Search results for: bacterial inoculum

Authors: Supanun Kangrang, Kraipat Cheenkachorn, Kittiphong Rattanaporn, Malinee Sriariyanun

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Rice straw is lignocellulosic biomass which can be utilized as substrate for the biogas production. However, due to the property and composition of rice straw, it is difficult to be degraded by hydrolysis enzymes. One of the pretreatment method that modifies such properties of lignocellulosic biomass is the application of lignocellulose-degrading microbial consortia. The aim of this study is to investigate the effect of microbial consortia to enhance biogas production. To select the high efficient consortium, cellulase enzymes were extracted and their activities were analyzed. The results suggested that microbial consortium culture obtained from cattle manure is the best candidate compared to decomposed wood and horse manure. A microbial consortium isolated from cattle manure was then mixed with anaerobic sludge and used as inoculum for biogas production. The optimal conditions for biogas production were investigated using response surface methodology (RSM). The tested parameters were the ratio of amount of microbial consortium isolated and amount of anaerobic sludge (MI:AS), substrate to inoculum ratio (S:I) and temperature. Here, the value of the regression coefficient R2 = 0.7661 could be explained by the model which is high to advocate the significance of the model. The highest cumulative biogas yield was 104.6 ml/g-rice straw at optimum ratio of MI:AS, ratio of S:I, and temperature of 2.5:1, 15:1 and 44°C respectively.

Keywords: lignocellulolytic biomass, microbial consortium, cellulase, biogas, Response Surface Methodology (RSM)

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Authors: Radka Obořilová, Hana Šimečková, Matěj Pastucha, Jan Přibyl, Petr Skládal, Ivana Mašlaňová, Zdeněk Farka

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Due to the spreading of antimicrobial resistance, alternative approaches to combat superinfections are being sought, both in the field of lysing agents and methods for studying bacterial lysis. A suitable alternative to antibiotics is phage therapy and enzybiotics, for which it is also necessary to study the mechanism of their action. Biosensor-based techniques allow rapid detection of pathogens in real time, verification of sensitivity to commonly used antimicrobial agents, and selection of suitable lysis agents. The detection of lysis takes place on the surface of the biosensor with immobilized bacteria, which has the potential to be used to study biofilms. An example of such a biosensor is surface plasmon resonance (SPR), which records the kinetics of bacterial lysis based on a change in the resonance angle. The bacteria are immobilized on the surface of the SPR chip, and the action of phage as the mass loss is monitored after a typical lytic cycle delay. Atomic force microscopy (AFM) is a technique for imaging of samples on the surface. In contrast to electron microscopy, it has the advantage of real-time imaging in the native conditions of the nutrient medium. In our case, Staphylococcus aureus was lysed using the enzyme lysostaphin and phage P68 from the familyPodoviridae at 37 ° C. In addition to visualization, AFM was used to study changes in mechanical properties during lysis, which resulted in a reduction of Young’s modulus (E) after disruption of the bacterial wall. Changes in E reflect the stiffness of the bacterium. These advanced methods provide deeper insight into bacterial lysis and can help to fight against bacterial diseases.

Keywords: biosensors, atomic force microscopy, surface plasmon resonance, bacterial lysis, staphylococcus aureus, phage P68

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Authors: Bernhard Widhalm, Cornelia Rieder-Gradinger, Ewald Srebotnik

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Pine wood (Pinus sylvestris L.) is the basic raw material for the production of Oriented Strand Boards (OSB) and the major source of volatile organic compounds, especially terpenes (like α- and β-pinene). To lower the total emission level of OSB, terpene metabolising microorganisms were therefore applied onto pine wood strands for the production of emission-reduced boards. Suitable microorganisms were identified during preliminary tests under laboratory conditions. At first, their terpene degrading potential was investigated in liquid culture, followed by laboratory tests using unsterile pine wood particles and strands. The main focus was laid on an adoptable terpene reduction in a short incubation time. An optimised bacterial mixture of Pseudomonas putida and Pseudomonas fluorescens showed the best results and was therefore used for further experiments on a larger scale. In an industry-compatible testing procedure, pine wood strands were incubated with the bacterial mixture for a period of 2 to 4 days. Incubation time was stopped by drying the strands. OSB were then manufactured from the pre-treated strands and emissions were measured by means of SPME/GC-MS analysis. Bacterial pre-treatment of strands resulted in a reduction of α-pinene- and β-pinene-emissions from OSB by 40% and 70%, respectively, even after only 2 days of incubation. The results of the investigation provide a basis for the application of microbial treatment within the industrial OSB production line, where shortest possible incubation times are required. For this purpose, the performance of the bacterial mixture will have to be further optimised.

Keywords: GC-MS, OSB, Pseudomonas sp., terpene degradation

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Authors: E. Osazee, L. U. Bashir

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This study was conducted at the Department of Biological Sciences, Usmanu Danfodiyo University, Sokoto, Nigeria, to determine the effects of different weights of cow dung on indigenous bacterial isolates in remediation of crude oil contaminated soils. The soil (1kg) was contaminated with 20g of crude oil and this was treated with three (40g, 80g and 120g) weights of cow dung. The soils were amended after two weeks of crude oil contamination. Soil samples were collected from the plastic bags for microbiological analyses. The isolates were cultured to test their ability to grow on crude oil. The ability of the isolates to utilize the crude oil was determined using media dilution technique. Bacteria such as Proteus mirabilis, Bacillus lacterosporus, Morganella morganii, Serratia marcescens and Bacillus alvei were isolated. The variables measured were heterotrophic bacterial populations, hydrocarbon utilizing bacterial populations and the percentage of crude oil degraded in the soils. Data collected were subjected to analysis of variance (ANOVA). Results obtained indicated that all the different weights of cow dung showed appreciable effect in crude oil decontamination. Based on the findings of the experiments, it could be deduced that 120g of cow dung promoted higher degradation of hydrocarbons. Thus, it should be recommended for remediation of crude oil contaminated soil in the study area.

