Search results for: microbial contamination
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1706

Search results for: microbial contamination

476 A Study on the Microbilogical Profile and Antibiotic Sensitivity Pattern of Bacterial Isolates Causing Urinary Tract Infection in Intensive Care Unit Patients in a Tertiary Care Hospital in Eastern India

Authors: Pampita Chakraborty, Sukumar Mukherjee

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The study was done to determine the microbiological profile and changing pattern of the pathogens causing UTI in the ICU patients. All the patients admitted to the ICU with urinary catheter insertion for more than 48hours were included in the study. Urine samples were collected in a sterile container with aseptic precaution using disposable syringe and was processed as per standards. Antimicrobial susceptibility test was done by Disc Diffusion method as per CLSI guidelines. A total of 100 urine samples were collected from ICU patients, out of which 30% showed significant bacterial growth and 7% showed growth of candida spp. Prevalence of UTI was more in female (73%) than male (27.%). Gram-negative bacilli 26(86.67%) were more common in our study followed by gram-positive cocci 4(13.33%). The most common uropathogens isolated were Escherichia coli 14 (46.67%), followed by Klebsiella spp 7(23.33%), Staphylococcus aureus 4(13.33%), Acinetobacter spp 3(10%), Enterococcus faecalis 1(3.33%) and Pseudomonas aeruginosa 1(3.33%). Most of the Gram-negative bacilli were sensitive to amikacin (80%) and nitrofurantoin (80%), where as all gram-positive organisms were sensitive to Vancomycin. A large number ESBL producers were also observed in this study. The study finding showed that E.coli is the predominant pathogen and has increasing resistance pattern to the commonly used antibiotics. The study proposes that the adherence to antibiotic policy is the key ingredients for successful outcome in ICU patients and also emphasizes that repeated evaluation of microbial characteristics and continuous surveillance of resistant bacteria is required for selection of appropriate antibiotic therapy.

Keywords: antimicrobial sensitivity, intensive care unit, nosocomial infection, urinary tract infection

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475 Evaluation of a Chitin Synthesis Inhibitor Novaluron in the Shrimp Palaemon Adspersus: Impact on Ecdysteroids and Chitin Contents

Authors: Hinda Berghiche, Hamida Benradia, Noureddine Soltani

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Pesticides are widely used in crop production and are known to induce a major contamination of ecosystems especially in aquatic environments. The leaching of a large amount of pollutants derived from agricultural activities (fertilizers, pesticides) might contaminate rivers which diverse into the likes and estuarine and coastal environments affecting several organisms such as crustacean species. In this context, there is searched for new selective insecticides with minimal toxic effects on the environment and human health such as growth insect regulators (GIRs). The current study aimed to examine the impact of novaluron (CE 20%), a potent benzoylphenylurea derivative insecticide on mosquito larvae, against non-target shrimp, Palaemon adspersus (Decapoda, Palaemonidae). The compound was tested at two concentrations (0.91 mg/L and 4.30 mg/L) corresponding respectively to the LC50 and LC90 determined against fourth-instar larvae of Culiseta longiareolata (Diptera, Culicidae). The molting hormone titer was determined in the haemolymph by an enzyme-immunoassay, while chitin was measured in peripheral integument at different stages during the molting cycle. Under normal conditions, the haemolymphatic ecdysteroid concentrations increased during the molting cycle to reach peak at stage D. In the treated series, we note absence of the peak at stage D and an increase at stages B, C and D as compared to the controls. Concerning the chitin amounts, we observe an increase from stage A to stage C followed by a decrease at stage D. Exposition of shrimps to novaluron resulted in a significant decrease of values at all molting stages with a dose-response effect. Thus, the insecticide can present secondary effects on this non-target arthropod species.

Keywords: toxicology, novaluron, crustacean, palaemon adspersus, ecdysteroids, cuticle, chitin

Procedia PDF Downloads 241
474 Unlocking the Potential of Neglected Cereal Resources Waste: Exploring Functional Properties of Algerian Pearl Millet Starch via Wet Milling and Ultrasound Techniques

Authors: Sarra Bouhallel, Sara Legbedj, Rima Messaoud, Sofia Saffarbatti

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In the context of global waste management and sustainable resource utilization, millets emerge as a vital yet underutilized cereal resource. Despite their exceptional nutritional profile and resilience to harsh environmental conditions, their potential remains largely untapped. This study aims to contribute to the valorization of seven Algerian pearl millet landraces (Pennisetum glaucum (L.) R. Br) from the southern region by focusing on the characterization of their starches. Utilizing both conventional wet milling, incorporating sodium azide as a microbial growth inhibitor, and a novel green technology—Ultrasound-assisted isolation, we explore avenues for enhancing the functional properties of these starches. Analysis of key functional properties such as swelling power and water solubility index reveals significant enhancements, particularly during heat treatment near the gelatinization temperature [70 - 80 °C]. Furthermore, our investigation into the influence of pre-treatment methods on isolated starches highlights the potential of Ultrasound-assisted isolation in reducing absorbency and water solubility compared to conventional methods. Through rigorous data analysis using SPSS software (Version 23), we ascertain the efficiency of Ultrasound-assisted isolation, underscoring its promising role in the valorization of pearl millet waste. This research not only sheds light on the functional properties of pearl millet starch but also underscores the imperative of sustainable waste management in harnessing the full potential of underutilized cereal resources.

Keywords: isolation, solubility, starch, swelling, ultrasound

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473 Comparative Correlation Investigation of Polynuclear Aromatic Hydrocarbons (PAHs) in Soils of Different Land Uses: Sources Evaluation Perspective

Authors: O. Onoriode Emoyan, E. Eyitemi Akporhonor, Charles Otobrise

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Polycyclic Aromatic Hydrocarbons (PAHs) are formed mainly as a result of incomplete combustion of organic materials during industrial, domestic activities or natural occurrence. Their toxicity and contamination of terrestrial and aquatic ecosystem have been established. Though with limited validity index, previous research has focused on PAHs isomer pair ratios of variable physicochemical properties in source identification. The objective of this investigation was to determine the empirical validity of Pearson correlation coefficient (PCC) and cluster analysis (CA) in PAHs source identification along soil samples of different land uses. Therefore, 16 PAHs grouped as endocrine disruption substances (EDSs) were determined in 10 sample stations in top and sub soils seasonally. PAHs was determined the use of Varian 300 gas chromatograph interfaced with flame ionization detector. Instruments and reagents used are of standard and chromatographic grades respectively. PCC and CA results showed that the classification of PAHs along kinetically and thermodyanamically-favoured and those derived directly from plants product through biologically mediated processes used in source signature is about the predominance PAHs are likely to be. Therefore the observed PAHs in the studied stations have trace quantities of the vast majority of the sixteen un-substituted PAHs which may ultimately inhabit the actual source signature authentication. Type and extent of bacterial metabolism, transformation products/substrates, and environmental factors such as: salinity, pH, oxygen concentration, nutrients, light intensity, temperature, co-substrates and environmental medium are hereby recommended as factors to be considered when evaluating possible sources of PAHs.

