Search results for: bacterial inhibition

Authors: Alok Kumar, Hari Ram, Lebin Thomas, Ved Pal Singh

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Organic solvent tolerant amylases and proteases of microbial origin are in great demand for their application in transglycosylation of water-insoluble flavanoids and in peptide synthesizing reaction in organic media. Most of the amylases and proteases are unstable in presence of organic solvent. In the present work two different bacterial strains M-11 and VP-07 were isolated from the soil sample of a dairy farm in Delhi, India, for the efficient production of extracellular amylase and protease through their screening on starch agar (SA) and skimmed milk agar (SMA) plates, respectively. Both the strains (M-11 and VP-07) were identified based on morphological, biochemical and 16S rRNA gene sequencing methods. After analysis through Ez-Taxon software, the strains M-11 and VP-07 were found to have maximum pairwise similarity of 98.63% and 100% with Bacillus subtilis subsp. inaquosorum BGSC 3A28 and Bacillus anthracis ATCC 14578 and were therefore identified as Bacillus sp. UKS1 and Bacillus sp. UKS2, respectively. Time course study of enzyme activity and bacterial growth has shown that both strains exhibited typical sigmoid growth behavior and maximum production of amylase (180 U/ml) and protease (78 U/ml) by these strains (UKS1 and UKS2) was commenced during stationary phase of growth at 24 and 20 h, respectively. Thereafter, both amylase and protease were tested for their tolerance towards organic solvents and were found to be active as well stable in p-xylene (130% and 115%), chloroform (110% and 112%), isooctane (119% and 107%), benzene (121% and 104%), n-hexane (116% and 103%) and toluene (112% and 101%, respectively). Owing to such properties, these enzymes can be exploited for their potential application in industries for organic synthesis.

Keywords: amylase, enzyme activity, industrial applications, organic solvent tolerant, protease

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Authors: Antonia Stergiou

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Background: Due to the developmental trajectory of executive function in preschool age, the majority of existing studies investigating the association between acute physical activity and cognitive control have focused on adolescents and adult population. Aim- The aim of this study was to investigate the possible effects of physical activity on the inhibitory control of pre-school children. Methods: This is a prospectively designed study that was conducted in a primary school in Bristol in June 2015. The total number of subjects was n=61 and 20 trials of a modified Eriksen Flanker Task were completed before and after a 30-minutes session of moderate exercise (including both 5 minutes of warm up and cool down). For each test a pre- and post-test assessment took place that included both congruent and incongruent trials. The congruent trials were considered as the control condition and the incongruent trials as those that measure inhibitory control (experimental condition). At the end of the assessment, the participants were instructed to choose the face that described their current feelings between three options (happy, neutral, sad). Results: There was a trend for increased accuracy following moderate exercise, but there was statistical significance (p > .05). However, there was statistically significant improvement in the reaction time following the same type of exercise (p = .005). Face board assessment revealed positive emotions after 30 minutes of moderate exercise. Conclusions: The current study supports findings from previous studies related to the benefits of physical activity on the children’s inhibitory control and provides evidence of those benefits in even younger ages. Further research should take place considering each child individually. Implementation of those findings could result in an improved curriculum in schools with additional time spent on physical education courses.

Keywords: cognitive control, inhibition, physical activity, pre-school children

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Authors: Sangyeop Shin, D. C. M. Kulatunga, S. H. S. Dananjaya, Chamilani Nikapitiya, Jehee Lee, Mahanama De Zoysa

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Saprolegniasis is one of the most devastating fungal diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated fungi belong to the member of S. parasitica, supported by its typical fungal features including cotton-like whitish mycelium, zoospores (primary and secondary) and phylogenetic analysis with internal transcribed spacer (ITS) region. Pathogenicity of isolated S. parasitica was developed in embryo, larvae, juvenile and adult zebrafish as a disease model. Up regulation of host genes encoding ZfTnf-α, Zfc-Rel, ZfIl-12, ZfLyz-c, Zfβ-def, and ZfHsp-70 was identified in zebrafish larvae after experimental challenge of S. parasitica showing the host immune responses against the S. parasitica. Survival of the juveniles upon fungal infection might be due to the increased immune protection in the host. Investigation of antifungal susceptibility of S. parasitica with natural lawsone (2-hydroxy-1,4-naphthoquinone) revealed the minimum inhibitory concentration (MIC) and percentage inhibition of radial growth (PIRG %) as 200 µg/mL and 31.8%, respectively. Lawsone was able to change the membrane permeability, and cause irreversible damage and disintegration to the cellular membranes of S. parasitica which might have effect on fungi growth inhibition. Moreover, the mycelium exposed to lawsone (MIC level) changed the transcriptional responses of S. parasitica genes. Overall results indicate that lawsone could be a potential and novel anti-S. parasitica agent for controlling S. parasitica infection.

Keywords: host-pathogen interactions, lawsone, rainbow trout, Saprolegnia parasitica, Saprolegniasis, zebrafish

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Authors: Tsegay Teklebrhan Gebremariam, Zhiliang, Arjan Jonker

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The study investigated that using corn gluten (CG) instead of cornmeal (CM) increased dietary sulfur shifted H₂ metabolism from methanogenesis to alternative sink and modulated microbiome in the rumen as well as hindgut segments of goats. Ruminal fermentation, CH₄ emissions and microbial abundance in goats (n = 24). The experiment was performed using a randomized block design with two dietary treatments (CM and CG with 400 g/kg DM each). Goats in CG increased sulfur, NDF and CP intake and decreased starch intake as compared with those in CM. Goats that received CG diet had decreased dissolved hydrogen (dH₂) (P = 0.01) and dissolved methane yield and emission (dCH₄) (P = 0.001), while increased dH₂S both in the rumen and hindgut segments than those fed CM. Goats fed CG had higher (p < 0.01) gene copies of microbiota and cellulolytic bacteria, whereas starch utilizing bacterial species were less in the rumen and hindgut than those fed CM. Higher (P < 0.05) methanogenic diversity and abundances of Methanimicrococcus and Methanomicrobium were observed in goats that consumed CG, whilst containing lower Methanobrevibacter populations than those receiving CM. The study suggested that goats fed corn gluten improved the gene copies of microbiota and fibrolytic bacterial species while reducing starch utilizing species in the rumen and hindgut segments as compared with that fed cornmeal. Goats consuming corn gluten had a more enriched methanogenic diversity and reduced Methanobrevibacter, a contributor to CH₄ emissions, as compared with goats fed CM. Corn gluten could be used as an alternative feed to decrease the enteric CH₄ emission in ruminant production.

