolites such as mycotoxins that contaminate plants and food products, leading to serious health issues for humans and animals and causing significant losses in global food production. In addition, mycotoxins represent a significant technological concern for the malting and brewing industries, as they may affect the quality and safety of raw materials (barley and malt) and final products (beer). Moreover, this situation is worsening due to the highly variable climatic conditions that favor microbial development and the societal desire to reduce the use of phytosanitary products, including fungicides. In this complex environmental, regulatory and economic context for the French barley-malt-beer industry, this project aims to develop an innovative biocontrol process by using technological bacteria, isolated from infection-resistant barley cultures, that are able to reduce the development of spoilage fungi and the associated mycotoxin production. The experimental approach consists of i) coculturing bacterial and pathogenic fungal strains in solid and liquid media to access the growth kinetics of these microorganisms and to evaluate the impact of these bacteria on fungal growth and mycotoxin production; then ii) the results will be used to carry out a micro-malting process in order to develop the aforementioned process, and iii) the technological and sanitary properties of the generated barley malts will finally be evaluated in order to validate the biocontrol process developed. The process is expected to make it possible to guarantee, with controlled costs, an irreproachable hygienic and technological quality of the malt, despite the increasingly complex and variable conditions for barley production. Thus, the results will not only make it possible to maintain the dominant world position of the French barley-malt chain but will also allow it to conquer emerging markets, mainly in Africa and Asia. The use of this process will also contribute to the reduction of the use of phytosanitary products in the field for barley production while reducing the level of contamination of malting plant effluents. Its environmental impact would therefore be significant, especially considering that barley is the fourth most-produced cereal in the world.

Keywords: barley, pathogenic fungi, mycotoxins, malting, bacterial biocontrol

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Authors: Valeria Velasco, Ana M. Bonilla, José L. Vergara, Alcides Lofa, Jorge Campos, Pedro Rojas-García

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Staphylococcus aureus is both commensal bacterium and opportunistic pathogen that can cause different diseases in humans and can rapidly develop antimicrobial resistance. Since this bacterium has the ability to colonize the nares and skin of humans and animals, there is a risk of contamination of food in different steps of the food chain supply. Emerging strains have been detected in food-producing animals and meat, such as methicillin-resistant S. aureus (MRSA). The aim of this study was to determine the prevalence and oxacillin susceptibility of S. aureus in the pork chain supply in Chile and to suggest some natural antimicrobials for control. A total of 487 samples were collected from pigs (n=332), carcasses (n=85), and retail pork meat (n=70). Presumptive S. aureus colonies were isolated by selective enrichment and culture media. The confirmation was carried out by biochemical testing (Api® Staph) and molecular technique PCR (detection of nuc and mecA genes, associated with S. aureus and methicillin resistance, respectively). The oxacillin (β-lactam antibiotic that replaced methicillin) susceptibility was assessed by minimum inhibitory concentration (MIC) using the Epsilometer test (Etest). A preliminary assay was carried out to test thymol, carvacrol, oregano essential oil (Origanum vulgare L.), Maqui or Chilean wineberry extract (Aristotelia chilensis (Mol.) Stuntz) as anti-staphylococcal agents using the disc diffusion method at different concentrations. The overall prevalence of S. aureus in the pork chain supply reached 33.9%. A higher prevalence of S. aureus was determined in carcasses (56.5%) than in pigs (28.3%) and pork meat (32.9%) (P ≤ 0.05). The prevalence of S. aureus in pigs sampled at farms (40.6%) was higher than in pigs sampled at slaughterhouses (23.3%) (P ≤ 0.05). The contamination of no packaged meat with S. aureus (43.1%) was higher than in packaged meat (5.3%) (P ≤ 0.05). The mecA gene was not detected in S. aureus strains isolated in this study. Two S. aureus strains exhibited oxacillin resistance (MIC ≥ 4µg/mL). Anti-staphylococcal activity was detected in solutions of thymol, carvacrol, and oregano essential oil at all concentrations tested. No anti-staphylococcal activity was detected in Maqui extract. Finally, S. aureus is present in the pork chain supply in Chile. Although the mecA gene was not detected, oxacillin resistance was found in S. aureus and could be attributed to another resistance mechanism. Thymol, carvacrol, and oregano essential oil could be used as anti-staphylococcal agents at low concentrations. Research project Fondecyt No. 11140379.

Keywords: antimicrobials, mecA gen, nuc gen, oxacillin susceptibility, pork meat

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Authors: Maria Ostroukhova, Zhanneta Zalutskaya, Elena Ermilova