Keywords: comparative correlation, kinetically and thermodynamically-favored PAHs, pearson correlation coefficient, cluster analysis, sources evaluation

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472 Micropropagation of Rhododendron tomentosum (Ledum palustre): An Endangered Plant of Scientific Interest as the Example of Ex Situ Conservation

Authors: Anna Jesionek, Aleksandra Szreniawa-Sztajnert, Zbigniew Jaremicz, Adam Kokotkiewicz, Natalia Filipowicz, Renata Ochocka, Bozena Zabiegala, Maria Luczkiewicz

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Rhododendron tomentosum (formerly Ledum palustre), an evergreen shrub grows in peaty soils in northern Europe, Asia and North America. In Poland, it is classified as an endangered species not only due to the drainage of wetlands, but also to the excessive collection of this repellent plant by human. The other valuable biological properties of R. tomentosum, used for years in folk medicine, include anti-inflammatory, analgesic and anti-microbial activity, conditioned by the essential oil content. Taking into account the importance of biodiversity and the potential therapeutic application, it was decided to establish, for the first time, the micropropagation protocol for R. tomentosum, for ex-situ conservation of this endangered species as well as to obtain the continuous source of in vivo and in-vitro plant material for further studies. This object was achieved by the selection of the explant and the media, which were modified within the scope of mineral composition, sugar content, pH and the growth regulators. As a result, the four-stage micropropagation protocol for R. tomentosum was specified, including shoot multiplication, elongation, rooting and ex-vitro adaptation. The genetic identification of the examined species and the compatibility of progeny plants with maternal ones was tested with molecular biology methods. Moreover, during the research process, the chemical composition of initial and regenerated plant and in vitro shoots was controlled in terms of volatile fraction by phytochemical analysis (GC and TLC methods). The correctness of the micropropagation procedure was confirmed by both types of studies.

Keywords: ex situ conservation, Ledum palustre, micropropagation, Rhododendron tomentosum

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471 Isolation and Identification of Microorganisms from Marine-Associated Samples under Laboratory Conditions

Authors: Sameen Tariq, Saira Bano, Sayyada Ghufrana Nadeem

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The Ocean, which covers over 70% of the world's surface, is wealthy in biodiversity as well as a rich wellspring of microorganisms with huge potential. The oceanic climate is home to an expansive scope of plants, creatures, and microorganisms. Marine microbial networks, which incorporate microscopic organisms, infections, and different microorganisms, enjoy different benefits in biotechnological processes. Samples were collected from marine environments, including soil and water samples, to cultivate the uncultured marine organisms by using Zobell’s medium, Sabouraud’s dextrose agar, and casein media for this purpose. Following isolation, we conduct microscopy and biochemical tests, including gelatin, starch, glucose, casein, catalase, and carbohydrate hydrolysis for further identification. The results show that more gram-positive and gram-negative bacteria. The isolation process of marine organisms is essential for understanding their ecological roles, unraveling their biological secrets, and harnessing their potential for various applications. Marine organisms exhibit remarkable adaptations to thrive in the diverse and challenging marine environment, offering vast potential for scientific, medical, and industrial applications. The isolation process plays a crucial role in unlocking the secrets of marine organisms, understanding their biological functions, and harnessing their valuable properties. They offer a rich source of bioactive compounds with pharmaceutical potential, including antibiotics, anticancer agents, and novel therapeutics. This study is an attempt to explore the diversity and dynamics related to marine microflora and their role in biofilm formation.

Keywords: marine microorganisms, ecosystem, fungi, biofilm, gram-positive, gram-negative

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470 Changes on Some Physical and Chemical Properties of Red Beetroot Juice during Ultrasound Pretreatment

Authors: Serdal Sabanci, Mutlu Çevik, Derya Tezcan, Cansu Çelebi, Filiz Içier

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Ultrasound is defined as sound waves having frequencies higher than 20 kHz, which is greater than the limits of the human hearing range. In recent years, ultrasonic treatment is an emerging technology being used increasingly in the food industry. It is applied as an alternative technique for different purposes such as microbial and enzyme inactivation, extraction, drying, filtration, crystallization, degas, cutting etc. Red beetroot (Beta vulgaris L.) is a root vegetable which is rich in mineral components, folic acid, dietary fiber, anthocyanin pigments. In this study, the application of low frequency high intensity ultrasound to the red beetroot slices and red beetroot juice for different treatment times (0, 5, 10, 15, 20 min) was investigated. Ultrasonicated red beetroot slices were also squeezed immediately. Changes on colour, betanin, pH and titratable acidity properties of red beetroot juices (the ultrasonicated juice (UJ) and the juice from ultrasonicated slices (JUS)) were determined. Although there was no significant difference statistically in the changes of color value of JUS samples due to ultrasound application (p>0.05), the color properties of UJ samples ultrasonicated for low durations were statistically different from raw material (p<0.05). The difference between color values of UJ and raw material disappeared (p>0.05) as the ultrasonication duration increased. The application of ultrasound to red beet root slices adversely affected and decreased the betanin content of JUS samples. On the other hand, the betanin content of UJ samples increased as the ultrasonication duration increased. Ultrasound treatment did not affect pH and titratable acidity of red beetroot juices statistically (p>0.05). The results suggest that ultrasound technology is the simple and economical technique which may successfully be employed for the processing of red beetroot juice with improved color and betanin quality. However, further investigation is still needed to confirm this.

Keywords: red beetroot, ultrasound, color, betanin

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469 Fertigation Use in Agriculture and Biosorption of Residual Nitrogen by Soil Microorganisms

Authors: Irina Mikajlo, Jakub Elbl, Helena Dvořáčková, Antonín Kintl, Jindřich Kynický, Martin Brtnický, Jaroslav Záhora

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Present work deals with the possible use of fertigation in agriculture and its impact on the availability of mineral nitrogen (Nmin) in topsoil and subsoil horizons. The aim of the present study is to demonstrate the effect of the organic matter presence in fertigation on microbial transformation and availability of mineral nitrogen forms. The main investigation reason is the potential use of pre-treated waste water, as a source of organic carbon (Corg) and residual nutrients (Nmin) for fertigation. Laboratory experiment has been conducted to demonstrate the effect of the arable land fertilization method on the Nmin availability in different depths of the soil with the usage of model experimental containers filled with soil from topsoil and podsoil horizons that were taken from the precise area. Tufted hairgrass (Deschampsia caespitosa) has been chosen as a model plant. The water source protection zone Brezova nad Svitavou has been a research area where significant underground reservoirs of drinking water of the highest quality are located. From the second half of the last century local sources of drinking water show nitrogenous compounds increase that get here almost only from arable lands. Therefore, an attention of the following text focuses on the fate of mineral nitrogen in the complex plant-soil. Research results show that the fertigation application with Corg in a combination with mineral fertilizer can reduce the amount of Nmin leached from topsoil horizon of agricultural soils. In addition, some plants biomass production reduce may occur.