Keywords: dissolved gasses, methanogenesis, microbial community, metagenomics

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Authors: Kara Terki Ibtissem, Hassaine Hafida, Bellifa Samia

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Introduction: Staphylococcus aureus is one of the species that are most frequently isolated from urinary catheters. The ability to produce a biofilm is an important step in the pathogenesis of these staphylococci; biofilm formation is strongly dependent on the environmental conditions it also depends on the different parameters these biofilms are subjected to. Antiseptics, including ethanol and betadine, are used in clinical practice for disinfection and infection prevention. Recent studies, however, demonstrate that disinfectants may enhance biofilm production in Staphylococci. Methods: In this study, 48 staphylococcus aureus isolated from urinary catheters at the University Hospital Center of Sidi Bel Abbes (in Northwestern Algeria) were analyzed to detect the formation of biofilm by culture on Red Congo Agar (RCA), the Tube Method (TM) and tissue Culture Plate (TCP) techniques, this last was also used to investigate the effect of ethanol and Betadine on the preformed biofilm In a second time to know which environment is most favorable to the formation of the biofilm we perform a statistical test based on the student test by the software R. Results: It has been found that 23 strains produced a bacterial slime on the Congo red medium, 5 strains produced a biofilm by the tube method, 2 of which are highly productive. In addition, 7 strains produced a biofilm on polystyrene micro-plates; this number was higher in the presence of ethanol 70% and ethanol 90% with 19 and 11 biofilm-producing strains, respectively. On the other hand, no biofilm was formed in the presence of Betadine. Conclusion: It is important to examine the response of biofilms following an imposed external constraint, such as disinfectants, in order to develop new strategies to combat bacterial biofilms but also to better control their formation.

Keywords: staphylococcus aureus, biofilm, urinary catheter, ethanol

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Authors: Atin Adhikari, Sushma Kurella, Pratik Banerjee, Nabanita Mukherjee, Yamini M. Chandana Gollapudi, Bushra Shah

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Different sharp instruments, drilling machines, and high speed rotary instruments are routinely used in dental clinics during dental cleaning. Therefore, these cleaning procedures release a lot of oral microorganisms including bacteria in clinic air and may cause significant occupational bioaerosol exposure risks for dentists, dental hygienists, patients, and dental clinic employees. Two major goals of this study were to quantify volumetric airborne concentrations of bacteria and to assess overall microbial activity in this type of occupational environment. The study was conducted in several dental clinics of southern Georgia and 15 dental cleaning procedures were targeted for sampling of airborne bacteria and testing of overall microbial activity in settled dusts over clinic floors. For air sampling, a Biostage viable cascade impactor was utilized, which comprises an inlet cone, precision-drilled 400-hole impactor stage, and a base that holds an agar plate (Tryptic soy agar). A high-flow Quick-Take-30 pump connected to this impactor pulls microorganisms in air at 28.3 L/min flow rate through the holes (jets) where they are collected on the agar surface for approx. five minutes. After sampling, agar plates containing the samples were placed in an ice chest with blue ice and plates were incubated at 30±2°C for 24 to 72 h. Colonies were counted and converted to airborne concentrations (CFU/m3) followed by positive hole corrections. Most abundant bacterial colonies (selected by visual screening) were identified by PCR amplicon sequencing of 16S rRNA genes. For understanding overall microbial activity in clinic floors and estimating a general cleanliness of the clinic surfaces during or after dental cleaning procedures, ATP levels were determined in swabbed dust samples collected from 10 cm2 floor surfaces. Concentration of ATP may indicate both the cell viability and the metabolic status of settled microorganisms in this situation. An ATP measuring kit was used, which utilized standard luciferin-luciferase fluorescence reaction and a luminometer, which quantified ATP levels as relative light units (RLU). Three air and dust samples were collected during each cleaning procedure (at the beginning, during cleaning, and immediately after the procedure was completed (n = 45). Concentrations at the beginning, during, and after dental cleaning procedures were 671±525, 917±1203, and 899±823 CFU/m3, respectively for airborne bacteria and 91±101, 243±129, and 139±77 RLU/sample, respectively for ATP levels. The concentrations of bacteria were significantly higher than typical indoor residential environments. Although an increasing trend for airborne bacteria was observed during cleaning, the data collected at three different time points were not significantly different (ANOVA: p = 0.38) probably due to high standard deviations of data. The ATP levels, however, demonstrated a significant difference (ANOVA: p <0.05) in this scenario indicating significant change in microbial activity on floor surfaces during dental cleaning. The most common bacterial genera identified were: Neisseria sp., Streptococcus sp., Chryseobacterium sp., Paenisporosarcina sp., and Vibrio sp. in terms of frequencies of occurrences, respectively. The study concluded that bacterial exposure in dental clinics could be a notable occupational biohazard, and appropriate respiratory protections for the employees are urgently needed.

Keywords: bioaerosols, hospital hygiene, indoor air quality, occupational biohazards

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Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

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In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: trichoderma spp., meloidogyne javanica, Fusarium oxysporum f.sp.radicis lycopersici, biocontrol

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Authors: Nijole Savickiene, Antonella Di Sotto, Gabriela Mazzanti, Rasa Starselskyte, Silvia Di Giacomo, Annabella Vitalone

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Plant lectins are non-enzymic and non-immune origin proteins that specifically recognize and bind to various sugar structures and possess the activity to agglutinate cells and/or precipitate polysaccharides and glycoconjugates. The emerging evidences showed that plant lectins contribute not only to tumour cell recognition but also to cell adhesion and localization, to signal transduction, to mitogenic cytotoxicity and apoptosis. Among chitin-binding lectins, the Urtica dioica agglutinin (UDA), which is a complex of different isoforms, has been poorly studied for its biological activity. In this context and according to the increasing interest for lectins as novel antitumor drugs, present paper aimed at evaluating the potential antimutagenic activity of a lectin-like glycoprotein-enriched fraction from aerial part of Urtica dioica L. Aim: to evaluate the potential chemopreventive properties of a protein - lectin fraction from the aerial part of Urtica dioica. Materials and methods: Protein – lectin fraction has been tested for the antimutagenic activity in bacteria (50–800 mg/plate; Ames test by the preincubation method) and for the cytotoxicity on human hepatoma HepG2 cells (0.06–2 mg/mL; 24 and 48 h incubation). Results: Protein – lectin fraction from stinging nettle was not cytotoxic on HepG2 cells up to 2 mg/mL; conversely, it exhibited a strong antimutagenic activity against the mutagen 2-aminoanthracene (2AA) in all strains tested (maximum inhibition of 56.78 and 61% in TA98, TA100, and WP2uvrA strains, respectively, at 800 mg/plate). Discussion and conclusions: Protein – lectin fraction from Urtica dioica L. possesses antimutagenic and radical scavenging properties. Being 2AA a pro-carcinogenic agent, we hypothesize that the antimutagenicity of it can be due to the inhibition of CYP450-isoenzymes, involved in the mutagen bioactivation.

Keywords: lectins, antimutagenicity, chemoprevention, Urtica dioica

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Authors: Sylwia K. Naliwajko, Justyna Moskwa, Patryk Nowakowski, Renata Markiewicz-Zukowska, Krystyna Gromkowska-Kepka, Anna Puscion-Jakubik, Maria H. Borawska

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Amygdalin is found in many fruit seeds, including apricot, peach, quince, apples, and almonds. Amygdalin (also named vitamin B17), as well as its sources, are commonly used as an alternative therapy or prevention of cancer. The potential activity of amygdalin is related to its enzymatic degradation to the hydrogen cyanide. Hydrogen cyanide is a toxic substance that causes liver and nerves damage, fever, coma or even death. Amygdalin is much better tolerated after intravenous than oral administration. The aim of this study was to examine the influence of amygdalin on glioblastoma multiforme cell lines. Three glioblastoma multiforme cell lines – U87MG, T98, LN18 were incubated (48 h) with amygdalin in concentrations 100, 250, 500, 1000 and 2000 µg/mL. The MTT (Thiazolyl Blue Tetrazolium Bromide) test and DNA binding test by [3H]-thymidine incorporation were used to determine the anti-proliferative activity of amygdalin. The secretion of metalloproteinases (MMP2 and MMP-9) from U87MG cells was estimated by gelatin zymography. The statistical analysis was performed using Statistica v. 13.0 software. The data was presented as a % of control. Amygdalin did not show significant inhibition of viability of all the glioblastoma cells in concentrations 100, 250, 500, 1000 µg/mL. In 2000 µg/mL there were significant differences compared to the control, but inhibition of viability was less than 20% (more than 80% of control). The average viability of U87MG cells was 92,0±4%, T98G: 85,8±3% and LN18: 94,7±2% of the control. There was no dose-response viability, and IC50 value was not recognized. DNA binding in U87MG cells was not inhibited (109,0±3 % of control). After treatment with amygdalin, we observed significantly increased secretion of MMP2 and MMP9 in U87MG cells (130,3±14% and 112,0±5% of control, respectively). Our results suggest that amygdalin has no anticancer activity in glioblastoma cell lines.