Keywords: crude oil, cow dung, amendment, bioremediation, decontamination

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Authors: I. Kamika, S. Azizi, M. Tekere

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Nanotechnology promises significant improvements of advanced materials and manufacturing techniques with a vast range of applications, which are critical for the future competitiveness of national industries. The manipulations and productions of materials, whilst, controlling the optical properties and surface area to a nanosize scale enabled a birth of a new field known as nanotechnology. However, their rapidly developing industry raises concerns about the environmental impacts of nanoparticles, as their effects on functional bacterial community in wastewater treatment remain unclear. The present research assessed the impact of cerium Oxide nanoparticles (nCeO) on the bacterial microbiome of an activated sludge system, which influenced its performance of this system on nutrient removal. Out of 15875 reads sequenced, a total of 13133 reads were non-chimeric. The wastewater samples were more dominant to the unclassified bacteria (51.07% of bacteria community) followed with the classified bacteria (48.93). Proteobacteria was the most dominant phylum in both classified and unclassified bacteria, whereas 18% of bacteria could even not be assigned a phylum and remained unclassified suggesting hitherto vast untapped microbial diversity. The bacterial operational taxonomic units (OTUs) ranged from 1014 to 2629 over the experimental period. The denitrification related species including Diaphorobacter species, Thauera species and those in the Sphaerotilus and Leptothrix group were found to be inhibited in a high concentration of CeO-NP. The diversity indices suggested that the bacterial community inhabiting the wastewater samples were less diverse as the concentration of CeO increases. The canonical correspondence analysis (CCA) results highlighted that the bacterial community variance had the strongest relationship with water temperature, conductivity, pH, and dissolved oxygen (DO) content as well as nCeO. The results provided the relationships between the microbial community and environmental variables in the wastewater samples.

Keywords: bacterial community, next generation, cerium oxide, wastewater, activated sludge, nanoparticles, nanotechnology

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Authors: Ilknur Tuncer, Nihayet Bizsel

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The scarcity of research in bacterial diversity and antimicrobial resistance in coastal environments as in Turkish coasts leads to difficulties in developing efficient monitoring and management programs. In the present study, biogeochemical analysis of sediments and antimicrobial susceptibility analysis of bacteria in Izmir Bay, eastern Aegean Sea under high anthropogenic pressure were aimed in summer period when anthropogenic input was maximum and at intertidal zone where the first terrigenious contact occurred for aquatic environment. Geochemical content of the intertidal zone of Izmir Bay was firstly illustrated such that total and organic carbon, nitrogen and phosphorus contents were high and the grain size distribution varied as sand and gravel. Bacterial diversity and antibiotic resistance were also firstly given for Izmir Bay. Antimicrobially assayed isolates underlined the multiple resistance in the inner, middle and outer bays with overall 19% high MAR (multiple antibiotic resistance) index. Phylogenetic analysis of 16S rRNA gene sequences indicated that 67 % of isolates belonged to the genus Bacillus and the rest included the families Alteromonadaceae, Bacillaceae, Exiguobacteriaceae, Halomonadaceae, Planococcaceae, and Staphylococcaceae.

Keywords: bacterial phylogeny, multiple antibiotic resistance, 16S rRNA genes, Izmir Bay, Aegean Sea

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Authors: M. J. Norashirene, Y. Zakiah, S. Nurdiana, I. Nur Hilwani, M. H. Siti Khairiyah, M. J. Muhamad Arif

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In this study, six bacterial isolates of a slightly thermophilic organism from the Sg. Klah hot spring, Malaysia were successfully isolated and designated as M7T55D1, M7T55D2, M7T55D3, M7T53D1, M7T53D2 and M7T53D3 respectively. The bacterial isolates were screened for their cellulose hydrolytic ability on Carboxymethlycellulose agar medium. The isolated bacterial strains were identified morphologically, biochemically and molecularly with the aid of 16S rDNA sequencing. All of the bacteria showed their optimum growth at a slightly alkaline pH of 7.5 with a temperature of 55°C. All strains were Gram-negative, non-spore forming type, strictly aerobic, catalase-positive and oxidase-positive with the ability to produce thermostable cellulase. Based on BLASTn results, bacterial isolates of M7T55D2 and M7T53D1 gave the highest homology (97%) with similarity to Tepidimonas ignava while isolates M7T55D1, M7T55D3, M7T53D2 and M7T53D3 showed their closest homology (97%-98%) with Tepidimonas thermarum. These cellulolytic thermophiles might have a commercial potential to produce valuable thermostable cellulase.

Keywords: cellulase, cellulolytic, thermophiles, 16S rDNA gene

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Authors: Florin Leon, Silvia Curteanu

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The Bacterial Foraging Optimization (BFO) algorithm is inspired by the behavior of bacteria such as Escherichia coli or Myxococcus xanthus when searching for food, more precisely the chemotaxis behavior. Bacteria perceive chemical gradients in the environment, such as nutrients, and also other individual bacteria, and move toward or in the opposite direction to those signals. The application example considered as a case study consists in establishing the dependency between the reaction yield of hydrogels based on polyacrylamide and the working conditions such as time, temperature, monomer, initiator, crosslinking agent and inclusion polymer concentrations, as well as type of the polymer added. This process is modeled with a neural network which is included in an optimization procedure based on BFO. An experimental study of BFO parameters is performed. The results show that the algorithm is quite robust and can obtain good results for diverse combinations of parameter values.