Keywords: fertigation, fertilizers, mineral nitrogen, soil microorganisms

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468 Remodeling of Gut Microbiome of Pakistani Expats in China After Intermittent Fasting/Ramadan Fasting

Authors: Hafiz Arbab Sakandar

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Time-restricted intermittent fasting (TRIF) impacts host’s physiology and health. Plenty of health benefits have been reported for TRIF in animal models. However, limited studies have been conducted on humans especially in underdeveloped economies. Here, we designed a study to investigate the impact of TRIF/Ramadan fasting (16:8) on the modulation of gut-microbiome structure, metabolic pathways, and predicted metabolites and explored the correlation among them at different time points (during and after the month of Ramadan) in Pakistani Expats living in China. We observed different trends of Shannon-Wiener index in different subjects; however, all subjects showed substantial change in bacterial diversity with the progression of TRIF. Moreover, the changes in gut microbial structure by the end of TRIF were higher vis-a-vis in the beginning, significant difference was observed among individuals. Additionally, metabolic pathways analysis revealed that amino acid, carbohydrate and energy metabolism, glycan biosynthesis metabolism of cofactors and vitamins were significantly affected by TRIF. Pyridoxamine, glutamate, citrulline, arachidonic acid, and short chain fatty acid showed substantial difference at different time points based on the predicted metabolism. In conclusion, these results contribute to further our understanding about the key relationship among, dietary intervention (TRIF), gut microbiome structure and function. The preliminary results from study demonstrate significant potential for elucidating the mechanisms underlying gut microbiome stability and enhancing the effectiveness of microbiome-tailored interventions among the Pakistani populace. Nonetheless, extensive, and rigorous large-scale research on the Pakistani population is necessary to expound on the association between diet, gut microbiome, and overall health.

Keywords: gut microbiome, health, fasting, functionality

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467 Sensitivity of Staphylococcus aureus Isolated from Subclinical Bovine Mastitis to Ciprofloxacin in Dairy Herd in Tabriz during 2013

Authors: Alireza Jafarzadeh, Samad Mosaferi, Mansour Khakpour

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Mastitis is an inflammation of the parenchyma of mammary gland regardless of the causes. Mastitis is characterized by a range of physical and chemical changes in the glandular tissue. The most important change in milk includes discoloration, the presence of clots and large number of leucocytes. There is swelling, heat, pain and edema in mammary gland in many clinical cases. Positive coagulase S. aureus is a major pathogen of the bovine mammary gland and a common cause of contagious mastitis in cattle. The aim of this study was to evaluate the outbreaks of Staphylococcus aureus mastitis. This study is conducted in ten dairy herds about one thousand cows. After doing CMT and identifying infected cows, the milk samples obtained from infected teats and transported to microbiological laboratories. After microbial culture of milk samples and isolating S. aureus, antimicrobial, sensitivity test was performed with disk diffusion method by ciprofloxacin, co-amoxiclav, erythromycin, penicillin, oxytetracyclin, sulfonamides, lincomycin and cefquinome. The study defined that the outbreak of subclinical positive coagulase Staphylococcus mastitis in dairy herd was 13.11% (5.6% S. aureus and 7.51% S. intermedicus). The antimicrobial sensitivity test shown that 87.23% of Staphylococcus aureus isolated from bovine mastitis in dairy herd was susceptible to ciprofloxacin, 93.9% to cefquinome, 4.67% to co-amoxiclav, 12.16% to erythromycin 86.11% to sulfonamides (co-trimoxazole), 3.35% lincomycin, 12.7% to oxytetracyclin and 5.98% to penicillin. Results of present defined that ciprofloxacin has a great effect on Staphylococcus aureus isolated from subclinical bovine mastitis dairy herd. It seems that cefquinome sulfonamides has a great effect on isolated Staphylococcus aureus in vivo.

Keywords: ciprofloxacin, mastitis, Staphylococcus aureus, dairy herd

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466 TAXAPRO, A Streamlined Pipeline to Analyze Shotgun Metagenomes

Authors: Sofia Sehli, Zainab El Ouafi, Casey Eddington, Soumaya Jbara, Kasambula Arthur Shem, Islam El Jaddaoui, Ayorinde Afolayan, Olaitan I. Awe, Allissa Dillman, Hassan Ghazal

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The ability to promptly sequence whole genomes at a relatively low cost has revolutionized the way we study the microbiome. Microbiologists are no longer limited to studying what can be grown in a laboratory and instead are given the opportunity to rapidly identify the makeup of microbial communities in a wide variety of environments. Analyzing whole genome sequencing (WGS) data is a complex process that involves multiple moving parts and might be rather unintuitive for scientists that don’t typically work with this type of data. Thus, to help lower the barrier for less-computationally inclined individuals, TAXAPRO was developed at the first Omics Codeathon held virtually by the African Society for Bioinformatics and Computational Biology (ASBCB) in June 2021. TAXAPRO is an advanced metagenomics pipeline that accurately assembles organelle genomes from whole-genome sequencing data. TAXAPRO seamlessly combines WGS analysis tools to create a pipeline that automatically processes raw WGS data and presents organism abundance information in both a tabular and graphical format. TAXAPRO was evaluated using COVID-19 patient gut microbiome data. Analysis performed by TAXAPRO demonstrated a high abundance of Clostridia and Bacteroidia genera and a low abundance of Proteobacteria genera relative to others in the gut microbiome of patients hospitalized with COVID-19, consistent with the original findings derived using a different analysis methodology. This provides crucial evidence that the TAXAPRO workflow dispenses reliable organism abundance information overnight without the hassle of performing the analysis manually.

Keywords: metagenomics, shotgun metagenomic sequence analysis, COVID-19, pipeline, bioinformatics

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465 Interaction Effects of Dietary Ginger, Zingiber Officinale, on Plasma Protein Fractions in Rainbow Trout, Oncorhynchus Mykiss

Authors: Ali Taheri Mirghaed, Sara Ahani, Ashkan Zargar, Seyyed Morteza Hoseini

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Diseases are the major challenges in intensive aquaculture that cause significant annual losses. Antibiotic-therapy is a common way to control bacterial disease in fish, and oxytetracycline (OTC) is the only oral antibiotic in aquaculture approved FDA. OTC has been found to have negative effects on fish, such as oxidative stress and immune-suppression, thus, it is necessary to mitigate such effects. Medicinal herbs have various benefits on fish, including antioxidant, immunostimulant, and anti-microbial effects. Therefore, we hypothesized if dietary ginger meal (GM) interacts with dietary OTC by monitoring plasma protein fractions in rainbow trout. The study was conducted as a 2 × 2 factorial design, including diets containing 0 and 1% GM and 0 and 1.66 % OTC (corresponding to 100 mg/kg fish biomass per day). After ten days treating the fish (60 g individual weight) with these feeds, blood samples were taken from al treatments (n =3). Plasma was separated by centrifugation, and protein fractions were determined by electrophoresis. The results showed that OTC and GM had interaction effects on total protein (P<0.001), albumin (P<0.001), alpha-1 fraction (P=0.010), alpha-2 fraction (P=0.001), beta-2 fraction (P=0.014), and gamma fraction (P<0.001). Beta-1 fraction was significantly (P=0.030) affected by dietary GM. GM decreased plasma total protein, albumin, and beta-2 but increased beta-1 fraction. OTC significantly decreased total protein (P<0.001), albumin (P=0.001), alpha-2 fraction (P<0.001), beta-2 fraction (P=0.004), and gamma fraction (P<0.001) but had no significant effects on alpha-1 and beta-1 fractions. Dietary GM inhibited/suppressed the effects of dietary OTC on the plasma total protein and protein fractions. In conclusion, adding 1% GM to diet can mitigate the negative effects of dietary OTC on plasma proteins. Thus, GM may boost health of rainbow trout during the period of medication with OTC.

