Search results for: bacterial

Authors: Carlos R. Duharte Rodríguez, Ibrain Ceballo Acosta, Carmen B. Busoch Morlán, Angel Regueiro Gómez, Annet Martinez Hernández

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This work shows the designing and characterization of a prototype of laboratory incubator as support of research in Microbiology, in particular during studies of bacterial growth in biological samples, with the help of optic methods (Turbidimetry) and electrometric measurements of bioimpedance. It shows the results of simulation and experimentation of the design proposed for the canals of measurement of the variables: temperature and humidity, with a high linearity from the adequate selection of sensors and analogue components of every channel, controlled with help of a microcontroller AT89C51 (ATMEL) with adequate benefits for this type of application.

Keywords: microbiology, bacterial growth, incubation station, microorganisms

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Authors: O. Baghriche, A. Zertal, C. Pulgarin, J. Kiwi, R. Sanjines

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High-Power Impulse Magnetron Sputtering (HIPIMS) is a technology that belongs to the field of Ionized PVD of thin films. This study shows the first complete report on ultrathin TiO2/Ag nano-particulate films sputtered by highly ionized pulsed plasma magnetron sputtering (HIPIMS) leading to fast bacterial loss of viability. The Ag and the TiO2/Ag sputtered films induced complete Escherichia coli inactivation in the dark, which was not observed in the case of TiO2. When Ag was present, the bacterial inactivation was accelerated under low intensity solar simulated light and this has implications for a potential for a practical technology. The design, preparation, testing and surface characterization of these innovative films are described in this study. The HIPIMS sputtered composite films present an appreciable savings in metals compared to films obtained by conventional sputtering methods. HIPIMS sputtering induces a strong interaction with the rugous polyester 3-D structure due to the higher fraction of the Ag-ions (M+) attained in the magnetron chamber. The immiscibility of Ag and TiO2 in the TiO2/Ag films is shown by High Angular Dark Field (HAADF) microscopy. The ionization degree of the film forming species is significantly increased and film growth is assisted by an intense ion flux. Reports have revealed the significant enhancement of the film properties as the HIPIMS technology is used. However, a decrease of the deposition rate, as compared to the conventional DC magnetron sputtering Pulsed (DCMSP) process is commonly observed during HIPIMS.

Keywords: E. coli, HIPIMS, inactivation bacterial, sputtering

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Authors: Chidambaram Thamaraiselvan, Carlos Dosoretz

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This work will explore the influence of low voltage electric field, both alternating (AC) and direct (DC) currents, on biofouling control to highly electrically conductive self-supporting carbon nanotubes (CNT) membranes at conditions which encourage bacterial growth. A mutant strain of Pseudomonas putida S12 was used a model bacterium. The antibiofouling studies were performed with flow-through mode connecting an electric circuit in resistive mode. Major emphasis was placed on AC due to its ability of repulsing and inactivating bacteria. The observations indicate that an AC potential >1500 mV, 1 kHz frequency, 100 Ω external resistance on ground side and pulse wave above the offset (+0.45) almost completely prevented attachment of bacteria (>98.5%) and bacterial inactivation (95.3±2.5%). Findings suggest that at the conditions applied, direct electron transfer might be dominant in a decrease of cell viability. AC resulted more effective than DC, both in terms of biofouling reduction compared to cathodic DC and in terms of cell inactivation compared to anodic DC. This electrically polarized CNT membranes offer a viable antibiofouling strategy to hinder biofouling and simplify membrane care during filtration.

Keywords: bacterial attachment, biofouling control, low electric potential, water treatment

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Authors: Surasak Siripornadulsil, Nutt Poomai, Wilailak Siripornadulsil

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Due to a high ethanol demand, the approach for effective ethanol production is important and has been developed rapidly worldwide. Several agricultural wastes are highly abundant in celluloses and the effective cellulose enzymes do exist widely among microorganisms. Accordingly, the cellulose degradation using microbial cellulose to produce a low-cost substrate for ethanol production has attracted more attention. In this study, the cellulose producing bacterial strain has been isolated from rich straw and identified by 16S rDNA sequence analysis as Acinetobacter sp. KKU44. This strain is able to grow and exhibit the cellulose activity. The optimal temperature for its growth and cellulose production is 37 °C. The optimal temperature of bacterial cellulose activity is 60 °C. The cellulose enzyme from Acinetobacter sp. KKU44 is heat-tolerant enzyme. The bacterial culture of 36 h. showed highest cellulose activity at 120 U/mL when grown in LB medium containing 2% (w/v). The capability of Acinetobacter sp. KKU44 to grow in cellulosic agricultural wastes as a sole carbon source and exhibiting the high cellulose activity at high temperature suggested that this strain could be potentially developed further as a cellulose degrading strain for a production of low-cost substrate used in ethanol production.

Keywords: cellulose enzyme, bagasse, rice straw, rice husk, acinetobacter sp. KKU44

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Authors: Adjebli Ahmed, Messis Abdelaziz, Ayeche Riad, Tighilet Karim, Talbi Melissa, Smaili Yanis, Lehri Mokrane, Louardiane Mustapha

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Colorectal cancer is one of the most common types of cancer worldwide, and its incidence has been increasing in recent years. Data and fecal samples from colorectal cancer patients were collected at the Amizour Public Hospital's oncology department (Bejaia, Algeria). Microbiological and cohort study were conducted at the Biological Engineering of Cancers laboratory at the Faculty of Medicine of the University of Bejaia. All the data showed that patients aged between 50 and 70 years were the most affected by colorectal cancer, while the age categories of [30-40] and [40-50] were the least affected. Males were more likely to be at risk of contracting colorectal cancer than females. The most common types of colorectal cancer among the studied population were sigmoid cancer, rectal cancer, transverse colon cancer, and ascending colon cancer. The hereditary factor was found to be more dominant than other risk factors. Bacterial identification revealed the presence of certain pathogenic and opportunistic bacterial genera, such as E. coli, K. pneumoniae, Shigella sp, and Streptococcus group D. These results led us to conclude that dysbiosis of the intestinal microbiome is strongly present in colorectal cancer patients at the EPH of Amizour.

