Search results for: Escherichia coli bacteria

Authors: V. Baweja, K. K. Gupta, V. Dubey, C. Keshavam

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Antimicrobial activity of six indigenous plants such as Tulsi Ocimum sanctum, Neem Azadirachta indica, Aloe vera, Turmeric Curcuma longa, Lantana Lantana camara, and Clove Syzygium aromaticum was assessed against the gut microbiota of the dengue fever mosquito Aedes aegypti, keeping in view that the presence of midgut bacteria may affect the ability of the vector to transmit pathogens. Eleven different types of bacterial clones were isolated from the midgut of lab-reared fourth instar larvae of Aedes aegypti and were grown on LB agar medium at an optimum temperature of 25 ºC. Identification of these bacteria was done on the basis of their colony characteristic such as colony size, shape, opacity, elevation, consistency, and growth. Light microscopic studies of the gut microbiota revealed dominance of Gram-negative cocci over gram positive cocci and bacilli and Gram-negative bacilli. Identification of species was done by chemical characterization of the colonies. Crude extracts of all test plants were screened for their antimicrobial activities against gut microbiota by disc diffusion assay. The zone of exclusion seen after 24 hr of incubation in different assays revealed the most potent antibacterial activities in neem followed by clove and turmeric. Lantana and Aloe vera were least effective.

Keywords: plant extract, aedes, dengue, antimicrobial activity

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Authors: Parmjit S. Panesar, Rupinder Kaur, Ram S. Singh

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Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hydrolyzed milk and transgalactosylation reaction for the synthesis of prebiotics such as lactulose and galactooligosaccharides. These prebiotics have various nutritional and technological benefits. Although, the enzyme is naturally present in almonds, peaches, apricots and other variety of fruits and animals, the extraction of enzyme from these sources increases the cost of enzyme. Therefore, focus has been shifted towards the production of low cost enzyme from the microorganisms such as bacteria, yeast and fungi. As compared to yeast and bacteria, fungal β-galactosidase is generally preferred as being extracellular and thermostable in nature. Keeping the above in view, the present study was carried out for the isolation of the β-galactosidase producing fungal strain from the food as well as the agricultural wastes. A total of more than 100 fungal cultures were examined for their potential in enzyme production. All the fungal strains were screened using X-gal and IPTG as inducers in the modified Czapek Dox Agar medium. Among the various isolated fungal strains, the strain exhibiting the highest enzyme activity was chosen for further phenotypic and genotypic characterization. The strain was identified as Rhizomucor pusillus on the basis of 5.8s RNA gene sequencing data.

Keywords: beta-galactosidase, enzyme, fungal, isolation

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Authors: Virve Pääkkönen, Heidi M. Cuffaro, Leo Tjäderhane

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Objectives: Odontoblasts are the outermost cell layer of dental pulp and form the dentin. Importance of bacterial products, e.g. lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria and lipopolysaccharide (LPS), a cell wall component of Gram-negative bacteria, have been indicated in the pathogenesis of pulpitis. Gram-positive bacteria are more prevalent in superficial carious lesion while the amount gram-negative is higher in the deep lesions. Objective of this study was to investigate the effect of these bacterial products on inflammatory response of pulp tissue. Interleukins (IL) were of special interest. Various ILs have been observed in the dentin-pulp complex of carious tooth in vivo. Methods: Tissue culture method was used for testing the effect of LTA and LPS on human odontoblasts. Enzymatic isolation technique was used to extract living odontoblasts for cell cultures. DNA microarray and quantitative PCR (qPCR) were used to characterize the changes in the expression profile of the tissue cultured odontoblasts. Laser microdissection was used to cut healthy and affected dentin and odontoblast layer directly under carious lesion for experiments. Cytokine array detecting 80 inflammatory cytokines was used to analyze the protein content of conditioned culture media as well as dentin and odontoblasts from the carious teeth. Results: LPS caused increased gene expression IL-1α, and -8 and decrease of IL-1β, 12 , -15 and -16 after 1h treatment, while after 24h treatment decrease of IL-8, -11 and 23 mRNAs was observed. LTA treatment caused cell death in the tissue cultured odontoblasts but in in the cell culture but not in cell culture. Cytokine array revealed at least 2-fold down-regulation of IL-1β, -10 and -12 in response to LPS treatment. Cytokine array of odontoblasts of carious teeth, as well as LPS-treated tissue-cultured odontoblasts, revealed increased protein amounts of IL-16, epidermal growth factor (EGF), angiogenin and IGFBP-1 as well as decreased amount of fractalkine. In carious dentin, increased amount of IL-1β, EGF and fractalkine was observed, as well as decreased level of GRO-1 and HGF. Conclusion: LPS caused marked changes in the expression of inflammatory cytokines in odontoblasts. Similar changes were observed in the odontoblasts cut directly under the carious lesion. These results help to shed light on the inflammatory processes happening during caries.

Keywords: inflammation, interleukin, lipoteichoic acid, odontoblasts

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Authors: Allan Kiptanui Kimisto, Geoffrey Odhiambo Ongondo, Anastasia Wairimu Muia, Cyrus Ndungu Kimani

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The major challenge to proper sewage sludge treatment processes is the poor understanding of sludge microbiome diversities. This study applied the whole-genome. shotgun metagenomics technique to profile the microbial composition of sewage sludge in two active digestion lagoons at the Nyeri-Kangemi Wastewater Treatment Plant in Nyeri County, Kenya. Total microbial community DNA was extracted from samples using the available ZymoBIOMICS™ DNA Miniprep Kit and sequenced using Shotgun metagenomics. Samples were analyzed using MG-RAST software (Project ID: mgp100988), which allowed for comparing taxonomic diversity before β-diversities studies for Bacteria, Archaea and Eukaryotes. The study identified 57 phyla, 145 classes, 301 orders, 506 families, 963 genera, and 1980 species. Bacteria dominated the microbes and comprised 28 species, 51 classes, 110 orders, 243 families, 597 genera, and 1518 species. The Bacteroides(6.77%) were dominant, followed by Acinetobacter(1.44%) belonging to the Gammaproteobacteria and Acidororax (1.36%), Bacillus (1.24%) and Clostridium (1.02%) belonging to Betaproteobacteria. Archaea recorded 5 phyla, 13 classes, 19 orders, 29 families, 60 genera,and87 species, with the dominant genera being Methanospirillum (16.01%), methanosarcina (15.70%), and Methanoregula(14.80%) and Methanosaeta (8.74%), Methanosphaerula(5.48%) and Methanobrevibacter(5.03%) being the subdominant group. The eukaryotes were the least in abundance and comprised 24 phyla, 81 classes, 301 orders, 506 families, 963 genera, and 980 species. Arabidopsis (4.91%) and Caenorhabditis (4.81%) dominated the eukaryotes, while Dityostelium (3.63%) and Drosophila(2.08%) were the subdominant genera. All these microbes play distinct roles in the anaerobic treatment process of sewage sludge. The local sludge microbial composition and abundance variations may be due to age difference differences between the two digestion lagoons in operation at the plant and the different degradation rales played by the taxa. The information presented in this study can help in the genetic manipulation or formulation of optimal microbial ratios to improve their effectiveness in sewage sludge treatment. This study recommends further research on how the different taxa respond to environmental changes over time and space.

