Search results for: microbial isolates

Authors: Prachi Choudhary, Jai Gopal Sharma

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Microbial cultures determine the type of organism, its abundance in the tested sample, or both. It is one of the primary diagnostic methods of microbiology. It is used to determine the cause of infectious disease by letting the agent multiply in a predetermined medium. Different bacteria produce colonies that may be very distinct from the bacterial species that produced them. To culture any pathogen or microorganism, we should first know about the types of media used in microbiology for culturing. Sometimes sub culturing is also done in various microorganisms if some mixed growth is seen in culture. Nearly 3 types of culture media based on consistency – solid, semi-solid, and liquid (broth) media; are further explained in the report. Then, The Five I's approach is a method for locating, growing, observing, and characterizing microorganisms, including inoculation and incubation. Isolation, inspection, and identification. For identification of bacteria, we have to culture the sample like urine, sputum, blood, etc., on suitable media; there are different methods of culturing the bacteria or microbe like pour plate method, streak plate method, swabbing by needle, pipetting, inoculation by loop, spreading by spreader, etc. After this, we see the bacterial growth after incubation of 24 hours, then according to the growth of bacteria antibiotics susceptibility test is conducted; this is done for sensitive antibiotics or resistance to that bacteria, and also for knowing the name of bacteria. Various methods like the dilution method, disk diffusion method, E test, etc., do antibiotics susceptibility tests. After that, various medicines are provided to the patients according to antibiotic sensitivity and resistance.

Keywords: inoculation, incubation, isolation, antibiotics suspectibility test, characterizing

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Authors: Tomasz Borowski, Dawid Sołoducha, Rafał Rakoczy, Marian Kordas

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The application of magnetic field is essential for a wide range of technologies or processes (i.e., magnetic hyperthermia, bioprocessing). From the practical point of view, bioprocess control is often limited to the regulation of temperature at constant values favourable to microbial growth. The main aim of this study is to determine the effect of various types of electromagnetic fields (i.e., static or alternating) on the heat transfer in a self-designed magnetically assisted reactor. The experimental set-up is equipped with a measuring instrument which controlled the temperature of the liquid inside the container and supervised the real-time acquisition of all the experimental data coming from the sensors. Temperature signals are also sampled from generator of magnetic field. The obtained temperature profiles were mathematically described and analyzed. The parameters characterizing the response to a step input of a first-order dynamic system were obtained and discussed. For example, the higher values of the time constant means slow signal (in this case, temperature) increase. After the period equal to about five-time constants, the sample temperature nearly reached the asymptotic value. This dynamical analysis allowed us to understand the heating effect under the action of various types of electromagnetic fields. Moreover, the proposed mathematical description can be used to compare the influence of different types of magnetic fields on heat transfer operations.

Keywords: heat transfer, magnetically assisted reactor, dynamical analysis, transient function

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Authors: Pampita Chakraborty, Sukumar Mukherjee

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This study was done to determine the prevalence of nosocomial infections in the ICU and to identify the common microorganisms causing these infections and their antimicrobial sensitivity pattern. Nosocomial infection or hospital-acquired infection is a localized or a systemic condition resulting from an adverse reaction to the presence of infectious agents. Nosocomial infections are not present or incubating when the patient is admitted to hospital or other health care facility. They are caused by pathogens that easily spread through the body. Many hospitalized patients have compromised immune systems, so they are less able to fight off infections. These infections occur worldwide, both in the developed and developing the world. They are a significant burden to patients and public health. They are a major cause of death and increased morbidity in hospitalized patients, which is a matter of serious concern today. This study was done during the period of one year (2012-2013) in the ICU of the tertiary care hospital in eastern India. Prevalence of nosocomial infection was determined; site of infection and the pattern of microorganisms were identified along with the assessment of antibiotic susceptibility profile. Patients who developed an infection after 48 hours of admission to the ICU were included in the study. A total of 324 ICU patients were analyzed, of these 79 patients were found to have developed a nosocomial infection (24.3% prevalence). Urinary tract infection was found to be more predominant followed by respiratory tract infection and soft tissue infection. The most frequently isolated microorganism was E. coli, Pseudomonas aeruginosa, Klebsiella pneumoniae followed by other organisms respectively. Antibiotic susceptibility test of these isolates was done against commonly used antibiotics. Patients admitted to the ICU are especially susceptible to nosocomial infections. Despite adequate antimicrobial treatment, nosocomial ICU infections can significantly affect ICU stay and can cause an increase in patient’s morbidity and mortality. Adherence to infection protocol, proper monitoring and the judicious use of antibiotics are important in preventing such infections on a regular basis.

Keywords: antibiotic susceptibility, intensive care unit, nosocomial infection, nosocomial pathogen

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Authors: Salma H. Abu Hafsa, A. Mendonca, B. Brehm-Stecher, A. A. Hassan, S. A. Ibrahim

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Enterococci are important inhabitants of the animal intestine and are widely used in probiotic products. A probiotic strain is expected to possess several desirable properties in order to exert beneficial effects. Therefore, the objective of this study was to isolate and characterize strains of Enterococcus sp. from chicken cecal and fecal samples to determine potential probiotic properties. Enterococci were isolated from thirty one chicken cecal and fecal samples collected from a local farm. In vitro studies were performed to assess antibacterial activity (using agar well diffusion and cell free supernatant broth technique against Salmonella enterica serotype Enteritidis), susceptibility to antibiotics (amoxycillin, cotrimoxazole, chloramphenicol, cefuroxime, ceftriaxone, ciprofloxacin, and nalidixic acid), survival in acidic conditions, resistance to bile salts, and their survival during simulated gastric juice conditions at pH 2.5. Isolates were identified using biochemical and molecular assays (API 50 CHL, and API ZYM kits followed by 16S rDNA gene sequence analysis). Two strains were identified, of which, Enteroccocus faecium was capable of inhibiting the growth of S. enteritidis and was susceptible to a wide range of antibiotics. In addition, the isolated strain exhibited significant resistance under highly acidic conditions (pH=2.5) for 8 hours and survived well in bile salt at 0.2% for 24 hours and showing ability to survive in the presence of simulated gastric juice at pH 2.5. Based on these results, the E. faecium isolate fulfills some of the criteria to be considered as a probiotic strain and therefore, could be used as a feed additive with good potential for controlling S. enteritidis in chickens. However, in vivo studies are needed to determine the safety of the strain.