Keywords: bacterial foraging, hydrogels, modeling and optimization, neural networks

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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

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Authors: Ilma Robo, Saimir Heta, Gerhard Nokaj

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Introduction: The method of treatment and the treatment protocols of apical periodontitis are now known, but the ongoing debate remains on whether or not prescription antibiotics should be given to patients suffering from this type of pathology. In fact, as an indication for prescribing antibiotics, this type of pathology remains between clinical indication and contraindication. Material and Methods: This article is of the short-communication type and has the sole purpose of analyzing the clinical picture of apical periodontitis and the fact that the appearance and extent of this pathology in the periapex area passes the stage when the host or the immune cells of the organism of the affected individual, react against bacterial factors. Results: Determining whether or not to prescribe systemic antibiotics according to literature sources can be avoided. In some cases, research in this field about this pathology even indicates endodontic rinsers or irrigants, such as chlorhexidine, in typical cases, mainly in persistent apical periodontitis. Conclusions: In times when bacterial resistance is a hot topic in some fields of scientific research, it is important to divide dental pathologies of bacterial origin into those when systemic antibiotic prescriptions must be given and those when every clinical issue is resolved only with endodontic root canal treatment. Even certain sources of published literature show the specifics of the most effective antibiotics against the bacterial flora causing the pathology of apical periodontitis.

Keywords: endodontic treatment, apical periodontitis, antibiotics, chlorhexidine

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Authors: B. D. Nkosi, T. F. Mutavhatsindi, J. J. Baloyi, R. Meeske, T. M. Langa, I. M. M. Malebana, M. D. Motiang

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Potato hash (PH), a by-product from food production industry, contains 188.4 g dry matter (DM)/kg and 3.4 g water soluble carbohydrate (WSC)/kg DM, and was mixed with wheat bran (70:30 as is basis) to provide 352 g DM/kg and 315 g WSC/kg DM. The materials were ensiled with or without silage additives in 1.5L anaerobic jars (3 jars/treatment) that were kept at 25-280 C for 3 months. Four types of silages were produced which were: control (no additive, denoted as T1), celluclast enzyme (denoted as T2), emsilage bacterial inoculant (denoted as T3) and silosolve bacterial inoculant (denoted as T4). Three jars per treatment were opened after 3 months of ensiling for the determination of nutritive values, fermentation characteristics and aerobic stability. Aerobic stability was done by exposing silage samples to air for 5 days. The addition of enzyme (T2) was reduced (P<0.05) silage pH, fiber fractions (NDF and ADF) while increasing (P < 0.05) residual WSC and lactic acid (LA) production, compared to other treatments. Silage produced had pH of < 4.0, indications of well-preserved silage. Bacterial inoculation (T3 and T4) improved (P < 0.05) aerobic stability of the silage, as indicated by increased number of hours and lower CO2 production, compared to other treatments. However, the aerobic stability of silage was worsen (P < 0.05) with the addition of an enzyme (T2). Further work to elucidate these effects on nutrient digestion and growth performance on ruminants fed the silage is needed.

Keywords: by-products, digestibility, feeds, inoculation, ruminants, silage

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Authors: Vanessa De Carvalho, Chad MacPherson, Julien Tremblay, Julie Champagne, Stephanie-Anne Girard

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Background: The interactions between bacteria and fungi in the human vaginal microbiome are fundamental to the concept of health and disease. The means by which the microbiota and mycobiota interact is still poorly understood and further studies are necessary to properly characterize this complex ecosystem. The aim of this study was to select a DNA extraction method capable of recovering high qualities of fungal and bacterial DNA from a single vaginal swab. Methods: 11 female volunteers ( ≥ 20 to < 55 years old) self-collected vaginal swabs in triplicates. Three commercial extraction kits: Masterpure Yeast Purification kit (Epicenter), PureLink™ Microbiome DNA Purification kit (Invitrogen), and Quick-DNA™ Fecal/Soil Microbe Miniprep kit (Zymo) were evaluated on the ability to recover fungal and bacterial DNA simultaneously. The extraction kits were compared on the basis of recovery, yield, purity, and the community richness of bacterial (16S rRNA - V3-V4 region) and fungal (ITS1) microbiota composition by Illumina MiSeq amplicon sequencing. Results: Recovery of bacterial DNA was achieved with all three kits while fungal DNA was only consistently recovered with Masterpure Yeast Purification kit (yield and purity). Overall, all kits displayed similar microbiota profiles for the top 20 OTUs; however, Quick-DNA™ Fecal/Soil Microbe Miniprep kit (Zymo) showed more species richness than the other two kits. Conclusion: In the present study, Masterpure Yeast purification kit proved to be a good candidate for purification of high quality fungal and bacterial DNA simultaneously. These findings have potential benefits that could be applied in future vaginal microbiome research. Whilst the use of a single extraction method would lessen the burden of multiple swab sampling, decrease laboratory workload and off-set costs associated with multiple DNA extractions, thoughtful consideration must be taken when selecting an extraction kit depending on the desired downstream application.