Keywords: shotgun metagenomics, sludge, bacteria, archaea, eukaryotes

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Authors: Livinus A. Obasi, Augustine N. Ajah

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Biotechnology in recent times has tried to develop a mechanism whereby sustainable electricity can be generated by the activity of microorganisms on waste and renewable biomass (often regarded as “negative value”) in a device called microbial fuel cell, MFC. In this paper, we established how the biocatalytic activities of bacteria on organic matter (substrates) produced some electrons with the associated removal of some water pollution parameters; Biochemical oxygen demand (BOD), chemical oxygen demand (COD) to the tune of 77.2% and 88.3% respectively from a petrochemical sanitary wastewater. The electricity generation was possible by conditioning the bacteria to operate anaerobically in one chamber referred to as the anode while the electrons are transferred to the fully aerated counter chamber containing the cathode. Power densities ranging from 12.83 mW/m² to 966.66 mW/m² were achieved using a dual-chamber starch membrane MFC experimental set-up. The maximum power density obtained in this research shows an improvement in the use of low cost MFC set up to achieve power production. Also, the level of organic matter removal from the sanitary waste water by the operation of this device clearly demonstrates its potential benefit in achieving an improved benign environment. The beauty of the MFCs is their potential utility in areas lacking electrical infrastructures like in most developing countries.

Keywords: bioelectricity, COD, microbial fuel cell, sanitary wastewater, wheat starch

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Authors: Benjamin Castillo, Luis Pastenes, Fernando Cordova

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The pathogen growth in animal source foods is a common problem in the food industry, causing monetary losses due to the spoiling of products or food intoxication outbreaks in the community. In this sense, the quality of the product is reflected by the population of deteriorating agents present in it, which are mainly bacteria. The factors which are likely associated with freshness in animal source foods are temperature and processing, storage, and transport times. However, the level of deterioration of products depends, in turn, on the characteristics of the bacterial population, causing the decomposition or spoiling, such as pH level and toxins. Knowing the growth dynamics of the agents that are involved in product contamination allows the monitoring for more efficient processing. This means better quality and reasonable costs, along with a better estimation of necessary time and temperature intervals for transport and storage in order to preserve product quality. The objective of this project is to design a secondary model that allows measuring the impact on temperature bacterial growth and the competition for pH adequacy and release of bacteriocins in order to describe such phenomenon and, thus, estimate food product half-life with the least possible risk of deterioration or spoiling. In order to achieve this objective, the authors propose an analysis of a three-dimensional ordinary differential which includes; logistic bacterial growth extended by the inhibitory action of bacteriocins including the effect of the medium pH; change in the medium pH levels through an adaptation of the Luedeking-Piret kinetic model; Bacteriocin concentration modeled similarly to pH levels. These three dimensions are being influenced by the temperature at all times. Then, this differential system is expanded, taking into consideration the variable temperature and the concentration of pulsed bacteriocins, which represent characteristics inherent of the modeling, such as transport and storage, as well as the incorporation of substances that inhibit bacterial growth. The main results lead to the fact that temperature changes in an early stage of transport increased the bacterial population significantly more than if it had increased during the final stage. On the other hand, the incorporation of bacteriocins, as in other investigations, proved to be efficient in the short and medium-term since, although the population of bacteria decreased, once the bacteriocins were depleted or degraded over time, the bacteria eventually returned to their regular growth rate. The efficacy of the bacteriocins at low temperatures decreased slightly, which equates with the fact that their natural degradation rate also decreased. In summary, the implementation of the mathematical model allowed the simulation of a set of possible bacteria present in animal based products, along with their properties, in various transport and storage situations, which led us to state that for inhibiting bacterial growth, the optimum is complementary low constant temperatures and the initial use of bacteriocins.

Keywords: bacterial growth, bacteriocins, mathematical modelling, temperature

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Authors: Paulina Zavistanaviciute, Vita Krungleviciute, Elena Bartkiene

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Lactic acid bacteria (LAB) are microbial supplements that increase the nutritional, therapeutic, and safety value of food and feed. Often LAB strains are incubated in an expensive commercially available de Man-Rogosa-Sharpe (MRS) medium; the cultures are centrifuged, and the cells are washing with sterile water. Potato juice and apple juice industry bio-products are industrial wastes which may constitute a source of digestible nutrients for microorganisms. Due to their low cost and good chemical composition, potato juice and apple juice production bio- products could have a potential application in LAB encapsulation. In this study, pure LAB (P. acidilactici and P. pentosaceus) were multiplied in a crushed potato juice and apple juice industry bio-products medium. Before using, bio-products were sterilized and filtered. No additives were added to mass, except apple juice industry bioproducts were diluted with sterile water (1/5; v/v). The tap of sterilised mass, and LAB cell suspension (5 mL), containing of 8.9 log10 colony-forming units (cfu) per mL of the P. acidilactici and P. pentosaceus was used to multiply the LAB for 72 h. The final colony number in the potato juice and apple juice bio- products substrate was on average 9.60 log10 cfu/g. In order to stabilize the LAB, several methods of dehydration have been tested: lyophilisation (MilrockKieffer Lane, Kingston, USA) and dehydration in spray drying system (SD-06, Keison, Great Britain). Into the spray drying system multiplied LAB in a crushed potato juice and apple juice bio-products medium was injected in peristaltic way (inlet temperature +60 °C, inlet air temperature +150° C, outgoing air temperature +80 °C, air flow 200 m3/h). After lyophilisation (-48 °C) and spray drying (+150 °C) the viable cell concentration in the fermented potato juice powder was 9.18 ± 0.09 log10 cfu/g and 9.04 ± 0.07 log10 cfu/g, respectively, and in apple mass powder 8.03 ± 0.04 log10 cfu/g and 7.03 ± 0.03 log10 cfu/g, respectively. Results indicated that during the storage (after 12 months) at room temperature (22 +/- 2 ºC) LAB count in dehydrated products was 5.18 log10 cfu/g and 7.00 log10 cfu/g (in spray dried and lyophilized potato juice powder, respectively), and 3.05 log10 cfu/g and 4.10 log10 cfu/g (in spray dried and lyophilized apple juice industry bio-products powder, respectively). According to obtained results, potato juice could be used as alternative substrate for P. acidilactici and P. pentosaceus cultivation, and by drying received powders can be used in food/feed industry as the LAB starters. Therefore, apple juice industry by- products before spray drying and lyophilisation should be modified (i. e. by using different starches) in order to improve its encapsulation.