Keywords: acid tolerance, antimicrobial activity, Enterococcus faecium, probiotic

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Authors: Nurul Hidayati

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Increasing public awareness of environmental pollution influences the search and development of technologies that help in clean up of organic and inorganic contaminants such as metals. Sludge waste of paper industries as toxic and hazardous material from specific source contains Pb, Zn, and Cu metal from waste soluble ink. An alternative and eco-friendly method of remediation technology is the use of biosurfactants and biosurfactant-producing microorganisms. Soil washing is among the methods available to remove heavy metal from sediments. The purpose of this research is to study effectiveness of biosurfactant with concentration = CMC for the removal of heavy metals, lead, zinc and copper in batch washing test under four different biosurfactant production by microbial origin. Pseudomonas putida T1(8), Bacillus subtilis 3K, Acinetobacter sp, and Actinobacillus sp was grown on mineral salt medium that had been already added with 2% concentration of molasses that it is a low cost application. The samples were kept in a shaker 120 rpm at room temperature for 3 days. Supernatants and sediments of sludge were separated by using a centrifuge and samples from supernatants were measured by atomic absorption spectrophotometer. The highest removal of Pb was up to 14,04% by Acinetobacter sp. Biosurfactant of Pseudomonas putida T1(8) have the highest removal for Zn and Cu up to 6,5% and 2,01% respectively. Biosurfactants have a role for removal process of the metals, including wetting, contact of biosurfactant to the surface of the sediments and detachment of the metals from the sediment. Biosurfactant has proven its ability as a washing agent in heavy metals removal from sediments, but more research is needed to optimize the process of removal heavy metals.

Keywords: biosurfactant, removal of heavy metals, sludge waste, paper industries

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Authors: Anika Pallapothu

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Periodontitis, a severe gum disease, poses a significant threat to the integrity of bone and soft tissues supporting teeth, primarily initiated by bacterial accumulation around the gum line. Conventional treatments like scaling/root planning and plaque removal are widely employed, but integrating modern technologies such as nanotechnology holds promise for innovative therapeutic approaches. This study explores the utilization of silver nanoparticles encapsulated within cellulose nanofiber (CNF) and mesoporous bioactive glass hydrogel matrices for periodontitis management. Silver nanoparticles exhibit potent antimicrobial properties by disrupting microbial cell membranes, inducing reactive oxygen species (ROS) generation, and interfering with vital cellular processes like ATP production and nucleic acid synthesis. Mesoporous bioactive glass, renowned for its high surface area, osteoconductive, and bioactivity, presents a favorable platform for pharmaceutical applications. Incorporating CNF enhances the properties of the hydrogel due to its biocompatibility, biodegradability, and water absorption capacity. The proposed composite material is anticipated to exert beneficial effects in periodontitis treatment by demonstrating antibacterial and anti-inflammatory activities, offering a promising avenue for future therapeutic interventions.

Keywords: periodontitis, cellulose nanofibers, silver nanoparticles, mesoporous bioactive glass, antibacterial activity, anti-inflammatory activity

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Authors: Abdul Walusansa

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The aim of this study was to assess the level of microbial pollution along Nakibiso stream. The study was carried out in polluted waters of Nakibiso stream, originating from Mbale municipality and running through ADRA Estates to Namatala Wetlands in Eastern Uganda. Four sites along the stream were selected basing on the activities of their vicinity. A total of 120 samples were collected in sterile bottles from the four sampling locations of the stream during the wet and dry seasons of the year 2011. The samples were taken to the National water and Sewerage Cooperation Laboratory for Analysis. Membrane filter technique was used to test for Erischerichia coli. Nitrogen, Phosphorus, pH, dissolved oxygen, electrical conductivity, total suspended solids, turbidity and temperature were also measured. Results for Nitrogen and Phosphorus for sites; 1, 2, 3 and 4 were 1.8, 8.8, 7.7 and 13.8 NH4-N mg/L; and 1.8, 2.1, 1.8 and 2.3 PO4-P mg/L respectively. Basing on these results, it was estimated that farmers use 115 and 24 Kg/acre of Nitrogen and Phosphorus respectively per month. Taking results for Nitrogen, the same amount of Nutrients in artificial fertilizers would cost $ 88. This shows that reuse of wastewater has a potential in terms of nutrients. The results for E. coli for sites 1, 2, 3 and 4 were 1.1 X 107, 9.1 X 105, 7.4 X 105, and 3.4 X 105 respectively. E. coli hence decreased downstream with statistically significant variations between sites 1 and 4. Site 1 had the highest mean E.coli counts. The bacterial contamination was significantly higher during the dry season when more water was needed for irrigation. Although the water had the potential for reuse in farming, bacterial contamination during both seasons was higher than 103 FC/100ml recommended by WHO for unrestricted Agriculture.

