Search results for: pathogenic bacteria
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1706

Search results for: pathogenic bacteria

1676 Comparison between Two Groups of Pathogenic Bacteria under Different Essential Oil Extract of Ocimum basilicum L.

Authors: A. M. Daneshian Moghaddam, J. Shayegh, J. Dolghari Sharaf

Abstract:

This study was conducted to assessment the antibacterial activities of different part of basil essential oil on the standard gram-negative bacteria include Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, and gram-positive ones including Bacillus cereus, Staphylococcus aureus, and Listeria monocytogen. The basil essential oil was provided from two part of plant (leaf and herb) at the two different developmental stage. The antibacterial properties of basil essential oil was studied Also agar disk diffusion, minimal inhibition concentration (MIC) and minimum bactericidal concentration (MBC) were detected. The results of agar disk diffusion tests showed the inhibition zones as follow: Listeria monocytogen 17.11-17.42 mm, St. aureus 29.20-30.56 mm, B. cereus 14.73-16.06 mm, E. coli 21.60-23.58 mm, Salmonella typhi 21.63-24.80 mm and for P. aeruginosa the maximum inhibition zones were seen on leaf essential oil. From the herb part of basil almost similar results were obtained: Listeria monocytogen 17.02-17.67 mm, St. aureus 29.60-30.41 mm, B. cereus 10.66-16.11 mm, E. coli 17.48-23.54 mm, Salmonella typhi 21.58-21.64 mm and for P. aeruginosa the maximum inhibition zones were seen. The MICs for gram-positive bacteria were as: B. cereus ranging 36-18 μg/mL, S. aureus 18 μg/mL, Listeria monocytogen 18-36 μg/mL and for gram-negative bacteria of E. coli, Salmonella typhi and P. aeruginosa were 18-9 μg/mL.

Keywords: basil (Ocimum basilicum) essential oil, gram-positive and gram negative bacteria, antibacterial activity, MIC, MBC

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1675 Signal Amplification Using Graphene Oxide in Label Free Biosensor for Pathogen Detection

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

Abstract:

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

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

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1674 Evaluation of Antimicrobial Activity of Different Dithiolethiones

Authors: Zehour Rahmani, Messouda Dekmouche, Mohamed Hadjadj, Mokhtar Saidi

Abstract:

In the last decades of the nineteenth century, the study of disease – causing microorganisms became concentrated on bacteria and largely institutionalized. In earlier years, the scientists interested in bacteria had originally been chemists like Pasteur, physicists like Tyndall, or botanists like Cohn and ward. For this reason, the objective of this research was to evaluate the potential of some dithiolethiones on standard microorganism strains as well as multi-drug resistant bacteria, which were isolated from hospitals. Recent studies have demonstrated, that several dithiolethione compounds, particularly (3H-1,2-dithiole-3-thione), exhibit the biological activities against several bacteria.

Keywords: bacteria, dithiolethiones, microorganism, potential

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1673 Combained Cultivation of Endemic Strains of Lactic Acid Bacteria and Yeast with Antimicrobial Properties

Authors: A. M. Isakhanyan, F. N. Tkhruni, N. N. Yakimovich, Z. I. Kuvaeva, T. V. Khachatryan

Abstract:

Introduction: At present, the simbiotics based on different genera and species of lactic acid bacteria (LAB) and yeasts are used. One of the basic properties of probiotics is presence of antimicrobial activity and therefore selection of LAB and yeast strains for their co-cultivation with the aim of increasing of the activity is topical. Since probiotic yeast and bacteria have different mechanisms of action, natural synergies between species, higher viability and increasing of antimicrobial activity might be expected from mixing both types of probiotics. Endemic strains of LAB Enterococcus faecium БТK-64, Lactobaccilus plantarum БТK-66, Pediococcus pentosus БТK-28, Lactobacillus rhamnosus БТK-109 and Kluyveromyces lactis БТX-412, Saccharomycopsis sp. БТX- 151 strains of yeast, with probiotic properties and hight antimicrobial activity, were selected. Strains are deposited in "Microbial Depository Center" (MDC) SPC "Armbiotechnology". Methods: LAB and yeast strains were isolated from different dairy products from rural households of Armenia. The genotyping by 16S rRNA sequencing for LAB and 26S RNA sequencing for yeast were used. Combined cultivation of LAB and yeast strains was carried out in the nutrient media on the basis of milk whey, in anaerobic conditions (without shaker, in a thermostat at 37oC, 48 hours). The complex preparations were obtained by purification of cell free culture broth (CFC) broth by the combination of ion-exchange chromatography and gel filtration methods. The spot-on-lawn method was applied for determination of antimicrobial activity and expressed in arbitrary units (AU/ml). Results. The obtained data showed that at the combined growth of bacteria and yeasts, the cultivation conditions (medium composition, time of growth, genera of LAB and yeasts) affected the display of antimicrobial activity. Purification of CFC broth allowed obtaining partially purified antimicrobial complex preparation which contains metabiotics from both bacteria and yeast. The complex preparation inhibited the growth of pathogenic and conditionally pathogenic bacteria, isolated from various internal organs from diseased animals and poultry with greater efficiency than the preparations derived individually alone from yeast and LAB strains. Discussion. Thus, our data shown perspectives of creation of a new class of antimicrobial preparations on the basis of combined cultivation of endemic strains of LAB and yeast. Obtained results suggest the prospect of use of the partially purified complex preparations instead antibiotics in the agriculture and for food safety. Acknowledgments: This work was supported by the RA MES State Committee of Science and Belarus National Foundation for Basic Research in the frames of the joint Armenian - Belarusian joint research project 13РБ-064.

Keywords: co-cultivation, antimicrobial activity, biosafety, metabiotics, lactic acid bacteria, yeast

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1672 Correlation between Copper Uptake and Decrease of Copper (Hypocupremia) in Burn Patients-Infected Pseudomonas aeruginosa

Authors: Khaled M. Khleifat

Abstract:

Pseudomonas aeruginosa was isolated from infected burn patients and characterized by standard biochemical tests. The in vitro copper uptake was compared between this isolated pathogenic strain and two non-pathogenic control strains of Gram-positive bacteria Bacillusthuringiensis strain Israelisas well as Gram-negative bacteria Enterobacter aerogenes. Maximum copper uptake of 470 ppm/g biomass was obtained by P. aeruginosa strain, while the control strains B. thuringiensis and Enterobacter aerogenes had copper uptake of 350 and 383 ppm/g biomass, respectively. However, the lowest copper uptake (60 ppm/g biomass) was observed with another control the saprophytic strain Pseudomonas (Shewanella) putrefaciens. A further investigation regarding the effect of copper toxicity on bacterial growth, gave an MIC score of 600 ppm for P. aeruginosa strain compared to 460 and 300 ppm for the two Gram positive and Gram negative control strains, respectively. In tandem with these in vitro findings, blood analysis on burn patients infected with P. aeruginosa has indicated a selective decrease of copper (hypocupremia) and ceruloplasmin plasma levels. The iron metabolism was also affected by this copper deprivation leading to a similar decrease in plasma levels of PCV, iron, total iron-binding capacity, and transferrin. All these hematological changes were significantly different (P < 0.05) from the matched group of non-infected burn patients. The observed hypocupremia in infected burn patients was attributed to demanding scavenger ability by P. aeruginosa strain for the copper of plasma.