Keywords: bio-products, encapsulation, lactic acid bacteria, sustainability

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Authors: Ozan Kahraman, Hao Feng

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Manothermosonication (MTS), which consists of the simultaneous application of heat and ultrasound under moderate pressure (100-700 kPa), is one of the technologies which destroy microorganisms and inactivates enzymes. Transmission electron microscopy (TEM) is a microscopy technique in which a beam of electrons is transmitted through an ultra-thin specimen, interacting with the specimen as it passes through it. The environmental scanning electron microscope or ESEM is a scanning electron microscope (SEM) that allows for the option of collecting electron micrographs of specimens that are "wet," uncoated. These microscopy techniques allow us to observe the processing effects on the samples. This study was conducted to investigate the effects of MTS and HTST treatments on the morphology of apple-carrot juices by using TEM and ESEM microscopy. Apple-carrot juices treated with HTST (72 0C, 15 s), MTS 50 °C (60 s, 200 kPa), and MTS 60 °C (30 s, 200 kPa) were observed in both ESEM and TEM microscopy. For TEM analysis, a drop of the solution dispersed in fixative solution was put onto a Parafilm ® sheet. The copper coated side of the TEM sample holder grid was gently laid on top of the droplet and incubated for 15 min. A drop of a 7% uranyl acetate solution was added and held for 2 min. The grid was then removed from the droplet and allowed to dry at room temperature and presented into the TEM. For ESEM analysis, a critical point drying of the filters was performed using a critical point dryer (CPD) (Samdri PVT- 3D, Tousimis Research Corp., Rockville, MD, USA). After the CPD, each filter was mounted onto a stub and coated with gold/palladium with a sputter coater (Desk II TSC Denton Vacuum, Moorestown, NJ, USA). E.Coli O157:H7 cells on the filters were observed with an ESEM (Philips XL30 ESEM-FEG, FEI Co., Eindhoven, The Netherland). ESEM (Environmental Scanning Electron Microscopy) and TEM (Transmission Electron Microscopy) images showed extensive damage for the samples treated with MTS at 50 and 60 °C such as ruptured cells and breakage on cell membranes. The damage was increasing with increasing exposure time.

Keywords: MTS, HTST, ESEM, TEM, E.COLI O157:H7

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Authors: Temitope D. Awobusuyi, Eric O. Amonsou, Muthulisi Siwela, Oluwatosin A. Ijabadeniyi

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Provitamin A-biofortified maize has been developed to alleviate Vitamin A deficiency; a major public health problem in developing countries. Amahewu, a non-alcoholic fermented maize based beverage is produced using white maize, which is deficient in Vitamin A. In this study, the suitable processing conditions for the production of amahewu using provitamin A-biofortified maize and the microbial quality of the processed products were evaluated. Provitamin A-biofortified amahewu was produced with reference to traditional processing method. Processing variables were Inoculum types (Malted provitamin A maize, Wheat bran, and lactobacillus mixed starter culture with either malted provitamin A or wheat bran) and concentration (0.5 %, 1 % and 2 %). A total of four provitamin A-biofortified amahewu products after fermentation were subjected to different storage conditions: 4ᴼC, 25ᴼC and 37ᴼC. pH and TTA were monitored throughout the storage period. Sample of provitamin A-biofortified amahewu were plated and observed every day for 5 days to assess the presence of Aerobic and Anaerobic spore formers, E.coli, Lactobacillus and Mould. The addition of starter culture substantially reduced the fermentation time (6 hour, pH 3.3) compared to those with no addition of starter culture (24 hour pH 3.5). It was observed that Lactobacillus were present from day 0 for all the storage temperatures. The presence of aerobic spore former and mould were observed on day 3. E.coli and Anaerobic spore formers were not present throughout the storage period. These microbial growth were minimal at 4ᴼC while 25ᴼC had higher counts of growth with 37ᴼC having the highest colony count. Throughout the storage period, pH of provitamin A-biofortified amahewu was stable. Provitamin A-biofortified amahewu stored under refrigerated condition (4ᴼC) had better storability compared to 25ᴼC and 37ᴼC. The production and microbial quality of provitamin A-biofortified amahewu might be important in combating Vitamin A Deficiency.

Keywords: biofortification, fermentation, maize, vitamin A deficiency

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Authors: Sarisha Devnath, Oluwatosin A. Ijabadeniyi

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In recent years milk contamination has become a major problem in households; due to the likely occurrence of bacteria, even after the milk has been processed. One such genus of bacteria causing unwanted growth is Bacillus. This research project looks at the presence of spore formers in processed milk from household refrigerators and the effect of pasteurization and high temperature on Bacillus spores activation. 24 samples each of UHT milk and pasteurised milk from 24 households were sampled for the presence of spore formers. While anaerobic spore formers were not found in any of the samples, the average aerobic spore formers in UHT milk and pasteurized milk however were 5.77 cfu/ml and 5.88 cfu/ml respectively. After sequencing, it was detected that the mixed culture contained Bacillus cereus, for both pasteurised and UHT milk samples. For the activation study, raw milk samples were collected and subjected to four different temperatures; 65˚C, 72˚C, 80˚C, 100˚C respectively. Samples were stored for 7 days at 5˚C and 10˚C and analysed daily. The average aerobic spore formers in raw milk for samples stored at 5˚C range between 4.67-6.00 cfu/ml while it ranges between 4.84-6.00 cfu/ml at 10˚C, signifying that the high temperatures could have resulted in germination of dominant spores. Statistical analysis conducted on these results indicated a significant difference between the numbers of colonies present at the different treatment temperatures the bacterium was exposed to. This work showed that household milk may constitute public health risk furthermore; pasteurization and higher temperatures may not be effective to remove aerobic spore formers because of Bacillus spores activation.

Keywords: sporeformers, bacillus, spores, activation, milk

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Authors: Mohammed Al-Bloushi, Sanghyun Jeong, Torove Leiknes

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Biofouling in the open recirculating cooling water systems may cause biological corrosion, which can reduce the performance, increase the energy consummation and lower heat exchange efficiencies of the cooling tower. Seawater cooling towers are prone to biofouling due to the presences of organic and inorganic compounds in the seawater. The availability of organic and inorganic nutrients, along with sunlight and continuous aeration of the cooling tower contributes to an environment that is ideal for microbial growth. Various microorganisms (algae, fungi, and bacteria) can grow in a cooling tower system under certain environmental conditions. The most commonly being used method to control the biofouling in the cooling tower is the addition of biocides such as chlorination. In this study, algae containing diatom and green algae were added to the cooling tower basin, and its viability was monitored in the recirculating cooling seawater loop as well as in the cooling tower basin. Continuous addition of biocides was employed in pilot-scale seawater cooling towers, and it was operated continuously for 2 months. Three different types of oxidizing biocides, namely chlorine, chlorine dioxide and ozone, were tested. The results showed that all biocides were effective in keeping the biological growth to the minimum regardless of algal addition. Amongst the biocides, ozone could reduce 99% of total live cells of bacteria and algae, followed by chlorine dioxide at 97%, while the conventional chlorine showed only 89% reduction in the bioactivities.