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The alternative oxidase (AOX) mediates cyanide-resistant respiration, which bypasses proton-pumping complexes III and IV of the cytochrome pathway to directly transfer electrons from reduced ubiquinone to molecular oxygen. In Chlamydomonas reinhardtii, AOX is a monomeric protein that is encoded by two genes of discrete subfamilies, AOX1 and AOX2. Although AOX has been proposed to play essential roles in stress tolerance of organisms, the role of subfamily AOX2 is largely unknown. In C. reinhardtii, AOX2 was initially identified as one of constitutively low expressed genes. Like other photosynthetic organisms C. reinhardtii cells frequently experience periods of hypoxia. To examine AOX2 transcriptional regulation and role of AOX2 in hypoxia adaptation, real-time PCR analysis and artificial microRNA method were employed. Two experimental approaches have been used to induce the anoxic conditions: dark-anaerobic and light-anaerobic conditions. C. reinhardtii cells exposed to the oxygen deprivation have shown increased AOX2 mRNA levels. By contrast, AOX1 was not an anoxia-responsive gene. In C. reinhardtii, a subset of genes is regulated by transcription factor CRR1 in anaerobic conditions. Notable, the AOX2 promoter region contains the potential motif for CRR1 binding. Therefore, the role of CRR1 in the control of AOX2 transcription was tested. The CRR1-underexpressing strains, that were generated and characterized in this work, exhibited low levels of AOX2 transcripts under anoxic conditions. However, the transformants still slightly induced AOX2 gene expression in the darkness. These confirmed our suggestions that darkness is a regulatory stimulus for AOX genes in C. reinhardtii. Thus, other factors must contribute to AOX2 promoter activity under dark-anoxic conditions. Moreover, knock-down of CRR1 caused a complete reduction of AOX2 expression under light-anoxic conditions. These results indicate that (1) CRR1 is required for AOX2 expression during hypoxia, and (2) AOX2 gene is regulated by CRR1 together with yet-unknown regulatory factor(s). In addition, the AOX2-underexpressing strains were generated. The analysis of amiRNA-AOX2 strains suggested a role of this alternative oxidase in hypoxia adaptation of the alga. In conclusion, the results reported here show that C. reinhardtii AOX2 gene is stress inducible. CRR1 transcriptional factor is involved in the regulation of the AOX2 gene expression in the absence of oxygen. Moreover, AOX2 but not AOX1 functions under oxygen deprivation. This work was supported by Russian Science Foundation (research grant № 16-14-10004).

Keywords: alternative oxidase 2, artificial microRNA approach, chlamydomonas reinhardtii, hypoxia

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Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

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Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

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Authors: Klementina Puto, Ermelinda Nexhipi, Evi Llaka

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Dairy by-products are a fairly good environment for microorganisms due to their composition for their growth. Microbial populations have a significant impact in the production of cheese, butter, yogurt, etc. in terms of their organoleptic quality and at the same time some also cause their breakdown. In this paper, the microbiological contamination of soft cheese, butter and yogurt produced in the country (domestic) and imported is assessed, as an indicator of hygiene with impact on public health. The study was extended during September 2018-June 2019 and was divided into three periods, September-December, January-March, and April-June. During this study, a total of 120 samples were analyzed, of which 60 samples of cheese and butter locally produced, and 60 samples of imported soft cheese and butter productions. The microbial indicators analyzed are Staphylococcus aureus and E. coli. Analyzes have been conducted at the Food Safety Laboratory (FSIV) in Tirana in accordance with EU Regulation 2073/2005. Sampling was performed according to the specific international standards for these products (ISO 6887 and ISO 8261). Sampling and transport of samples were done under sterile conditions. Also, coding of samples was done to preserve the anonymity of subjects. After the analysis, the country's soft cheese products compared to imports were more contaminated with S. aureus and E. coli. Meanwhile, the imported butter samples that were analyzed, resulted within norms compared to domestic ones. Based on the results, it was concluded that the microbial quality of samples of cheese, butter and yogurt analyzed remains a real problem for hygiene in Albania. The study will also serve business operators in Albania to improve their work to ensure good hygiene on the basis of the HACCP plan and to provide a guarantee of consumer health.

Keywords: consumer, health, dairy, by-products, microbial

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Authors: Nassima Behidj-Benyounes, Thoraya Dahmene, Nadjiba Chebout

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This work examines the study of the antimicrobrial effect of alkaloids extracted from the seeds of Peganum harmala L (Zygophyllaceae). This natural substance is extracted by using different solvents (aqueous, ethanolic, and hexane). The evaluation of the antimicrobial activity has only dealt with alkaloids. The antimicrobial effect of alkaloids is evaluated on several microorganisms. It has been tested on eight bacterial strains. The extract has been studied by using two yeasts. Finally, three molds have been studied. It should be noted that these agents are characterized by a high frequency of contamination and pathogenicity. Through this study, we note that Staphylococcus aureus, Saccharomyces cerievisae and E. coli are very sensitive in respect of the ethanol extract. Pseudomonas aerogenosa and Penicillium sp. are resistant to this extract. The other microorganisms are moderately sensitive. The study of the antimicrobial activity of different extracts of the Harmel has shown an optimal activity with the ethanol extract.

Keywords: Peganum harmala L., seeds, alkaloids, bacteria, fungi, yeast, antimicrobial activity

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Authors: Fiston Masudi Tambwe, Lwanga Charles, Arfang Badji, Unzimai Innocent

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The development of maize varieties combining Provitamin A (PVA), high yield, and Striga resistance is an effective and affordable strategy to contribute to food security in sub-Saharan Africa, where maize is a staple food crop. There has been limited research on introgressing PVA genes into Striga-resistant maize genotypes. The objectives of this study were to: i) determine the mode of gene action controlling PVA carotenoid accumulation in Striga-resistant maize, ii) identify Striga-resistant maize hybrids with high PVA content and stable yield, and iii) validate the presence of PVA functional markers in offspring. Six elite, Striga-resistant inbred females were crossed with six high-PVA inbred males in a North Carolina Design II and their offspring were evaluated in four environments, following a 5x8 alpha lattice design with four hybrid checks. Results revealed that both additive and non-additive gene action control carotenoid accumulation in the present study, with a predominance of non-additive gene effects for PVA. Hybrids STR1004xCLHP0352 and STR1004xCLHP0046 - identified as Striga-resistant because they supported fewer Striga plants – were the highest-yielding genotypes with a moderate PVA concentration of 5.48 and 5.77 µg/g, respectively. However, those two hybrids were not stable in terms of yield across all environments. Hybrid STR1007xCLHP0046, however, supported fewer Striga plants, had a yield of 4.52 T/ha, a PVA concentration of 4.52 µg/g, and was also stable. Gel-based marker systems of CrtRB1 and LCYE were used to screen the hybrids and favorable alleles of CrtRB1 primers were detected in 20 hybrids, confirming good levels of PVA carotenoids. Hybrids with favorable alleles of LCYE had the highest concentration of non-PVA carotenoids. These findings will contribute to the development of high-yielding PVA-rich maize varieties in Uganda.