Keywords: E. coli, nitrogen, phosphorus, water reuse, waste water

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Authors: Adrian Ionut Nicoara, Denisa Ionela Ene, Alina Maria Holban, Cristina Busuioc

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At present, the development of materials with biomedical applications is a domain of interest that will produce a full series of benefits in engineering and medicine. In this sense, it is required to use a natural material, and this paper is focused on the development of a composite material based on bacterial cellulose – hydroxyapatite and silver nanoparticles with applications in hard tissue. Bacterial cellulose own features like biocompatibility, non-toxicity character and flexibility. Moreover, the bacterial cellulose can be conjugated with different forms of active silver to possess antimicrobial activity. Hydroxyapatite is well known that can mimic at a significant level the activity of the initial bone. The material was synthesized by using an ultrasound probe and finally characterized by several methods. Thereby, the morphological properties were analyzed by using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). Because the synthesized material has medical application in restore the tissue and to fight against microbial invasion, the samples were tested from the biological point of view by evaluating the biodegradability in phosphate-buffered saline (PBS) and simulated body fluid (SBF) and moreover the antimicrobial effect was performed on Gram-positive bacterium Staphylococcus aureus, Gram-negative bacterium Escherichia coli, and fungi Candida albicans. The results reveal that the obtained material has specific characteristics for bone regeneration.

Keywords: bacterial cellulose, biomaterials, hydroxyapatite, scaffolds materials

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Authors: A. Mohammed, A. Babatunde

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This study investigated the integrated removal of heavy metals, organic matter and nitrogen from landfill leachate using a novel laboratory scale constructed wetland system. The main objectives of this study were: (i) to assess the overall effectiveness of the constructed wetland system for treating landfill leachate; (ii) to examine the interactions and impact of key leachate constituents (heavy metals, organic matter and nitrogen) on the overall removal dynamics and efficiency. The constructed wetland system consisted of four stages operated in tidal flow and anoxic conditions. Results obtained from 215 days of operation have demonstrated extraordinary heavy metals removal up to 100%. Analysis of the physico- chemical data reveal that the controlling factors for metals removal were the anoxic condition and the use of the novel media (dewatered ferric sludge which is a by-product of drinking water treatment process) as the main substrate in the constructed wetland system. Results show that the use of the ferric sludge enhanced heavy metals removal and brought more flexibility to simultaneous nitrification and denitrification which occurs within the microbial flocs. Furthermore, COD and NH₄-N were effectively removed in the system and this coincided with enhanced aeration in the 2nd and 3rd stages of the constructed wetland system. Overall, the results demonstrated that the ferric dewatered sludge constructed wetland system would be an effective solution for integrated removal of pollutants from landfill leachates.

Keywords: constructed wetland, ferric dewatered sludge, heavy metals, landfill leachate

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Authors: Neha Sharma, Kanthi K. Kondepudi, Naveen Gupta

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Phytases are phytate specific phosphatases catalyzing the step-wise dephosphorylation of phytate, which acts as an anti-nutritional factor in food due to its strong binding capacity to minerals. In recent years microbial phytases have been explored for improving nutritional quality of food. But the major limitation is acceptability of phytases from these microorganisms. Therefore, efforts are being made to isolate organisms which are generally regarded as safe for human consumption such as Lactic Acid Bacteria (LAB). Phytases from these organisms will have an edge over other phytase sources due to its probiotic attributes. Only few LAB have been reported to give phytase activity that too is generally seen as intracellular. LAB producing extracellular phytase will be more useful as it can degrade phytate more effectively. Moreover, enzyme from such isolate will have application in food processing also. Only few species of Lactobacillus producing extracellular phytase have been reported so far. This study reports the isolation of a probiotic strain of Lactobacillus fermentum spp KA1 which produces extracellular phytase. Conditions for the optimal production of phytase have been optimized and the enzyme production resulted in an approximately 13-fold increase in yield. The phytate degradation potential of extracellular phytase in rice bran has been explored and conditions for optimal degradation were optimized. Under optimal conditions, there was 43.26% release of inorganic phosphate and 6.45% decrease of phytate content.

Keywords: Lactobacillus, phytase, phytate reduction, rice bran

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Authors: Arun Kharate, Sarata Kumar Sahu, Susen Kumar Panda, Niranjan Sahoo, H. K. Panda

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In the present study, an attempt has been made for isolation and identification of feline panleukopaenia virus (FPLV) from wild felids of Nandankanan zoo, Odisha, India, along with prevalence study of FPLV. Fecal samples collected from wild felids (26 tigers, 22 lions, 5 leopards, 3 hyenas, 1 jaguar, 2 foxes and 1 wild cat) were subjected to hemagglutinnation test and fluorescent antibody test. In hemagglutinnation test 13 (50%) samples from tiger, 14 (63.63%) samples from lions, 1 (20%) sample from leopards, 1 (50%) from fox, 3 (100%) samples from hyenas and 1 (100%) sample from wild cat were positive. On fluorescent antibody test (FAT), 15 (57.69%) samples from tiger, 18 (81.81%) from lions, 2 (40%) from leopards, 1 (50%) from fox, 3 (100%) from hyenas and 1 (100%) from wild cat were positive. FPLV was isolated using MDBK cell line and preliminary characterization was done on the basis of characteristic cytopathic effect. The virus samples were quantified through titration in MDBK cells. Serological confirmation of FPLV isolates was carried out by HI test, micro-SNT and indirect-ELISA. Physico-chemical characters like pH and temperature resistance along molecular identification using specific FPLV primers was carried out. Seroprevalence study of 36 serum samples employing HI test, micro SNT and indirect-ELISA revealed prevalence of 38.8, 44.4 and 72.2% respectively. During study period an adult tigress and a tiger cub died suspected of feline panleukopenia. The necropsy findings in both animals showed hemorrhagic gastroenteritis. The cytological examination revealed presence of intranuclear inclusion bodies in the intestinal epithelial cells. Spleen, mesenteric lymph node and intestine were positive for feline panleukopenia by FAT. The investigation revealed that feline panleukopenia was prevalent in wild felines of Nandankanan zoo.