Keywords: algae, biocide, biofouling, seawater cooling tower

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Authors: Agampodi Promoda Perera, Yong Shin, Mi Kyoung Park

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The successful detection of pathogenic bacteria in blood provides important information for early detection, diagnosis and the prevention and treatment of infectious diseases. Silicon microring resonators are refractive-index-based optical biosensors that provide highly sensitive, label-free, real-time multiplexed detection of biomolecules. We demonstrate the technique of using GO (graphene oxide) to enhance the signal output of the silicon microring optical sensor. The activated carboxylic groups in GO molecules bind directly to single stranded DNA with an amino modified 5’ end. This conjugation amplifies the shift in resonant wavelength in a real-time manner. We designed a capture probe for strain Staphylococcus aureus of 21 bp and a longer complementary target sequence of 70 bp. The mismatched target sequence we used was of Streptococcus agalactiae of 70 bp. GO is added after the complementary binding of the probe and target. GO conjugates to the unbound single stranded segment of the target and increase the wavelength shift on the silicon microring resonator. Furthermore, our results show that GO could successfully differentiate between the mismatched DNA sequences from the complementary DNA sequence. Therefore, the proposed concept could effectively enhance sensitivity of pathogen detection sensors.

Keywords: label free biosensor, pathogenic bacteria, graphene oxide, diagnosis

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Authors: Benguerine Zohra, Merzak Siham, Bouziane Cheimaa, Si Tayeb Fatima, Jou Siham, Belkessam

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Introduction: Antibiotics, widely produced and consumed in large quantities, have proven problematic due to various types of side effects. The development of bacterial resistance to currently available antibiotics has made the search for new antibacterial agents necessary. One alternative strategy to combat antibiotic-resistant bacteria is the use of natural antimicrobial substances such as plant extracts. The objective of this study is to provide an overview of the antibacterial effects of a plant native to the Middle East and Mediterranean regions, Salvia officinalis (sage). Materials and Methods: This review article was conducted by searching studies in the PubMed, Scopus, JSTOR, and SpringerLink databases. The search terms were "Salvia officinalis L." and "antibacterial effects." Only studies that met our inclusion criteria (in English, antibacterial effects of Salvia officinalis L., and primarily dating from 2012 to 2023) were accepted for further review. Results and Discussion: The initial search strategy identified approximately 78 references, with only 13 articles included in this review. The synthesis of the articles revealed that several data sources confirm the antimicrobial effects of S. officinalis. Its essential oil and alcoholic extract exhibit strong bactericidal and bacteriostatic effects against both Gram-positive and Gram-negative bacteria. Conclusion: The significant value of the extract, oil, and leaves of S. officinalis calls for further studies on the other useful and unknown properties of this multi-purpose plant.

Keywords: salvia officinalis, literature review, antibacterial, effects

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Authors: Adriana Sosa, Susana Rincon, Chérif Ben, Diana Cabañas, Juan E. Ruiz, Alejandro Zepeda

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The complex chemical composition (mixtures of ammonium and recalcitrant compounds) of the effluents from the chemical, pharmaceutical and petrochemical industries represents a challenge in their biological treatment. This treatment involves nitrification process that can suffer an inhibition due to the presence of aromatic compounds giving as a result the decrease of the process efficiency. The inhibitory effects on nitrification in the presence of aromatic compounds have already been studied; however a few studies have considered the presence of phenolic compounds in the form of mixtures, which is the form that they are present in real context. For this reason, we realized a kinetic study on the nitrifying process in the presence of different concentrations of a mixture of phenol, o-cresol, m-cresol and p-cresol (0 - 320 mg C/L) in a sequencing batch reactor (SBR). Firstly, the nitrifying process was evaluated in absence of the phenolic mixture (control 1) in a SBR with 2 L working volume and 176 mg/L of nitrogen of microbial protein. Total oxidation of initial ammonium (efficiency; ENH4+ of 100 %) to nitrate (nitrifying yield; YNO3- of 0.95) were obtained with specific rates of ammonium consumption (qN-NH4+) and nitrate production (qN-NO3-) (of 1.11 ± 0.04 h-1 and 0.67 h-1 ± 0.11 respectively. During the phase of acclimation with 40 mg C/L of the phenolic mixture, an inhibitory effect on the nitrifying process was observed, provoking a decrease in ENH4+ and YNO3- (11 and 54 % respectively) as well as in the specific rates (89 y 46 % respectively), being the ammonia oxidizing bacteria (BAO) the most affected. However, in the next cycles without the phenolic mixture (control 2), the nitrifying consortium was able to recover its nitrifying capacity (ENH4+ = 100% and YNO3-=0.98). Afterwards the SBR was fed with 10 mg C/L of the phenolic mixture, obtaining and ENH4+ of 100%, YNO3- and qN-NH4+ 0.62 ± 0.006 and 0.13 ± 0.004 respectively, while the qN-NO3- was 0.49 ± 0.007. Moreover, with the increase of the phenolic concentrations (10-160 mg C/L) and the number of cycles the nitrifying consortium was able to oxidize the ammonia with ENH4+ of 100 % and YNO3- close to 1. However a decrease in the values of the nitrification specific rates and increase in the oxidation in phenolic compounds (70 to 94%) were observed. Finally, in the presence of 320 mg C/L, the nitrifying consortium was able to simultaneously oxidize the ammonia (ENH4+= 100%) and the phenolic mixture (p-cresol>phenol>m-cresol>o-cresol) being the o-cresol the most recalcitrant compound. In all the experiments the use of a SBR allowed a respiratory adaptation of the consortium to oxidize the phenolic mixture achieving greater adaptation of the nitrite-oxidizing bacteria (NOB) than in the ammonia-oxidizing bacteria (AOB).

Keywords: cresol, inhibition, nitrification, phenol, sequencing batch reactor

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Authors: Jung-En Kuan, Whei-Fen Wu

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In this study, a symbiotic bacteria from yellow mealworm's (Tenebrio molitor) mid-gut was isolated with characteristics of growth on minimal-tributyrin medium. After a PCR-amplification of its 16s rDNA, the resultant nucleotide sequences were then analyzed by schemes of the phylogeny trees. Accordingly, it was designated as Pseudomonas nitroreducens D-01. Next, by searching the lipolytic enzymes in its protein data bank, one of those potential lipolytic α/β hydrolases was identified, again using PCR-amplification and nucleotide-sequencing methods. To construct an expression of this lipolytic gene in plasmids, the target-gene primers were then designed, carrying the C-terminal his-tag sequences. Using the vector pET21a, a recombinant lipolytic hydrolase D gene with his-tag nucleotides was successfully cloned into it, of which the lipolytic D gene is under a control of the T7 promoter. After transformation of the resultant plasmids into Eescherichia coli BL21 (DE3), an IPTG inducer was used for the induction of the recombinant proteins. The protein products were then purified by metal-ion affinity column, and the purified proteins were found capable of forming a clear zone on tributyrin agar plate. Shortly, its enzyme activities were determined by degradation of p-nitrophenyl ester(s), and the substantial yellow end-product, p-nitrophenol, was measured at O.D.405 nm. Specifically, this lipolytic enzyme efficiently targets p-nitrophenyl butyrate. As well, it shows the most reactive activities at 40°C, pH 8 in potassium phosphate buffer. In thermal stability assays, the activities of this enzyme dramatically drop when the temperature is above 50°C. In metal ion assays, MgCl₂ and NH₄Cl induce the enzyme activities while MnSO₄, NiSO₄, CaCl₂, ZnSO₄, CoCl₂, CuSO₄, FeSO₄, and FeCl₃ reduce its activities. Besides, NaCl has no effects on its enzyme activities. Most organic solvents decrease the activities of this enzyme, such as hexane, methanol, ethanol, acetone, isopropanol, chloroform, and ethyl acetate. However, its enzyme activities increase when DMSO exists. All the surfactants like Triton X-100, Tween 80, Tween 20, and Brij35 decrease its lipolytic activities. Using Lineweaver-Burk double reciprocal methods, the function of the enzyme kinetics were determined such as Km = 0.488 (mM), Vmax = 0.0644 (mM/min), and kcat = 3.01x10³ (s⁻¹), as well the total efficiency of kcat/Km is 6.17 x10³ (mM⁻¹/s⁻¹). Afterwards, based on the phylogenetic analyses, this lipolytic protein is classified to type IV lipase by its homologous conserved region in this lipase family.