Keywords: microbiome, colorectal cancer, risk factors, bacterial identification

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Authors: Recep Keşli, Cengiz Demir, Onur Türkyılmaz

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Objective: The aim of this study was to determine the prevalence of Mycoplasma hominis and Ureaplasma urealyticum as the causative agents in men with non-gonococcal urethtritis, and anti-bacterial resistance rates. Methods: The Study was carried out in the two Medical Microbiology Laboratories belonging to: Konya Education and Research Hospital and ANS Practice and Research Hospital, Afyon Kocatepe University, between January 2012 and December 2015. Urethral samples were obtained from patients by using a swab. Mycoplasma hominis and Ureaplasma urealyticum were detected by using Mycoplasma IST-2 kit (bio-Mérieux, Marcy l'Étoile, France). Neisseria gonorrhoea was excluded by Gram staining and culture methods. Results: Of all the one hundred and eighty-eight male patients with urethritis, forty M. hominis and forty two U. urealyticum were detected. Resistance rates of M. hominis strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 5 %, 65 %, 25 %, 5 %, 80 %, 20 %, 20 %, 20 %, 5 %, respectively. Resistance rates of U. urealyticum strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 4.7 %, 66.6 %, 23.8 %, 4.75 %, 81 %, 19 %, 19 %, 4.7 % respectively. No resistance was detected against to josamycin, for both the strains. Conclusions: It was concluded that; ciprofloxacin and ofloxacin had the weakest; josamycin, doxycycline, and tetracycline had the strongest in vitro anti-bacterial activity, for treatment of the NGU. So josamycin, doxycycline, and tetracycline should be preferred as the first choice of anti-bacterial agents, for treatment of the patients with non-gonococcal male urethritis.

Keywords: antimicrobial resistance, Mycoplasma hominis, non-gonococcal urethritis, Ureaplasma urealyticum

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Authors: Ahmed F. Azmy, Amal E. Saafan, Tamer M. Essam, Magdy A. Amin, Shaban H. Ahmed

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Bacterial strains capable of degradation of malathion from the domestic sewage were isolated by an enrichment culture technique. Three bacterial strains were screened and identified as Acinetobacter baumannii (AFA), Pseudomonas aeruginosae (PS1),andPseudomonas mendocina (PS2) based on morphological, biochemical identification and 16S rRNA sequence analysis. Acinetobacter baumannii AFA was the most efficient malathion degrading bacterium, so used for further biodegradation study. AFA was able to grow in mineral salt medium (MSM) supplemented with malathion (100 mg/l) as a sole carbon source, and within 14 days, 84% of the initial dose was degraded by the isolate measured by high performance liquid chromatography. Strain AFA could also degrade other organophosphorus compounds including diazenon, chlorpyrifos and fenitrothion. The effect of different culture conditions on the degradation of malathion like inoculum density, other carbon or nitrogen sources, temperature and shaking were examined. Degradation of malathion and bacterial cell growth were accelerated when culture media were supplemented with yeast extract, glucose and citrate. The optimum conditions for malathion degradation by strain AFA were; an inoculum density of 1.5x 1012CFU/ml at 30°C with shaking. A specific polymerase chain reaction primers were designed manually using multiple sequence alignment of the corresponding carboxylesterase enzymes of Acinetobacter species. Sequencing result of amplified PCR product and phylogenetic analysis showed low degree of homology with the other carboxylesterase enzymes of Acinetobacter strains, so we suggested that this enzyme is a novel esterase enzyme. Isolated bacterial strains may have potential role for use in bioremediation of malathion contaminated.

Keywords: Acinetobacter baumannii, biodegradation, malathion, organophosphate pesticides

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Authors: A. Hamieh, Z. Olama, H. Holail

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Drinking Water Distribution Systems provide opportunities for microorganisms that enter the drinking water to develop into biofilms. Antimicrobial agents, mainly chlorine, are used to disinfect drinking water, however, there are not yet standardized disinfection strategies with reliable efficacy and development of novel anti-biofilm strategies is still of major concern. In the present study the ability of Lactobacillus acidophilus and Streptomyces sp. cell free supernatants to inhibit the bacterial biofilm formation in Drinking Water Distribution System in Lebanon was investigated. Treatment with cell free supernatants of Lactobacillus acidophilus and Streptomyces sp. at 20% concentration resulted in average biofilm inhibition (52.89 and 39.66% respectively). A preliminary investigation about the mode of action of biofilm inhibition revealed that cell free supernatants showed no bacteriostatic or bactericidal activity against all the tested isolates. Pre-coating wells with supernatants revealed that Lactobacillus acidophilus cell free supernatant inhibited average biofilm formation (62.53%) by altering the adhesion of bacterial isolates to the surface, preventing the initial attachment step, which is important for biofilm production.