Keywords: Feline panleukopenia, fluorescent antibody test, hemagglutination test, indirect-ELISA, Nandankanan zoo

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Authors: Gaofetoge Lenetha, Malerato Moloi, Ntsoaki Malebo

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Microbial contamination of ready-to-eat foods and beverages sold by street vendors has become an important public health issue. In developing countries including South Africa, health risks related to such kinds of foods are thought to be common. Thus, this study assessed knowledge, attitude and practices of street food vendors. Street vendors in the city of Mangaung Metro were investigated in order to assess their knowledge, attitudes and handling practices. A semi-structured questionnaire and checklist were used in interviews to determine the status of the vending sites and associa. ted food-handling practices. Data was collected by means of a face-to-face interview. The majority of respondents were black females. Hundred percent (100%) of the participants did not have any food safety training. However, street vendors showed a positive attitude towards food safety. Despite the positive attitude, vendors showed some non-compliance when it comes to handling food. During the survey, it was also observed that the vending stalls lack basic infrastructures like toilets and potable water that is currently a major problem. This study indicates a need for improvements in the environmental conditions at these sites to prevent foodborne diseases. Moreover, based on the results observed food safety and food hygiene training or workshops for street vendors are highly recommended.

Keywords: food hygiene, foodborne illnesses, food safety, Street foods

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Authors: Pello Alfonso-Muniozguren, Madeleine Bussemaker, Ralph Chadeesingh, Caryn Jones, David Oakley, Judy Lee, Devendra Saroj

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Current industrialized livestock agriculture is growing every year leading to an increase in the generation of wastewater that varies considerably in terms of organic content and microbial population. Therefore, suitable wastewater treatment methods are required to ensure the wastewater quality meet regulations before discharge. In the present study, a combined lab scale activated sludge-filtration-ozonation system was used to treat a pre-treated abattoir wastewater. A hydraulic retention time of 24 hours and a solid retention time of 13 days were used for the activated sludge process, followed by a filtration step (4-7 µm) and using ozone as tertiary treatment. An average reduction of 93% and 98% was achieved for Chemical Oxygen Demand (COD) and Biological Oxygen Demand (BOD), respectively, obtaining final values of 128 mg/L COD and 12 mg/L BOD. For the Total Suspended Solids (TSS), the average reduction increased to 99% in the same system, reducing the final value down to 3 mg/L. Additionally, 98% reduction in Phosphorus (P) and a complete inactivation of Total Coliforms (TC) was obtained after 17 min ozonation time. For Total Viable Counts (TVC), a drastic reduction was observed with 30 min ozonation time (6 log inactivation) at an ozone dose of 71 mg O3/L. Overall, the combined process was sufficient to meet discharge requirements without further treatment for the measured parameters (COD, BOD, TSS, P, TC, and TVC).

Keywords: abattoir waste water, activated sludge, ozone, waste water treatment

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Authors: Surena V.

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Background and Aim: Periodontal diseases are among the most common infectious diseases all around the world, even in developed countries. Considering the increased rate of microbial resistance to antibiotics and the chemical side effects of antibiotics and antiseptics used for the treatment of periodontal disease, there is a need for an alternative antimicrobial agent with fewer complications. Medicinal herbs have recently become popular as antimicrobial and preventive agents. This study aimed to assess the antibacterial effects of hydroalcoholic extracts of Lawsonia inermis, Malva sylvestris and Boswellia serrata on Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans). Materials and Methods: Hydroalcoholic extracts of the three medicinal plants were obtained by the maceration technique and A. actinomycetemcomitans was cultured. The antimicrobial efficacy of the three medicinal plants was compared with that of 0.2% chlorhexidine (CHX) according to the CLSI protocol using agar disc diffusion and broth microdilution techniques. All tests were repeated three times. Results: Hydroalcoholic extracts of all three plants had antimicrobial activity against A. actinomycetemcomitans. The minimum inhibitory concentration (MIC) of Lawsonia inermis, Malva sylvestris, and Boswellia serrata was 78.1, 156.2, and 1666 µg/mL with no significant difference between them. The MIC of CHX was 3.33 µg/mL, which was significantly higher than that of Boswellia serrata extract. Conclusion: Given that, further in vivo studies confirm other properties of these extracts and their safety in terms of cytotoxicity and mutagenicity, hydroalcoholic extracts of Lawsonia inermis and Malva sylvestris may be used in mouthwashes or local delivery systems to affect periodontal biofilm.

Keywords: actinobacilus actinomycetem commitans, lawsonia inermis, malva sylvestris, boswellia serrata

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Authors: Abu Salim Mustafa

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Brucellosis is a zoonotic disease endemic in Kuwait. Human brucellosis can be caused by several Brucella species with Brucella melitensis causing the most severe and Brucella abortus the least severe disease. Furthermore, relapses are common after successful chemotherapy of patients. The classical biochemical methods of culture and serology for identification of Brucellae provide information about the species and serotypes only. However, to differentiate between relapse and reinfection/epidemiological investigations, the identification of genotypes using molecular methods is essential. In this study, four molecular methods [16S rRNA gene sequencing, real-time PCR, enterobacterial repetitive intergenic consensus (ERIC)-PCR and multilocus variable-number tandem-repeat analysis (MLVA)-16] were evaluated for the identification and typing of 75 strains of Brucella isolated in Kuwait. The 16S rRNA gene sequencing suggested that all the strains were B. melitensis and real-time PCR confirmed their species identity as B. melitensis. The ERIC-PCR band profiles produced a dendrogram of 75 branches suggesting each strain to be of a unique type. The cluster classification, based on ~ 80% similarity, divided all the ERIC genotypes into two clusters, A and B. Cluster A consisted of 9 ERIC genotypes (A1-A9) corresponding to 9 individual strains. Cluster B comprised of 13 ERIC genotypes (B1-B13) with B5 forming the largest cluster of 51 strains. MLVA-16 identified all isolates as B. melitensis and divided them into 71 MLVA-types. The cluster analysis of MLVA-16-types suggested that most of the strains in Kuwait originated from the East Mediterranean Region, a few from the African group and one new genotype closely matched with the West Mediterranean region. In conclusion, this work demonstrates that B. melitensis, the most pathogenic species of Brucella, is prevalent in Kuwait. Furthermore, MLVA-16 is the best molecular method, which can identify the Brucella species and genotypes as well as determine their origin in the global context. Supported by Kuwait University Research Sector grants MI04/15 and SRUL02/13.