Keywords: enzyme, esterase, lipotic hydrolase, type IV

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Authors: Muhammad Younis Laghari

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Global climate change has had many effects on the aquatic environment, and the major issue is pollution. Along with the other pollutants, there are a significant number of human microbial pathogens that pollute the water bodies. Another concern about the water quality is that the major aquatic resources bring water-borne pathogens and other related diseases. These resources include industrial effluent, untreated domestic sewage, acid mine drainage, etc. However, these water discharges through various routes may have treatment to eliminate the pathogenic microbes. Therefore, it is essential to control the leakage from sewer systems, residential discharge, and agricultural run-off. These pathogenic microbes have been implicated in the lives of water health (fishes), which is harmful and causes diseases. Mostly, the mortality of aquatic species results because of catastrophic floods due to poor water waste treatment and sanitation that introduce pathogenic bacteria into rivers. Pathogens survive in rivers and remain poorly known but essential to control water-borne diseases. The presence of bacteria in watercourses is diverse and constitutes a complicated subject. Many species are autochthonous and play an important role in aquatic ecosystems, while many others arise from untreated or poorly treated waste from industrial and domestic sources. Further, more investigation is required to know the induction of water-borne pathogens in various water resources and the potential impacts of water resource development on pathogen contamination.

Keywords: microbial pathogens, contamination, water resources, river water body

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Authors: Mona Alharbi

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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.

Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis

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Authors: Mohammad Hajjartabar, Tahereh Kermani Ranjbar

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P. aeruginosa EF2 was isolated and identified from human infection sources before in our previous study. The present study was performed to determine the characteristics and activity role of bioflocculant produced by the bacterium in flocculation of the wastewater active sludge treatment. The bacterium was inoculated and then was grown in an orbital shaker at 250 rpm for 5 days at 35 °C under TSB and peptone water media. After incubation period, culture broths of the bacterial strain was collected and washed. The concentration of the bacteria was adjusted. For the extraction of the bacterial bioflocculant, culture was centrifuged at 6000 rpm for 20 min at 4 °C to remove bacterial cells. Supernatant was decanted and pellet containing bioflocculant was dried at 105 °C to a constant weight according to APHA, 2005. The chemical composition of the extracted bioflocculant from the bacterial sample was then analyzed. Wastewater active sludge sample obtained from aeration tank from one of wastewater treatment plants in Tehran, was first mixed thoroughly. After addition of bioflocculant, improvements in floc density were observed with an increase in bioflocculant. The results of this study strongly suggested that the extracted bioflucculant played a significant role in flocculation of the wastewater sample. The use of wild bacteria and nutrient regulation techniques instead of genetic manipulation opens wide investigation area in the future to improve wastewater treatment processes. Also this may put a new path in front of us to attain and improve the more effective bioflocculant using the purified microbial culture in wastewater treatment.

Keywords: wastewater treatment, P. aeruginosa, sludge treatment

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Authors: Maria Luisa de la Torre, Javier Aroba, Jose Miguel Davila, Aguasanta M. Sarmiento

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Steel is one of the most widely used materials in polymetallic sulfide mining installations. One of the main problems suffered by these facilities is the economic losses due to the corrosion of this material, which is accelerated and aggravated by the contact with acid waters generated in these mines when sulfides come into contact with oxygen and water. This generation of acidic water, in turn, is accelerated by the presence of acidophilic bacteria. In order to gain a more detailed understanding of this corrosion process and the interaction between steel and acidic water, a laboratory experiment was carried out in which carbon steel plates were introduced into four different solutions for 27 days: distilled water (BK), which tried to assimilate the effect produced by rain on this material, an acid solution from a mine with a high Fe2+/Fe3+ (PO) content, another acid solution of water from another mine with a high Fe3+/Fe2+ (PH) content and, finally, one that reproduced the acid mine water with a high Fe2+/Fe3+ content but in which there were no bacteria (ST). Every 24 hours, physicochemical parameters were measured and water samples were taken to carry out an analysis of the dissolved elements. The results of these measurements were processed using an explainable AI model based on fuzzy logic. It could be seen that, in all cases, there was an increase in pH, as well as in the concentrations of Fe and, in particular, Fe(II), as a consequence of the oxidation of the steel plates. Proportionally, the increase in Fe concentration was higher in PO and ST than in PH because Fe precipitates were produced in the latter. The rise of Fe(II) was proportionally much higher in PH and, especially in the first hours of exposure, because it started from a lower initial concentration of this ion. Although to a lesser extent than in PH, the greater increase in Fe(II) also occurred faster in PO than in ST, a consequence of the action of the catalytic bacteria. On the other hand, Cu concentrations decreased throughout the experiment (with the exception of distilled water, which initially had no Cu, as a result of an electrochemical process that generates a precipitation of Cu together with Fe hydroxides. This decrease is lower in PH because the high total acidity keeps it in solution for a longer time. With the application of an artificial intelligence tool, it has been possible to evaluate the effects of steel corrosion in mining environments, corroborating and extending what was obtained by means of classical statistics. Acknowledgments: This work has been supported by MCIU/AEI/10.13039/501100011033/FEDER, UE, throughout the project PID2021-123130OB-I00.