Keywords: biofilm, cell free supernatant, distribution system, drinking water, lactobacillus acidophilus, streptomyces sp, adhesion

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Authors: Musadiq Idris, Sajjad Ali, Syed A. Khaliq, Umer Farooq

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Postmortem record of 217 captive ungulates including Black-buck (n=31), Chinkara (n=20), Hog deer (n=116), Spotted deer (n=35), Red Deer n=(04), and Rusa deer (n=11) submitted to the Veterinary Research Institute, Lahore, Pakistan was analyzed to determine the primary cause of mortality in these animals. The submissions included temporal distribution from Government wildlife captive farms, zoo, and private ownerships, over a three year period (2007-2009). The most common cause of death was found to be trauma (20.27%), followed by parasitic diseases (15.67%), bacterial diseases (11.98%), stillbirths (9.21%), snakebites (2.76%), gut affections (2.30%), neoplasia (1.38%) and starvation (0.92%). The exact cause of death could not be determined in 77 of 217 animals. Pneumonia (8.29%) and tuberculosis (3.69%) were the most common bacterial diseases. Analyses for parasitic infestation revealed tapeworms to be highest (11.05%), followed by roundworms (8.29%) and hemoparasitism (5.07%) (babesiosis and theileriosis). The mortality rate in young ungulates was lower as compared to adults (32.26% and 67.74%). Gender wise data presented higher mortality in females (55.30%) compared to males (44.70%). In conclusion, highest mortality factor in captive ungulates was trauma, followed by parasitic and bacterial infestations/infections of tapeworms and pneumonia, respectively. Furthermore, necropsies provided substantial information on etiology of death and other related epidemiological aspects.

Keywords: age, epidemiology, gender, mortality, ungulates

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Authors: Muhammad Awais Afzal, Lorena Tuchscherr De Hauschopp, Christian Hübner

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Introduction: Autophagy is an evolutionarily conserved lysosome-dependent degradation pathway, which can be induced by extrinsic and intrinsic stressors in living systems to adapt to fluctuating environmental conditions. In the context of inflammatory stress, autophagy contributes to the elimination of invading pathogens, the regulation of innate and adaptive immune mechanisms, and regulation of inflammasome activity as well as tissue damage repair. Lysosomes can be recycled from autolysosomes by the process of autophagic lysosome reformation (ALR), which depends on the presence of several proteins including Spatacsin. Thus ALR contributes to the replenishment of lysosomes that are available for fusion with autophagosomes in situations of increased autophagic turnover, e.g., during bacterial infections, inflammatory stress or sepsis. Objectives: We aimed to assess whether ALR plays a role for cell survival in an in-vitro bacterial infection model. Methods: Mouse embryonic fibroblasts (MEFs) were isolated from wild-type mice and Spatacsin (Spg11-/-) knockout mice. Wild-type MEFs and Spg11-/- MEFs were infected with Staphylococcus aureus (multiplication of infection (MOI) used was 10). After 8 and 16 hours of infection, cell viability was assessed on BD flow cytometer through propidium iodide intake. Bacterial intake by cells was also calculated by plating cell lysates on blood agar plates. Results: in-vitro infection of MEFs with Staphylococcus aureus showed a marked decrease of cell viability in ALR deficient Spatacsin knockout (Spg11-/-) MEFs after 16 hours of infection as compared to wild-type MEFs (n=3 independent experiments; p < 0.0001) although no difference was observed for bacterial intake by both genotypes. Conclusion: Suggesting that ALR is important for the defense of invading pathogens e.g. S. aureus, we observed a marked increase of cell death in an in-vitro infection model in cells with compromised ALR.

Keywords: autophagy, autophagic lysosome reformation, bacterial infections, Staphylococcus aureus

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

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

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

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Authors: Benjamin Castillo, Luis Pastenes, Fernando Cerdova

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The contamination of food by microbial agents is a common problem in the industry, especially regarding the elaboration of animal source products. Incorrect manipulation of the machinery or on the raw materials can cause a decrease in production or an epidemiological outbreak due to intoxication. In order to improve food product quality, different methods have been used to reduce or, at least, to slow down the growth of the pathogens, especially deteriorated, infectious or toxigenic bacteria. These methods are usually carried out under low temperatures and short processing time (abiotic agents), along with the application of antibacterial substances, such as bacteriocins (biotic agents). This, in a controlled and efficient way that fulfills the purpose of bacterial control without damaging the final product. Therefore, the objective of the present study is to design a secondary mathematical model that allows the prediction of both the biotic and abiotic factor impact associated with animal source food processing. In order to accomplish this objective, the authors propose a three-dimensional differential equation model, whose components are: bacterial growth, release, production and artificial incorporation of bacteriocins and changes in pH levels of the medium. These three dimensions are constantly being influenced by the temperature of the medium. Secondly, this model adapts to an idealized situation of cross-contamination animal source food processing, with the study agents being both the animal product and the contact surface. Thirdly, the stochastic simulations and the parametric sensibility analysis are compared with referential data. The main results obtained from the analysis and simulations of the mathematical model were to discover that, although bacterial growth can be stopped in lower temperatures, even lower ones are needed to eradicate it. However, this can be not only expensive, but counterproductive as well in terms of the quality of the raw materials and, on the other hand, higher temperatures accelerate bacterial growth. In other aspects, the use and efficiency of bacteriocins are an effective alternative in the short and medium terms. Moreover, an indicator of bacterial growth is a low-level pH, since lots of deteriorating bacteria are lactic acids. Lastly, the processing times are a secondary agent of concern when the rest of the aforementioned agents are under control. Our main conclusion is that when acclimating a mathematical model within the context of the industrial process, it can generate new tools that predict bacterial contamination, the impact of bacterial inhibition, and processing method times. In addition, the mathematical modeling proposed logistic input of broad application, which can be replicated on non-meat food products, other pathogens or even on contamination by crossed contact of allergen foods.