Keywords: Brucella, ERIC-PCR, MLVA-16, RT-PCR, 16S rRNA gene sequencing

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Authors: Abed Hannane, Rouag Noureddine

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The present study aims to evaluate the association of fresh garlic (Allium sativum L.) and storage at 4°C in preserving the microbiological, nutritional, and sanitary quality of Merguez-type sausages prepared and sold locally from meat offal. The analysis focused on the evaluation of the microbiological quality of fifteen samples randomly taken from several butcheries in the wilaya of BBA, eastern Algeria. The bacteriological analysis revealed the presence of 6.88.10⁵ CFU/g of total aerobic bacteria, 5.39.10⁵ CFU/g of total coliforms, 2.23.10⁵ CFU/g of fecal coliforms, 2.43.103 CFU/g of Escherichia coli and 1.8.10⁵ CFU/g of coagulase-positive staphylococci, values higher than Algerian standards. The addition of fresh garlic as an antibacterial preservative at concentrations of 0.06, 0.12, 0.18, and 0.24 g/g to ground beef samples and stored in the refrigerator at 4°C for 15 days. The addition of garlic to Merguez made it possible to significantly reduce the presence of different bacterial groups during their refrigerated storage, compared to untreated meat, bringing it below the standards defined in the matter. Thus, the use of garlic as a food additive at a concentration of 0.12 g/g was sufficient to obtain levels according to Algerian standards equal to 1.8.10⁴ CFU/g of total aerobic bacteria, 9.48.10³ CFU/ g of total coliforms, 3.68.10³ UFC/g fecal coliforms, 4.56.10² UFC/g of E.coli 2.39.10⁴ UFC/g of coagulase-positive staphylococci. It is clear that thanks to the addition of garlic to Merguez, the sanitary quality has been improved by reducing the aerobic bacterial load and increasing the shelf life at 4°C.

Keywords: antimicrobial effect, garlic, sausage, storage

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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: Rimzhim Gupta, Bhanupriya Boruah, Jayant M. Modak, Giridhar Madras

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Microbial contamination is one of the major concerns in the field of water treatment. AOP (advanced oxidation processes) is well-established method to resolve the issue of removal of contaminants in water. A Z-scheme composite g-C₃N₄/CuWO₄ was synthesized by sol-gel method for the photocatalytic inactivation of a mixed population of Gram-positive bacteria (S. aureus) and Gram-negative bacteria (E. coli). The photoinactivation was observed for different types of bacteria in the same medium together and individually in the absence of the nutrients. The lattice structures and phase purities were determined by X-ray diffraction. For morphological and topographical features, scanning electron microscopy and transmission electron microscopy analyses were carried out. The band edges of the semiconductor (valence band and conduction band) were determined by ultraviolet photoelectron microscopy. The lifetime of the charge carriers and band gap of the semiconductors were determined by time resolved florescence spectroscopy and diffused reflectance spectroscopy, respectively. The effect of weight ratio of C₃N₄ and CuWO₄ was observed by performing photocatalytic experiments. To investigate the exact mechanism and major responsible radicals for photocatalysis, scavenger studies were performed. The rate constants and order of the inactivation reactions were obtained by power law kinetics. For E. coli and S. aureus, the order of reaction and rate constants are 1.15, 0.9 and 1.39 ± 0.03 (CFU/mL)⁻⁰.¹⁵ h⁻¹, 47.95 ± 1.2 (CFU/mL)⁰.¹ h⁻¹, respectively.

Keywords: z-scheme, E. coli, S. aureus, sol-gel

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Authors: Eitan Elaad

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The current study aimed to examine lying and pretended fairness by police personnel in sharing situations. Forty Israeli police officers and 40 laypeople from the community, all males, self-assessed their lie-telling ability, rated the frequency of their lies, evaluated the acceptability of lying, and indicated using rational and intuitive thinking while lying. Next, according to the ultimatum game procedure, participants were asked to share 100 points with an imagined target, either a male policeman or a male non-policeman. Participants allocated points to the target person bearing in mind that the other person must accept or reject their offer. Participants' goal was to retain as many points as possible, and to this end, they could tell the target person that fewer than 100 points were available for distribution. We defined concealment or lying as the difference between the available 100 points and the sum of points designated for sharing. Results indicated that police officers lied less to their fellow police targets than non-police targets, whereas laypeople lied less to non-police targets than imagined police targets. The ratio between the points offered to the imagined target person and the points endowed by the participant as available for sharing defined pretended fairness.Enhanced pretended fairness indicates higher motivation to display fair sharing even if the fair sharing is fictitious. Police officers presented higher pretended fairness to police targets than laypeople, whereas laypeople set off more fairness to non-police targets than police officers. We discussed the results concerning occupation solidarity and loyalty among police personnel. Specifically, police work involves uncertainty, danger and risk, coercive authority, and the use of force, which isolates the police from the community and dictates strong bonds of solidarity between police personnel. No wonder police officers shared more points (lied less) to fellow police targets than non-police targets. On the other hand, police legitimacy or the belief that the police are acting honestly in the best interest of the citizens constitutes citizens' attitudes toward the police. The relatively low number of points shared for distribution by laypeople to police targets indicates difficulties with the legitimacy of the Israeli police.