Keywords: bacterial vaginosis, DNA extraction, microbiota, mycobiota, vagina, vulvovaginal candidiasis, women’s health

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Authors: Sihame Amakrane, Zineb Ouahdi, Mohammed Salah, Said Belaaouad

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\This research explores the efficacy of novel pyrimidine derivatives on bacterial strains such as Escherichia coli, Staphylococcus aureus, and Myccobacterium tuberculosis, utilizing bending energy calculations. Of the 25 compounds examined, 13 displayed potent activity against all the bacterial strains under study, exhibiting bending energy measurements between -7.4 and -10.7 kcal/mol. The -7.4 kcal/mol value corresponds to the bending energy of the SA12 and SA13 compounds with the 2xct protein (Staphylococcus aureus), whereas the -10.7 kcal/molis linked with the bending energy of SA6 and SA11 compounds with the 6GAV protein (Myccobacterium tuberculosis). Further research will involve a QSAR (Quantitative Structure-Activity Relationship) study aimed at constructing a reliable model to combat the aforementioned bacterial strains and a molecular dynamics study to evaluate the stability of ligand-protein complexes.

Keywords: docking, QSAR, bending energy, e. coli

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Authors: Diana Larisa Vladoiu, Vasile Ostafe, Adriana Isvoran

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Molecular docking calculations reveal that pesticides provide favorable interactions with the bacterial chitinases. Pesticides interact with both hydrophilic and aromatic residues involved in the active site of the enzymes, their positions partially overlapping the substrate and the inhibitors locations. Molecular docking outcomes, in correlation with experimental literature data, suggest that the pesticides may be degraded or having an inhibitor effect on the activity of these enzymes, depending of the application dose and rate.

Keywords: chitinases, inhibition, molecular docking, pesticides

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Authors: H. Agbaria, A. Salman, M. Huleihel, G. Beck, D. H. Rich, S. Mordechai, J. Kapelushnik

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Viral and bacterial infections are responsible for variety of diseases. These infections have similar symptoms like fever, sneezing, inflammation, vomiting, diarrhea and fatigue. Thus, physicians may encounter difficulties in distinguishing between viral and bacterial infections based on these symptoms. Bacterial infections differ from viral infections in many other important respects regarding the response to various medications and the structure of the organisms. In many cases, it is difficult to know the origin of the infection. The physician orders a blood, urine test, or 'culture test' of tissue to diagnose the infection type when it is necessary. Using these methods, the time that elapses between the receipt of patient material and the presentation of the test results to the clinician is typically too long ( > 24 hours). This time is crucial in many cases for saving the life of the patient and for planning the right medical treatment. Thus, rapid identification of bacterial and viral infections in the lab is of great importance for effective treatment especially in cases of emergency. Blood was collected from 50 patients with confirmed viral infection and 50 with confirmed bacterial infection. White blood cells (WBCs) and plasma were isolated and deposited on a zinc selenide slide, dried and measured under a Fourier transform infrared (FTIR) microscope to obtain their infrared absorption spectra. The acquired spectra of WBCs and plasma were analyzed in order to differentiate between the two types of infections. In this study, the potential of FTIR microscopy in tandem with multivariate analysis was evaluated for the identification of the agent that causes the human infection. The method was used to identify the infectious agent type as either bacterial or viral, based on an analysis of the blood components [i.e., white blood cells (WBC) and plasma] using their infrared vibrational spectra. The time required for the analysis and evaluation after obtaining the blood sample was less than one hour. In the analysis, minute spectral differences in several bands of the FTIR spectra of WBCs were observed between groups of samples with viral and bacterial infections. By employing the techniques of feature extraction with linear discriminant analysis (LDA), a sensitivity of ~92 % and a specificity of ~86 % for an infection type diagnosis was achieved. The present preliminary study suggests that FTIR spectroscopy of WBCs is a potentially feasible and efficient tool for the diagnosis of the infection type.

Keywords: viral infection, bacterial infection, linear discriminant analysis, plasma, white blood cells, infrared spectroscopy

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Authors: M. A. El-Khateeb

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The growth of the olive oil production in Saudi Arabia peculiarly in Al Jouf region in recent years has been accompanied by an increase in the discharge of associated processing waste. Olive mill waste is produced throughout the extraction of oil from the olive fruit using the traditional mill and press process. Deterioration of the environment due to olive mill disposal wastes is a serious problem. When olive mill waste disposed into the soil, it affects soil quality, soil micro flora, and also toxic to plants. The aim of this work is to isolate microorganism (bacterial or fungal strains) from OMW capable of degrading phenols. Olive mill wastewater, olive mill waste and soil (beside oil production mill) contaminated with olive waste were used for isolation of phenol tolerant microorganisms. Four strains (two fungal and two bacterial) were isolated from olive mill waste. The isolated strains were Candida tropicalis and Phanerochaete chrysosporium (fungal strains) and Bacillus sp. and Rhodococcus sp. (bacterial strains). These strains were able to degrade phenols and could be used for bioremediation of olive mill waste.