Keywords: Pseudomonas aeruginosa, hypocupremia, correlation, PCV

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1671 Evaluation of Antimicrobial Properties of Lactic Acid Bacteria of Enterococcus Genus

Authors: Kristina Karapetyan, Flora Tkhruni, Tsovinar Balabekyan, Arevik Israyelyan, Tatyana Khachatryan

Abstract:

The ability of the lactic acid bacteria (LAB) to prevent and cure a variety of diseases, their protective role against infections and colonization of pathogenic microorganisms in the digestive tract, has lead to the coining of the term probiotics or pro-life. LAB inhibiting the growth of pathogenic and food spoilage microorganisms, maintaining the nutritive quality and improving the shelf life of foods. They have also been used as flavor and texture producers. Enterococcus strains have been used for treatment of diseases such as diarrhea or antibiotic associated diarrhea, inflammatory pathologies that affect colon such as irritable bowel syndrome, or immune regulation, diarrhea caused by antibiotic treatments. The obtaining and investigation of biological properties of proteinoceous antibiotics, on the basis of probiotic LAB shown, that bacteriocins, metabiotics, and peptides of LAB represent bactericides have a broad range of activity and are excellent candidates for development of new prophylactic and therapeutic substances to complement or replace conventional antibiotics. The genotyping by 16S rRNA sequencing for LAB were used. Cell free culture broth (CFC) broth was purified by the Gel filtration method on the Sephadex Superfine G 25 resin. Antimicrobial activity was determined by spot-on-lawn method and expressed in arbitrary units (AU/ml). The diversity of multidrug-resistance (MDR) of pathogenic strains to antibiotics, most widely used for treatment of human diseases in the Republics of Armenia and Nagorno Karabakh were examined. It was shown, that difference of resistance of pathogens to antibiotics depends on their isolation sources. The influences of partially purified antimicrobial preparations (AMP), obtained from the different strains of Enterococcus genus on the growth of MDR pathogenic bacteria were investigated. It was shown, that bacteriocin containing partially purified preparations, obtained from different strains of Enterococcus faecium and durans species, possess bactericidal or bacteriostatic activity against antibiotic resistant intestinal, spoilage and food-borne pathogens such as Listeria monocytogenes, Staphylococcus aureus, E. coli, and Salmonella. Endemic strains of LAB, isolated from Matsoni made from donkey, buffalo and goat milk, shown broad spectrum of activity against food spoiling microorganisms, moulds and fungi, such as Salmonella sp., Esherichia coli, Aspergillus and Penicillium species. Highest activity against MDR pathogens shown bacteria, isolated from goat milk products. High stability of the investigated strains of the genus Enerococcus, isolated from samples of matsun from different regions of Nagorno-Karabakh (NKR) to the antibiotics was shown. The obtained data show high stability of the investigated different strains of the genus Enerococcus. The high genetic diversity in Enterococcus group suggests adaptations for specific mutations in different environments. Thus, endemic strains of LAB are able to produce bacteriocins with high and different inhibitory activity against broad spectrum of microorganisms isolated from different sources and belong to different taxonomic group. Prospect of the use of certain antimicrobial preparations against pathogenic strains is obvious. These AMP can be applied for long term use against different etiology antibiotic resistant pathogens for prevention or treatment of infectional diseases as an alternative to antibiotics.

Keywords: antimicrobial biopreparation, endemic lactic acid bacteria, intra-species diversity, multidrug resistance of pathogens

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1670 Association between Copper Uptake and Decrease of Copper (hypocupremia) in Burn Patients-Infected Pseudomonas aeruginosa

Authors: Khaled Khleifat, Muayyad Abboud, Amjad Khleifat, Humodi Saeed

Abstract:

In this study, Pseudomonas aeruginosa was isolated from infected burn patients and characterized by standard biochemical tests. The in vitro copper uptake was compared between this isolated pathogenic strain and two non-pathogenic control strains of Gram positive bacteria Bacillusthuringiensis strain Israelisas well as Gram negative bacteria Enterobacter aerogenes. Maximum copper uptake of 470 ppm/g biomass was obtained by P. aeruginosa strain, while the control strains B. thuringiensis andEnterobacter aerogenes had copper uptake of 350 and 383 ppm/g biomass, respectively. However, the lowest copper uptake (60 ppm/g biomass) was observed with another control the saprophytic strain Pseudomonas (Shewanella) putrefaciens. A further investigation regarding the effect of copper toxicity on bacterial growth, gave an MIC score of 600 ppm for P. aeruginosa strain compared to 460 and 300 ppm for the two Gram positive and Gram negative control strains, respectively. In tandem with these in vitro findings, blood analysis on burn patients infected with P. aeruginosa has indicated a selective decrease of copper (hypocupremia) and ceruloplasmin plasma levels. The iron metabolism was also affected by this copper deprivation leading to a similar decrease in plasma levels of PCV, iron, total iron binding capacity, and transferrin. All these hematological changes were significantly different (P < 0.05) from the matched group of non-infected burn patients. The observed hypocupremia in infected burn patients was attributed to demanding scavenger ability by P. aeruginosa strain for the copper of plasma.

Keywords: pseudomonas, Cu uptake, burn patients, biosorption

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1669 Development of Antimicrobial Properties Nutraceuticals: Gummy Candies with Addition of Bovine Colostrum, Essential Oils and Probiotics

Authors: E. Bartkiene, M. Ruzauskas, V. Lele, P. Zavistanaviciute, J. Bernatoniene, V. Jakstas, L. Ivanauskas, D. Zadeike, D. Klupsaite, P. Viskelis, J. Bendoraitiene, V. Navikaite-Snipaitiene, G. Juodeikiene

Abstract:

In this study, antimicrobial nutraceuticals; gummy candies (GC) from bovine colostrum (BC), essential oils (EOs), probiotic lactic acid bacteria (PLAB), and their combinations, were developed. For antimicrobial GC preparation, heteropolysaccharide (agar) was used. The antimicrobial properties of EOs (Eugenia caryophyllata, Thymus vulgaris, Citrus reticulata L., Citrus paradisi L.), BC, L. paracasei LUHS244, L. plantarum LUHS135, and their combinations against pathogenic bacteria strains (Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, Salmonella enterica, Escherichia coli, Proteus mirabilis, and Pseudomonas aeruginosa) were evaluated. The highest antimicrobial properties by EO’s (Eugenia caryophyllata and Thymus vulgaris) were established. The optimal ingredients composition for antimicrobial GC preparation was established, which incorporate the BC fermented with L. paracasei LUHS244 in combination with Thymus vulgaris or Eugenia caryophyllata. These ingredients showed high inhibition properties of all tested pathogenic strains (except Pseudomonas aeruginosa). Antimicrobial GC formula consisting of thyme EO (up to 0.2%) and fermented BC (up to 3%), and for taste masking, mandarin or grapefruit EOs (up to 0.2%) was used. Developed GC high overall acceptability and antimicrobial properties, thus, antimicrobial GC could be a preferred form of nutraceuticals. This study was fulfilled with the support of the LSMU-KTU joint project.

Keywords: antimicrobial activity, bovine colostrum, essential oil, gummy candy, probiotic

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1668 Probiotic Properties of Lactic Acid Bacteria Isolated from Fermented Food

Authors: Wilailak Siripornadulsil, Siriyanapat Tasaku, Jutamas Buahorm, Surasak Siripornadulsil

Abstract:

The objectives of this study were to isolate LAB from various sources, dietary supplement, Thai traditional fermented food, and freshwater fish and to characterize their potential as probiotic cultures. Out of 1,558 isolates, 730 were identified as LAB based on isolation on MRS agar supplemented with a bromocresol purple indicator and CaCO3 and gram-positive, catalase and oxidase negative characteristics. Eight isolates showed the potential probiotic properties including tolerance to acid, bile salt and heat, proteolytic, amylolytic and lipolytic activities and oxalate-degrading capability. They all showed the antimicrobial activity against some Gram-negative and Gram-positive pathogenic bacteria. Based on 16S rDNA sequence analysis, they were identified as Enterococcus faecalis BT2 and MG30, Leconostoc mesenteroides SW64 and Pediococcus pentosaceous BD33, CF32, NP6, PS34 and SW5. The health beneficial effects and food safety will be further investigated and developed as a probiotic or protective culture used in Nile tilapia belly flap meat fermentation.