Keywords: amygdalin, anticancer, cell line, glioblastoma

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Authors: Pardis Abolghasemi, Benyamin Hatamsaz

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Background: Diabetic nephropathy is a serious health problem described by specific kidney structure and functional disturbance. Renoprotective effects of the stem cells secretase have been shown in many kidney diseases. The aim is to evaluate the capability of human Wharton’s jelly mesenchymal stem cells conditioned media (hWJMSCs-CM) to alleviate DN in streptozotocin (STZ)-induced diabetes. Methods: Diabetic nephropathy was induced by injection of STZ (60 mg/kg, IP) in twenty rats. Conditioned media was extracted from hWJMSCs at third passages. At week 8, diabetic rats were divided into two groups: treated (hWJMSCs-CM, 500 μl/rat for three weeks, IP) and not treated (DN). In the 11th week, three groups (control, DN and DN+hWJMSCs-CM) were kept in metabolic cages and urine was collected for 24h. Blood pressure (BP) and heart rate (HR) were continuously recorded. The serum samples were maintained for measuring BUN, Cr and angiotensin-converting enzyme (ACE) activity. The left kidney was kept at -80°C for ACE activity assessment. The right kidney and pancreas were used for histopathologic evaluation. Result: Diabetic nephropathy was detected by microalbuminuria and increased albumin/creatinine ratio, as well as the pancreas and renal structural disturbance. Glomerular filtration rate, BP and HR increased in the DN group. The ACE activity was elevated in the serum and kidneys of the DN group. Administration of hWJMSCs-CM modulated the renal functional and structural disturbance and decreased the ACE activity. Conclusion: Conditioned media was extracted from hWJMSCs may have a Renoprotective effect in diabetic nephropathy. This may happen through regulation of ACE activity and renin-angiotensin system inhibition.

Keywords: diabetic nephropathy, mesenchymal stem cells, immunomodulation, anti-inflammation

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Authors: E. Heintz, H. J. van Lent, K. Glass, J. Lim

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The trend for sodium reduction in food products is clear. Following the World Health Organization (WHO) publication on sodium usage and intake, several countries have introduced initiatives to reduce food-related sodium intake. As salt is a common food preservative, this trend motivates the formulation of a suitable additive with comparable benefits of shelf life extension and microbial safety. Organic acid derivatives like acetates are known as generic microbial growth inhibitors and are commonly applied as additives to meet food safety demands. However, modern consumers have negative perceptions towards -synthetic-derived additives and increasingly prefer natural alternatives. Vinegar, for example, is a well-known natural fermentation product used in food preservation. However, the high acidity of vinegar often makes it impractical for direct use in meat products and a neutralized form would be desirable. This research demonstrates the efficacy of powdered vinegar (Provian DV) in inhibiting Listeria and spoilage organisms (LAB) to increase safety and shelf life of meat products. For this, the efficacy of Provian DV was compared to the efficacy of Provian K, a commonly used sodium free acetate-based preservative, which is known for its inhibition against Listeria. Materials & methods— Cured pork hams: Ingredients: Pork ham muscle, water, salt, dextrose, sodium tripolyphosphate, carrageenan, sodium nitrite, sodium erythorbate, and starch. Targets: 73-74% moisture, 1.75+0.1% salt, and pH 6.4+0.1. Treatments: Control (no antimicrobials), Provian®K 0.5% and 0.75%, Provian®DV 0.5%, 0.65%, 0.8% and 1.0%. Meat formulations in casings were cooked reaching an internal temperature of 73.9oC, cooled overnight and stored for 4 days at 4oC until inoculation. Inoculation: Sliced products were inoculated with approximately 3-log per gram of a cocktail of L. monocytogenes (including serotypes 4b, 1/2a and 1/2b) or LAB-cocktail (C. divergens and L. mesenteroides). Inoculated slices were vacuum packaged and stored at 4oC and 7°C. Samples were incubated 28 days (LAB) or 12 weeks (L. monocytogenes) Microbial analysis: Microbial populations were enumerated in rinsate obtained after adding 100ml of sterile Butterfield’s phosphate buffer to each package and massaging the contents externally by hand. L. monocytogenes populations were determined on triplicate samples by surface plating on Modified Oxford agar whereas LAB plate counts were determined on triplicate samples by surface plating on All Purpose Tween agar with 0.4% bromocresol purple. Proximate analysis: Triplicate non-inoculated ground samples were analyzed for the moisture content, pH, aw, salt, and residual nitrite. Results—The results confirmed the no growth of Listeria on cured ham with 0.5% Provian K stored at 4°C and 7°C for 12 weeks, whereas the no-antimicrobial control showed a 1-log increase within two weeks. 0.5% Provian DV demonstrated similar efficacy towards Listeria inhibition at 4°C while 0.65% Provian DV was required to match the Listeria control at 7°C. 0.75% Provian K and 1% Provian DV were needed to show inhibition of the LAB for 4 weeks at both temperatures. Conclusions—This research demonstrated that it is possible to increase safety and shelf life of cured ready-to-eat ham using preservatives that meet current food trends, like sodium reduction and natural origin.

Keywords: food safety, natural preservation, listeria control, shelf life extension

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Authors: Shittu Bashirat, Shittu Mujeeb, Oluremi Adeolu, Orisadare Olayiwola, Jikeme Osameke, Bello Lateef

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Septicemia in neonates refers to generalized bacterial infection documented by positive blood culture in the first 28 days of life and is one of the leading causes of neonatal mortality in sub-Sahara Africa. Thrombocytopenia in newborns is a result of increased platelet consumption; sepsis was found to be the most common risk factor. The objective of the study was to determine if there are organism-specific platelet responses among the 2 groups of bacterial agents: Gram-positive and Gram-negative bacteria, and also to examine the association of platelet count and prothrombin time with neonatal septicemia. 232 blood samples were collected for this study. The blood culture was performed using Bactec 9050, an instrumented blood culture system. The platelet count and prothrombin time were performed using Abacus Junior 5 hematology analyzer and i-STAT 1 analyzer respectively. Of the 231 neonates hospitalized with clinical sepsis, blood culture reports were positive in 51 cases (21.4%). Klebsiella spp. (35.3%) and Staphylococcus aureus (27.5%) were the most common Gram-negative and Gram-positive isolates respectively. Thrombocytopenia was observed in 30 (58.8%) of the neonates with septicemia. Of the 9 (17.6%) patients with severe thrombocytopenia, seven (77.8%) had Klebsiella spp. septicemia. Out of the 21(63.6%) of thrombocytopenia produced by Gram-negative isolate, 17 (80.9) had increased prothrombin time. In conclusion, Gram-negative organisms showed the highest cases of severe thrombocytopenia and prolonged PT. This study has helped to establish a disturbance in hemostatic systems in neonates with septicemia. Further studies, however, may be required to assess other hemostasis parameters in order to understand their interaction with the infectious organisms in neonates.