Keywords: bacteriocins, cross-contamination, mathematical model, temperature

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

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

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

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Authors: Uraib Sharaha, Guy Beck, Joseph Kapelushnik, Adam H. Agbaria, Itshak Lapidot, Shaul Mordechai, Ahmad Salman, Mahmoud Huleihel

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Infectious diseases cause a significant burden on the public health and the economic stability of societies all over the world for several centuries. A reliable detection of the causative agent of infection is not possible based on clinical features, since some of these infections have similar symptoms, including fever, sneezing, inflammation, vomiting, diarrhea, and fatigue. Moreover, physicians usually encounter difficulties in distinguishing between viral and bacterial infections based on symptoms. Therefore, there is an ongoing need for sensitive, specific, and rapid methods for identification of the etiology of the infection. This intricate issue perplex doctors and researchers since it has serious repercussions. In this study, we evaluated the potential of the mid-infrared spectroscopic method for rapid and reliable identification of bacterial and viral infections based on simple peripheral blood samples. Fourier transform infrared (FTIR) spectroscopy is considered a successful diagnostic method in the biological and medical fields. Many studies confirmed the great potential of the combination of FTIR spectroscopy and machine learning as a powerful diagnostic tool in medicine since it is a very sensitive method, which can detect and monitor the molecular and biochemical changes in biological samples. We believed that this method would play a major role in improving the health situation, raising the level of health in the community, and reducing the economic burdens in the health sector resulting from the indiscriminate use of antibiotics. We collected peripheral blood samples from young 364 patients, of which 93 were controls, 126 had bacterial infections, and 145 had viral infections, with ages lower than18 years old, limited to those who were diagnosed with fever-producing illness. Our preliminary results showed that it is possible to determine the infectious agent with high success rates of 82% for sensitivity and 80% for specificity, based on the WBC data.

Keywords: infectious diseases, (FTIR) spectroscopy, viral infections, bacterial infections.

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Authors: Chamilani Nikapitiya, Mahanama De Zoysa, Jehee Lee

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Most of the bacterial diseases are associated with high mortalities in aquaculture species and causing huge economic losses. Different approaches have been taken to prevent or control of bacterial diseases including use of vaccines, probiotics, chemotherapy, water quality management, etc. Antibiotics are widely applying as chemotherapy to control bacterial diseases, however, it has been shown that frequent use of antibiotics is favored to develop multi-drug resistance bacteria. Therefore, phages and phage encoded lytic proteins are known to be one of the most promising alternatives for antibiotics to avoid the emergence of antibiotic-resistant bacteria. We isolated and characterized the two lytic phages, namely pAh-1 and pAs-1 against pathogenic Aeromonas hydrophila and Aeromonas salmonicida, respectively. Morphological characteristics were analyzed by Transmission electron microscopy (TEM) and host strain specificities were tested with Aeromonas and other closely related bacterial strains. TEM analysis revealed that both pAh-1 and pAsm-1 are composed of an icosahedral head and a segmented tail, and we suggest that, they are new members of Myoviridae family. Genome sizes of isolated phages were estimated by restriction enzyme digestion of genomic DNA using selected endonucleases followed by agarose gel electrophoresis. Estimated genome size of pAh-1 and pAs-1 were approximately 64 Kbp and 120 Kbp, respectively. Both pAh-1 and pAs-1 have shown narrow host specificity. Moreover, protective effects of phage therapy against fish pathogenic A. hydrophila were investigated in zebrafish model. The survival rate was 40% higher when zebrafish received intra-peritoneal injection (i.p.) of pAh-1 were simultaneously challenge A. hydrophila (2 x 106 CFU/fish) compared to that without phage treatment. Overall results suggest that both pAh-1 and pAs-1 can be used as a potential phage therapy to control Aeromonas infections in aquaculture.

Keywords: Aeromonas infections, antibiotic resistance, bacteriophage, bio-control, lytic phage

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Authors: Thobela Conco, Sheena Kumari, Chika Nnadozie, Mahmoud Nasr, Thor A. Stenström, Mushal Ali, Arshad Ismail, Faizal Bux

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The discharge of antibiotics and its residues into the wastewater treatment plants (WWTP’s) create a conducive environment for the development of antibiotic resistant pathogens. This presents a risk of potential dissemination of antibiotic resistant pathogens and antibiotic resistance genes into the environment. It is, therefore, necessary to study the level of antibiotic resistance genes (ARG’s) among bacterial pathogens that proliferate in biological wastewater treatment systems. In the current study, metagenomic and meta-transcriptomic sequences of samples collected from the influents, secondary effluents and post chlorinated effluents of three wastewater treatment plants treating domestic and hospital effluents in Durban, South Africa, were analyzed for profiling of ARG’s among bacterial pathogens. Results show that a variety of ARG’s, mostly, aminoglycoside, β-lactamases, tetracycline and sulfonamide resistance genes were harbored by diverse bacterial genera found at different stages of treatment. A significant variation in diversity of pathogen and ARGs between the treatment plant was observed; however, treated final effluent samples from all three plants showed a significant reduction in bacterial pathogens and detected ARG’s. Both pre- and post-chlorinated samples showed the presence of mobile genetic elements (MGE’s), indicating the inefficiency of chlorination to remove of ARG’s integrated with MGE’s. In conclusion, the study showed the wastewater treatment plant efficiently caused the reduction and removal of certain ARG’s, even though the initial focus was the removal of biological nutrients.