Keywords: probiotic, lactic acid bacteria, pathogen, protective culture

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1667 Antibacterial Effects of Some Medicinal and Aromatic Plant Extracts on Pathogenic Bacteria Isolated from Pear Orchards

Authors: Kubilay Kurtulus Bastas

Abstract:

Bacterial diseases are very destructive and cause economic losses on pears. Promising plant extracts for the management of plant diseases are environmentally safe, long-lasting and extracts of certain plants contain alkaloids, tannins, quinones, coumarins, phenolic compounds, and phytoalexins. In this study, bacteria were isolated from different parts of pear exhibiting characteristic symptoms of bacterial diseases from the Central Anatolia, Turkey. Pathogenic bacteria were identified by morphological, physiological, biochemical and molecular methods as fire blight (Erwinia amylovora (39%)), bacterial blossom blast and blister bark (Pseudomonas syringae pv. syringae (22%)), crown gall (Rhizobium radiobacter (1%)) from different pear cultivars, and determined virulence levels of the pathogens with pathogenicity tests. The air-dried 25 plant material was ground into fine powder and extraction was performed at room temperature by maceration with 80% (v/v) methanol/distilled water. The minimum inhibitory concentration (MIC) values were determined by using modified disc diffusion method at five different concentrations and streptomycin sulphate was used as control chemical. Bacterial suspensions were prepared as 108 CFU ml⁻¹ densities and 100 µl bacterial suspensions were spread to TSA medium. Antimicrobial activity was evaluated by measuring the inhibition zones in reference to the test organisms. Among the tested plants, Origanum vulgare, Hedera helix, Satureja hortensis, Rhus coriaria, Eucalyptus globulus, Rosmarinus officinalis, Ocimum basilicum, Salvia officinalis, Cuminum cyminum and Thymus vulgaris showed a good antibacterial activity and they inhibited the growth of the pathogens with inhibition zone diameter ranging from 7 to 27 mm at 20% (w/v) in absolute methanol in vitro conditions. In vivo, the highest efficacy was determined as 27% on reducing tumor formation of R. radiobacter, and 48% and 41% on reducing shoot blight of E. amylovora and P. s. pv. syringae on pear seedlings, respectively. Obtaining data indicated that some plant extracts may be used against the bacterial diseases on pome fruits within sustainable and organic management programs.

Keywords: bacteria, eco-friendly management, organic, pear, plant extract

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1666 Environmental Health Risk Assessment of Hospital Wastewater in Enugu Urban, Nigeria

Authors: C. T. Eze, I. N. E. Onwurah

Abstract:

An important hydrogeologic problem in areas of high faults formations is high environmental health hazard occasioned by microbial and heavy metals contamination of ground waters. Consequently, we examined the microbial load and heavy metals concentration of hospital wastewater discharged into the environment at Park Lane General Hospital Enugu Urban, Nigeria. The microbial counts, characteristics and frequency of occurrences of the isolated microorganisms were determined by cultural, morphological and biochemical characteristics using established procedure while the varying concentrations of the identified heavy metals were determined using the spectrophotometric method. The microbiological analyses showed a mean total aerobic bacteria counts from 13.7 ± 0.65 × 107 to 22.8 ± 1.14 ×1010 CFU/ml, mean total anaerobic bacteria counts from 6.0 ± 1.6 × 103 to 1.7 ± 0.41 ×104 CFU/ml and mean total fungal counts from 0 ± 0 to 2.3 ± 0.16 × 105 CFU/ml. The isolated micro-organisms which included both pathogenic and non-pathogenic organisms were Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Bacillus subtilis, Proteus vulgaris, Klesbsiella pneumonia and bacteriodes sp. The only fungal isolate was Candida albican. The heavy metals identified in the leachate were Arsenic, Cadmium, Lead, Mercury and Chromium and their concentrations ranged from 0.003 ± 0.00082 to 0.14 ± 0.0082 mg/l. These values were above WHO permissible limits while others fall within the limits. Therefore, hospital waste water can pose the environmental health risk when not properly treated before discharge, especially in geologic formations with high fault formations.