Keywords: ginger, plasma protein electrophoresis, dietary additive, rainbow trout

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464 The Importance of Storage Period on Biogas Potential of Cattle Manure

Authors: Seongwon Im, Jimin Kim, Kyeongcheol Kim, Dong-Hoon Kim

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Cattle manure (CM) produced from farmhas been utilized to soils for increasing crop production owing to high nutrients content and effective microorganisms. Some cities with the concentrated activity of livestock industry have suffered from environmental problems, such as odorous gas emissions and soil and water pollution, caused by excessive use of compost. As an alternative option, the anaerobic digestion (AD) process can be utilized, which can reduce the volume of organic waste but also produce energy. According to Korea-Ministry of Trade, Industry, and Energy (KMTIE), the energy potential of CM via biogas production was estimated to be 0.8 million TOE per year, which is higher than that of other organic wastes. However, limited energy is recovered since useful organic matter, capable of converting to biogas, may be degraded during the long storage period (1-6 months).In this study, the effect of storage period on biogas potential of CM was investigated. Compared to fresh CM (VS 14±1 g/L, COD 205±5 g/L, TKN 7.4±0.8 g/L, NH4+-N 1.5±0.1), old CM has higher organic (35-37%) and nitrogen content (50-100%) due to the drying process during storage. After stabilization period, biogas potential of 0.09 L CH4/g VS was obtained in R1 (old CM supplement) at HRT of 150-100 d, and it was decreased further to 0.06 L CH4/g VS at HRT of 80 d. The drop of pH and organic acids accumulation were not observed during the whole operation of R1. Ammonia stripping and pretreatment of CM were found to be not effective to increase CH4 yield. On the other hand, a sudden increase of biogas potential to 0.19-0.22 L CH4/g VS was achieved in R2 after changing feedstock to fresh CM. The expected reason for the low biogas potential of old CM might be related with the composition of organic matters in CM. Easily biodegradable organic matters in the fresh CM were contained in high concentration, butthey were removed by microorganisms during storing CM in a farm, resulting low biogas yield. This study implies that fresh storage is important to make AD process applicable for CM.

Keywords: storage period, cattle manure, biogas potential, microbial analysis

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463 Isolation and Expansion of Human Periosteum-Derived Mesenchymal Stem Cells in Defined Serum-Free Culture Medium

Authors: Ainur Mukhambetova, Miras Karzhauov, Vyacheslav Ogay

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Introduction: Mesenchymal stem cells (MSCs) have the capacity to be differentiated into several cell lineages and are a promising source for cell therapy and tissue engineering. However, currently most MSCs culturing protocols use media supplemented with fetal bovine serum (FBS), which limits their application in clinic due to the possibility of zoonotic infections, contamination and immunological reactions. Consequently, formulating effective serum free culture medium becomes one of the important problems in contemporary cell biotechnology. Objectives: The aim of this study was to define an optimal serum-free medium for culturing of periosteum derived MSCs. Materials and methods: The MSCs were extracted from human periosteum and transferred to the culture flasks pretreated with CELLstart™. Immunophenotypic characterization, proliferation and in vitro differentiation of cells grown on STEM PRO® MSC SFM were compared to the cells cultured in the standard FBS containing media. Chromosome analysis and flow cytometry were also performed. Results: We have shown that cells were grown on STEM PRO® MSC SFM retained all the morphological, immunophenotypic (CD73, CD90, CD105, vimentin and Stro-1) and cell differentiation characteristics specific to MSCs. Chromosome analysis indicated no anomalies in the chromosome structure. Flow cytometry showed a high expression of cell adhesion molecules CD44 (98,8%), CD90 (97,4%), CD105 (99,1%). In addition, we have shown that cell is grown on STEM PRO® MSC SFM have higher proliferation capacity compared to cell expanded on standard FBS containing the medium. Conclusion: We have shown that STEM PRO® MSC SFM is optimal for culturing periosteum derived human MSCs which subsequently can be safely used in cell therapy.

Keywords: cell technologies, periosteum-derived MSCs, regenerative medicine, serum-free medium

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462 Phylogenetic Differential Separation of Environmental Samples

Authors: Amber C. W. Vandepoele, Michael A. Marciano

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Biological analyses frequently focus on single organisms, however many times, the biological sample consists of more than the target organism; for example, human microbiome research targets bacterial DNA, yet most samples consist largely of human DNA. Therefore, there would be an advantage to removing these contaminating organisms. Conversely, some analyses focus on a single organism but would greatly benefit from the additional information regarding the other organismal components of the sample. Forensic analysis is one such example, wherein most forensic casework, human DNA is targeted; however, it typically exists in complex non-pristine sample substrates such as soil or unclean surfaces. These complex samples are commonly comprised of not just human tissue but also microbial and plant life, where these organisms may help gain more forensically relevant information about a specific location or interaction. This project aims to optimize a ‘phylogenetic’ differential extraction method that will separate mammalian, bacterial and plant cells in a mixed sample. This is accomplished through the use of size exclusion separation, whereby the different cell types are separated through multiple filtrations using 5 μm filters. The components are then lysed via differential enzymatic sensitivities among the cells and extracted with minimal contribution from the preceding component. This extraction method will then allow complex DNA samples to be more easily interpreted through non-targeting sequencing since the data will not be skewed toward the smaller and usually more numerous bacterial DNAs. This research project has demonstrated that this ‘phylogenetic’ differential extraction method successfully separated the epithelial and bacterial cells from each other with minimal cell loss. We will take this one step further, showing that when adding the plant cells into the mixture, they will be separated and extracted from the sample. Research is ongoing, and results are pending.

Keywords: DNA isolation, geolocation, non-human, phylogenetic separation

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461 Evaluation of Elements Impurities in Drugs According to Pharmacopoeia by use FESEM-EDS Technique

Authors: Rafid Doulab

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Elemental Impurities in the Pharmaceuticals industryis are indispensable to ensure pharmaceuticalssafety for 24 elements. Although atomic absorption and inductively coupled plasma are used in the U.S Pharmacopeia and the European Pharmacopoeia, FESEM with energy dispersive spectrometers can be applied as an alternative analysis method for quantitative and qualitative results for a variety of elements without chemical pretreatment, unlike other techniques. This technique characterizes by shortest time, with more less contamination, no reagent consumption, and generation of minimal residue or waste, as well as sample preparations time limiting, with minimal analysis error. Simple dilution for powder or direct analysis for liquid, we analyzed the usefulness of EDS method in testing with field emission scanning electron microscopy (FESEM, SUPRA 55 Carl Zeiss Germany) with an X-ray energy dispersion (XFlash6l10 Bruker Germany). The samples analyzed directly without coating by applied 5µ of known concentrated diluted sample on carbon stub with accelerated voltage according to sample thickness, the result for this spot was in atomic percentage, and by Avogadro converted factor, the final result will be in microgram. Conclusion and recommendation: The conclusion of this study is application of FESEM-EDS in US pharmacopeia and ICH /Q3D guideline to reach a high-precision and accurate method in element impurities analysis of drugs or bulk materials to determine the permitted daily exposure PDE in liquid or solid specimens, and to obtain better results than other techniques, by the way it does not require complex methods or chemicals for digestion, which interfere with the final results with the possibility of to keep the sample at any time for re analysis. The recommendation is to use this technique in pharmacopeia as standard methods like inductively coupled plasma both ICP-AES, ICP-OES, and ICP-MS.