Keywords: bioremediation, bacteria, fungi, Sakaka

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Authors: Thalía Moreno-García Malo, Santiago Torres-Ríos, María G. González-Cruz, María M. Hernández-Arroyo, Sergio R. Trejo-Estrada

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Agave atrovirens is the main source of agave sap, the raw material for the production of pulque, an artisanal fermented beverage, traditional since prehispanic times in the highlands of central Mexico. Agave sap is rich in glucose, sucrose and fructooligosaccharides, and strongly differs from agave syrup from A. tequilana, which is mostly a high molecular weight fructan. Agave sap is converted into pulque by a highly diverse microbial community which includes bacteria, yeast and even filamentous fungi. The bacterial diversity has been recently studied. But the composition of consortia derived from directed enrichments differs sharply from the whole fermentative consortium. Using classical microbiology methods, and selective liquid and solid media formulations, either bacterial or fungal consortia were developed and analyzed. Bacterial consortia able to catabolize specific prebiotic saccharides were selected and preserved for future developments. Different media formulations, selective for bacterial genera such as Bifidobacterium, Lactobacillus, Pediococcus, Lactococcus and Enterococcus were also used. For yeast, specific media, osmotic pressure and unique carbon sources were used as selective agents. Results show that most groups are represented in the enrichment cultures; although very few are recoverable from the whole consortium in artisanal pulque. Diversity and abundance vary among consortia. Potential bacterial probiotics obtained from agave sap and agave juices show tolerance to hydrochloric acid, as well as strong antimicrobial activity.

Keywords: Agave, pulque, microbial consortia, prebiotic activity

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Authors: Sang Guen Kim, Sib Sankar Giri, Jin Woo Jun, Saekil Yun, Hyoun Joong Kim, Sang Wha Kim, Jung Woo Kang, Se Jin Han, Se Chang Park

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Emerging of the super bacteria, bacteriophages are considered to be as an alternative to antibiotics. As the demand of phage increased, economical and large production of phage is becoming one of the critical points. For the therapeutic use, what is important is to eradicate the pathogenic bacteria as fast as possible, so higher concentration of phages is generally needed for effective therapeutic function. On the contrary, for the maximum production, bacteria work as a phage producing factory. As a microbial cell factory, bacteria is needed to last longer producing the phages without eradication. Consequently, killing the bacteria fast has a negative effect on large production. In this study, Multiplicity of Infection (MOI) was manipulated based on initial bacterial inoculation and used phage pAh-6C which has therapeutic effect against Aeromonas hydrophila. 1, 5 and 10 percent of overnight bacterial culture was inoculated and each bacterial culture was co-cultured with the phage of which MOI of 0.01, 0.0001, and 0.000001 respectively. Simply changing the initial MOI as well as bacterial inoculation concentration has regulated the production quantity of the phage without any other changes to culture conditions. It is anticipated that this result can be used as a foundational data for mass production of lytic bacteriophages which can be used as the therapeutic bio-control agent.

Keywords: bacteriophage, multiplicity of infection, optimization, Aeromonas hydrophila

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Authors: Ghazala Yaqub, Zubi Sadiq, Almas Hamid, Saira Iqbal

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This paper is the very first report of three dual action hybrids synthesized by covalent linkage of carbazole based novel antibacterial compounds with efflux pump inhibitors i.e., indole acetic acid/gallic acid. Novel carbazole based antibacterial compounds were prepared first and then these were covalently linked with efflux pump inhibitors which leads to the successful formation of hybrids. All prepared compounds were evaluated for their bacterial cell killing capability against Escherichia coli, Staphylococcus aureus, Pasteurella multocida and Bacillus subtilis. Compound were effective against all tested bacterial strains at different concentrations. But when these compounds were linked with efflux pump inhibitors they showed dramatic enhancement in their bacterial cell killing potential and minimum inhibitory concentration of all hybrids ranges from 7.250 µg/mL to 0.0283 µg/mL.

Keywords: antimicrobial assay, carbazole, dual action hybrids, efflux pump inhibitors

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Authors: Satish Babu Rajulapati

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The removal of toxic and heavy metal contaminants from wastewater streams and industrial effluents is one of the most important environmental issues being faced world over. In the present study three bacterial cultures tolerating high concentrations of chromium were isolated from the soil and wastewater sample collected from the tanneries located in Warangal, Telangana state. The bacterial species were identified as Bacillus sp., Staphylococcus sp. and pseudomonas sp. Preliminary studies were carried out with the three bacterial species at various operating parameters such as pH and temperature. The results indicate that pseudomonas sp. is the efficient one in the uptake of Cr(VI). Further, detailed investigation of Pseudomonas sp. have been carried out to determine the efficiency of removal of Cr(VI). The various parameters influencing the biosorption of Cr(VI) such as pH, temperature, initial chromium concentration, innoculum size and incubation time have been studied. Response Surface Methodology (RSM) was applied to optimize the removal of Cr(VI). Maximum Cr(VI) removal was found to be 85.72% Cr(VI) atpH 7, temperature 35 °C, initial concentration 67mg/l, inoculums size 9 %(v/v) and time 60 hrs.

Keywords: Staphylococcus sp, chromium, RSM, optimization, Cr(IV)

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Authors: Luz Stella Cadavid-Rodriguez, Viviana E. Castro-Lopez