Keywords: carbon steel, corrosion, acid mine drainage, artificial intelligence, fuzzy logic

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Authors: Amanuel Bekuma, Rebecca Swift, Sarah Jackson, Ben Biddulph

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Economic loss to frost damage is increasing over the past years in the Western Australian Wheatbelt. Agronomic, genetic, and climatic works have still found a weak correlation between temperature and frost damage. One possibility that has not been explored within the Australian cropping system is whether ice nucleation active bacteria (INB) either present in situ on crop residue or introduced by rainfall could be responsible for the increased sensitivity of cereal plants to frost at different stages of development. This study investigated upper and lower leaf canopy, stubble, and soil as a potential site of ice nucleation activity (INA) and tracked the changes in INA during the plant development. We found that older leaves of wheat are the primary sites of ice nucleation (-4.7 to -6.3°C) followed by stubble (-5.7 to -6.7°C) which increases the risk of frost damage during heading and flowering (the most susceptible stages). However, healthy and green upper canopy leaves (flag and flag-2) and the soil have lower INA (< -11°C) during the frost-sensitive stage of wheat. We anticipate the higher INA on the stubble and older leaves to be due to the presence of biologically active ice-nucleating bacteria (INB), known to cause frost injury to sensitive plants at -5°C. Stubble retained or applied during the growing season further exacerbates additional frost risk by potentially increasing the INB load. The implications of the result for stubble and frost risk management in a frost-prone landscape will be discussed.

Keywords: frost, ice-nucleation-activity, stubble, wheat

Procedia PDF Downloads 138

Authors: Zsolt Szucs, Viktor Kelemen, Son Le Thai, Magdolna Csavas, Erzsebet Roth, Gyula Batta, Annelies Stevaert, Evelien Vanderlinden, Aniko Borbas, Lieve Naesens, Pal Herczegh

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The approval of modern glycopeptide antibiotics such as dalbavancin and oritavancin which have excellent activity against Gram-positive bacteria, encouraged our research group to prepare semisynthetic compounds from several members of glycopeptides by various chemical methods. Derivatives from the aglycone of ristocetin, eremomycin, vancomycin and a pseudoaglycon of teicoplanin have been synthesized in a systematic manner. Interestingly, some of the aglycoristocetin derivatives displayed noteworthy anti-influenza activity. More recently our group has been focusing on the modifications of one of the pseudoaglycons of teicoplanin. The reaction of N-ethoxycarbonyl maleimide derivatives with the primary amino function, the copper-catalysed azide-alkyne click reaction and the sulfonylation of the N-terminus were utilized to obtain systematic series of compounds. All substituents provide a more lipophilic character to the new molecules compared to the parent antibiotics, which is known to be favourable for activity against resistant bacteria. Lipoglycopeptides are also known to have antiviral properties, which has been predominantly studied on HIV by others. The structure-activity relationship study of our compounds revealed the influence of a few structural elements on biological activity. In many cases, minimal changes in lipophilicity and structure produced great differences in efficacy and cytotoxicity. In vitro experiments showed that these compounds are not only active against glycopeptide resistant Gram-positive bacteria but in several cases they prevent the infection of cell cultures by different strains of influenza viruses. This is probably related to the inhibition of the viral entry into the host cell nucleus, of which the exact mechanism is unknown. In some instances, reasonably low concentrations were sufficient to observe this effect. Several derivatives were highly cytotoxic at the same time, but some of them displayed a good selectivity index. The antiviral properties of the compounds are not restricted to influenza viruses e.g., some of them showed good activity against Human Coronavirus 229E. This work could potentially lead to the development of antiviral drugs which possess the crucial structural motifs that are needed for antiviral activity, while missing those which contribute to the antibacterial effect.

Keywords: antiviral, glycopeptide, semisynthetic, teicoplanin

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Authors: Magdalena Smoktunowicz, Renata Wawrzyniak, Malgorzata Waleron, Krzysztof Waleron

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Pectobacterium spp. were previously classified into the Erwinia genus founded in 1917 to unite at that time all Gram-negative, fermentative, nonsporulating and peritrichous flagellated plant pathogenic bacteria. After work of Waldee (1945), on Approved Lists of Bacterial Names and bacteriology manuals in 1980, they were described either under the species named Erwinia or Pectobacterium. The Pectobacterium genus was formally described in 1998 of 265 Pectobacterium strains. Currently, there are 21 species of Pectobacterium bacteria, including Pectobacterium betavasculorum since 2003, which caused soft rot on sugar beet tubers. Based on the biochemical experiments carried out for this, it is known that these bacteria are gram-negative, catalase-positive, oxidase-negative, facultatively anaerobic, using gelatin and causing symptoms of soft rot on potato and sugar beet tubers. The mere fact of growing on sugar beet may indicate a metabolism characteristic only for this species. Metabolomics, broadly defined as the biology of the metabolic systems, which allows to make comprehensive measurements of metabolites. Metabolomics, in combination with genomics, are complementary tools for the identification of metabolites and their reactions, and thus for the reconstruction of metabolic networks. The aim of this study was to apply the GC-MS-based untargeted metabolomics to study the metabolism of P. betavasculorum in different growing conditions. The metabolomic profiles of biomass and biomass media were determined. For sample preparation the following protocol was used: extraction with 900 µl of methanol: chloroform: water mixture (10: 3: 1, v: v) were added to 900 µl of biomass from the bottom of the tube and up to 900 µl of nutrient medium from the bacterial biomass. After centrifugation (13,000 x g, 15 min, 4oC), 300µL of the obtained supernatants were concentrated by rotary vacuum and evaporated to dryness. Afterwards, two-step derivatization procedure was performed before GC-MS analyses. The obtained results were subjected to statistical calculations with the use of both uni- and multivariate tests. The obtained results were evaluated using KEGG database, to asses which metabolic pathways are activated and which genes are responsible for it, during the metabolism of given substrates contained in the growing environment. The observed metabolic changes, combined with biochemical and physiological tests, may enable pathway discovery, regulatory inference and understanding of the homeostatic abilities of P. betavasculorum.

Keywords: GC-MS chromatograpfy, metabolomics, metabolism, pectobacterium strains, pectobacterium betavasculorum

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Authors: Xiuguo Zhao, He Li, Alireza Yazdani, Xiaoning Zheng, Xinxi Xu

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Wound infected poses a serious threat to the surgery on the patient during the process of surgery. Understanding the bacteria-carrying particles (BCPs) transportation and deposition in the open surgical wound model play essential role in protecting wound against being infected. Therefore BCPs transportation and deposition in the surgical wound model were investigated using force-coupling method (FCM) based computational fluid dynamics. The BCPs deposition in the wound was strongly associated with BCPs diameter and concentration. The results showed that the rise on the BCPs deposition was increasing not only with the increase of BCPs diameters but also with the increase of the BCPs concentration. BCPs deposition morphology was impacted by the combination of size distribution, airflow patterns and model geometry. The deposition morphology exhibited the characteristic with BCPs deposition on the sidewall in wound model and no BCPs deposition on the bottom of the wound model mainly because the airflow movement in one direction from up to down and then side created by laminar system constructing airflow patterns and then made BCPs hard deposit in the bottom of the wound model due to wound geometry limit. It was also observed that inertial impact becomes a main mechanism of the BCPs deposition. This work may contribute to next study in BCPs deposition limit, as well as wound infected estimation in surgical-site infections.