Keywords: pharmacopoeia, FESEM-EDS, element impurities, atomic concentration

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460 Utilization of Extracted Spirogyra sp. Media Fermented by Gluconacetobacter Xylinum for Cellulose Production as Raw Material for Paper Product

Authors: T. S. Desak Ketut, A.n. Isna, A.a. Ayu, D. P. Ririn, Suharjono Hadiatullah

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The requirement of paper from year to year rise rapidly. The raising of cellulose requirement in paper production caused increasing of wood requirement with the effect that limited forest areal because of deforestation. Alternative cellulose that can be used for making paper is microbial cellulose. The objective of this research are to know the effectivity fermentation media Spirogyra sp. by Gluconacetobacter xylinum for cellulose production as material for the making of paper and to know effect composition bacterial cellulose composite product of Gluconacetobacter xylinum in Spirogyra sp. The method, was used, is as follow, 1) the effect assay from variation composition of fermentation media to bacterial cellulose production by Gluconacetobacter xylinum. 2) The effect assay of composition bacterial cellulose fermentation producted by Gluconacetobacter xylinum in extracted Spirogyra media to paper quality. The result of this research is variation fermentation media Spirogyra sp. affect to production of cellulose by Gluconacetobacter xylinum. Thus, result showed by the highest value and significantly different in thickness parameter, dry weight and wet weight of nata in sucrose concentration 7,5 % and urea 0,75 %. Composition composite of bacterial cellulose from fermentation product by Gluconacetobacter xylinum in media Spirogyra sp. affect to paper quality from wet nata and dry nata. Parameters thickness, weight, water absorpsion, density and gramatur showed highest result in sucrose concentration 7,5 % and urea concentration 0,75 %, except paper density from dry nata had highest result in sucrose and urea concentration 0%.

Keywords: cellulose, fermentation media, , Gluconacetobacter xylinum, paper, Spirogyra sp.

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459 Identification and Characterization of Oil-Degrading Bacteria from Crude Oil-Contaminated Desert Soil in Northeastern Jordan

Authors: Mohammad Aladwan, Adelia Skripova

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Bioremediation aspects of crude oil-polluted fields can be achieved by isolation and identification of bacterial species from oil-contaminated soil in order to choose the most active isolates and increase the strength of others. In this study, oil-degrading bacteria were isolated and identified from oil-contaminated soil samples in northeastern Jordan. The bacterial growth count (CFU/g) was between 1.06×10⁵ and 0.75×10⁹. Eighty-two bacterial isolates were characterized by their morphology and biochemical tests. The identified bacterial genera included: Klebsiella, Staphylococcus, Citrobacter, Lactobacillus, Alcaligenes, Pseudomonas, Hafnia, Micrococcus, Rhodococcus, Serratia, Enterobacter, Bacillus, Salmonella, Mycobacterium, Corynebacterium, and Acetobacter. Molecular identification of a universal primer 16S rDNA gene was used to identify four bacterial isolates: Microbacterium esteraromaticum strain L20, Pseudomonas stutzeri strain 13636M, Klebsilla pneumoniae, and uncultured Klebsilla sp., known as new strains. Our results indicate that their specific oil-degrading bacteria isolates might have a high strength of oil degradation from oil-contaminated sites. Staphylococcus intermedius (75%), Corynebacterium xerosis (75%), and Pseudomonas fluorescens (50%) showed a high growth rate on different types of hydrocarbons, such as crude oil, toluene, naphthalene, and hexane. In addition, monooxygenase and catechol 2,3-dioxygenase were detected in 17 bacterial isolates, indicating their superior hydrocarbon degradation potential. Total petroleum hydrocarbons were analyzed using gas chromatography for soil samples. Soil samples M5, M7, and M8 showed the highest levels (43,645, 47,805, and 45,991 ppm, respectively), and M4 had the lowest level (7,514 ppm). All soil samples were analyzed for heavy metal contamination (Cu, Cd, Mn, Zn, and Pb). Site M7 contains the highest levels of Cu, Mn, and Pb, while Site M8 contains the highest levels of Mn and Zn. In the future, these isolates of bacteria can be used for the cleanup of oil-contaminated soil.

Keywords: bioremediation, 16S rDNA gene, oil-degrading bacteria, hydrocarbons

Procedia PDF Downloads 110
458 Effect of Processing Parameters on the Physical Properties of Pineapple Pomace Based Aquafeed

Authors: Oluwafemi Babatunde Oduntan, Isaac A. Bamgboye

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The solid waste disposal and its management from pineapple juice processing constitute environmental contamination affecting public health. The use of this by-product called pomace has potentials to reduce cost of aquafeed. Pineapple pomace collected after juice extraction was dried and milled. The interactive effects of feeding rate (1.28, 1.44 and 1.60kg/min), screw speed (305, 355 and 405rpm), moisture content (16, 19 and 22%), temperatures (60, 80, 100 and 120°C), cutting speed (1300, 1400 and 1500rpm), pomace inclusion ratio (5, 10, 15, 20%) and open surface die (50, 75 and 100%) on the extrudate physical properties (bulk density, unit density, expansion ratio, durability and floatability) were investigated using optimal custom design (OCD) matrix and response surface methodology. The predicted values were found to be in good agreement with the experimental values for, expansion ratio, durability and floatability (R2 = 0.7970; 0.9264; 0.9098 respectively) with the exceptions of unit density and bulk density (R2 = 0.1639; 0.2768 respectively). All the extrudates showed relatively high floatability, durability. The inclusion of pineapple pomace produced less expanded and more compact textured extrudates. Results indicated that increased in the value of pineapple pomace, screw speed, feeding rate decreased unit density, bulk density, expansion ratio, durability and floatability of the extrudate. However, increasing moisture content of feed mash resulted in increase unit density and bulk density. Addition of extrusion temperature and cutting speed increased the floatability and durability of extrudate. The proportion of pineapple pomace in aquafeed extruded product was observed to have significantly lower effect on the selected responses.

Keywords: aquafeed, extrusion, physical properties, pineapple pomace, waste

Procedia PDF Downloads 258
457 Simultaneous Saccharification and Fermentation for D-Lactic Acid Production from Dried Distillers Grains with Solubles

Authors: Nurul Aqilah Mohd Zaini, Afroditi Chatzifragkou, Dimitris Charalampopoulos

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D-Lactic acid production is gaining increasing attention due to the thermostable properties of its polymer, Polylactic Acid (PLA). In this study, D-lactic acid was produced in microbial cultures using Lactobacillus coryniformis subsp. torquens as D-lactic acid producer and hydrolysates of Dried Distillers Grains with Solubles (DDGS) as fermentation substrate. Prior to fermentation, DDGS was first alkaline pretreated with 5% (w/v) NaOH, for 15 minutes (121oC/ ~16 psi). This led to the generation of DDGS solid residues, rich in carbohydrates and especially cellulose (~52%). The carbohydrate-rich solids were then subjected to enzymatic hydrolysis with Accellerase® 1500. For Separate Hydrolysis and Fermentation (SHF), enzymatic hydrolysis was carried out at 50oC for 24 hours, followed by fermentation of D-lactic acid at 37oC in controlled pH 6. The obtained hydrolysate contained 24 g/l glucose, 5.4 g/l xylose and 0.6 g/l arabinose. In the case of Simultaneous Saccharification and Fermentation (SSF), hydrolysis and fermentation were conducted in a single step process at 37oC in pH 5. The enzymatic hydrolysis of DGGS pretreated solids took place mostly during lag phase of L. coryniformis fermentation, with only a small amount of glucose consumed during the first 6 h. When exponential phase was started, glucose generation reduced as the microorganism started to consume glucose for D-lactic acid production. Higher concentrations of D-lactic acid were produced when SSF approach was applied, with 28 g/l D-lactic acid after 24 h of fermentation (84.5% yield). In contrast, 21.2 g/l D-lactic acid were produced when SHF was used. The optical pu rity of D-lactic acid produced from both experiments was 99.9%. Besides, approximately 2 g/l acetic acid was also generated due to lactic acid degradation after glucose depletion in SHF. SSF was proved an efficient towards DDGS ulilisation and D-lactic acid production, by reducing the overall processing time, yielding sufficient D-lactic acid concentrations without the generation of fermentation by-products.