Keywords: gene action, stability, striga resistance, provitamin A markers, beta-carotene hydroxylase 1, CrtRB1, beta-carotene, beta-cryptoxanthin, lycopene epsilon cyclase, LCYE

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Authors: Ana C. Sousa, Beatriz C. Santos, Fátima N. Serralha

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The Tagus estuary is heavily affected by industrial and urban activities, making bioremediation studies crucial for environmental preservation. Fuel contamination in the area can arise from various anthropogenic sources, such as oil spills from shipping, fuel storage and transfer operations, and industrial discharges. These pollutants can cause severe harm to the ecosystem and the organisms, including humans, that inhabit it. Nonetheless, there are always natural organisms with the ability to resist these pollutants and transform them into non-toxic or harmless substances, which defines the process of bioremediation. Exploring the microbial communities existing in soil and their capacity to break down hydrocarbons has the potential to enhance the development of more efficient bioremediation approaches. The aim of this investigation was to explore the existence of hydrocarbonoclastic microorganisms in six locations within the Tagus estuary, three on the north bank: Trancão River, Praia Fluvial do Cais das Colinas and Praia de Algés, and three on the south bank: Praia Fluvial de Alcochete, Praia Fluvial de Alburrica, and Praia da Trafaria. In all studied locations, native microorganisms of the genus Pseudomonas were identified. The bioremediation rate of common hydrocarbons like gasoline, hexane, and toluene was assessed using the redox indicator 2,6-dichlorophenolindophenol (DCPIP). Effective hydrocarbon-degrading bacterial strains were identified in all analyzed areas, despite adverse environmental conditions. The highest bioremediation rates were achieved for gasoline (68%) in Alburrica, hexane (65%) in Algés, and toluene (79%) in Algés. Generally, the bacteria demonstrated efficient degradation of hydrocarbons added to the culture medium, with higher rates of aerobic biodegradation of hydrocarbons observed. These findings underscore the necessity for further in situ studies to better comprehend the relationship between native microbial communities and the potential for pollutant degradation in soil.

Keywords: biodegradability rate, hydrocarbonoclastic microorganisms, soil bioremediation, tagus estuary

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Authors: Funmilola O. Omoya, Joseph O. Obameso, Titus A. Olukibiti

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This study was carried out to reveal the bacterial composition of aerosol in the studied abattoirs. Bacteria isolated were characterized according to microbiological standards. Factors such as temperature and distance were considered as variable in this study. The isolation was carried out at different temperatures such as 27oC, 31oC and 29oC and at various distances of 100meters and 200meters away from the slaughter sites. Result obtained showed that strains of Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Lactobacillus alimentarius and Micrococcus sp. were identified. The total viable counts showed that more microorganisms were present in the morning while the least viable count of 388 cfu was recorded in the evening period of this study. This study also showed that more microbial loads were recorded the further the distance is to the slaughter site. Conclusively, the array of bacteria isolated suggests that abattoir sites may be a potential source of pathogenic organisms to commuters if located within residential environment.

Keywords: abattoir, aerosol, bacterial composition, environment

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Authors: Somaye Eskandary

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Polycyclic aromatic hydrocarbons (PAHs) are present in crude oil and its derivatives contaminate soil and also increase carcinogen and mutagen contamination, which is a concern for researchers. Land farming is one of the methods that remove pollutants from the soil by native microorganisms. It seems that this technology is cost-effective, environmentally friendly and causes less debris problem to be disposed. This study aimed to refine the polycyclic aromatic hydrocarbons from oil-contaminated soil using the land farming method. In addition to examine the concentration of polycyclic aromatic hydrocarbons by GC-FID, some characteristics such as soil microbial respiration and dehydrogenase, peroxidase, urease, acid and alkaline phosphatase enzyme concentration were also measured. The results showed that after land farming process the concentrations of some polycyclic aromatic hydrocarbons dropped to 50 percent. The results showed that the enzyme concentration is reduced by reducing the concentration of hydrocarbons and microbial respiration. These results emphasize the process of land farming for removal of polycyclic aromatic hydrocarbons from soil by indigenous microorganisms.

Keywords: soil contamination, gas chromatography, native microorganisms, soil enzymes, microbial respiration, carcinogen

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Authors: Pei-Lin Yueh, Bor-Yann Chen, Chuan-Chung Hsueh

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This first-attempt study revealed that decolorized intermediates of azo dyes could act as redox mediators to assist wastewater (WW) decolorization due to enhancement of electron-transport phenomena. Electrochemical impedance spectra indicated that hydroxyl and amino-substituent(s) were functional group(s) as redox-mediator(s). As azo dyes are usually multiple benzene rings structured, their derived decolorized intermediates are likely to play roles of electron shuttles due to lower barrier of energy gap for electron shuttling. According to cyclic voltammetric profiles, redox-mediating characteristics of decolorized intermediates of azo dyes (e.g., RBu171, RR198, RR141, and RBk5) were clearly disclosed. With supplementation of biodecolorized metabolites of RR141 and 198, decolorization performance of could be evidently augmented. This study also suggested the optimal modes of microbial fuel cell (MFC)-assisted WW decolorization would be plug-flow or batch mode of operation with no mix. Single chamber-MFCs would be more favourable than double chamber MFCs due to non-mixing contacting reactor scheme for operation.