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Fish waste (FW) has a high content of potentially biodegradable components, so it is amenable to be digested anaerobically. In this line, anaerobic digestion (AD) of FW has been studied for biogas production. Nevertheless, intermediate products such as volatile fatty acids (VFA), generated during the acidogenic stage, have been scarce investigated, even though they have a high potential as a renewable source of carbon. In the literature, there are few studies about the Inoculum-Substrate (I/S) ratio on acidogenesis. On the other hand, it is well known that pH is a critical factor in the production of VFA. The optimum pH for the production of VFA seems to change depending on the substrate and can vary in a range between 5.25 and 11. Nonetheless, the literature about VFA production from protein-rich waste, such as FW, is scarce. In this context, it is necessary to deepen on the determination of the optimal operating conditions of acidogenic fermentation for VFA production from protein-rich waste. Therefore, the aim of this research was to optimize the volatile fatty acid production from artisanal fishing waste, studying the effect of pH and the I/S ratio on the acidogenic process. For this research, the inoculum used was a methanogenic sludge (MS) obtained from a UASB reactor treating wastewater of a slaughterhouse plant, and the FW was collected in the port of Tumaco (Colombia) from the local artisanal fishers. The acidogenic fermentation experiments were conducted in batch mode, in 500 mL glass bottles as anaerobic reactors, equipped with rubber stoppers provided with a valve to release biogas. The effective volume used was 300 mL. The experiments were carried out for 15 days at a mesophilic temperature of 37± 2 °C and constant agitation of 200 rpm. The effect of 3 pH levels: 5, 7, 9, coupled with five I/S ratios, corresponding to 0.20, 0.15, 0.10, 0.05, 0.00 was evaluated taking as a response variable the production of VFA. A complete randomized block design was selected for the experiments in a 5x3 factorial arrangement, with two repetitions per treatment. At the beginning and during the process, pH in the experimental reactors was adjusted to the corresponding values of 5, 7, and 9 using 1M NaOH or 1M H2SO4, as was appropriated. In addition, once the optimum I/S ratio was determined, the process was evaluated at this condition without pH control. The results indicated that pH is the main factor in the production of VFA, obtaining the highest concentration with neutral pH. By reducing the I/S ratio, as low as 0.05, it was possible to maximize VFA production. Thus, the optimum conditions found were natural pH (6.6-7.7) and I/S ratio of 0.05, with which it was possible to reach a maximum total VFA concentration of 70.3 g Ac/L, whose major components were acetic acid (35%) and butyric acid (32%). The findings showed that the acidogenic fermentation of FW is an efficient way of producing VFA and that the operating conditions can be simple and economical.

Keywords: acidogenesis, artisanal fishing waste, inoculum to substrate ratio, volatile fatty acids

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Authors: E. Abustam, M. Yusuf, H. M. Ali, M. I. Said, F. N. Yuliati

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The aim of this study was to improve the durability and quality of Bali beef (M. Longissimus dorsi) and broiler carcass through the addition of liquid smoke as a natural preservative. This study was using Longissimus dorsi muscle from male Bali beef aged 3 years, broiler breast and thigh aged 40 days. Three types of meat were marinated in liquid smoke with concentrations of 0, 5, and 10% for 30 minutes at the level of 20% of the sample weight (w/w). The samples were storage at 2-5°C for 1 month. This study designed as a factorial experiment 3 x 3 x 4 based on a completely randomized design with 5 replications; the first factor was meat type (beef, chicken breast and chicken thigh); the 2nd factor was liquid smoke concentrations (0, 5, and 10%), and the 3rd factor was storage duration (1, 2, 3, and 4 weeks). Parameters measured were TBA value, total bacterial colonies, water holding capacity (WHC), shear force value both before and after cooking (80°C – 15min.), and cooking loss. The results showed that the type of meat produced WHC, shear force value, cooking loss and TBA differed between the three types of meat. Higher concentration of liquid smoke, the WHC, shear force value, TBA, and total bacterial colonies were decreased; at a concentration of 10% of liquid smoke, the total bacterial colonies decreased by 57.3% from untreated with liquid smoke. Longer storage, the total bacterial colonies and WHC were increased, while the shear force value and cooking loss were decreased. It can be concluded that a 10% concentration of liquid smoke was able to maintain fat oxidation and bacterial growth in Bali beef and chicken breast and thigh.

Keywords: Bali beef, chicken meat, liquid smoke, meat quality

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

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

Keywords: abattoir, aerosol, bacterial composition, environment

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Authors: Gizem Buldum, Alexander Bismarck, Athanasios Mantalaris

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Cellulose is the most abundant biopolymer on earth and it is currently being utilised in a multitude of industrial applications. Over the last 30 years, attention has been paid to the bacterial cellulose (BC), since BC exhibits unique physical, chemical and mechanical properties when compared to plant-based cellulose, including high purity and biocompatibility. Although Acetobacter xylinum is the most efficient producer of BC, it’s long doubling time results in insufficient yields of the cellulose production. This limits widespread and continued use of BC. In this study, E. coli BL21 (DE3) or E. coli HMS cells are selected as host organisms for the expression of bacterial cellulose synthase operon (bcs) of A.xylinum. The expression system is created based on pET-Duet1 and pCDF plasmid vectors, which carry bcs operon. The results showed that all bcs genes were successfully transferred and expressed in E.coli strains. The expressions of bcs proteins were shown by SDS and Native page analyses. The functionality of the bcs operon was proved by congo red binding assay. The effect of culturing temperature and the inducer concentration (IPTG) on cell growth and plasmid stability were monitored. The percentage of plasmid harboring cells induced with 0.025 mM IPTG was obtained as 85% at 22˚C in the end of 10-hr culturing period. It was confirmed that the high output cellulose production machinery of A.xylinum can be transferred into other organisms.