Keywords: antibiotic resistance, mobile genetic elements, wastewater, wastewater treatment plants

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Authors: Zeina Bourhane, Anders Lanzen, Christine Cagnon, Olfa Ben Said, Cristiana Cravo-Laureau, Robert Duran

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The anthropogenic pressure in coastal areas increases dramatically with the exploitation of environmental resources. Biomonitoring coastal areas are crucial to determine the impact of pollutants on bacterial communities in soils and sediments since they provide important ecosystem services. However, relevant biomonitoring tools allowing fast determination of the ecological status are yet to be defined. Microbial ecology approaches provide useful information for developing such microbial monitoring tools reporting on the effect of environmental stressors. Chemical and microbial molecular approaches were combined in order to determine microbial bioindicators for assessing the ecological status of soil and river ecosystems around the Ichkeul Lake (Tunisia), an area highly impacted by human activities. Samples were collected along soil/river/lake continuums in three stations around the Ichkeul Lake influenced by different human activities at two seasons (summer and winter). Contaminant pressure indexes (PI), including PAHs (Polycyclic aromatic hydrocarbons), alkanes, and OCPs (Organochlorine pesticides) contents, showed significant differences in the contamination level between the stations with seasonal variation. Bacterial communities were characterized by 16S ribosomal RNAs (rRNA) gene metabarcoding. Although microgAMBI indexes, determined from the sequencing data, were in accordance with contaminant contents, they were not sufficient to fully explain the PI. Therefore, further microbial indicators are still to be defined. The comparison of bacterial communities revealed the specific microbial assemblage for soil, river, and lake sediments, which were significantly correlated with contaminant contents and PI. Such observation offers the possibility to define a relevant set of bioindicators for reporting the effects of human activities on the microbial community structure. Such bioindicators might constitute useful monitoring tools for the management of microbial communities in coastal areas.

Keywords: bacterial communities, biomonitoring, contamination, human impacts, microbial bioindicators

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Authors: Beatriz Suárez López, Gabriela Forman

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Kombucha (a symbiotic culture of bacteria and yeast) produces material capable of acquiring multiple shapes and textures that change significantly under different environment or temperature variations (e.g., when it is exposed to wet conditions), properties that may be explored in the scenic industry. This paper presents an analysis of its specific characteristics, exploring them as a non-conventional material for arts and performance. Costume Design uses surfaces as a powerful way of expression to represent concepts and stories; it may apply the unique features of nano bacterial cellulose (NBC) as assets in this artistic context. A mix of qualitative and quantitative (interventionist) methodology approaches were used -review of relevant literature to deepen knowledge on the research topic (crossing bibliography from different fields of studies: Biology, Art, Costume Design, etc.); as well as descriptive methods: laboratorial experiments, document quantities, observation to identify material properties and possibilities used to express a multiple narrative ideas, concepts and feelings. The results confirmed that NBC is an interactive and versatile material viable to be used in an alternative scenic context; its unique aesthetic and performative qualities, which change in contact to moisture, is a resource that can be used to show a visual and poetic impact on stage.

Keywords: biotechnological materials, contemporary dance, costume design, nano bacterial cellulose, performing arts

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Authors: Saleh Alkarri, Melinda Frame, Dimple Sharma, John Cairney, Lee Maddan, Jin H. Kim, Jonathan O. Rayner, Teresa M. Bergholz, Muhammad Rabnawaz

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Reported herein is an investigation of anti-bacterial and anti-virus properties, mode of action of Mg(OH)₂ and copper-infused Mg(OH)₂ nanoplatelets (NPs) on melt-compounded and thermally embossed polypropylene (PP) surfaces. The anti-viral activity for the NPs was studied in aqueous liquid suspensions against SARS-CoV-2, and the mode of action was investigated on neat NPs and PP samples that were thermally embossed with NPs. Anti-bacterial studies for melt-compounded NPs in PP confirmed approximately 1 log reduction of E. coli populations in 24 h, while for thermally embossed NPs, an 8 log reduction of E. coli populations was observed. In addition, the NPs exhibit anti-viral activity against SARS-CoV-2. Fluorescence microscopy revealed that reactive oxygen species (ROS) is the main mode of action through which Mg(OH)₂ and Cu-Infused Mg(OH)₂act against microbes. Plastics with anti-microbial surfaces from where biocides are non-leachable are highly desirable. This work provides a general fabrication strategy for developing anti-microbial plastic surfaces.

Keywords: anti-microbial activity, E. coli K-12 MG1655, anti-viral activity, SARS-CoV-2, copper-infused magnesium hydroxide, non-leachable, ROS, compounding, surface embossing, dyes

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Authors: Mhuji Kilonzo, Patrick Ndakidemi, Musa Chacha

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Objective: To evaluate antibacterial activity from four selected medicinal plants namely Mystroxylon aethiopicum, Lonchocarpus capassa, Albizia anthelmentica and Myrica salicifolia used for management of bacterial infection in Tanzania. Methods: Minimum Inhibitory Concentration (MIC) of plants extracts against the tested bacterial species was determined by using 96 wells microdilution method. In this method, 50 μL of nutrient broth were loaded in each well followed by 50 μL of extract (100 mg/mL) to make a final volume of 100 μL. Subsequently, 50 μL were transferred from first rows of each well to the second rows and the process was repeated down the columns to the last wells from which 50 μL were discarded. Thereafter, 50 μL of the selected bacterial suspension were added to each well thus making a final volume of 100 μL. The lowest concentration which showed no bacterial growth was considered as MIC. Results: It was revealed that L. capassa leaf ethyl acetate extract exhibited antibacterial activity against Salmonella kisarawe and Salmonella typhi with MIC values of 0.39 and 0.781 mg/mL respectively. Likewise, L. capassa root bark ethyl acetate extracts inhibited growth of S. typhi and E. coli with MIC values of 0.39 and 0.781 mg/mL respectively. The M. aethiopicum leaf and root bark chloroform extracts displayed antibacterial activity against S. kisarawe and S. typhi respectively with MIC value of 0.781 mg/mL. The M. salicifolia stem bark ethyl acetate exhibited antibacterial activity against P. aeruginosa with MIC value of 0.39 mg/mL whereas the methanolic stem and root bark of the same plant inhibited the growth of Proteus mirabilis and Klebsiella pneumoniae with MIC value of 0.781 mg/mL. Conclusion: It was concluded that M. aethiopicum, L. capassa, A. anthelmentica and M. salicifolia are potential source of antibacterial agents. Further studies to establish structures of antibacterial and evaluate active ingredients are recommended.