Keywords: neonates, septicemia, thrombocytopenia, prolonged prothrombin time, platelet count

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Authors: Anish Shah, Steve A. Wakelin, Derrick Moot, Aurélie Laugraud, Hayley J. Ridgway

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A mixed pasture system of ryegrass-clover is used in New Zealand, where clovers are generally inoculated with commercially available strains of rhizobia. The community of rhizobia living in the soil and the way in which they interact with the plant are affected by different biotic and abiotic factors. In general, bacterial richness and diversity in soil varies by soil pH. pH also affects cell physiology and acts as a master variable that controls the wider soil physiochemical conditions such as P availability, Al release and micronutrient availability. As such, pH can have both primary and secondary effects on soil biology and processes. The aim of this work was to investigate the effect of soil pH on the genetic diversity and metabolic profile of Rhizobium leguminosarum strains nodulating clover. Soils were collected from 12 farms across New Zealand which had a pH(water) range of between 4.9 and 7.5, with four acidic (pH 4.9 – 5.5), four ‘neutral’ (5.8 – 6.1) and four alkaline (6.5 – 7.5) soils. Bacteria were recovered from nodules of Trifolium repens (white clover) and T. subterraneum (subterranean clover) grown in the soils. The strains were cultured and screened against a range of pH-amended media to demonstrate whether they were adapted to pH levels similar to their native soils. The strains which showed high relative growth at a given pH (~20% of those isolated) were selected for metabolic and taxonomic profiling. The Omnilog (Biolog Inc., Hayward, CA) phenotype array was used to perform assays on carbon (C) utilisation for selected strains. DNA was extracted from the strains which had differing C utilisation profiles and PCR products for both forward and reverse primers were sequenced for the following genes: 16S rRNA, recA, nodC, nodD and nifH (symbiotic).

Keywords: bacterial diversity, clover, metabolic and taxonomic profiling, pH adaptation, rhizobia

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Authors: Ajay Krishnamurthy, Mariyappan Mahesh Kumar, Sellamuthu Periyar Selvam

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Fruits are considered as functional foods due to the presence of various bioactive compounds available such as polyphenols, which are beneficial to health when consumed as part of our diet. The primary objective of this study was to analyze the various functional groups present in ethanolic extracts of Hylocereus undatus and Garcinia indica and also measure their antibacterial and antioxidant potential respectively thereby affirming its nutraceutical potential. To fulfill our objective, a Fourier - transform Infrared Spectroscopy (FTIR) was conducted for functional group analysis, Total Phenolic Content and DPPH free radical scavenging activity for measuring it anti-oxidant potential and agar-well diffusion assay for antibacterial potential. On careful observation and analysis of the spectrum it was found that both the fruit extracts contain similar compounds viz. Phenols, Alkanes, Alkenes, Aldehydes, Ketones, Carboxylic Acid and Amines. Total phenolic content of H.undatus and G.indica was estimated to be (26.85 ± 1.84 mg GAE/100g) and (32.84 ± 1.63 mg GAE/100g) respectively which corresponds to an inhibition of 84% and 81% respectively. H.undatus shows an inhibition of (3.4 ± 2.1mm) in gram-positive and (4.2 ± 2.24mm) in gram-negative organism on the other hand G.indica shows (2.1 ± 0.98mm) in gram-positive and (3.1 ± 1.44mm) in gram negative. The presence of such diverse compounds in the fruits helps us to understand the necessity for the inclusion of fruits in our daily diet and also helps the pharmaceutical industry in realizing the importance of exotic fruits as a potential nutraceutical.

Keywords: DPPH, fourier-transform infrared spectroscopy (FTIR), Hylocereus undatus, Garcinia indica

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Authors: Irina M. Kolesnikova, Andrey M. Gaponov, Sergey A. Roumiantsev, Tatiana V. Grigoryeva, Dilyara R. Khusnutdinova, Dilyara R. Kamaldinova, Alexander V. Shestopalov

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Background. Obese patients have unequal risks of metabolic disorders. It is accepted to distinguish between metabolically healthy obesity (MHO) and metabolically unhealthy obesity (MUHO). MUHO patients have a high risk of metabolic disorders, insulin resistance, and diabetes mellitus. Among the other things, the gut microbiota also contributes to the development of metabolic disorders in obesity. Obesity is accompanied by significant changes in the gut microbial community. In turn, bacterial translocation from the intestine is the basis for the blood microbiome formation. The aim was to study the features of the blood microbiome in patients with various metabolic types of obesity. Patients, materials, methods. The study included 116 healthy donors and 101 obese patients. Depending on the metabolic type of obesity, the obese patients were divided into subgroups with MHO (n=36) and MUHO (n=53). Quantitative and qualitative assessment of the blood microbiome was based on metagenomic analysis. Blood samples were used to isolate DNA and perform sequencing of the variable v3-v4 region of the 16S rRNA gene. Alpha diversity indices (Simpson index, Shannon index, Chao1 index, phylogenetic diversity, the number of observed operational taxonomic units) were calculated. Moreover, we compared taxa (phyla, classes, orders, and families) in terms of isolation frequency and the taxon share in the total bacterial DNA pool between different patient groups. Results. In patients with MHO, the characteristics of the alpha-diversity of the blood microbiome were like those of healthy donors. However, MUHO was associated with an increase in all diversity indices. The main phyla of the blood microbiome were Bacteroidetes, Firmicutes, Proteobacteria, and Actinobacteria. Cyanobacteria, TM7, Thermi, Verrucomicrobia, Chloroflexi, Acidobacteria, Planctomycetes, Gemmatimonadetes, and Tenericutes were found to be less significant phyla of the blood microbiome. Phyla Acidobacteria, TM7, and Verrucomicrobia were more often isolated in blood samples of patients with MUHO compared with healthy donors. Obese patients had a decrease in some taxonomic ranks (Bacilli, Caulobacteraceae, Barnesiellaceae, Rikenellaceae, Williamsiaceae). These changes appear to be related to the increased diversity of the blood microbiome observed in obesity. An increase of Lachnospiraceae, Succinivibrionaceae, Prevotellaceae, and S24-7 was noted for MUHO patients, which, apparently, is explained by a magnification in intestinal permeability. Conclusion. Blood microbiome differs in obese patients and healthy donors at class, order, and family levels. Moreover, the nature of the changes is determined by the metabolic type of obesity. MUHO linked to increased diversity of the blood microbiome. This appears to be due to increased microbial translocation from the intestine and non-intestinal sources.