Keywords: bacterial cellulose, biopolymer, recombinant expression system, production

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Authors: Hardik Goswami, Gayatri Swarnakar

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-Mastitis disease has been known as one of the most costly diseases of dairy cattle and observed as an inflammatory disease of cow and buffalo udder. Mastitis badly affected animal health, quality of milk and economics of milk production along with cause’s great economic loss. Bacteria have been representing the most common etiological agents of mastitis. The antibiotic sensitivity test was important to attain accurate treatment of mastitis. The aim of present research work was to explore prevalence and antibiotic susceptibility pattern of bacterial isolates recovered from cow and buffalo clinical mastitis milk sample. During the period of April 2010 to April 2014, total 1487 clinical mastitis milk samples of cow and buffalo were tested to check the prevalence of mastitis causing bacterial isolates. Milk samples were collected aseptically from the udder at the time of morning milking. The most prevalent bacterial isolates were Staphylococcus aureus (24.34%) followed by coliform bacteria (15.87%), coagulase negative Staphylococcus aureus (13.85%), non-coliform bacteria (13.05%), mixed infection (12.51%), Streptococcus spp. (10.96%). Out of 1487, 140 (9.42%) mastitis milk samples showed no growth on culture media. Identification of bacteria made on the basis of Standard Microbial features and procedures. Antibiotic susceptibility of bacterial isolates was investigated by Kirby-Bauer disk diffusion method. In vitro Antibiotic susceptibility test of bacterial isolates revealed higher sensitivity to Gentamicin (74.6%), Ciprofloxacin (62.1%) and Amikacin (59.4%). The lower susceptibility was shown to Amoxicillin (21.6%), Erythromycin (26.4%) and Ceftizoxime (29.9%). Antibiotic sensitivity pattern revealed Gentamicin are the possible effective antibiotic against the major prevalent mastitis pathogens. Present research work would be helpful in increase production, quality and quantity of milk, increase annual income of dairy owners and improve health of cow and buffaloes.

Keywords: antibiotic, buffalo, cow, mastitis, prevalence

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Authors: Aldi Nel, Cliff L. W. Jones, Justin O. G. Kemp, Peter J. Britz

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Kelp Ecklonia maxima is included in formulated abalone feeds in South Africa, but its effect on abalone growth, feed utilisation efficiency and gut-bacterial communities has not previously been investigated. An eight-month on-farm growth trial with sub-adult Haliotis midae (~43 mm shell length) fed graded levels of kelp in formulated feeds was conducted. Kelp inclusion (0.44–3.54 % of pellet dry mass) promoted faster growth (65.7 – 74.5 % total mass gain), with better feed and protein conversions (FCR: 1.4 – 1.8; PER 2.3 – 2.7), compared to abalone fed the non-supplemented feed (52.3% total mass gain; FCR: 2.1; PER 1.9; p < 0.001). The gut-bacterial communities of abalone fed kelp-supplemented feed (0.88 % of pellet dry mass) were subsequently compared with that of abalone fed a non-supplemented control diet. Abalone gut-bacterial DNA was sequenced using 16S rRNA pyrosequencing and sequences were clustered into operational taxonomic units (OTUs) at a 97 % similarity level. A supplementary 16S rRNA denaturing gradient gel electrophoresis (DGGE) analysis was conducted. The dominant OTUs differed in terms of their relative abundances, with that of an autochthonous Mollicutes strain being significantly higher (p = 0.03) in the guts of abalone fed kelp-supplemented feed. The DGGE band patterns displayed a higher within-group variability of dominant bacterial strains for abalone fed the control diet, suggesting that dietary inclusion of kelp, which is rich in fermentable polysaccharides, promotes a balanced gut-bacterial community. This may contribute to the better feed utilisation and growth in abalone fed kelp-supplemented feeds.

Keywords: abfeed, digestion, macroalgae, mariculture

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Authors: Islam Zeb

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Water of poor quality has a potential of probable contamination and a way to spread pollutant in the field and surrounding environment. A number of comprehensive reviews articles have been published which highlight irrigation water as a source of pathogenic microorganisms and heavy metals toxicity that leads to chronic diseases in human. Here a study was plan to determine the microbial status of irrigation water collected from various location of district Swat in various months. The analyses were carried out at Environmental Horticulture Laboratory, Department of Horticulture, The University of Agriculture Peshawar, during the year 2018 – 19. The experiment was laid out in Randomized Complete Block Design (RCBD) with two factors and three replicates. Factor A consist of different locations, and factor B represent various months. The results of microbial status for various locations in irrigation water showed the highest value for Total Bacterial Count, Enterobacteriacea, E. coli, Salmonella, and Listeria (9.05, 8.54, 6.01, 5.84, and 5.03 log cfu L-1 respectively) for samples collected from mingora location, whereas the lowest values for Total Bacterial Count, Enterobacteriacea, E. coli, Salmonella and Listeria (6.70, 6.38, 4.47, 4.42 and 3.77 log cfu L-1 respectively) were observed for matta location. Data for various months showed maximum Total Bacterial Count, Enterobacteriacea, E. coli, Salmonella, and Listeria (12.01, 11.70, 8.46, 8.41, and 6.88 log cfu L-1, respectively) were noted for the irrigation water samples collected in May/June whereas the lowest range for Total Bacterial Count, Enterobacteriacea, E. coli, Salmonella and Listeria (4.41, 4.08, 2.61, 2.55 and 3.39 log cfu L-1 respectively) were observed in Jan/Feb. A significant interaction was found for all the studied parameters it was concluded that maximum bacterial groups were recorded in the months of May/June from Mingora location, it might be due to favorable weather condition.