Keywords: redox mediators, dye decolorization, bioelectricity generation, microbial fuel cells

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Authors: Nurhajati Hakim, Kiki Amalia, A. Agustian, H. Hermansah, Y. Yulnafatmawita

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Ultisols require greater amounts of fertilizer application compared to other soils and susceptible to erosion. Unfortunately, the price of synthetic fertilizers has increased over time during the years, making them unaffordable for most Indonesian farmers. While terrace technique to control erosion very costly.Over the last century, efforts to reduce reliance on synthetic agro-chemicals fertilizers and erosion control have recently focused on Tithonia diversifolia as a fertilizer alternative, and as hedgerow plant to control erosion. Generally known by its common name of tree marigold or Mexican sunflower, this plant has attracted considerable attention for its prolific production of green biomass, rich in nitrogen, phosphorous and potassium (NPK). In pot experiments has founded some microbial such as Mycorrhizal, Azotobacter, Azospirillum, phosphate solubilizing bacterial (PSB) and fungi (PSF) are expected to play an important role in biomass production and high nutrient uptake of this plant. This issue of importance was pursued further in the following investigation in field condition. The aim of this study was to determine the type of microbial combination suitable for Tithonia cultivation as hedgerow plants in Ultisols which have higher biomass production and nutrient content, and decline soil erosion. The field experiment was conducted with 6 treatments in a randomized block design (RBD) using 3 replications. The treatments were: Tithonia rhizosphere without microbial inoculated (A); Inokulanted by Mycorrhizal + Azotobacter + Azospirillium (B); Mycorrhizal + PSF (C); Mycorrhizal + PSB(D); Mycorrhizal + PSB + PSF(E);and without hedgerow Tithonia (F).The microbial substrates were inoculated into the Tithonia rhizosphere in the nursery. The young Tithonia plants were then planted as hedgerow on Ultisols in the experimental field for 8 months, and pruned once every 2 months. Soil erosion were collected every rainy time. The differences between treatments were statistically significant by HSD test at the 95% level of probability. The result showed that treatment C (mycorrhizal + PSB) was the most effective, and followed by treatment D (mycorrhizal + PSF) in producing higher Tithonia biomass about 8 t dry matter 2000 m-2 ha-1 y-1 and declined soil erosion 71-75%.

Keywords: hedgerow tithonia, microbial inoculants, organic fertilizer, soil erosion control

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Authors: Lali Kutateladze, Tamar Urushadze, Tamar Dudauri, Besarion Metreveli, Nino Zakariashvili, Izolda Khokhashvili, Maya Jobava

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Lignocellulosic waste streams from agriculture, paper and wood industry are renewable, plentiful and low-cost raw materials that can be used for large-scale production of liquid and gaseous biofuels. As opposed to prevailing multi-stage biotechnological processes developed for bioconversion of cellulosic substrates to ethanol where high-cost cellulase preparations are used, Consolidated Bioprocessing (CBP) offers to accomplish cellulose and xylan hydrolysis followed by fermentation of both C6 and C5 sugars to ethanol in a single-stage process. Syntrophic microbial consortium comprising of anaerobic, thermophilic, cellulolytic, and saccharolytic bacteria in the genus Clostridia with improved ethanol productivity and high tolerance to fermentation end-products had been proposed for achieving CBP. 65 new strains of anaerobic thermophilic cellulolytic and saccharolytic Clostridia were isolated from different wetlands and hot springs in Georgia. Using new isolates, fermentation of mechanically pretreated wheat straw and corn stalks was done under oxygen-free nitrogen environment in thermophilic conditions (T=550C) and pH 7.1. Process duration was 120 hours. Liquid and gaseous products of fermentation were analyzed on a daily basis using Perkin-Elmer gas chromatographs with flame ionization and thermal detectors. Residual cellulose, xylan, xylose, and glucose were determined using standard methods. Cellulolytic and saccharolytic bacteria strains degraded mechanically pretreated herbaceous cellulosic wastes and fermented glucose and xylose to ethanol, acetic acid and gaseous products like hydrogen and CO2. Specifically, maximum yield of ethanol was reached at 96 h of fermentation and varied between 2.9 – 3.2 g/ 10 g of substrate. The content of acetic acid didn’t exceed 0.35 g/l. Other volatile fatty acids were detected in trace quantities.