Keywords: blood microbiome, blood bacterial DNA, obesity, metabolically healthy obesity, metabolically unhealthy obesity

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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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Authors: B. Samuel Raj, Solomon R. D. Jebakumar

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Microbial fuel cells (MFCs) are an emerging and promising method for achieving sustainable energy since they can remove contaminated organic matter and simultaneously generate electricity. Our approach was driven in three different ways like Bacterial fuel cell, Soil Microbial fuel cell (Soil MFC) and Plant Microbial fuel cell (Plant MFC). Bacterial MFC: Sulphate reducing bacteria (SRB) were isolated and identified as the efficient electricigens which is able to produce ±2.5V (689mW/m2) and it has sustainable activity for 120 days. Experimental data with different MFC revealed that high electricity production harvested continuously for 90 days 1.45V (381mW/m2), 1.98V (456mW/m2) respectively. Biofilm formation was confirmed on the surface of the anode by high content screening (HCS) and scanning electron Microscopic analysis (SEM). Soil MFC: Soil MFC was constructed with low cost and standard Mudwatt soil MFC was purchased from keegotech (USA). Vermicompost soil (V1) produce high energy (± 3.5V for ± 400 days) compared to Agricultural soil (A1) (± 2V for ± 150 days). Biofilm formation was confirmed by HCS and SEM analysis. This finding provides a method for extracting energy from organic matter, but also suggests a strategy for promoting the bioremediation of organic contaminants in subsurface environments. Our Soil MFC were able to run successfully a 3.5V fan and three LED continuously for 150 days. Plant MFC: Amaranthus candatus (P1) and Triticum aestivium (P2) were used in Plant MFC to confirm the electricity production from plant associated microbes, four uniform size of Plant MFC were constructed and checked for energy production. P2 produce high energy (± 3.2V for 40 days) with harvesting interval of two times and P1 produces moderate energy without harvesting interval (±1.5V for 24 days). P2 is able run 3.5V fan continuously for 10days whereas P1 needs optimization of growth conditions to produce high energy.

Keywords: microbial fuel cell, biofilm, soil microbial fuel cell, plant microbial fuel cell

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Authors: Sudeshna Mukherjee, Leena Fageria, R. Venkataramana Dilip, Rajdeep Chowdhury, Jitendra Panwar

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Metal nanoparticles in recent years have gained importance in cancer therapy due to their enhanced permeability retention effect. Among various nanomaterials, silver nanoparticles (AgNPs) have received considerable attention due to their unique properties like conductivity, chemical stability, relative lower toxicity and outstanding therapeutic potential, such as anti-inflammatory, antimicrobial and anti-cancerous activities. In this study, we took a greener approach to synthesize silver nanoparticle from fungus and analyze its effects on both epithelial and mesenchymal derived cancer cells. Much research has been done on nanoparticle-induced apoptosis, but little is known about its role in autophagy. In our study, the silver nanoparticles were seen to induce autophagy which was analyzed by studying the expression of several autophagy markers like, LC3B-II and ATG genes. Monodansylcadaverine (MDC) assay also revealed the induction of autophagy upon treatment with AgNPs. Inhibition of autophagy by chloroquine resulted in increased cell death suggesting autophagy as a survival strategy adopted by the cells. In parallel to autophagy induction, silver nanoparticles induced ROS accumulation. Interestingly, autophagy inhibition by chloroquine increased ROS level, resulting in enhanced cell death. We further analyzed MAPK signaling upon AgNP treatment. It was observed that along with autophagy, activation of JNK signaling served as pro-survival while ERK signaling served as a pro-death signal. Our results provide valuable insights into the role of autophagy upon AgNP exposure and provide cues to probabilistic strategies to effectively sensitize cancer cells.

Keywords: autophagy, JNK signalling, reactive oxygen species, silver nanoparticles

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Authors: Muhammad Naeem Chaudhry, Fahim Nawaz, Rana Nauman Shabbir

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New strategies aimed at increasing the resilience of crop plants to the negative effects of climate change represent important research priorities of plant scientists. The use of soil microorganisms to alleviate abiotic stresses like drought has gained particular importance in recent past. A field experiment was planned to investigate the effect of phosphorous solubilizing bacteria on yield and seed quality of Camelina (Camelina sativa L.) under water deficit conditions. The study was conducted at Agronomic Research Farm, University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur, during 4th week of November, 2013. The available seeds of Camelina sativa were inoculated with two bacterial strains (pseudomonas and Bacillus spp.) and grown under various water stress levels i.e. D0, (four irrigations), D3 (three irrigation), D2 (two irrigations), and D1 (one irrigation). The results revealed that drought stress significantly reduced the plant growth and yield, consequently reducing protein contents and oil concentration in camelina. The exposure to drought stress decreased plant height (16%), plant population (27%), number of fertile branches (41-59%), number of pods per plant (35%) and seed per pod (33%). Drought stress also exerted a negative impact on yield characteristics by reducing the 1000-seed weight (65%), final seed yield (52%), biological yield (22%) and harvest index (39%) of camelina. However, the inoculation of seeds with Pseudomonas and Bacillus spp. promoted the plant growth characterized by increased plant height and enhanced plant population. It was noted that inoculation of seeds with Pseudomonas resulted in the maximum plant population (113.4 cm), primary branches (19 plant-1), and number of pods (664 plant-1), whereas Bacillus inoculation resulted in maximum plant height (113.4 cm), seeds per pod (15.9), 1000-seed weight (1.85 g), and seed yield (3378.8 kg ha-1). Moreover, the inoculation with Bacillus also significantly improved the quality attributes of camelina and gave 3.5% and 2.1% higher oil contents than Pseudomonas and control (no-inoculation), respectively. Similarly, the same strain also resulted in maximum protein contents (33.3%). Our results confirmed the hypothesis that inoculation of seeds with phosphorous solubilizing bacterial strains is an effective, viable and environment-friendly approach to improve yield and quality of camelina under water deficit conditions. However, further studies are suggested to investigate the physiological and molecular processes, stimulated by bacterial strains, for increasing drought tolerance in food crops.

Keywords: Camelina, drought stress, phosphate solubilizing bacteria, seed quality

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Authors: Jung-Il Kang, Jeon Eon Park, Yu-Jin Moon, Young-Seok Ahn, Eun-Sook Yoo, Hee-Kyoung Kang

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This study was conducted to evaluate the effect of Undariopsis peterseniana, a seaweed native to Jeju Island, Korea, on the growth of hair. The dermal papilla cells (DPCs) have known to regulate hair growth cycle and length of hair follicle through interact with epithelial cells. When immortalized vibrissa DPCs were treated with the U. peterseniana extract, the U. peterseniana extract significantly increased the proliferation of DPCs. The effect of U. peterseniana extract on the growth of vibrissa follicles was also examined. U. peterseniana extract significantly increased the hair-fiber lengths of the vibrissa follicles. Hair loss is partly caused by dihydrotestosterone (DHT) binding to androgen receptor in hair follicles, and the inhibition of 5α-reductase activity can prevent hair loss through the decrease of DHT level. The U. peterseniana extract inhibited 5α-reductase activity. Minoxidil, a potent hair-growth agent, can induce proliferation in NIH3T3 fibroblasts by opening KATP channels. We thus examined the proliferative effects of U. peterseniana extract in NIH3T3 fibroblasts. U. peterseniana extract significantly increased the proliferation of NIH3T3 fibroblasts. Tetraethylammonium chloride (TEA), a K+ channel blocker, inhibited U. peterseniana-induced proliferation in NIH3T3 fibroblasts. These results suggest that U. peterseniana could have the potential to treat alopecia through the proliferation of DPCs, the inhibition of 5α-reductase activity and the opening of KATP channels. [Acknowledgement] This research was supported by The Leading Human Resource Training Program of Regional Neo industry through the National Research Foundation of Korea(NRF) funded by the Ministry of Science, ICT and future Planning (2016H1D5A1908786).