Keywords: DDGS, alkaline pretreatment, SSF, D-lactic acid

Procedia PDF Downloads 324
456 Permeable Bio-Reactive Barriers to Tackle Petroleum Hydrocarbon Contamination in the Sub-Antarctic

Authors: Benjamin L. Freidman, Sally L. Gras, Ian Snape, Geoff W. Stevens, Kathryn A. Mumford

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Increasing transportation and storage of petroleum hydrocarbons in Antarctic and sub-Antarctic regions have resulted in frequent accidental spills. Migrating petroleum hydrocarbon spills can have a significant impact on terrestrial and marine ecosystems in cold regions, as harsh environmental conditions result in heightened sensitivity to pollution. This migration of contaminants has led to the development of Permeable Reactive Barriers (PRB) for application in cold regions. PRB’s are one of the most practical technologies for on-site or in-situ groundwater remediation in cold regions due to their minimal energy, monitoring and maintenance requirements. The Main Power House site has been used as a fuel storage and power generation area for the Macquarie Island research station since at least 1960. Soil analysis at the site has revealed Total Petroleum Hydrocarbon (TPH) (C9-C28) concentrations as high as 19,000 mg/kg soil. Groundwater TPH concentrations at this site can exceed 350 mg/L TPH. Ongoing migration of petroleum hydrocarbons into the neighbouring marine ecosystem resulted in the installation of a ‘funnel and gate’ PRB in November 2014. The ‘funnel and gate’ design successfully intercepted contaminated groundwater and analysis of TPH retention and biodegradation on PRB media are currently underway. Installation of the PRB facilitates research aimed at better understanding the contribution of particle attached biofilms to the remediation of groundwater systems. Bench-scale PRB system analysis at The University of Melbourne is currently examining the role biofilms play in petroleum hydrocarbon degradation, and how controlled release nutrient media can heighten the metabolic activity of biofilms in cold regions in the presence of low temperatures and low nutrient groundwater.

Keywords: groundwater, petroleum, Macquarie island, funnel and gate

Procedia PDF Downloads 350
455 Repeated Batch Cultivation: A Novel Empty and Fill Strategy for the Enhanced Production of a Biodegradable Polymer, Polyhydroxy Alkanoate by Alcaligenes latus

Authors: Geeta Gahlawat, Ashok Kumar Srivastava

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In the present study, a simple drain and fill protocol strategy of repeated batch was adopted for enhancement in polyhydroxyalkanoates (PHAs) production using alcaligenes latus DSM 1124. Repeated batch strategy helped in increasing the longevity of otherwise decaying culture in the bioreactor by supplementing fresh substrates during each cycle of repeated-batch. The main advantages of repeated batch are its ease of operation, enhancement of culture stability towards contamination, minimization of pre-culture effects and maintenance of organism at high growth rates. The cultivation of A. latus was carried out in 7 L bioreactor containing 4 L optimized nutrient medium and a comparison with the batch mode fermentation was done to evaluate the performance of repeated batch in terms of PHAs accumulation and productivity. The statistically optimized medium recipe consisted of: 25 g/L Sucrose, 2.8 g/L (NH4)2SO4, 3.25 g/L KH2PO4, 3.25 g/L Na2HPO4, 0.2 g/L MgSO4, 1.5 mL/L trace element solution. In this strategy, 20% (v/v) of the culture broth was removed from the reactor and supplemented with an equal volume of fresh medium when sucrose concentration inside the reactor decreased below 8 g/L. The fermenter was operated for three repeated batch cycles and fresh nutrient feeding was done at 27 h, 48 h, and 60 h. Repeated batch operation resulted in a total biomass of 27.89 g/L and PHAs concentration 20.55 g/L at the end of 69 h which was a marked improvement as compared to batch cultivation (8.71 g/L biomass and 6.24 g/L PHAs). This strategy demonstrated 3.3 fold and 1.8 fold increase in PHAs concentration and volumetric productivity, respectively as compared to batch cultivation. Repeated batch cultivation strategy had also the benefit of avoiding non-productive time period required for cleaning, refilling and sterilization of bioreactor, thereby increasing the overall volumetric productivity and making the entire process cost-effective too.

Keywords: alcaligenes, biodegradation, polyhydroxyalkanoates, repeated batch

Procedia PDF Downloads 358
454 Biogenic Amines Production during RAS Cheese Ripening

Authors: Amr Amer

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Cheeses are among those high-protein-containing foodstuffs in which enzymatic and microbial activities cause the formation of biogenic amines from amino acids decarboxylation. The amount of biogenic amines in cheese may act as a useful indicator of the hygienic quality of the product. In other words, their presence in cheese is related to its spoilage and safety. Formation of biogenic amines during Ras cheese (Egyptian hard cheese) ripening was investigated for 4 months. Three batches of Ras cheese were manufactured using Egyptian traditional method. From each batch, Samples were collected at 1, 7, 15, 30, 60, 90 and 120 days after cheese manufacture. The concentrations of biogenic amines (Tyramine, Histamine, Cadaverine and Tryptamine) were analyzed by high performance liquid chromatography (HPLC). There was a significant increased (P<0.05) in Tyramine levels from 4.34± 0.07 mg|100g in the first day of storage till reached 88.77± 0.14 mg|100g at a 120-day of storage. Also, Histamine and Cadaverine levels had the same increased pattern of Tyramine reaching 64.94± 0.10 and 28.28± 0.08 mg|100g in a 120- day of storage, respectively. While, there was a fluctuation in the concentration of Tryptamine level during ripening period as it decreased from 3.24± 0.06 to 2.66± 0.11 mg|100g at 60-day of storage then reached 5.38±0.08 mg|100g in a 120- day of storage. Biogenic amines can be formed in cheese during production and storage: many variables, as pH, salt concentration, bacterial activity as well as moisture, storage temperature and ripening time, play a relevant role in their formation. Comparing the obtained results with the recommended standard by Food and Drug Administration "FDA" (2001), High levels of biogenic amines in various Ras cheeses consumed in Egypt exceeded the permissible value (10 mg%) which seemed to pose a threat to public health. In this study, presence of high concentrations of biogenic amines (Tyramine, Histamine, cadaverine and Tryptamine) in Egyptian Ras cheeses reflects the bad hygienic conditions under which they produced and stored. Accordingly, the levels of biogenic amines in different cheeses should be come in accordance with the safe permissible limit recommended by FDA to ensure human safety.