Keywords: contamination, irrigation water, microbes, SWAT, various months

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

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

Keywords: admixture, organisms, porosity, strength

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Authors: H. Mayton, X. Yan, A. G. Taylor

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Infested tomato seeds were used to investigate the influence of Xanthomonas euvesicatoria on germination and seed to seedling transmission in a controlled environment and greenhouse assays in an effort to develop effective seed treatments and characterize seed borne transmission of bacterial leaf spot of tomato. Bacterial leaf spot of tomato, caused by four distinct Xanthomonas species, X. euvesicatoria, X. gardneri, X. perforans, and X. vesicatoria, is a serious disease worldwide. In the United States, disease prevention is expensive for commercial growers in warm, humid regions of the country, and crop losses can be devastating. In this study, four different infested tomato seed lots were extracted from tomato fruits infected with bacterial leaf spot from a field in New York State in 2017 that had been inoculated with X. euvesicatoria. In addition, vacuum infiltration at 61 kilopascals for 1, 5, 10, and 15 minutes and seed soaking for 5, 10, 15, and 30 minutes with different bacterial concentrations were used to artificially infest seed in the laboratory. For controlled environment assays, infested tomato seeds from the field and laboratory were placed othe n moistened blue blotter in square plastic boxes (10 cm x 10 cm) and incubated at 20/30 ˚C with an 8/16 hour light cycle, respectively. Infested tomato seeds from the field and laboratory were also planted in small plastic trays in soil (peat-lite medium) and placed in the greenhouse with 24/18 ˚C day and night temperatures, respectively, with a 14-hour photoperiod. Seed germination was assessed after eight days in the laboratory and 14 days in the greenhouse. Polymerase chain reaction (PCR) using the hrpB7 primers (RST65 [5’- GTCGTCGTTACGGCAAGGTGGTG-3’] and RST69 [5’-TCGCCCAGCGTCATCAGGCCATC-3’]) was performed to confirm presence or absence of the bacterial pathogen in seed lots collected from the field and in germinating seedlings in all experiments. For infested seed lots from the field, germination was lowest (84%) in the seed lot with the highest level of bacterial infestation (55%) and ranged from 84-98%. No adverse effect on germination was observed from artificially infested seeds for any bacterial concentration and method of infiltration when compared to a non-infested control. Germination in laboratory assays for artificially infested seeds ranged from 82-100%. In controlled environment assays, 2.5 % were PCR positive for the pathogen, and in the greenhouse assays, no infected seedlings were detected. From these experiments, X. euvesicatoria does not appear to adversely influence germination. The lowest rate of germination from field collected seed may be due to contamination with multiple pathogens and saprophytic organisms as no effect of artificial bacterial seed infestation in the laboratory on germination was observed. No evidence of systemic movement from seed to seedling was observed in the greenhouse assays; however, in the controlled environment assays, some seedlings were PCR positive. Additional experiments are underway with green fluorescent protein-expressing isolates to further characterize seed to seedling transmission of the bacterial leaf spot pathogen in tomato.

Keywords: bacterial leaf spot, seed germination, tomato, Xanthomonas euvesicatoria

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Authors: Sesethu G. Njokweni, Marelize Botes, Emile W. H. Van Zyl

Abstract:

An alternative strategy to the production of bioethanol is by examining the degradability of biomass in a natural system such as the rumen of mammals. This anaerobic microbial community has higher cellulolytic activities than microbial communities from other habitats and degrades cellulose to produce volatile fatty acids (VFA), methane and CO₂. VFAs have the potential to serve as intermediate products for electrochemical conversion to hydrocarbon fuels. In vitro mimicking of this process would be more cost-effective than bioethanol production as it does not require chemical pre-treatment of biomass, a sterile environment or added enzymes. The strategies of the carboxylate platform and the co-cultures of a bovine ruminal microbiota from cannulated cows were combined in order to investigate and optimize the bioconversion of agricultural biomass (apple and grape pomace, citrus pulp, sugarcane bagasse and triticale straw) to high value VFAs as intermediates for biofuel production in a consolidated bioprocess. Optimisation of reactor conditions was investigated using five different ruminal inoculum concentrations; 5,10,15,20 and 25% with fixed pH at 6.8 and temperature at 39 ˚C. The ANKOM 200/220 fiber analyser was used to analyse in vitro neutral detergent fiber (NDF) disappearance of the feedstuffs. Fresh and cryo-frozen (5% DMSO and 50% glycerol for 3 months) rumen cultures were tested for the retainment of fermentation capacity and durability in 72 h fermentations in 125 ml serum vials using a FURO medical solutions 6-valve gas manifold to induce anaerobic conditions. Fermentation of apple pomace, triticale straw, and grape pomace showed no significant difference (P > 0.05) in the effect of 15 and 20 % inoculum concentrations for the total VFA yield. However, high performance liquid chromatographic separation within the two inoculum concentrations showed a significant difference (P < 0.05) in acetic acid yield, with 20% inoculum concentration being the optimum at 4.67 g/l. NDF disappearance of 85% in 96 h and total VFA yield of 11.5 g/l in 72 h (A/P ratio = 2.04) for apple pomace entailed that it was the optimal feedstuff for this process. The NDF disappearance and VFA yield of DMSO (82% NDF disappearance and 10.6 g/l VFA) and glycerol (90% NDF disappearance and 11.6 g/l VFA) stored rumen also showed significantly similar degradability of apple pomace with lack of treatment effect differences compared to a fresh rumen control (P > 0.05). The lack of treatment effects was a positive sign in indicating that there was no difference between the stored samples and the fresh rumen control. Retaining of the fermentation capacity within the preserved cultures suggests that its metabolic characteristics were preserved due to resilience and redundancy of the rumen culture. The amount of degradability and VFA yield within a short span was similar to other carboxylate platforms that have longer run times. This study shows that by virtue of faster rates and high extent of degradability, small scale alternatives to bioethanol such as rumen microbiomes and other natural fermenting microbiomes can be employed to enhance the feasibility of biofuels large-scale implementation.

Keywords: agricultural wastes, carboxylate platform, rumen microbiome, volatile fatty acids

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