Keywords: hair growth, Undariopsis peterseniana, vibrissa follicles, dermal papilla cells, 5α-reductase, KATP channels

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Authors: Sebaa H., Cherifi F., Djabeur Abderrezak M.

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Argania spinosa, Sapotaceae sole representative in Algeria and Morocco; hence it is endemic in these regions. However, it is a recognised oil, forage, and timber tree highly adapted to aridity. The exploitation of the argan fruits produces considerable amounts of under or related products. These products, such as the endocarps of a fruit, recuperated after the use of kernels to extract oil. This research studies in detail the contents of total phenolic content was determined by Folin Ciocalteu reagent and Flavonoids by aluminum chloride colorimetric assay). Antioxidant activity of extracts was expressed as the percentage of DPPH radical inhibition and IC50 values (μg/mL). Antimicrobial activity evaluated using agar disk diffusion method against reference Pseudomonas aeruginosa ATTC 27453, Escherichia coli ATCC 23922. Immature endocarps showed a higher polyphenol content than mature endocarps. The total phenolic content in immature endocarps was found to vary from 983,75+ /- 0.45 to 980,1 +/- 0.43 mg gallic acid equivalents/g dry weight, whereas in mature endocarps, the polyphenol content ranged from 100,58 mg/g +/- 0.42 to 105 +/- 0.55% mg gallic acid equivalent / g dry weight. The flavonoid content was 16.5 mg equivalent catechin/g dry weight and 9.81mg equivalent catechin /g dry weight for immature and mature endocarp fruits, respectively. DPPH assay of the endocarps extract yielded a half-maximal effective concentration (IC50) value in the immature endocarps (549.33 μg/mL) than in mature endocarps (322 μg/mL). This result can be attributed to the higher phenolics and flavonoid compounds in the immature endocarps. Methanol extract of immature endocarps exhibited antibacterial activity against E.colie (inhibition zone, 11mm).

Keywords: antioxidant activity, antimicrobial activity, total phenolic content, DPPH assay

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Authors: Shahrukh Ahmad, Purnendu Bose

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Algal-Bacterial photobioreactors (ABPBRs) have emerged as a promising technology for sustainable biomass production and wastewater treatment. Nutrient removal is seldom done in sewage treatment plants and large volumes of wastewater which still have nutrients are being discharged and that can lead to eutrophication. That is why ABPBR plays a vital role in wastewater treatment. However, improving the efficiency of ABPBR remains a significant challenge. This study aims to enhance ABPBR efficiency by focusing on two key aspects: nutrient removal and cost-effective optimization of the light source. By integrating nutrient removal and cost analysis for light source optimization, this study proposes practical strategies for improving ABPBR efficiency. To reduce organic carbon and convert ammonia to nitrates, domestic wastewater from a 130 MLD sewage treatment plant (STP) was aerated with a hydraulic retention time (HRT) of 2 days. The treated supernatant had an approximate nitrate and phosphate values of 16 ppm as N and 6 ppm as P, respectively. This supernatant was then fed into the ABPBR, and the removal of nutrients (nitrate as N and phosphate as P) was observed using different colored LED bulbs, namely white, blue, red, yellow, and green. The ABPBR operated with a 9-hour light and 3-hour dark cycle, using only one color of bulbs per cycle. The study found that the white LED bulb, with a photosynthetic photon flux density (PPFD) value of 82.61 µmol.m-2 .sec-1 , exhibited the highest removal efficiency. It achieved a removal rate of 91.56% for nitrate and 86.44% for phosphate, surpassing the other colored bulbs. Conversely, the green LED bulbs showed the lowest removal efficiencies, with 58.08% for nitrate and 47.48% for phosphate at an HRT of 5 days. The quantum PAR (Photosynthetic Active Radiation) meter measured the photosynthetic photon flux density for each colored bulb setting inside the photo chamber, confirming that white LED bulbs operated at a wider wavelength band than the others. Furthermore, a cost comparison was conducted for each colored bulb setting. The study revealed that the white LED bulb had the lowest average cost (Indian Rupee)/light intensity (µmol.m-2 .sec-1 ) value at 19.40, while the green LED bulbs had the highest average cost (INR)/light intensity (µmol.m-2 .sec-1 ) value at 115.11. Based on these comparative tests, it was concluded that the white LED bulbs were the most efficient and costeffective light source for an algal photobioreactor. They can be effectively utilized for nutrient removal from secondary treated wastewater which helps in improving the overall wastewater quality before it is discharged back into the environment.

Keywords: algal bacterial photobioreactor, domestic wastewater, nutrient removal, led bulbs

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Authors: Varsha Shinde, Belle Damodara Shenoy

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Tarballs are crude oil remnants found in oceans after long term weathering process and are a global concern since several decades as potential marine pollutant. Being complicated in structure microbial remediation of tarballs in natural environment is a slow process. They are rich in high molecular weight alkanes and poly aromatic hydrocarbons which are resistant to microbial attack and other environmental factors, therefore remain in environment for long time. However, it has been found that many bacteria and fungi inhabit on tarballs for nutrients and shelter. Many of them are supposed to be oil degraders, while others are supposed to be getting benefited by byproducts formed during hydrocarbon metabolism. Thus tarballs are forming special interesting ecological niche of microbes. This work aimed to study diversity of bacteria and fungi from tarballs and to see their potential application in crude oil degradation. The samples of tarballs were collected from Betul beach of south Goa (India). Different methods were used to isolate culturable fraction of bacteria and fungi from it. Those were sequenced for 16S rRNA gene and ITS for molecular level identification. The 16S rRNA gene sequence analysis revealed the presence of 13 bacterial genera/clades (Alcanivorax, Brevibacterium, Bacillus, Cellulomonas, Enterobacter, Klebsiella, Marinobacter, Nitratireductor, Pantoea, Pseudomonas, Pseudoxanthomonas, Tistrella and Vibrio), while the ITS sequence analysis placed the fungi in 8 diverse genera/ clades (Aspergillus, Byssochlamys, Monascus, Paecilomyces, Penicillium, Scytalidium/ Xylogone, Talaromyces and Trichoderma). All bacterial isolates were screened for oil degradation capacity. Potential strains were subjected to crude oil degradation experiment for quantification. Results were analyzed by GC-MS-MS.

Keywords: bacteria, biodegradation, crude oil, diversity, fungi, tarballs

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Authors: Endang Purwati, Ely Vebriyanti, R. Puji Hartini, Hendri Purwanto

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This study aimed to determine the effect of addition cinnamon extract (Cinnamomum burmannii) in making goat milk yogurt product isolates dadih (Pediococcus pentosaceus) to antioxidant activity and cholesterol levels. The method of research was the experimental method by using a Randomized Block Design (RBD), which consists of 5 treatments with 4 groups as replication. Treatment in this study was used of cinnamon extract as A (0%), B (1%), C (2%), D (3%), E (4%) in a goat’s milk yoghurt. This study was used 4200 ml of Peranakan Etawa goat’s milk and 80 ml of cinnamon extract. The variable analyzed were water content, pH value, total lactic acid bacterial colonies, antioxidant activity and cholesterol levels. The average water content ranged from 81.2-85.56%. Mean pH values rang between 4.74–4.30. Mean total lactic acid bacteria colonies ranged from 3.87 x 10⁸ - 7.95 x 10⁸ CFU/ml. The average of the antioxidant activity ranged between 10.98%-27.88%. Average of cholesterol levels ranged from 14.0 mg/ml–17.5 mg/ml. The results showed that the addition of cinnamon extract in making goat milk yoghurt product isolates dadih (Pediococcus pentosaceus) significantly different (P < 0.05) to water content, pH value, total lactic acid bacterial colonies, antioxidant activity and cholesterol levels. In conclusion, the study shows that using of cinnamon extract 4% is the best in making goat milk yoghurt.