Keywords: Ras cheese, biogenic amines, tyramine, histamine, cadaverine

Procedia PDF Downloads 426
453 Bio-Mimetic Foam Fractionation Technology for the Treatment of Per- and PolyFluoroAlkyl Substances (PFAS) in Contaminated Water

Authors: Hugo Carronnier, Wassim Almouallem, Eric Branquet

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Per- and polyfluoroalkyl Substances (PFAS) are a group of man-made refractory compounds that have been widely used in a variety of industrial and commercial products since the 1940s, leading to contamination of groundwater and surface water systems. They are persistent, bioaccumulative and toxic chemicals. Foam fractionation is a potential remedial technique for treating PFAS-contaminated water, taking advantage of the high surface activity to remove them from the solution by adsorption onto the surface of the air bubbles. Nevertheless, traditional foam fractionation technology developed for PFAS is challenging and found to be ineffective in treating the less surface-active compounds. Different chemicals were the subject of investigation as amendments to achieve better removal. However, most amendments are toxic, expensive and complicated to use. In this situation, patent-pending PFAS technology overcomes these challenges by using rather biological amendments. Results from the first laboratory trial showed remarkable results using a simple and cheap BioFoam Fractionation (BioFF) process based on biomimetics. The study showed that the BioFF process is effective in removing greater than 99% of PFOA (C8), PFOS (C8), PFHpS (C7) and PFHxS (C6) in PFAS-contaminated water. For other PFAS such as PFDA (C10) and 6:2 FTAB, a slightly less stable removal between 94% and 96% was achieved while between 34% and 73% removal efficiency was observed for PFBA (C4), PFBS (C4), PFHxA (C6), and Gen-X. In sum, the advantages of the BioFF presented as a low-waste production, a cost and energy-efficient operation and the use of a biodegradable amendment requiring no separation step after treatment, coupled with these first findings, suggest that the BioFF process is a highly applicable treatment technology for PFAS contaminated water. Additional investigations are currently carried on in order to optimize the process and establish a promising strategy for on-site PFAS remediation.

Keywords: PFAS, treatment, foam fractionation, contaminated amendments

Procedia PDF Downloads 65
452 A contribution to Phytochemical and Biological Studies of Ailanthus Alitssima Swingle Cultivated in Egypt

Authors: Ahmed Samy Elnoby

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Ailanthus altissima native to Asia which belongs to the family Simaroubaceae was subjected to phytochemical screening and biological investigations. Phytochemical screening revealed the presence of carbohydrates, tannins, sterols, flavonoids and traces of saponins. In addition, quantitative determination of phenolics and flavonoid content were performed. The antimicrobial activity of methanolic extract of the leaves was determined against gram-positive, gram-negative bacteria in addition to fungi using a modified Kirby-Bauer disc diffusion method that was compared with standard discs ampicillin which acts as an antibacterial agent and amphotericin B which acts as an antifungal agent. A high potency was observed against gram-positive bacteria mainly staphylococcus aureus, gram-negative bacteria mainly Escherichia coli and showed no potency against fungi mainly Aspergillus flavus and candida albicans. On the other hand, the antioxidant activity of the extract was determined by 1, 1-diphenyl-2- diphenyl-2-picryl-hydrazil (DPPH). A very low potency was shown by using DPPH for the antioxidant effect so IC50 = 0 ug/ml, IC90 =0 ug /ml and remark gave 47.2 % at 100 ug/ml which is very weak. Cytotoxic activity was determined by using MTT assay (3-4, 5-Dimethylthiazol-2-yl)-2, 5-Diphenyltetrazolium Bromide) against MCF7 (Human Caucasian breast adenocarcinoma) cell line. A moderate potency was shown by using MCF7 cell line for cytotoxic effect so LC50= 90.2 ug/ml, LC90=139.9 ug/ml and the remark gave 55.2% at 100 ug/ml which is of moderate activity so, Ailanthus altissima can be considered to be a promising antimicrobial agent from natural origin.

Keywords: Ailanthus altissima, TLC, HPLC, anti-microbial activity, antifungal activity, antioxidant, cytotoxic activity

Procedia PDF Downloads 164
451 Evaluation of Biological Seed Coating Technology On-Field Performance of Wheat in Regenerative Agriculture and Conventional Systems

Authors: S. Brain, P. J. Storer, H. Strydom, Z. M. Solaiman

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Increasing farmer awareness of soil health, the impact of agricultural management practices, and the requirement for high-quality agricultural produce are major factors driving the rapid adoption of biological seed treatments - currently valued globally at USD 1.5 billion. Biological seed coatings with multistrain plant beneficial microbial technology have the capability to affect plant establishment, growth, and development positively. These beneficial plant microbes can potentially increase soil health, plant yield, and nutrition – acting as bio fertilisers, rhizoremediators, phytostimulators, and stress modulators, and can ultimately reduce the overall use of agrichemicals. A field trial was conducted on MACE wheat in the central wheat belt of Western Australia to evaluate a proprietary seed coating technology (Langleys Bio-EnergeticTM Microbe blend (BMB)) on a conventional program (+/- BMB microbes) and a Regenerative Biomineral fertiliser program (+/- BMB microbes). The Conventional (+BMB) and Biomineral (+BMB) treated plants had no fungicide treatments and had no disease issues. Control (No fertiliser, No microbes), Conventional (No Microbes), and Biomineral (No Microbes) were treated with fungicides (seed dressing and foliar). From the research findings, compared to control and no microbe treatments, both the Conventional (+ BMB) and Biomineral (+ BMB) showed significant increases in Soil Carbon (SOC), Seed germination, nutrient use efficiency (NUE) of nitrogen, phosphate and mineral nutrients, grain mineral nutrient uptake, protein %, hectolitre weight, and fewer screenings, yield, and gross margins.

Keywords: biological seed coating, biomineral fertiliser, plant nutrition, regenerative and conventional agriculture

Procedia PDF Downloads 66
450 Characterization of the Microorganisms Associated with Pleurotus ostractus and Pleurotus tuber-Regium Spent Mushroom Substrate

Authors: Samuel E. Okere, Anthony E. Ataga

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Introduction: The microbial ecology of Pleurotus osteratus and Pleurotus tuber–regium spent mushroom substrate (SMS) were characterized to determine other ways of its utilization. Materials and Methods: The microbiological properties of the spent mushroom substrate were determined using standard methods. This study was carried out at the Microbiology Laboratory University of Port Harcourt, Rivers State, Nigeria. Results: Quantitative microbiological analysis revealed that Pleurotus osteratus spent mushroom substrate (POSMS) contained 7.9x10⁵ and 1.2 x10³ cfu/g of total heterotrophic bacteria and total fungi count respectively while Pleurotus tuber-regium spent mushroom substrate (PTSMS) contained 1.38x10⁶ and 9.0 x10² cfu/g of total heterotrophic bacteria count and total fungi count respectively. The fungi species encountered from Pleurotus tuber-regium spent mushroom substrate (PTSMS) include Aspergillus and Cladosporum species, while Aspergillus and Penicillium species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). However, the bacteria species encountered from Pleurotus tuber-regium spent mushroom substrate include Bacillus, Acinetobacter, Alcaligenes, Actinobacter, and Pseudomonas species while Bacillus, Actinobacteria, Aeromonas, Lactobacillus and Aerococcus species were encountered from Pleurotus osteratus spent mushroom substrate (POSMS). Conclusion: Therefore based on the findings from this study, it can be concluded that spent mushroom substrate contain microorganisms that can be utilized both in bioremediation of oil-polluted soils as they contain important hydrocarbon utilizing microorganisms such as Penicillium, Aspergillus and Bacillus species and also as sources of plant growth-promoting rhizobacteria (PGPR) such as Pseudomonas and Bacillus species which can induce resistance on plants. However, further studies are recommended, especially to molecularly characterize these microorganisms.