Keywords: lying, fairness, police solidarity, police legitimacy, sharing, ultimatum game

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Authors: Monday O. Ale, Adebukola A. Akintade, Olasunbo O. Orungbemi

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Fermentation as an important operation in the processing of cocoa beans is now affected by the recent climate change across the globe. The major requirement for effective fermentation is the ability of the material used to retain sufficient heat for the required microbial activities. Apart from the effects of climate on the rate of heat retention, the materials used for fermentation plays an important role. Most Farmers still restrict fermentation activities to the use of traditional methods. Improving on cocoa fermentation in this era of climate change makes it necessary to work on other materials that can be suitable for cocoa fermentation. Therefore, the objective of this study was to determine the effects of fermentation techniques on the quality of cocoa beans. The materials used in this fermentation research were heap-leaves (traditional), stainless steel, plastic tin, plastic basket and wooden box. The period of fermentation varies from zero days to 10 days. Physical and chemical tests were carried out for variables in quality determination in the samples. The weight per bean varied from 1.0-1.2 g after drying across the samples and the major color of the dry beans observed was brown except with the samples from stainless steel. The moisture content varied from 5.5-7%. The mineral content and the heavy metals decreased with increase in the fermentation period. A wooden box can conclusively be used as an alternative to heap-leaves as there was no significant difference in the physical features of the samples fermented with the two methods. The use of a wooden box as an alternative for cocoa fermentation is therefore recommended for cocoa farmers.

Keywords: fermentation, effects, fermentation materials, period, quality

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Authors: Mahmoud Huleihel, George Abu-Aqil, Manal Suleiman, Klaris Riesenberg, Itshak Lapidot, Ahmad Salman

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Urinary tract infections (UTIs) are considered to be the most common bacterial infections worldwide, which are caused mainly by Escherichia (E.) coli (about 80%). Klebsiella pneumoniae (about 10%) and Pseudomonas aeruginosa (about 6%). Although antibiotics are considered as the most effective treatment for bacterial infectious diseases, unfortunately, most of the bacteria already have developed resistance to the majority of the commonly available antibiotics. Therefore, it is crucial to identify the infecting bacteria and to determine its susceptibility to antibiotics for prescribing effective treatment. Classical methods are time consuming, require ~48 hours for determining bacterial susceptibility. Thus, it is highly urgent to develop a new method that can significantly reduce the time required for determining both infecting bacterium at the species level and diagnose its susceptibility to antibiotics. Fourier-Transform Infrared (FTIR) spectroscopy is well known as a sensitive and rapid method, which can detect minor molecular changes in bacterial genome associated with the development of resistance to antibiotics. The main goal of this study is to examine the potential of FTIR spectroscopy, in tandem with machine learning algorithms, to identify the infected bacteria at the species level and to determine E. coli susceptibility to different antibiotics directly from patients' urine in about 30minutes. For this goal, 1600 different E. coli isolates were isolated for different patients' urine sample, measured by FTIR, and analyzed using different machine learning algorithm like Random Forest, XGBoost, and CNN. We achieved 98% success in isolate level identification and 89% accuracy in susceptibility determination.

Keywords: urinary tract infections (UTIs), E. coli, Klebsiella pneumonia, Pseudomonas aeruginosa, bacterial, susceptibility to antibiotics, infrared microscopy, machine learning

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Authors: Shobha U. Jadhav, R. S. Saler

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Onion is most Important Vegetable crop grown throughout the world. Onion suffers from pest and fungal diseases but the fungicides cause pollution and disturb microbial balance of soil. Under integrated fungal disease management programme cost effective and eco- friendly component like plant extract are used to control plant pathogens. Alternaria porri, Fusarium oxysporium, Stemphylium vesicarium are soil borne pathogens of onion. Effect of three different plant extract (Datura metel, Pongamia pinnata, Ipomoea palmata) at five different concentration Viz, 10,25,50,75 and 100 percentage on these pathogens was studied by food poisoning techniquie. Detura metal gave 94.73% growth of Alternaria porri at 10% extract concentraton and 26.31% growth in 100% extract concentration. As compared to Fusarium oxysporium, and Stemphylium vesicarium, Alternaria porri give good inhibitory response. In Pongamia pinnata L. at 10% extract concentration 84.21% growth and at 100% extract concentration 36.84% growth of Stemphylium vesicarium was observed. Stemphylium vesicarium give good in inhibitory response as compared to Alternaria porri and Fusarium oxysporium. Ipomoea palmata in 10% extract concentration 92% growth and in 100% extract concentration 40% growth of Fusarium oxysporium was recorded. Fusarium oxysporium give good inhibitory response as compared to Alternaria porri and, Stemphylium vesicarium.

Keywords: pathogen, onion, plant extract, Allium cepa L.

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Authors: Sidra Pervez, Afsheen Aman, Shah Ali Ul Qader

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The importance of attaining an optimum performance of an enzyme is often a question of devising an effective method for its immobilization. Cross-linked enzyme aggregate (CLEAs) is a new approach for immobilization of enzymes using carrier free strategy. This method is exquisitely simple (involving precipitation of the enzyme from aqueous buffer followed by cross-linking of the resulting physical aggregates of enzyme molecules) and amenable to rapid optimization. Among many industrial enzymes, amyloglucosidase is an important amylolytic enzyme that hydrolyzes alpha (1→4) and alpha (1→6) glycosidic bonds in starch molecule and produce glucose as a sole end product. Glucose liberated by amyloglucosidase can be used for the production of ethanol and glucose syrups. Besides this amyloglucosidase can be widely used in various food and pharmaceuticals industries. For production of amyloglucosidase on commercial scale, filamentous fungi of genera Aspergillus are mostly used because they secrete large amount of enzymes extracellularly. The current investigation was based on isolation and identification of filamentous fungi from genus Aspergillus for the production of amyloglucosidase in submerged fermentation and optimization of cultivation parameters for starch saccharification. Natural isolates were identified as Aspergillus niger KIBGE-IB36, Aspergillus fumigatus KIBGE-IB33, Aspergillus flavus KIBGE-IB34 and Aspergillus terreus KIBGE-IB35 on taxonomical basis and 18S rDNA analysis and their sequence were submitted to GenBank. Among them, Aspergillus fumigatus KIBGE-IB33 was selected on the basis of maximum enzyme production. After optimization of fermentation conditions enzyme was immobilized on CLEA. Different parameters were optimized for maximum immobilization of amyloglucosidase. Data of enzyme stability (thermal and Storage) and reusability suggested the applicability of immobilized amyloglucosidase for continuous saccharification of starch in industrial processes.