Keywords: Albizia anthelmentica, Lonchocarpus capassa, Mystroxylon aethiopicum, Myrica salicifolia

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Authors: F. A. Ogundolie, F. Okogue, D. V. Adegunloye

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Greywater samples were obtained from three restaurants in the Federal University of Technology; Akure coded SSR, MGR and GGR. Fungi isolates obtained include Rhizopus stolonifer, Aspergillus niger, Mucor mucedo, Aspergillus flavus, Saccharomyces cerevisiae. Of these fungi isolates obtained, R. stolonifer, A. niger and A. flavus showed significant degradation ability on grey water and was used for this research. A simple bioreactor was constructed using biodegradation process in purification of waste water samples. Waste water undergoes primary treatment; secondary treatment involves the introduction of the isolated organisms into the waste water sample and the tertiary treatment which involved the use of filter candle and the sand bed filtration process to achieve the end product without the use of chemicals. A. niger brought about significant reduction in both the bacterial load and the fungi load of the greywater samples of the three respective restaurants with a reduction of (1.29 × 108 to 1.57 × 102 cfu/ml; 1.04 × 108 to 1.12 × 102 cfu/ml and 1.72 × 108 to 1.60 × 102 cfu/ml) for bacterial load in SSR, MGR and GGR respectively. Reduction of 2.01 × 104 to 1.2 × 101; 1.72 × 104 to 1.1 × 101, and 2.50 × 104 to 1.5 × 101 in fungi load from SSR, MGR and GGR respectively. Result of degradation of these selected waste water by the fungi showed that A. niger was probably more potent in the degradation of organic matter and hence, A. niger could be used in the treatment of wastewater.

Keywords: Aspergillus niger, greywater, bacterial, fungi, microbial load, bioreactor, biodegradation, purification, organic matter and filtration

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Authors: Yantao Li, Jun Zheng

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Food poison caused by consumption of contaminated food, especially seafood, is one of most serious public health threats worldwide. Vibrio parahaemolyticus is emerging bacterial pathogen and the leading cause of human gastroenteritis associated with food poison, especially in the southern coastal region of China. To successfully cause disease in host, bacterial pathogens need to overcome the host-derived stresses encountered during infection. One of the toxic chemical species elaborated by the host is nitric oxide (NO). NO is generated by acidified nitrite in the stomach and by enzymes of the inducible NO synthase (iNOS) in the host cell, and is toxic to bacteria. Bacterial pathogens have evolved some mechanisms to battle with this toxic stress. Such mechanisms include genes to sense NO produced from immune system and activate others to detoxify NO toxicity, and genes to repair the damage caused by toxic reactive nitrogen species (RNS) generated during NO toxic stress. However, little is known about the NO resistance in V. parahaemolyticus. In this study, a transposon coupled with next generation sequencing (Tn-seq) technology will be utilized to identify genes for NO resistance in V. parahaemolyticus. Our strategy will include construction the saturating transposon insertion library, transposon library challenging with NO, next generation sequencing (NGS), bioinformatics analysis and verification of the identified genes in vitro and in vivo.

Keywords: vibrio parahaemolyticus, nitric oxide, tn-seq, virulence

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Authors: Sofia Papadimitriou

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INTRODUCTION: The differential diagnosis between acute viral and bacterial infections is an important cost-effectiveness parameter at the stage of the treatment process in order to achieve the maximum benefits in therapeutic intervention by combining the minimum cost to ensure the proper use of antibiotics.The discovery of sensitive and robust molecular diagnostic tests in response to the role of the host in infections has enhanced the accurate diagnosis and differentiation of infections. METHOD: The study used a sample of six independent blood samples (total=756) which are associated with human proteins-proteins, each of which at the transcription stage expresses a different response in the host network between viral and bacterial infections.Τhe individual blood samples are subjected to a sequence of computer filters that identify a gene panel corresponding to an autonomous diagnostic score. The data set and the correspondence of the gene panel to the diagnostic patents a new Bangalore -Viral Bacterial (BL-VB). FINDING: We use a biomarker based on the blood of 10 genes(Panel-VB) that are an important prognostic value for the detection of viruses from bacterial infections with a weighted average AUROC of 0.97(95% CL:0.96-0.99) in eleven independent samples (sets n=898). We discovered a base with a patient score (VB 10 ) according to the table, which is a significant diagnostic value with a weighted average of AUROC 0.94(95% CL: 0.91-0.98) in 2996 patient samples from 56 public sets of data from 19 different countries. We also studied VB 10 in a new cohort of South India (BL-VB,n=56) and found 97% accuracy in confirmed cases of viral and bacterial infections. We found that VB 10 (a)accurately identifies the type of infection even in unspecified cases negative to the culture (b) shows its clinical condition recovery and (c) applies to all age groups, covering a wide range of acute bacterial and viral infectious, including non-specific pathogens. We applied our VB 10 rating to publicly available COVID 19 data and found that our rating diagnosed viral infection in patient samples. RESULTS: Τhe results of the study showed the diagnostic power of the biomarker VB 10 as a diagnostic test for the accurate diagnosis of acute infections in recovery conditions. We look forward to helping you make clinical decisions about prescribing antibiotics and integrating them into your policies management of antibiotic stewardship efforts. CONCLUSIONS: Overall, we are developing a new property of the RNA-based biomarker and a new blood test to differentiate between viral and bacterial infections to assist a physician in designing the optimal treatment regimen to contribute to the proper use of antibiotics and reduce the burden on antimicrobial resistance, AMR.