Keywords: antioxidant, cholesterol, cinnamon, Pediococcus pentosaceus, yoghurt

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Authors: Driouiche Ali, Karmal Ilham

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The Agadir region is characterized by a dry climate, ranging from arid attenuated by oceanic influences to hyper-arid. The water mobilized in the agricultural sector of greater Agadir is 95% of underground origin and comes from the water table of Chtouka. The rest represents the surface waters of the Youssef Ben Tachfine dam. These waters are intended for the irrigation of 26880 hectares of modern agriculture. More than 120 boreholes and wells are currently exploited. Their depth varies between 10 m and 200 m and the unit flow rates of the boreholes are 5 to 50 l/s. A drop in the level of the water table of about 1.5 m/year, on average, has been observed during the last five years. Farmers are thus called upon to improve irrigation methods. Thus, localized or drip irrigation is adopted to allow rational use of water. The importance of this irrigation system is due to the fact that water is applied directly to the root zone and its compatibility with fertilization. However, this irrigation system faces a thorny problem which is the clogging of pipes and drippers. This leads to a lack of uniformity of irrigation over time. This so-called scaling phenomenon, the consequences of which are harmful (cleaning or replacement of pipes), leads to considerable unproductive expenditure. The objective set by this work is the search for green inhibitors likely to prevent this phenomenon of scaling. This study requires a better knowledge of these waters, their physico-chemical characteristics and their scaling power. Thus, using the "LCGE" controlled degassing technique, we initially evaluated, on pure calco-carbonic water at 30°F, the scaling-inhibiting power of some available plant extracts in our region of Souss-Massa. We then carried out a comparative study of the efficacy of these green inhibitors. The action of the most effective green inhibitor on real agricultural waters was then studied.

Keywords: green inhibitors, localized irrigation, plant extracts, scaling inhibition

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Authors: Ellana Boada, Eric Santos-Clotas, Alba Cabrera-Codony, Maria Martin, Lluis Baneras, Frederic Gich

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Volatile methylsiloxanes (VMS) are a group of manmade silicone compounds widely used in household and industrial applications that end up on the biogas produced through the anaerobic digestion of organic matter in landfills and wastewater treatment plants. The presence of VMS during the biogas energy conversion can cause damage on the engines, reducing the efficiency of this renewable energy source. Non regenerative adsorption onto activated carbon is the most widely used technology to remove siloxanes from biogas, while new trends point out that biotechnology offers a low-cost and environmentally friendly alternative to conventional technologies. The first objective of this research was to enrich, isolate and identify bacterial species able to grow using siloxane molecules as a sole carbon source: anoxic wastewater sludge was used as initial inoculum in liquid anoxic enrichments, adding D4 (as representative siloxane compound) previously adsorbed on activated carbon. After several months of acclimatization, liquid enrichments were plated onto solid media containing D4 and thirty-four bacterial isolates were obtained. 16S rRNA gene sequencing allowed the identification of strains belonging to the following species: Ciceribacter lividus, Alicycliphilus denitrificans, Pseudomonas aeruginosa and Pseudomonas citronellolis which are described to be capable to degrade toxic volatile organic compounds. Kinetic assays with 8 representative strains revealed higher cell growth in the presence of D4 compared to the control. Our second objective was to characterize the community composition and diversity of the microbial community present in the enrichments and to elucidate whether the isolated strains were representative members of the community or not. DNA samples were extracted, the 16S rRNA gene was amplified (515F & 806R primer pair), and the microbiome analyzed from sequences obtained with a MiSeq PE250 platform. Results showed that the retrieved isolates only represented a minor fraction of the microorganisms present in the enrichment samples, which were represented by Alpha, Beta, and Gamma proteobacteria as dominant groups in the category class thus suggesting that other microbial species and/or consortia may be important for D4 biodegradation. These results highlight the need of additional protocols for the isolation of relevant D4 degraders. Currently, we are developing molecular tools targeting key genes involved in siloxane biodegradation to identify and quantify the capacity of the isolates to metabolize D4 in batch cultures supplied with a synthetic gas stream of air containing 60 mg m⁻³ of D4 together with other volatile organic compounds found in the biogas mixture (i.e. toluene, hexane and limonene). The isolates were used as inoculum in a biotrickling filter containing lava rocks and activated carbon to assess their capacity for siloxane removal. Preliminary results of biotrickling filter performance showed 35% of siloxane biodegradation in a contact time of 14 minutes, denoting that biological siloxane removal is a promising technology for biogas upgrading.

Keywords: bacterial cultivation, biogas upgrading, microbiome, siloxanes

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Authors: Vivek Bhat, Rohini Kelkar, Sanjay Biswas

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Introduction: Cancer patients are at increased risk of bacterial infections. This may due to the disease process itself, the effect of chemotherapeutic drugs or invasive procedures such as catheterization. A wide variety of bacteria including some emerging pathogens are increasingly being reported from these patients. The incidence of multidrug-resistant organisms particularly in the Gram negative group is also increasing, with higher resistance rates seen to cephalosporins, β-lactam/β-lactam inhibitor combinations, and the carbapenems. This study documents the bacteriological spectrum of infections and their resistance patterns in cancer patients. Methods: This study includes all bacterial isolates recovered from infections cancer patients over a period of 18 months. Samples included Blood cultures, Pus/wound swabs, urine, tissue biopsies, body fluids, catheter tips and respiratory specimens such as sputum and bronchoalveolar lavage (BAL). All samples were processed in the microbiology laboratory as per standard laboratory protocols. Organisms were identified to species level and antimicrobial susceptibility testing was performed manually by the disc diffusion technique or in the Vitek-2 (Biomereux, France) instrument. Interpretations were as per Clinical laboratory Standards Institute (CLSI) guidelines. Results: A total of 1150 bacterial isolates were cultured from 884 test samples during the study period. Of these 227 were Gram-positive and 923 were Gram-negative organisms. Staphylococcus aureus (99 isolates) was the commonest Gram-positive isolate followed by Enterococcus (79) and Gr A Streptococcus (30). Among the Gram negatives, E. coli (304), Pseudomonas aeruginosa (201) and Klebsiella pneumoniae (190) were the most common. Of the Staphylococcus aureus isolates 27.2% were methicillin resistant. Only 5.06% enterococci were vancomycin resistant. High rates of resistance to cefotaxime and ciprofloxacin were seen amongst E. coli (84.8% & 83.55%) and Klebsiella pneumoniae (71 & 62.1%) respectively. Resistance to carbapenems (meropenem) was high at 70% in Acinetobacter spp.; however all isolates were sensitive to colistin. Among the aminoglycosides, amikacin retained good efficacy against Escherichia coli (82.9%) and Pseudomonas aeruginosa (78.1%). Occasional isolates of emerging pathogens such as Chryseobacterium indologens, Roseomonas, and Achromobacter xyloxidans were also recovered. Conclusion: The common infections in cancer patients include respiratory, wound, tract infections and sepsis. The commonest isolates include Staphylococcus aureus, Enterococci, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. There is a high level of resistance to the commonly used antibiotics among Gram-negative organisms.