Keywords: anaerobic bacteria, cellulosic wastes, Clostridia sp, ethanol

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Authors: Amine Alaoui Sanae, Tagjdid Reda, Loutfi Chafiqa, Melloul Merouane, Laloui Aziz, Touil Nadia, El Fahim, E. Mostafa

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Background: Rotavirus group A (RVA) strains with G8P[14] specificities are usually detected in calves and goats. However, these strains have been reported globally in humans and have often been characterized as originating from zoonotic transmissions, particularly in area where ruminants and humans live side-by-side. Whether human P[14] genotypes are two-way and can be transmitted to animal species remains to be established. Here we describe VP4 deduced amino-acid relationships of three Moroccan P[14] genotypes originating from different species and the receptiveness of an alpine goat to a human G8P[14] through an experimental infection. Material/methods: the human MA31 RVA strain was originally identified in a four years old girl presenting an acute gastroenteritis hospitalized at the pediatric care unit in Rabat Hospital in 2011. The virus was isolated and propagated in MA104 cells in the presence of trypsin. Ch_10S and 8045_S animal RVA strains were identified in fecal samples of a 2-week-old native goat and 3-week-old calf with diarrhea in 2011 in Bouaarfa and My Bousselham respectively. Genomic RNAs of all strains were subjected to a two-step RT-PCR and sequenced using the consensus primers VP4. The phylogenetic tree for MA31, Ch_10S and 8045_S VP4 and a set of published P[14] genotypes was constructed using MEGA6 software. The receptivity of MA31 strain by an eight month-old alpine goat was assayed. The animal was orally and intraperitonally inoculated with a dose of 8.5 TCID50 of virus stock at passage level 3. The shedding of the virus was tested by a real time RT-PCR assay. Results: The phylogenetic tree showed that the three Moroccan strains MA31, Ch_10S and 8045_S VP4 were highly related to each other (100% similar at the nucleotide level). They were clustered together with the B10925, Sp813, PA77 and P169 strains isolated in Belgium, Spain and Italy respectively. The Belgian strain B10925 was the most closely related to the Moroccan strains. In contrast, the 8045_S and Ch_10S strains were clustered distantly from the Tunisian calf strain B137 and the goat strain cap455 isolated in South Africa respectively. The human MA31 RVA strain was able to induce bloody diarrhea at 2 days post infection (dpi) in the alpine goat kid. RVA virus shedding started by 2 dpi (Ct value of 28) and continued until 5 dpi (Ct value of 25) with a concomitant elevation in the body temperature. Conclusions: Our study while limited to one animal, is the first study proving experimentally that a human P[14] genotype causes diarrhea and virus shedding in the goat. This result reinforce the potential role of inter- species transmission in generating novel and rare rotavirus strains such G8P[14] which infect humans.

Keywords: interspecies transmission, rotavirus, goat, human

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Authors: Karthick Gopalan, Selvamohan Thankiah

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Biosurfactants are lipid compounds produced by microbes, which are amphipathic molecules consisting of hydrophophic and hydrophilic domains. In the present investigation, ten bacterial strains were isolated from petroleum oil contaminated sites near petrol bunk. Oil collapsing test, haemolytic activity were used as a criteria for primary isolation of biosurfactant producing bacteria. In this study, all the bacterial strains gave positive results. Among the ten strains, two were observed as good biosurfactant producers, they utilize the diesel as a sole carbon source. Optimization of biosurfactant producing bacteria isolated from petroleum oil contaminated sites was carried out using different parameters such as, temperature (20ºC, 25ºC, 30ºC, 37ºC and 45ºC), pH (5,6,7,8 & 9) and nitrogen sources (ammonium chloride, ammonium carbonate and sodium nitrate). Biosurfactants produced by bacteria were extracted, dried and quantified. As a result of optimization of parameters the suitable values for the production of more amount of biosurfactant by the isolated bacterial species was observed as 30ºC (0.543 gm/lt) in the pH 7 (0.537 gm/lt) with ammonium nitrate (0.431 gm/lt) as sole carbon source.

Keywords: isolation and identification, biosurfactant, microorganism, bioaugmentation

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Authors: Checkfaith I. Aizebeoje, Temitope O. Lawal, Bolanle A. Adeniyi

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The overuse and abuse of antibiotics in treating zoonotic infections in humans and opportunistic infections in rabbit has contributed to the increase in antimicrobial drug resistance, therefore, an alternative to antibiotics is needed in treating these infections. The study was carried out to determine the antimicrobial activity of lactic acid bacteria (LAB) isolated from rabbit’s faeces against multidrug-resistant (MDR) pathogens isolated from the same rabbit. Twelve faecal samples and twelve swabs from fur samples were randomly collected aseptically from apparently healthy rabbits from Ajibode, Ibadan and University of Ibadan research farm in Ibadan, Oyo state, Nigeria. Lactic acid bacteria and multidrug-resistant pathogens were isolated using appropriate agar media and identified by partial sequencing of the 16SrRNA gene. Antibiotic susceptibility pattern of isolated bacteria and LAB were determined by the agar diffusion method. The antibacterial activity of the LAB against the test pathogens was determined using the agar overlay and agar diffusion methods. The pathogens Myroides gitamensis, Citrobacter rodentium, Acinetobacter johnsonii, Enterobacter oryzendophyticus and Serratia marcescens as well as twenty-eight (28) species of LAB belonging to Acetobacter and Lactobacillus genera were identified and characterized. Lactobacillus plantarum had the highest (60.71%) occurrence of the LAB. Viable cells and cell free supernatant (CFS) of isolated LAB inhibited the growth of the test organisms with the largest zone of inhibition (40 mm) produced by Lactobacillus plantarum against Citrobacter rodentium. This study showed that LAB from rabbit possess considerable antibacterial activity against multidrug-resistant bacteria from the same environment.