Keywords: acute infections, antimicrobial resistance, biomarker, blood transcriptome, systems biology, classifier diagnostic score

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Authors: Raana Babadi Fathipour

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Biofilms, those intricate structures of microbial aggregation that emerge as microorganisms adhere to animate or inanimate surfaces, possess an innate capacity to shield their inhabitants from adversities within the environment whilst fortifying their endurance against antimicrobial agents. This remarkable aspect facilitates the persistence and virulence of said microorganisms, establishing biofilm formation as an integral component of bacterial survival mechanisms. However, should foodborne pathogens adopt this mode of existence, the potentiality for foodborne disease infections becomes alarmingly intensified—an alarming prospect that harbors significant public health hazards and engenders deleterious economic ramifications. Thus, due to these consequences lurking on the horizon, extensive research concentrating upon comprehending biofilms and devising efficacious removal strategies assumes a position imbued with paramount importance within the realm of the food industry. The problem of food waste resulting from spoilage in the food industry continues to present a widespread challenge to both environmental sustainability and the security of our food supplies. In this comprehensive analysis, we delve into the formation of bacterial biofilms, highlighting the specific issues they pose within the realm of food production. Additionally, we provide an overview of various types of common foodborne pathogens that tend to thrive in these biofilms. Furthermore, we summarize existing strategies aimed at tackling or managing detrimental bacterial biofilm growth. We also introduce contemporary approaches that show promise in terms of controlling this issue and highlight their potential for further advancement. Ultimately, our focus lies on outlining prospects for future development as they pertain specifically to combatting bacterial biofilms within the field.

Keywords: foodborne pathogens, food safety, biofilm, resistance, quorum-sensing

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Authors: Sungging Pintowantoro, Yuli Setiyorini, Rochman Rochim, Agung Purniawan

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The bacterial infections are frequent and undesired occurrences after bone fracture treatment. One approach to reduce the incidence of bone fracture infection is the additional of microbial agents into bone cement. In this study, the synthesis of bone cement from re-cycles biowaste was successfully conducted completed with anti-bacterial function. The re-cycle of biowaste using microwave assisted was done in our previous studies in order to produce some of powder (calcium carbonate, carbonated-hydroxyapatite and chitosan). The ratio of these powder combined with methylmethacrylate (MMA) as the matrix in bone cement were investigated using XRD, FTIR, SEM-EDX, hardness test and anti-bacterial test, respectively. From the XRD, FTIR and EDX were resulted the formation of carbonated-hydroxyapatite, calcium carbonate and chitosan. The morphology was revealed porous structure both C2H3K1L and C2H1K3L, respectively. The antibacterial activity was tested against Staphylococcus aureus (S. aureus) for 24 hours. The inhibition of S. aureus was clearly shown, the hollow zone was resulted in various distance 14.2mm, 7.5mm, and 7.7mm, respectively. The hardness test was depicted in various results, however, C2H1K3L can be achived 36.84HV which is closed to dry cancelous bone 35HV. In general, this study results was promising materials to use as bone cement materials.

Keywords: biomaterials, biowaste recycling, materials processing, microwave processing

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Authors: Sayaka Ono, Ryutaro Imai, Tomoko Ehara, Tetsuya Matsumoto, Hajime Matsumura

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Background: Wound pH affects a number of important factors in wound healing. We previously measured the pH value of the exudates collected from second-degree burns and found that the increase in pH was observed in the burn wounds in which colonized by Staphylococcus spp., and the increase in pH was evident prior to the clinical findings of local infection. To investigate the relationship between the changes of pH value and bacterial growth, we performed in vitro study using Pseudomonas aeruginosa and liquid medium as a locally infected wound equivalent model. Methods: Pseudomonas aeruginosa standard strain (ATCCR 10145TM) was cultured at 37 °C environment in Luria Broth Miller medium. The absorbance rate which means the amount of bacteria was measured by a microplate reader 2300EnSpireTM). The pH was measured using pH-indicator strips (MColorpHastTM). The statistical analysis was performed using the product-moment correlation coefficient of Pearson's. Results: The absorbance rate and pH value were increased along with culture period. There was a positive correlation between pH value and absorbance rate (n = 27, Pearson's r = 0.985). Moreover, there was a positive correlation between pH value and the culture period (n = 18, Pearson's r = 0.901). The bacteria was well growth in the media from pH 6.6 to pH 8.0 and the pH of culture media converged at 8 -9 along with the bacterial growth. Conclusion: From these results, we conclude that pH value of the wound is correlated with the number of viable bacteria and bacterial growth periods.

Keywords: colonization, potential of hydrogen, Pseudomonas aeruginosa, wound

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Authors: Muhammad Shahzad Tufail, Iram Liaqat, Umer Sohail Meer, Muhammad Ishtaiq, Muhammad Sattar