Keywords: aspergillus, immobilization, industrial processes, starch saccharification

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Authors: J. Herrero, D. Puigserver, I. Nijenhuis, K. Kuntze, J. M. Carmona

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In the transition zone between aquifers and basal aquitards, the perchloroethene (PCE)-pools are more recalcitrant than those elsewhere in the aquifer. Although biodegradation of chloroethenes occur in this zone, it is a slow process and a remediation strategy is needed. The aim of this study is to demonstrate that combined strategy of biostimulation and in situ chemical reduction (ISCR) is more efficient than the two separated strategies. Four different microcosm experiments with sediment and groundwater of a selected field site where an aged pool exists at the bottom of a transition zone were designed under i) natural conditions, ii) biostimulation with lactic acid, iii) ISCR with zero-value iron (ZVI) and under iv) a combined strategy with lactic acid and ZVI. Biotic and abiotic dehalogenation, terminal electron acceptor processes and evolution of microbial communities were determined for each experiment. The main results were: i) reductive dehalogenation of PCE-pools occurs under sulfate-reducing conditions; ii) biostimulation with lactic acid supports more pronounced reductive dehalogenation of PCE and trichloroethene (TCE), but results in an accumulation of 1,2-cis-dichloroethene (cDCE); iii) ISCR with ZVI produces a sustained dehalogenation of PCE and its metabolites iv) combined strategy of biostimulation and ISCR results in a fast dehalogenation of PCE and TCE and a sustained dehalogenation of cisDCE. These findings suggest that biostimulation and ISCR with ZVI are the most suitable strategies for a complete reductive dehalogenation of PCE-pools in the transition zone and further to enable the dissolution of dense non-aqueous phase liquids.

Keywords: aged PCE-pool, anaerobic microcosm experiment, biostimulation, in situ chemical reduction, natural attenuation

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Authors: Roxana Gheorghita, Gheorghe Gutt

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Environmental pollution due to non-degradation of packaging from the food and pharmaceutical industry is reaching increasingly alarming levels. The packaging used for food supplements is usually composed of successive layers of synthetic materials, conventional, glue, and paint. The situation is becoming more and more problematic as the population, according to statistics, uses food supplements more and more often. The solution can be represented by edible packaging, completely biodegradable, and compostable. The tested materials were obtained from biopolymers, agar, carrageenan, and alginate, in well-established quantities and plasticized with glycerol. Rosemary, thyme, and oregano essential oils have been added in varying proportions. The obtained films are completely water-soluble in hot liquids (with a temperature of about 80° C) and can be consumed with the product contained. The films were glossy, pleasant to the touch, thin (thicknesses between 32.8 and 52.8 μm), transparent, and with a pleasant smell, specific to the added essential oil. Tested for microbial evaluation, none of the films indicated the presence of E. coli, S. aureus, enterobacteria, coliform bacteria, yeasts, or molds. This aspect can also be helped by the low values of the water activity index (located between 0.546 and 0.576). The mechanical properties indicated that the material became more resistant with the addition of essential oil, the best values being recorded by the addition of oregano. The results obtained indicate the possibility of using biopolymer-based films with the addition of rosemary, thyme, and oregano essential oil, for wrapping food supplements, thus replacing conventional packaging, multilayer, impossible to sort and recycle.

Keywords: edible films, food supplements, oregano, rosemary, thyme

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Authors: Donatella Cimini, Sergio D’ambrosio, Saba Sadiq, Chiara Schiraldi

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Chondroitin sulfate (CS), as well as modified CS, and unsulfated chondroitin, are largely applied in research today. CS is a linear glycosaminoglycan normally present in cartilage-rich tissues and bones in the form of proteoglycans decorated with sulfate groups in different positions. CS is used as an effective non-pharmacological alternative for the treatment of osteoarthritis, and other potential applications in the biomedical field are being investigated. Some bacteria, such as E. coli K4, produce a polysaccharide that is a precursor of CS (unsulfated chondroitin). This work focused on the construction of integrative E. coli K4 recombinant strains overexpressing genes (kfoA, kfoF, pgm and galU in different combinations) involved in the biosynthesis of the nucleotide sugars necessary for polysaccharide synthesis. Strain growth and polymer production were evaluated using renewable waste materials as substrates in shake flasks and small-scale batch fermentation processes. Results demonstrated the potential to replace pure sugars with cheaper medium components to establish environmentally sustainable and cost-effective production routes for potential industrial development. In fact, although excellent fermentation results have been described so far by employing strains that naturally produce chondroitin-like polysaccharides on semi-defined media, there is still the need to reduce manufacturing costs by providing a cost-effective biotechnological alternative to currently used animal-based extraction procedures.