Keywords: antibacterial activities, cell-free supernatant, lactic acid bacteria; multidrug-resistant pathogens, rabbits’ faeces

Procedia PDF Downloads 110

Authors: Z. Abdessemed, N. Messaâdia, M. Houhamdi

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The PAH (Polycyclic Aromatic Hydrocarbons) represent a persistent source of pollution for oil field soils. Their degradation, essentially dominated by the aerobic bacterial and fungal flora, exhibits certain aspects for remediation of these soils microbial oxygenases have, as their substrates, a large range of PAH. The variety and the performance of these enzymes allow the initiation of the biodegradation of any PAH through many different metabolic pathways. These pathways are very important for the recycling of the PAH in the biosphere, where substances supposed indigestible by living organisms are rapidly transformed into simples compounds, directly assimilated by the intermediate metabolism of other microorganisms.

Keywords: polycyclic aromatic hydrocarbons, microbial oxygenases, biodegradation, metabolic pathways

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Authors: Frederick Otite Ighovojah

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Corrosion in the oil and gas industries is one of the most common causes of failure. Such failure includes leaks in above-ground storage tanks (AGST). The involvement of microorganisms in the corrosion process in AGST systems is often ignored, and this outlines the need to investigate the effect of microbial corrosion in oil and gas facilities. This study's methodology comprised gathering generated water samples from a nearby AGST oil facility that was operating, which were then equally divided into two batch reactors, 1 and 2. Each batch reactor was filled with five prepared X60 coupons using sterilized forceps. To provide nutrients for the microorganisms in batch reactor 1 during the test period, 2g of NPK 15- 15-15 fertilizer was added on a weekly basis. To kill the microorganisms and significantly lower their concentration in the generated water, 5ml of dissolved ozone (a biocide) with a 0.5ppm concentration was added to batch reactor 2. The weight loss measurement (WLM) was used to evaluate for corrosion. Coupons were removed from each batch reactor, and weight loss was measured at every interval of 336 hrs for 2016 hrs. The overall results obtained indicated that coupons from the batch 1 reactor showed a higher corrosion rate and higher mass loss, and this was due to the metabolic production of an aggressive compound in the medium.

Keywords: AGST, microbial corrosion, reactor, X60 steel

Procedia PDF Downloads 63

Authors: Sidra Shaw, Sarenna Shaw, Chae Joon Lee, Irimpan Mathews, Eric Koehn

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One of the biggest public health concerns of our time is increasing antimicrobial resistance. As of 2019, the CDC has documented more than 2.8 million serious antibiotic resistant infections in the United States. Currently, antibiotic resistant infections are directly implicated in over 750,000 deaths per year globally. On our current trajectory, British economist Jim O’Neill predicts that by 2050, an additional 10 million people (about half the population of New York) will die annually due to drug resistant infections. As a result, new biochemical pathways must be targeted to generate next generation antibiotic drugs that will be effective against drug resistant bacteria. One enticing target is the biosynthesis of DNA within bacteria, as few drugs interrupt this essential life process. Thymidylate synthase enzymes are essential for life as they catalyze the synthesis of a DNA building block, 2′-deoxythymidine-5′-monophosphate (dTMP). In humans, the thymidylate synthase enzyme (TSase) has been shown to be distinct from the flavin-dependent thymidylate synthase (FDTS) produced by many pathogenic bacteria. TSase and FDTS have distinct structures and mechanisms of catalysis, which should allow selective inhibition of FDTS over human TSase. Currently, C. diff is one of the most antibiotic resistant bacteria, and no drugs that target thymine biosynthesis exist for C. diff. Here we present the initial biochemical characterization of FDTS from C. diff. Specifically, we examine enzyme kinetics and binding features of this enzyme to determine the nature of interaction with ligands/inhibitors and understand the molecular mechanism of catalysis. This research will provide more insight into the targetability of the C. diff FDTS enzyme for novel antibiotic drugs.

Keywords: flavin-dependent thymidylate synthase, FDTS, clostridioides difficile, C. diff, antibiotic resistance, DNA synthesis, enzyme kinetics, binding features

Procedia PDF Downloads 74

Authors: Apotiola Z. O., Anyakorah C. I., Kuforiji O. O.

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Sensory and microbial properties of fresh and canned Calocybe indica (milky mushroom) were evaluated. The mushroom was grown under a controlled environment with hardwood (Cola nitida) and rice bran substrate (4:1) canned in a brine solution of salt and citric acid. Analysis was carried out using standard methods. The overall acceptability ranged between 5.62 and 6.50, with sample S30 adjudged the best. In all, significant differences p<0.01 exist in the panelist judgment. Thus, the incorporation of salt and citric acid at 3.5g and 1.5g, respectively, improved sensory attributes such as texture, aroma, color, and overall acceptability. There was no coliform and fungi growth on the samples throughout the storage period. The bacterial count, on the other hand, was observed only in the fifth and sixth week of the storage period which varied between 0.2 to 0.9 x 103 cfu/g. The highest value was observed in sample S20 of the sixth week of storage, while the lowest value was recorded in sample S30 of the sixth week of storage. Based on 16S rRNA gene sequencing, bacterial species were taxonomically confirmed as Bacillus thuringiensis. The percentile compositions and Sequence ID of the bacterial species in the mushroom was 90%.