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Nanotechnology is a vibrant field with numerous applications in many different branches of science and technology. Several methods are used to synthesize nanoparticles (NPs), which have multiple range of applications. Comparatively, the biogenic synthesis of NPs is a more economical and environmentally favourable method than the traditional chemical method. The current study aims to synthesize biogenically silver nanoparticles (AgNPs) using bacterial isolates. Four bacterial strains Escherichia coli (MT448673), Pseudomonas aeruginosa (MN900691), Bacillus subtilis (MN900684) and Bacillus licheniformis (MN900686) were used for the synthesis of AgNPs from silver nitrate (AgNO3) solution. The biofilm time kinetics of four bacterial isolates (P. aeruginosa, E. coli, B. licheniformis and B. subtilis) was analysed by incubating bacterial cultures at 37◦C in test tubes over a period of different time intervals i.e., 2, 3, 5 and 7 days following crystal violet staining method. All the four strains had ability to form strong biofilms between 48 to 72 hours of incubation. Two strains (B. subtilis and B. licheniformis) formed significant (p < 0.05) biofilm after 3 days of incubation period. The other two strains (E. coli and P. aeruginosa) showed strong biofilm formation after 2 days of incubation. Next, the antibiofilm activity of biogenically synthesized AgNPs (10 - 100 µgmL-1) was analysed against biofilm forming human pathogenic bacteria. Findings of the work revealed that 60-90% inhibition was observed at 60 µgmL-1 of AgNPs, while maximum inhibition (i.e.,100%) was found at highest concentration (90 µgmL-1). It was evident that highly significant (p < 0.05) decrease in biofilm formation was observed with increasing concentration of AgNPs.

Keywords: antibiofilm, biofilm formation, nanotechnology, pathogenic bacteria, silver nanoparticles

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Authors: Cho Achidi

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Tomato is a crucial food crop significantly contributes to global food and nutrition security. However, postharvest losses severely limit its role. Therefore, it is necessary to develop sustainable strategies to minimize these losses and improve the shelf-life of tomato fruits. One of the major concerns is bacterial infections, particularly by faecal coliform bacteria, which can cause food poisoning and illnesses like diarrhoea and dysentery. This study seeks to identify the presence of coliform bacteria on tomato fruits in fields and markets in Muea, Buea Municipality. The study also evaluated different management strategies to reduce the bacterial incidence and load on tomato fruits. A total of 200 fruits were sampled for both the coliform survey and shelf-life analysis. Ten farmers and traders provided samples, including asymptomatic and symptomatic tomato fruits. The samples designated for shelf-life analysis were treated with Aquatab, warm water, lemon, and onion. The results indicated that out of the 80 symptomatic samples collected, 12.5% contained faecal and total coliform species. Among the ten farms sampled, 14% were infected with coliform bacteria, with the highest infestation rate of 60% recorded in field 4. Furthermore, 15% of the asymptomatic tomato fruits were found to be infected by coliform bacteria. Regarding the management strategies, Aquatabs exhibited the highest efficacy in reducing the incidence of coliform bacteria on tomato fruits, followed by onion and lemon extracts. Although hot water treatment effectively removed bacteria from the fruits, damaging the cell wall negatively affected their shelf-life. Overall, this study emphasizes the severity of coliform bacterial pathogens in the Muea area, particularly their occurrence on asymptomatic tomatoes, which poses a significant concern for plant quarantine services. It also demonstrates potential options for mitigating this bacterial challenge.

Keywords: tomato, shelf-life analysis, food and nutrition security, coliform bbacteria

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Authors: Francis Gyapong, Denis Yar

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The use of cell phones has become an indispensable tool in the hospital's settings. Cell phones are used in hospitals without restrictions regardless of their unknown microbial load. However, the indiscriminate use of mobile devices, especially at health facilities, can act as a vehicle for transmitting pathogenic bacteria and other microorganisms. These potential pathogens become exogenous sources of infection for the patients and are also a potential health hazard for self and as well as family members. These are a growing problem in many health care institutions. Innovations in mobile communication have led to better patient care in diabetes, asthma, and increased in vaccine uptake via SMS. Notwithstanding, the use of cell phones can be a great potential source for nosocomial infections. Many studies reported heavy microbial contamination of cell phones among healthcare workers and communities. However, limited studies have been reported in our region on bacterial contamination on cell phones among healthcare workers. This study assessed microbial contamination of cell phones of health care workers (HCWs) at the Mampong Municipal Government Hospital (MMGH), Ghana. A cross-sectional design was used to characterize bacterial microflora on cell phones of HCWs at the MMGH. A total of thirty-five (35) swab samples of cell phones of HCWs at the Laboratory, Dental Unit, Children’s Ward, Theater and Male ward were randomly collected for laboratory examinations. A suspension of the swab samples was each streak on blood and MacConkey agar and incubated at 37℃ for 48 hours. Bacterial isolates were identified using appropriate laboratory and biochemical tests. Kirby-Bauer disc diffusion method was used to determine the antimicrobial sensitivity tests of the isolates. Data analysis was performed using SPSS version 16. All mobile phones sampled were contaminated with one or more bacterial isolates. Cell phones from the Male ward, Dental Unit, Laboratory, Theatre and Children’s ward had at least three different bacterial isolates; 85.7%, 71.4%, 57.1% and 28.6% for both Theater and Children’s ward respectively. Bacterial contaminants identified were Staphylococcus epidermidis (37%), Staphylococcus aureus (26%), E. coli (20%), Bacillus spp. (11%) and Klebsiella spp. (6 %). Except for the Children ward, E. coli was isolated at all study sites and predominant (42.9%) at the Dental Unit while Klebsiella spp. (28.6%) was only isolated at the Children’s ward. Antibiotic sensitivity testing of Staphylococcus aureus indicated that they were highly sensitive to cephalexin (89%) tetracycline (80%), gentamycin (75%), lincomycin (70%), ciprofloxacin (67%) and highly resistant to ampicillin (75%). Some of these bacteria isolated are potential pathogens and their presence on cell phones of HCWs could be transmitted to patients and their families. Hence strict hand washing before and after every contact with patient and phone be enforced to reduce the risk of nosocomial infections.

Keywords: mobile phones, bacterial contamination, patients, MMGH

Procedia PDF Downloads 69