Keywords: E. coli K4, chondroitin, microbial cell factories, glycosaminoglycans, renewable resources

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Authors: Dawit Chankaye

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Background: Azadirachta indica is the most versatile medicinal plant known as “the village pharmacy”. The plant is known for its broad spectrum of biological activity in India and various countries throughout history by many different human cultures. The present study was undertaken to determine the antimalarial and antidiabetic properties of the leaf extracts of A. indica grown in Ethiopia when treated in vivo. This work has also been concerned with determining essential oil composition and the antimicrobial activity of the plant in vitro. Methods: Leaf extracts were prepared using three different selected solvents. Standard and clinical isolates were treated with extracts of the leaves of A. indica using the agar well diffusion method. The antimalarial and antidiabetic tests were conducted in vivo in mice. Phytochemical screening was done using various chemical tests, and the volatile oil constituents were determined using gas chromatography-mass spectrometry (GC/MS). Results: In vivo antimalarial activity studies showed 85.23%, 69.01%, and 81.54% suppression of parasitemia for 70% ethanol, acetone, and water extracts, respectively. The extracts collected from the leaves also showed reduced blood sugar levels in alloxan-induced diabetic mice. In addition, the solvent extracts were shown to have an inhibitory effect on the growth of microorganisms under the study. The minimum inhibitory concentration (MIC) ranged from 850 to 1050 µg/ml. Notably, the phytochemical investigation of the ethanol extracts showed the presence of secondary metabolites. Seventeen compounds (mainly sesquiterpenes) that represent 75.45% of the essential oil were characterized by GC/MS analysis. Conclusion: Extracts examined in this study indicated that the leaf of A. indica grown in Ethiopia retained the biological activities demonstrating the extent equivalent to when it was grown in its natural habitat. In addition, phytochemical investigation and GC/MS analysis of volatile oil constituents showed comparable results to those presented in India and elsewhere.

Keywords: Azadirachta indica, vivo, antimalarial activity, antidiabetic activity, alloxan, mice, phytochemical

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Authors: Raana Babadi Fathipour

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Street foods hold a significant place in the tapestry of socioeconomic and cultural norms, beloved across the globe. Serving as a convenient and affordable option for city dwellers seeking nourishment, these culinary delights also serve as a vital source of income for vendors, particularly women. Additionally, street food acts as a mirror reflecting traditional local customs and practices, an element that draws tourists to experience the authenticity of a culture firsthand. Despite its many virtues, concerns have emerged regarding the microbiological safety of street food worldwide. Often prepared and sold in subpar conditions without proper oversight or regulation, street food has become synonymous with potential health risks. The presence of elevated levels of fecal indicator bacteria and various pathogens in these unregulated delicacies further perpetuates anxieties surrounding their consumption. This analysis delves into the intricate microbiological intricacies inherent in street food, shedding light on the pertinent safety concerns and prevalent pathogens. Additionally, it elaborates on the worldwide standing of this vital economic endeavor. Moreover, it advocates for the adoption of molecular detection techniques over conventional culture-based methods to gain a more comprehensive grasp of the true microbial risks posed by street cuisine. Acknowledgment marks the initial step towards resolving any given issue.

Keywords: foodborne pathogens, microbiological safety, street food, viruses

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Authors: Asato Takaishi, Masashi Nasuhara, Ayuko Itsuki, Kenichi Suga

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Stevia (Stevia rebaudiana Bertoni) is a composite plant native to Paraguay. Stevia sweetener is derived from a hot water extract of Stevia (Stevia extract), which has some effects such as histamine decomposition, antioxidative effect, and blood sugar level-lowering function. The steviol glycosides in the Stevia extract are considered to contribute to these effects. In addition, these effects increase by the fermentation. However, it takes a long time for fermentation of Stevia extract and the fermentation liquid sometimes decays during the fermentation process because natural fermentation method is used. The aim of this study is to perform the fermentation of Stevia extract in a shorter period, and to produce the fermentation liquid in stable quality. From the natural fermentation liquid of Stevia extract, the four strains of useful (good taste) microorganisms were isolated using dilution plate count method and some properties were determined. The base sequences of 16S rDNA and 28S rDNA revealed three bacteria (two Lactobacillus sp. and Microbacterium sp.) and one yeast (Issatchenkia sp.). This result has corresponded that several kinds of lactic bacterium such as Lactobacillus pentosus and Lactobacillus buchneri were isolated from Stevia leaves. Liquid chromatography/mass spectrometory (LC/MS/MS) and High-Performance Liquid Chromatography (HPLC) were used to determine the contents of steviol glycosides and neutral sugars. When these strains were cultured in the sterile Stevia extract, the steviol and stevioside were increased in the fermented Stevia extract. So, it was suggested that the rebaudioside A and the mixture of steviol glycosides in the Stevia extract were decomposed into stevioside and steviol by microbial metabolism.

Keywords: fermentation, lactobacillus, Stevia, steviol glycosides, yeast

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Authors: Chandra Sekhar Singh, P. Chakrapani, B. Arun Jyothi, A. Roja Rani

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The green synthesis of metallic nanoparticles (NPs) involves biocompatible ingredients under physiological conditions of temperature and pressure. Moreover, the biologically active molecules involved in the green synthesis of NPs act as functionalizing ligands, making these NPs more suitable for biomedical applications. Among the most important bioreductants are plant extracts, which are relatively easy to handle, readily available, low cost, and have been well explored for the green synthesis of other nanomaterials. Various types of metallic NPs have already been synthesized using plant extracts. They have wide applicability in various areas such as electronics, catalysis, chemistry, energy, and medicine. Metallic nanoparticles are traditionally synthesized by wet chemical techniques, where the chemicals used are quite often toxic and flammable. In our study, we were described a cost effective and environment friendly technique for green synthesis of silver nanoparticles from 1mM AgNO3 solution through the aqueous extract of Erythrina indica as reducing as well as capping agent. Nanoparticles were characterized using UV–Vis absorption spectroscopy, FTIR, XRD, X-ray diffraction, SEM and TEM analysis showed the average particle size of 30 nm as well as revealed their spherical structure. Further these biologically synthesized nanoparticles were found to be highly toxic against different human pathogens viz. two Gram positive namely Klebsiella pneumonia and Bacillus subtilis bacteria and two were Gram negative bacteria namely Staphylococcus aureus and Escherichia coli (E. coli). This is for the first time reporting that Erythrina indica plant extract was used for the synthesis of nanoparticles.

Keywords: silver nanoparticles, green synthesis, antibacterial activity, FTIR, TEM, SEM

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