Keywords: bacterial count, microbial property, sensory, sawdust, texture

Procedia PDF Downloads 42

Authors: S. S. Mahlangu, R. K. K. Mbaya, D. D. Delport, H. Van. Zyl

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The degrading effect due to bacterial growth on the structural integrity of concrete floor surfaces is predictable; this consequently cause development of surface micro cracks in which organisms penetrate through resulting in surface spalling. Hence, the need to develop mix design meeting the requirement of floor surfaces exposed to aggressive agent to improve certain material properties with good workability, extended lifespan and low cost is essential. In this work, tests were performed to examine the microbial activity on kitchen floor surfaces and the effect of adding admixtures. The biochemical test shows the existence of microorganisms (E.coli, Streptococcus) on newly casted structure. Of up to 6% porosity was reduced and improvement on structural integrity was observed upon adding mineral admixtures from the concrete mortar. The SEM result after 84 days of curing specimens, shows that chemical admixtures have significant role to enable retard bacterial penetration and good quality structure is achieved.

Keywords: admixture, organisms, porosity, strength

Procedia PDF Downloads 219

Authors: Muhammad Adeel Arshad, Shaukat Ali Bhatti, Faiz-ul Hassan

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Manipulating microbial ecosystem in the rumen is considered as an important strategy to optimize production efficiency in ruminants. In the past, antibiotics and synthetic chemical compounds have been used for the manipulation of rumen fermentation. However, since the non-therapeutic use of antibiotics has been banned, efforts are being focused to search out safe alternative products. In tropics, crop residues and forage grazing are major dietary sources for ruminants. Poor digestibility and utilization of these feedstuffs by animals is a limiting factor to exploit the full potential of ruminants in this area. Hence, there is a need to enhance the utilization of these available feeding resources. One of the potential strategies in this regard is the use of direct-fed microbes. Bacteria and fungi are mostly used as direct-fed microbes to improve animal health and productivity. Commonly used bacterial species include lactic acid-producing and utilizing bacteria (Lactobacillus, Streptococcus, Enterococcus, Bifidobacterium, and Bacillus) and fungal species of yeast are Saccharomyces and Aspergillus. Direct-fed microbes modulate microbial balance in the gastrointestinal tract through the competitive exclusion of pathogenic species and favoring beneficial microbes. Improvement in weight gain and feed efficiency has been observed as a result of feeding direct-fed bacteria. The use of fungi as a direct-fed microbe may prevent excessive production of lactate and harmful oxygen in the rumen leading to better feed digestibility. However, the mechanistic mode of action for bacterial or fungal direct-fed microbes has not been established yet. Various reports have confirmed an increase in dry matter intake, milk yield, and milk contents in response to the administration of direct-fed microbes. However, the application of a direct-fed microbe has shown variable responses mainly attributed to dosages and strains of microbes. Nonetheless, it is concluded that the inclusion of direct-fed microbes may mediate the rumen ecosystem to manage lactic acid production and utilization in both clinical and sub-acute rumen acidosis.

Keywords: microbes, roughages, rumen, feed efficiency, production, fermentation

Procedia PDF Downloads 114

Authors: Hamed A. El-Serehy, Khaled A. Al-Rasheid, Fahad A Al-Misned

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The meiofaunal population fluctuation, microbial dynamic and the composition of the sedimentary organic matter were investigated seasonally in the Egyptian shores along the northern part of Red Sea. Total meiofaunal population densities were extremely low with an annual average of 109 ±26 ind./10 cm2 and largely dominated by nematodes (on annual average from 52% to 94% of total meiofaunal density). The benthic microbial population densities ranged from 0.26±0.02 x 108 to 102.67±18.62 x 108/g dry sediment. Total sedimentary organic matter concentrations varied between 5.8 and 11.6 mg/g and the organic carbon, which was measured as summation of the carbohydrates, proteins and lipids, accounted for only a small fraction of being 32 % of the total organic matter. Chlorophyll a attained very low values and fluctuated between 2 and 11 µg/g. The very low chlorophyll a concentration in the Egyptian coasts along the Red Sea can suggest that the sedimentary organic matter along the Egyptian coasts is dominated by organic detrital and heterotrophic bacteria on one hand, and do not promote carbon transfer towards the higher trophic level on the other hand. However, the present study indicates that the existing of well diversified meiofaunal group, with a total of ten meiofaunal taxa, can serve as food for higher trophic levels in the Red Sea marine ecosystem.

Keywords: bacteria, meiofauna, intertidal sediments, Red Sea

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Authors: Asma Lamin, Anna H. Kaksonen, Ivan Cole, Paul White, Xiao-Bo Chen

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Microbiologically influenced corrosion (MIC) is a process in which microorganisms initiate, facilitate, or accelerate the electrochemical corrosion reactions of metallic components. Several reports documented that MIC accounts for about 20 to 40 % of the total cost of corrosion. Biofilm formation due to the presence of microorganisms on the surface of metal components is known to play a vital role in MIC, which can lead to severe consequences in various environmental and industrial settings. Quorum sensing (QS) system plays a major role in regulating biofilm formation and control the expression of some microbial enzymes. QS is a communication mechanism between microorganisms that involves the regulation of gene expression as a response to the microbial cell density within an environment. This process is employed by both Gram-positive and Gram-negative bacteria to regulate different physiological functions. QS involves production, detection, and responses to signalling chemicals, known as auto-inducers. QS controls specific processes important for the microbial community, such as biofilm formation, virulence factor expression, production of secondary metabolites and stress adaptation mechanisms. The use of QS inhibitors (QSIs) has been proposed as a possible solution to biofilm related challenges in many different applications. Although QSIs have demonstrated some strength in tackling biofouling, QSI-based strategies to control microbially influenced corrosion have not been thoroughly investigated. As such, our research aims to target the QS mechanisms as a strategy for mitigating MIC on metal surfaces in engineered systems.