Keywords: BCPs deposition, computational fluid dynamics, force-coupling method (FCM), numerical simulation, open surgical wound model

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Authors: Begna Tulu, Gobena Ameni

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Understanding the types of Mycobacterium bovis (M. bovis) strains circulating in a country and exploring its zoonotic potential has significant contribution in the effort to design control strategies. The main aim of this study was to review and compile the results of studies conducted on M. bovis genotyping and its zoonotic potential of M. bovis in Ethiopia. A systematic search and review of articles published on M. bovis strains in Ethiopia were made. PubMed and Google Scholar databases were considered for the search while the keywords used were 'Mycobacteria,' 'Mycobacterium bovis,' 'Bovine Tuberculosis' and 'Ethiopia.' Fourteen studies were considered in this review and a total of 31 distinct strains of M. bovis (N=211) were obtained; the most dominant strains were SB0133 (N=62, 29.4%), SB1176 (N=61, 28.9%), and followed by SB0134 and SB1476 each (N=18, 8.5%). The clustering rate of M. bovis strains was found to be 42.0%. On the other hand, 6 strains of M. bovis were reported from human namely; SB0665 (N=4), SB0303 (N=2), SB0982 (N=2), SB0133 (N=1), SB1176 (N=1), and 1 new strain. Similarly, a total of 8 strains (N=13) of M. tuberculosis bacteria were also identified from animal subjects; namely SIT149 (N=3), SIT1 (N=2), SIT1688 (n=2), SIT262 (N=2), SIT53 (N=1), SIT59 (N=1), and one new-Ethiopian strain. The result showed that the genetic diversity of M. bovis strains reported from Ethiopia are less diversified and highly clustered. And also the result underlines that there is an ongoing active transmission of M. bovis and M. tuberculosis between human and animals in Ethiopia because a significant number strains of both type of bacteria were reported from human and animals.

Keywords: mycobacterium bovis, Mycobacterium tuberculosis, zoonotic potential, genetic diversity, Ethiopia

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Authors: Ibrahim Siddig Hamid, Ikram Mohamed Eltayeb

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Trachyspermum ammi belongs to the family Apiaceae, is used traditionally for the treatment of gastrointestinal ailments, lack of appetite and bronchial problems as well used as antiseptic, antimicrobial, antipyretic, febrifugal and in the treatment of typhoid fever. Peganum harmala belongs to the family Zygophyllaceae it has been reported to have an antibacterial activity and used to treat depression and recurring fevers. It also used to kill algae, bacteria, intestinal parasites and molds. In Sudan, the combination of two plants are traditionally used for the treatment of bacillary dysentery. Bacillary dysentery is caused by one or more types of Shigella species bacteria mainly Shigella dysenteri and shigella flexneri. Bacillary dysentery is mainly found in hot countries like Sudan with poor hygiene and sanitation. Bacillary dysentery causes sudden onset of high fever and chills, abdominal pain, cramps and bloating, urgency to pass stool, weight loss, and dehydration and if left untreated it can lead to serious complications including delirium, convulsions and coma. A serious infection like this can be fatal within 24 hours. The objective of this study is to investigate the in vitro susceptibility of Sh. flexneri and Sh. dysenteriae to the T. ammi and P. harmala. T. ammi and P. harmala were extracted by 96% ethanol using Soxhlet apparatus. The antimicrobial activity of the extracts was investigated according to the disc diffusion method. The discs were prepared by soaking sterilized filter paper discs in 20 microliter of serially diluted solutions of each plant extract with the concentrations (100, 50, 25, 12.5, 6.25mg/dl) then placing them on Muller Hinton Agar plates that were inoculated with bacterial suspension separately, the plates were incubated for 24 hours at 37c and the minimum inhibitory concentration of the extract which was the least concentration of the extract to inhibit fungal growth was determined. The results showed the high antimicrobial activity of T. ammi extract with an average diameter zone ranging from 18-20 mm and its minimum inhibitory concentration was found to be 25 mg/ml against the two shigella species. P. harmala extract was found to have slight antibacterial effect against the two bacteria. This result justified the Sudanese traditional use of Trachyspermum ammi plant for the treatment of bacillary dysentery.

Keywords: harmala, peganum, shigella, trachyspermum

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Authors: Tiantian Min, Chuanxiang Cheng, Jin Yue

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The growth and reproduction of microorganisms in food packaging can cause food decay and foodborne diseases, which pose a serious threat to the health of consumers and even cause serious economic losses. Active food packaging containing antibacterial bioactive compounds is a promising strategy for extending the shelf life of products and maintaining the food quality, as well as reducing the food waste. However, most active packaging can only act as slow-release effect for antimicrobials, which causes the release rate of antimicrobials not match the growth rate of microorganisms. Stimuli-responsive active packaging materials based on biopolymeric substrates and bioactive substances that respond to some biological and non-biological trigger factors provide more opportunities for fresh food preservation. The biological stimuli factors such as relative humidity, pH and enzyme existed in the exudate secreted by microorganisms have been expected to design food packaging materials. These stimuli-responsive materials achieved accurate release or delivery of bioactive substances at specific time and appropriate dose. Recently, metal-organic-frameworks (MOFs) nanoparticles become attractive carriers to enhance the efficiency of bioactive compounds or drugs. Cellulose nanofibrils have been widely applied for film substrates due to their biodegradability and biocompatibility. The abundant hydroxyl groups in cellulose can be oxidized to carboxyl groups by TEMPO, making it easier to anchoring MOFs and to be further modification. In this study, a pH and enzyme dual-responsive CAR@ZIF-8/TOCNF/PE film was fabricated by in-situ growth of ZIF-8 nanoparticles onto TEMPO-oxidized cellulose (TOCNF) film and further coated with pectin (PE) for stabilization and controlled release of carvacrol (CAR). The enzyme triggered release of CAR was achieved owing to the degradation of pectin by pectinase secreted by microorganisms. Similarly, the pH-responsive release of CAR was attributed to the unique skeleton degradation of ZIF-8, further accelerating the release of CAR from the topological structure of ZIF-8. The composite film performed excellent crystallinity and adsorb ability confirmed by X-ray diffraction and BET analysis, and the inhibition efficiency against Escherichia coli, Staphylococcus aureus and Aspergillus niger reached more than 99%. The composite film was capable of releasing CAR when exposure to dose-dependent enzyme (0.1, 0.2, and 0.3 mg/mL) and acidic condition (pH = 5). When inoculated 10 μL of Aspergillus niger spore suspension on the equatorial position of mango and raspberries, this composite film acted as packaging pads effectively inhibited the mycelial growth and prolonged the shelf life of mango and raspberries to 7 days. Such MOF-TOCNF based film provided a targeted, controlled and sustained release of bioactive compounds for long-term antibacterial activity and preservation effect, which can also avoid the cross-contamination of fruits.