Keywords: characterization, microorganisms, mushroom, spent substrate

Procedia PDF Downloads 147
449 Scalable and Accurate Detection of Pathogens from Whole-Genome Shotgun Sequencing

Authors: Janos Juhasz, Sandor Pongor, Balazs Ligeti

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Next-generation sequencing, especially whole genome shotgun sequencing, is becoming a common approach to gain insight into the microbiomes in a culture-independent way, even in clinical practice. It does not only give us information about the species composition of an environmental sample but opens the possibility to detect antimicrobial resistance and novel, or currently unknown, pathogens. Accurately and reliably detecting the microbial strains is a challenging task. Here we present a sensitive approach for detecting pathogens in metagenomics samples with special regard to detecting novel variants of known pathogens. We have developed a pipeline that uses fast, short read aligner programs (i.e., Bowtie2/BWA) and comprehensive nucleotide databases. Taxonomic binning is based on the lowest common ancestor (LCA) principle; each read is assigned to a taxon, covering the most significantly hit taxa. This approach helps in balancing between sensitivity and running time. The program was tested both on experimental and synthetic data. The results implicate that our method performs as good as the state-of-the-art BLAST-based ones, furthermore, in some cases, it even proves to be better, while running two orders magnitude faster. It is sensitive and capable of identifying taxa being present only in small abundance. Moreover, it needs two orders of magnitude less reads to complete the identification than MetaPhLan2 does. We analyzed an experimental anthrax dataset (B. anthracis strain BA104). The majority of the reads (96.50%) was classified as Bacillus anthracis, a small portion, 1.2%, was classified as other species from the Bacillus genus. We demonstrate that the evaluation of high-throughput sequencing data is feasible in a reasonable time with good classification accuracy.

Keywords: metagenomics, taxonomy binning, pathogens, microbiome, B. anthracis

Procedia PDF Downloads 121
448 Antimicrobial Resistance Patterns of Campylobacter from Pig and Cattle Carcasses in Poland

Authors: Renata Szewczyk, Beata Lachtara, Kinga Wieczorek, Jacek Osek

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Campylobacter is recognized as the main cause of bacterial gastrointestinal infections in Europe. A main source of the pathogen is poultry and poultry meat; however, other animals like pigs and cattle can also be reservoirs of the bacteria. Human Campylobacter infections are often self-limiting but in some cases, macrolide and fluoroquinolones have to be used. The aim of this study was to determine antimicrobial resistance patterns (AMR) of Campylobacter isolated from pig and cattle carcasses. Between July 2009 and December 2015, 735 swabs from pig (n = 457) and cattle (n = 278) carcasses were collected at Polish slaughterhouses. All samples were tested for the presence of Campylobacter by ISO 10272-1 and confirmed to species level using PCR. The antimicrobial susceptibility of Campylobacter isolates was determined by a microbroth dilution method with six antimicrobials: gentamicin (GEN), streptomycin (STR), erythromycin (ERY), nalidixic acid (NAL), ciprofloxacin (CIP) and tetracycline (TET). It was found that 167 of 735 samples (22.7%) were contaminated with Campylobacter. The vast majority of them were of pig origin (134; 80.2%), whereas for cattle carcasses Campylobacter was less prevalent (33; 19.8%). Among positive samples C. coli was predominant species (123; 73.7%) and it was isolated mainly from pig carcasses. The remaining isolates were identified as C. jejuni (44; 26.3%). Antimicrobial susceptibility indicated that 22 out of 167 Campylobacter (13.2%) were sensitive to all antimicrobials used. Fourteen of them were C. jejuni (63.6%; pig, n = 6; cattle, n = 8) and 8 was C. coli (36.4%; pig, n = 4; cattle, n = 4). Most of the Campylobacter isolates (145; 86.8%) were resistant to one or more antimicrobials (C. coli, n = 115; C. jejuni, n = 30). Comparing the AMR for Campylobacter species it was found that the most common pattern for C. jejuni was CIP-NAL-TET (9; 30.0%), whereas CIP-NAL-STR-TET was predominant among C. coli (47; 40.9%). Multiresistance, defined as resistance to three or more classes of antimicrobials, was found in 57 C. coli strains, mostly obtained from pig (52 isolates). On the other hand, only one C. jejuni strain, isolated from cattle, showed multiresistance with pattern CIP-NAL-STR-TET. Moreover, CIP-NAL-STR-TET was characteristic for most of multiresistant C. coli isolates (47; 82.5%). For the remaining C. coli the resistance patterns were CIP-ERY-NAL-TET (7 strains; 12.3%) and for one strain of each patterns: ERY-STR-TET, CIP-STR-TET, CIP-NAL-GEN-STR-TET. According to the present findings resistance to erythromycin was observed only in 11 C. coli (pig, n = 10; cattle, n = 1). In conclusion, the results of this study showed that pig carcasses may be a serious public health concern because of contamination with C. coli that might features multiresistance to antimicrobials.

Keywords: antimicrobial resistance, Campylobacter, carcasses, multi resistance

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447 Assessment of Chromium Concentration and Human Health Risk in the Steelpoort River Sub-Catchment of the Olifants River Basin, South Africa

Authors: Abraham Addo-Bediako

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Many freshwater ecosystems are facing immense pressure from anthropogenic activities, such as agricultural, industrial and mining. Trace metal pollution in freshwater ecosystems has become an issue of public health concern due to its toxicity and persistence in the environment. Trace elements pose a serious risk not only to the environment and aquatic biota but also humans. Chromium is one of such trace elements and its pollution in surface waters and groundwaters represents a serious environmental problem. In South Africa, agriculture, mining, industrial and domestic wastes are the main contributors to chromium discharge in rivers. The common forms of chromium are chromium (III) and chromium (VI). The latter is the most toxic because it can cause damage to human health. The aim of the study was to assess the contamination of chromium in the water and sediments of two rivers in the Steelpoort River sub-catchment of the Olifants River Basin, South Africa and human health risk. The concentration of Cr was analyzed using inductively coupled plasma–optical emission spectrometry (ICP-OES). The concentration of the metal was found to exceed the threshold limit, mainly in areas of high human activities. The hazard quotient through ingestion exposure did not exceed the threshold limit of 1 for adults and children and cancer risk for adults and children computed did not exceed the threshold limit of 10-4. Thus, there is no potential health risk from chromium through ingestion of drinking water for now. However, with increasing human activities, especially mining, the concentration could increase and become harmful to humans who depend on rivers for drinking water. It is recommended that proper management strategies should be taken to minimize the impact of chromium on the rivers and water from the rivers should properly be treated before domestic use.

Keywords: land use, health risk, metal pollution, water quality

Procedia PDF Downloads 71