Keywords: quorum sensing, quorum quenching, biofilm, biocorrosion

Procedia PDF Downloads 71

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

Procedia PDF Downloads 146

Authors: Zorica Tomičić, Ružica Tomičić, Peter Raspor

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Growing resistance of pathogenic yeast Candida glabrata to many classes of antifungal drugs has stimulated efforts to discover new agents to combat a rising number of invasive C. glabrata infections, which deserves a great deal of concern due to the high mortality rate in immunocompromised populations. One promising strategy is the use of probiotic microorganisms, which, when administered in adequate amounts, confers a health benefit. A selected number of probiotic organisms, Saccharomyces boulardii among them, have been tested as potential biotherapeutic agents. The aim of this study was to investigate the effect of the probiotic yeast S. boulardii on the adhesion of clinical isolates of C. glabrata at different temperatures, pH values, and in the presence of three clinically important antifungal drugs, such as fluconazole, itraconazole and amphotericin B. The method used to assess adhesion was crystal violet staining. The selection of antimycotics concentrations used in the adhesion assay was based on minimum inhibitory concentrations (MICs) obtained by the preliminarily performed microdilution modification of the Reference method for broth dilution antifungal susceptibility testing of yeast (Clinical and Laboratory Standards Institute (CLSI), standard M27-A2). the results showed that despite the nonadhesiveness of S. boulardii cells, probiotic yeast significantly suppressed the adhesion of C. glabrata strains. Besides, at specific strain ratios, a slight stimulatory effect was observed in some C. glabrata strains, which highlights the importance of strain specificity and opens up further research interests. When environmental conditions are considered, temperature and pH significantly influenced co-culture adhesion of C. glabrata and S. boulardii. The adhesion of C. glabrata strains was relatively equally reduced over all tested temperature range (28°C, 37°C, 39°C and 42°C) in the presence of S. boulardii cells, while the adhesion of a few C. glabrata strains were significantly stimulated at 28°C and suppressed at 42°C. Further, the adhesion was highly dependent on pH, with the highest adherence at pH 4 and lowest at pH 8.5. It was observed that S. boulardii did not manage to suppress the adhesion of C. glabrata strains at high pH. Antimycotics on the other hand showed a greater impact, since S. boulardii failed to affect co-culture adhesion at higher antimycotics concentrations. As expected, exposure to various concentrations of amphotericin B significantly reduced the adherence ability of C.glabrata strains both in a single culture and co-culture with S. boulardii. Therefore, it can be speculated that S. boulardii could substitute the effect of antimycotics in a range concentrations and with specific type of strains. This would certainly change the view on the treatment of yeast infections in the future.

Keywords: adhesion, antimycotics, candida glabrata, saccharomyces boulardii

Procedia PDF Downloads 55

Authors: Muhammad Rafiullah Khan, Vanee Chonhenchob

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The effect of sodium alginate (1%) based edible coating incorporated natural essential oils; thymol, carvone and carvacrol as antimicrobial agents at different concentrations (0.1, 0.5 and 1.0 %) on the quality changes of fresh-cut pineapple were investigated. Pineapple dipped in distilled water was served as control. After coating, fruit were sealed in a modified atmosphere package (MAP) using high permeable film; and stored at 5 °C. Gas composition in package headspace, color values (L*, a*, b*, C*), TSS, pH, ethanol, browning, and microbial decay were monitored during storage. Oxygen concentration continuously decreased while carbon dioxide concentration inside all packages continuously increased over time. Color parameters (L*, b*, c*) decreased and a* values increased during storage. All essential oils significantly (p ≤ 0.05) prevented microbial growth than control. A significantly higher (p ≤ 0.05) ethanol content was found in the control than in all other treatments. Visible microbial growth, high ethanol, and low color values limited the shelf life to 6 days in control as compared to 9 days in all other treatments. Among all essential oils, thymol at all concentrations maintained the overall quality of the pineapple and could potentially be used commercially in fresh fruit industries for longer storage.

Keywords: essential oils, antibrowning agents, antimicrobial agents, modified atmosphere packaging, pineapple, microbial decay

Procedia PDF Downloads 37

Authors: Muhammad Rafiullah Khan, Vanee Chonhenchob

Abstract:

The effect of sodium alginate (1%) based edible coating incorporated natural essential oils, thymol, carvone, and carvacrol as antimicrobial agents at different concentrations (0.1, 0.5, and 1.0%) on the quality changes of fresh-cut pineapple was investigated. Pineapple dipped in distilled water was served as control. After coating, the fruit was sealed in a modified atmosphere package (MAP) using high permeable film and stored at 5°C. Gas composition in package headspace, color values (L*, a*, b*, C*), TSS, pH, ethanol, browning, and microbial decay were monitored during storage. Oxygen concentration continuously decreased while carbon dioxide concentration inside all packages continuously increased over time. Color parameters (L*, b*, c*) decreased, and a* values increased during storage. All essential oils significantly (p ≤ 0.05) prevented microbial growth than control. A significantly higher (p ≤ 0.05) ethanol content was found in the control than in all other treatments. Visible microbial growth, high ethanol, and low color values limited the shelf life to 6 days in control as compared to 9 days in all other treatments. Among all essential oils, thymol at all concentrations maintained the overall quality of the pineapple and could potentially be used commercially in fresh fruit industries for longer storage.

Keywords: essential oils, antibrowning agents, antimicrobial agents, modified atmosphere packaging, microbial decay, pineapple

Procedia PDF Downloads 37