Keywords: active food packaging, controlled release, fruit preservation, in-situ growth, stimuli-responsive

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Authors: Eusebio A. Jiménez-Arévalo, Deifilia Ahuatzi-Chacón, Juvencio Galíndez-Mayer, Cleotilde Juárez-Ramírez, Nora Ruiz-Ordaz

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Bendiocarb is a known toxic xenobiotic that presents acute and chronic risks for freshwater invertebrates and estuarine and marine biota; thus, the treatment of water contaminated with the insecticide is of concern. In this paper, a bacterial community with the capacity to grow in bendiocarb as its sole carbon and nitrogen source was isolated by enrichment techniques in batch culture, from samples of a composting plant located in the northeast of Mexico City. Eight cultivable bacteria were isolated from the microbial community, by PCR amplification of 16 rDNA; Pseudoxanthomonas spadix (NC_016147.2, 98%), Ochrobacterium anthropi (NC_009668.1, 97%), Staphylococcus capitis (NZ_CP007601.1, 99%), Bosea thiooxidans. (NZ_LMAR01000067.1, 99%), Pseudomonas denitrificans. (NC_020829.1, 99%), Agromyces sp. (NZ_LMKQ01000001.1, 98%), Bacillus thuringiensis. (NC_022873.1, 97%), Pseudomonas alkylphenolia (NZ_CP009048.1, 98%). NCBI accession numbers and percentage of similarity are indicated in parentheses. These bacteria were regarded as the isolated species for having the best similarity matches. The ability to degrade bendiocarb by the immobilized bacterial community in a packed bed biofilm reactor, using as support volcanic stone fragments (tezontle), was evaluated. The reactor system was operated in batch using mineral salts medium and 30 mg/L of bendiocarb as carbon and nitrogen source. With this system, an overall removal efficiency (η_bend) rounding 90%, was reached.

Keywords: bendiocarb, biodegradation, biofilm reactor, carbamate insecticide

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Authors: Yasaman Sanayei, Alireza Bahiraie

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This paper presents a systematic methodology based on the application of artificial neural networks for sequencing batch reactor (SBR). The SBR is a fill-and-draw biological wastewater technology, which is specially suited for nutrient removal. Employing reactive dye by Sphingomonas paucimobilis bacteria at sequence batch reactor is a novel approach of dye removal. The influent COD, MLVSS, and reaction time were selected as the process inputs and the effluent COD and BOD as the process outputs. The best possible result for the discrete pole parameter was a= 0.44. In orderto adjust the parameters of ANN, the Levenberg-Marquardt (LM) algorithm was employed. The results predicted by the model were compared to the experimental data and showed a high correlation with R2> 0.99 and a low mean absolute error (MAE). The results from this study reveal that the developed model is accurate and efficacious in predicting COD and BOD parameters of the dye-containing wastewater treated by SBR. The proposed modeling approach can be applied to other industrial wastewater treatment systems to predict effluent characteristics. Note that SBR are normally operated with constant predefined duration of the stages, thus, resulting in low efficient operation. Data obtained from the on-line electronic sensors installed in the SBR and from the control quality laboratory analysis have been used to develop the optimal architecture of two different ANN. The results have shown that the developed models can be used as efficient and cost-effective predictive tools for the system analysed.

Keywords: artificial neural network, COD removal, SBR, Sphingomonas paucimobilis

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Authors: Katarzyna Miśkiewicz, Magdalena Lasoń-Rydel, Małgorzata Krępska, Katarzyna Sieczyńska, Iwona Masłowska-Lipowicz, Katarzyna Ławińska

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The aim of the study was to analyse a selection of 12 pasteurised products made from fruit and vegetables, such as fruit juices, fruit drinks, jams, marmalades and jam produced by small regional factories as well as large food corporations. The research was carried out as part of the project "Innovative system of healthy and regional food distribution", funded by the Ministry of Education and Science (Poland), which aims to create an economically and organisationally strong agri-food industry in Poland through effective cooperation between scientific and socio-economic actors. The main activities of the project include support for the creation of new distribution channels for regional food products and their easy access to a wide group of potential customers while maintaining the highest quality standards. One of the key areas of the project is food quality analyses conducted to indicate the competitive advantage of regional products. Presented here are studies on the content of sugars, vitamin C, preservatives, synthetic colours, sweeteners, sodium and potassium, as well as studies on the microbiological purity of selected products made from fruit and vegetables. The composition of products made from fruit and vegetables varies greatly and depends on both the type of raw material and the way it is processed. Of the samples tested, fruit drinks contained the least amount of sugars, and jam and marmalade made by large producers and bought in large chain stores contained the most. However, the low sugar content of some fruit drinks is due to the presence of the sweetener sucralose in their composition. The vitamin C content of the samples varied, being higher in products where it was added during production. All products made in small local factories were free of food additives such as preservatives, sweeteners and synthetic colours, indicating their superiority over products made by large producers. Products made in small local factories were characterised by a relatively high potassium content. The microbiological purity of commercial products was confirmed - no Salmonella spp. were detected, and the number of mesophilic bacteria, moulds, yeasts, and β-glucuronidase-positive E. coli was below the limit of quantification.

Keywords: fruit and vegetable products, sugars, food additives, HPLC, ICP-OES

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Authors: Maria L. G. Lourenco, Keylla H. N. P. Pereira, Viviane Y. Hibaru, Fabiana F. Souza, Joao C. P. Ferreira, Simone B. Chiacchio, Luiz H. A. Machado

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Neonatal sepsis is a systemic response of acute infection by bacteria that may lead to high mortality in a litter. This study aims to report a case of sepsis by alpha-hemolytic Streptococcus and Mannheimia haemolytica in neonate dogs. A pregnant, mixed-breed bitch at approximately the 60th day of pregnancy was admitted to the Sao Paulo State University (UNESP) Veterinary Hospital, Botucatu, Sao Paulo, Brazil, and subjected to a c-section due to uterine atony and fetuses no heartbeats on the ultrasound examination. The mother presented leukopenia of 1.6 thousand leukocytes, and there was no other information regarding previous clinical history. Among the offspring, four were stillborn, and five were born alive. On clinical examination, neonates weighed between 312 and 384 grams. Reflexes were present, and the newborn's body temperature was between 89.9 ºF and 96.4 ºF. Neonates also presented clinical signs of neonatal infection: omphalitis, abdomen, and extremities with cyanotic color, hematuria, and diarrhea (meconium). Complementary tests revealed leukopenia. The presence of alpha hemolytic streptococcus and Mannheimia haemolytica was revealed in the bacterial culture. The bacteria were sensitive to cephalosporins and penicillin on the antibiogram. Treatment for sepsis was instituted with the drug ceftriaxone, at a dose of 50 mg per kilogram, administered intravenous (jugular vein). Subsequently administered subcutaneous, every 12 hours, for seven days. Heated fluid therapy was performed, with Ringer lactate, at a dose of 4 ml per 100 grams of weight, intravenous. Heating measures were instituted. Blood plasma was also administered, at a dose of 2 mL per 100 grams of weight, administered subcutaneous, as a source of passive immunity. A maternal milk substitute was instituted, and lactation was discontinued since the mother was unable to nurse due to the infection. The mother was neutered during the c-section and treated with ceftriaxone (50 mg/kg). After seven days, the newborns presented normal clinical signs and no alterations in the hemogram. Early diagnosis and intervention were essential for the survival of these patients.

Keywords: neonatal infection, puppies, bacteria, newborn

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