Keywords: bacteria, resistance, infection, cancer

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Authors: M. Marjaneh, M. Y. Narazah, H. Shahrul

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Array-based gene expression analysis is a powerful tool to profile expression of genes and to generate information on therapeutic effects of new anti-cancer compounds. Anti-apoptotic effect of thymoquinone was studied in MCF7 breast cancer cell line using gene expression profiling with cDNA micro array. The purity and yield of RNA samples were determined using RNeasyPlus Mini kit. The Agilent RNA 6000 Nano LabChip kit evaluated the quantity of the RNA samples. AffinityScript RT oligo-dT promoter primer was used to generate cDNA strands. T7 RNA polymerase was used to convert cDNA to cRNA. The cRNA samples and human universal reference RNA were labelled with Cy-3-CTP and Cy-5-CTP, respectively. Feature Extraction and GeneSpring software analysed the data. The single experiment analysis revealed involvement of 64 pathways with up-regulated genes and 78 pathways with down-regulated genes. The MAPK and p38-MAPK pathways were inhibited due to the up-regulation of PTPRR gene. The inhibition of p38-MAPK suggested up-regulation of TGF-ß pathway. Inhibition of p38 - MAPK caused up-regulation of TP53 and down-regulation of Bcl2 genes indicating involvement of intrinsic apoptotic pathway. Down-regulation of CARD16 gene as an adaptor molecule regulated CASP1 and suggested necrosis-like programmed cell death and involvement of caspase in apoptosis. Furthermore, down-regulation of GPCR, EGF-EGFR signalling pathways suggested reduction of ER. Involvement of AhR pathway which control cytochrome P450 and glucuronidation pathways showed metabolism of Thymoquinone. The findings showed differential expression of several genes in apoptosis pathways with thymoquinone treatment in estrogen receptor-positive breast cancer cells.

Keywords: cDNA microarray, thymoquinone, CARD16, PTPRR, CASP10

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Authors: Justine Garraud, Hervé Capiaux, Cécile Le Guern, Pierre Gaudin, Clémentine Lapie, Samuel Chaffron, Erwan Delage, Thierry Lebeau

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The repeated use of Bordeaux mixture (copper sulphate) and other chemical forms of copper (Cu) has led to its accumulation in wine-growing soils for more than a century, to the point of modifying the ecosystem of these soils. Phytoextraction of copper could progressively reduce the Cu load in these soils, and even to recycle copper (e.g. as a micronutrient in animal nutrition) by cultivating the extracting plants in the inter-row of the vineyards. Soil cleaning up usually requires several years because the chemical speciation of Cu in solution is mainly based on forms complexed with dissolved organic matter (DOM) that are not phytoavailable, unlike the "free" forms (Cu2+). Indeed, more than 98% of Cu in the solution is bound to DOM. The selection and inoculation of invineyardsoils in vineyard soils ofbacteria(bioaugmentation) able to degrade Cu-DOM complexes could increase the phytoavailable pool of Cu2+ in the soil solution (in addition to bacteria which first mobilize Cu in solution from the soil bearing phases) in order to increase phytoextraction performance. In this study, sevenCu-accumulating plants potentially usable in inter-row were tested for their Cu phytoextraction capacity in hydroponics (ray-grass, brown mustard, buckwheat, hemp, sunflower, oats, and chicory). Also, a bacterial consortium was tested: Pseudomonas sp. previously studied for its ability to mobilize Cu through the pyoverdine siderophore (complexing agent) and potentially to degrade Cu-DOM complexes, and a second bacterium (to be selected) able to promote the survival of Pseudomonas sp. following its inoculation in soil. Interaction network method was used based on the notions of co-occurrence and, therefore, of bacterial abundance found in the same soils. Bacteria from the EcoVitiSol project (Alsace, France) were targeted. The final step consisted of incoupling the bacterial consortium with the chosen plant in soil pots. The degradation of Cu-DOMcomplexes is measured on the basis of the absorption index at 254nm, which gives insight on the aromaticity of the DOM. The“free” Cu in solution (from the mobilization of Cu and/or the degradation of Cu-MOD complexes) is assessed by measuring pCu. Eventually, Cu accumulation in plants is measured by ICP-AES. The selection of the plant is currently being finalized. The interaction network method targeted the best positive interactions ofFlavobacterium sp. with Pseudomonassp. These bacteria are both PGPR (plant growth promoting rhizobacteria) with the ability to improve the plant growth and to mobilize Cu from the soil bearing phases (siderophores). Also, these bacteria are known to degrade phenolic groups, which are highly present in DOM. They could therefore contribute to the degradation of DOM-Cu. The results of the upcoming bacteria-plant coupling tests in pots will be also presented.

Keywords: complexes Cu-DOM, bioaugmentation, phytoavailability, phytoextraction

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Authors: Judit Perjessy, Zsolt Zalan, Ferenc Hegyi, Eniko Horvath-Szanics, Krisztina Takacs, Andras Nagy, Adel Klupacs, Erika Koppany-Szabo, Zhirong Wang, Kaituo Wang, Muying Du, Jianquan Kan

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The post-harvest diseases cause great economic losses in the fruit and vegetables; the prevention of these deterioration has great importance. Against the fungi, which cause most of the diseases, are extensively used the fungicides. However, there are increasing consumer concerns over the presence of pesticide residues in food. An alternative and in recent years, increasingly studied method for the prevention of the diseases is biocontrol, where antagonistic microorganisms are used for the control of fungi. The genera of Lactobacillus is well known and extensively studied, but its applicability as biocontrol agents in post-harvest preservation of fruit and vegetables is poorly investigated. However these bacteria can be found on the surface of the plants and have great antimicrobial activity. In our study we have investigated the chitinase activity, the antifungal effect and the applicability of several Lactobacillus strains to select potential biocontrol agents. We investigated the determination of the environmental parameters of a gene (encoding chitinase) expression and we also investigated the relationship between actual antifungal activity and potential chitinase activity. Mixed cultures were also developed to enhance the antifungal activity and determined the optimal mold spore and bacteria concentration ratio for the appropriate efficacy. Five Lactobacillus strains (L. acidophilus N2, L. delbrueckii subsp. bulgaricus B397, L. sp. 2231, L. sake subsp. sake 2471, L. buchneri 1145) possess chitinase-coding gene from the 43 investigated Lactobacillus strains. Proteins with similar molecular weight and separation properties like bacterial chitinases were detected from these strains, which also possess chitin-binding property. Nevertheless, they were inactive, lacks the chitinolytic activity. In point of the cumulative activity of inhibition, our results showed that certain strains were statistically significant in a positive direction compared to other strains, e.g., L. rhamnosus VT1 and L. Casey 154 have shown great general antifungal effect against 11 molds from the genera Penicillium and Botrytis and isolated from spoiled fruit and vegetables. Also, some mixed cultures (L. rhamnosus VT1 - L. Plantarum 299v) showed significant antifungal effects against the indigenous molds on the surface of apple fruit during the industrial storage experiment. Thus, they could be promising for post-harvest biopreservation.

Keywords: biocontrol, chitinase, Lactobacillus, post-harvest

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