Keywords: bacterial isolates, fungal isolates, heavy metals, hospital wastewater, microbial counts

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1665 Emergence of Fluoroquinolone Resistance in Pigs, Nigeria

Authors: Igbakura I. Luga, Alex A. Adikwu

Abstract:

A comparison of resistance to quinolones was carried out on isolates of Shiga toxin-producing Escherichia coliO157:H7 from cattle and mecA and nuc genes harbouring Staphylococcus aureus from pigs. The isolates were separately tested in the first and current decades of the 21st century. The objective was to demonstrate the dissemination of resistance to this frontline class of antibiotic by bacteria from food animals and bring to the limelight the spread of antibiotic resistance in Nigeria. A total of 10 isolates of the E. coli O157:H7 and 9 of mecA and nuc genes harbouring S. aureus were obtained following isolation, biochemical testing, and serological identification using the Remel Wellcolex E. coli O157:H7 test. Shiga toxin-production screening in the E. coli O157:H7 using the verotoxin E. coli reverse passive latex agglutination (VTEC-RPLA) test; and molecular identification of the mecA and nuc genes in S. aureus. Detection of the mecA and nuc genes were carried out using the protocol by the Danish Technical University (DTU) using the following primers mecA-1:5'-GGGATCATAGCGTCATTATTC-3', mecA-2: 5'-AACGATTGTGACACGATAGCC-3', nuc-1: 5'-TCAGCAAATGCATCACAAACAG-3', nuc-2: 5'-CGTAAATGCACTTGCTTCAGG-3' for the mecA and nuc genes, respectively. The nuc genes confirm the S. aureus isolates and the mecA genes as being methicillin-resistant and so pathogenic to man. The fluoroquinolones used in the antibiotic resistance testing were norfloxacin (10 µg) and ciprofloxacin (5 µg) in the E. coli O157:H7 isolates and ciprofloxacin (5 µg) in the S. aureus isolates. Susceptibility was tested using the disk diffusion method on Muller-Hinton agar. Fluoroquinolone resistance was not detected from isolates of E. coli O157:H7 from cattle. However, 44% (4/9) of the S. aureus were resistant to ciprofloxacin. Resistance of up to 44% in isolates of mecA and nuc genes harbouring S. aureus is a compelling evidence for the rapid spread of antibiotic resistance from bacteria in food animals from Nigeria. Ciprofloxacin is the drug of choice for the treatment of Typhoid fever, therefore widespread resistance to it in pathogenic bacteria is of great public health significance. The study concludes that antibiotic resistance in bacteria from food animals is on the increase in Nigeria. The National Food and Drug Administration and Control (NAFDAC) agency in Nigeria should implement the World Health Organization (WHO) global action plan on antimicrobial resistance. A good starting point can be coordinating the WHO, Office of International Epizootics (OIE), Food and Agricultural Organization (FAO) tripartite draft antimicrobial resistance monitoring and evaluation (M&E) framework in Nigeria.

Keywords: Fluoroquinolone, Nigeria, resistance, Staphylococcus aureus

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1664 The Antibacterial Efficacy of Gold Nanoparticles Derived from Gomphrena celosioides and Prunus amygdalus (Almond) Leaves on Selected Bacterial Pathogens

Authors: M. E. Abalaka, S. Y. Daniyan, S. O. Adeyemo, D. Damisa

Abstract:

Gold nanoparticles (AuNPs) have gained increasing interest in recent times. This is greatly due to their special features, which include unusual optical and electronic properties, high stability and biological compatibility, controllable morphology and size dispersion, and easy surface functionalization. In typical synthesis, AuNPs were produced by reduction of gold salt AuCl4 in an appropriate solvent. A stabilizing agent was added to prevent the particles from aggregating. The antibacterial activity of different sizes of gold nanoparticles was investigated against Staphylococcus aureus, Salmonella typhi and Pseudomonas pneumonia using the disk diffusion method in a Müeller–Hinton Agar. The Au-NPs were effective against all bacteria tested. That the Au-NPs were successfully synthesized in suspension and were used to study the antibacterial activity of the two medicinal plants against some bacterial pathogens suggests that Au-NPs can be employed as an effective bacteria inhibitor and may be an effective tool in medical field. The study clearly showed that the Au-NPs exhibiting inhibition towards the tested pathogenic bacteria in vitro could have the same effects in vivo and thus may be useful in the medical field if well researched into.

Keywords: gold nanoparticles, Gomphrena celesioides, Prunus amygdalus, pathogens

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1663 Evaluation of Antimicrobial Susceptibility Profile of Urinary Tract Infections in Massoud Medical Laboratory: 2018-2021

Authors: Ali Ghorbanipour

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The aim of this study is to investigate the drug resistance pattern and the value of the MIC (minimum inhibitory concentration)method to reduce the impact of infectious diseases and the slow development of resistance. Method: The study was conducted on clinical specimens collected between 2018 to 2021. identification of isolates and antibiotic susceptibility testing were performed using conventional biochemical tests. Antibiotic resistance was determined using kibry-Bauer disk diffusion and MIC by E-test methods comparative with microdilution plate elisa method. Results were interpreted according to CLSI. Results: Out of 249600 different clinical specimens, 18720 different pathogenic bacteria by overall detection ratio 7.7% were detected. Among pathogen bacterial were Gram negative bacteria (70%,n=13000) and Gram positive bacteria(30%,n=5720).Medically relevant gram-negative bacteria include a multitude of species such as E.coli , Klebsiella .spp , Pseudomonas .aeroginosa , Acinetobacter .spp , Enterobacterspp ,and gram positive bacteria Staphylococcus.spp , Enterococcus .spp , Streptococcus .spp was isolated . Conclusion: Our results highlighted that the resistance ratio among Gram Negative bacteria and Gram positive bacteria with different infection is high it suggest constant screening and follow-up programs for the detection of antibiotic resistance and the value of MIC drug susceptibility reporting that provide a new way to the usage of resistant antibiotic in combination with other antibiotics or accurate weight of antibiotics that inhibit or kill bacteria. Evaluation of wrong medication in the expansion of resistance and side effects of over usage antibiotics are goals. Ali ghorbanipour presently working as a supervision at the microbiology department of Massoud medical laboratory. Iran. Earlier, he worked as head department of pulmonary infection in firoozgarhospital, Iran. He received master degree in 2012 from Fergusson College. His research prime objective is a biologic wound dressing .to his credit, he has Published10 articles in various international congresses by presenting posters.

Keywords: antimicrobial profile, MIC & MBC Method, microplate antimicrobial assay, E-test

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1662 Antimicrobial Potential of Calendula officinalis Extracts on Flavobacterium columnare of Clarias gariepinus Fingerlings

Authors: Nelson Rotimi Osungbemiro, Sanni Rafiu Olugbenga, Abayomi Olufemi Olajuyigbe

Abstract:

Ninety Fingerlings of Clarias gariepinus were exposed to the pathogenic Flavobacterium columnare a Gram Negative bacteria responsible for high mortality in fish pond raised young fish (fries and fingerlings) of Clarias sp. in Southwestern Nigeria. After feeding with 40% crude protein pelletized fish feed for 5 days, the fishes were divided into two groups, one group was treated with extracts from Calendula officinalis flowers, while the second group was not treated (control). The results indicated that, at day 5, colony formation had been manifesting and at day 7, skin lesion occurred and at the 8th day, first mortality of fish occurred, and this continued steadily on the 9th-12th day when all the fishes were dead. Whereas, in the group that was treated with Calendula sp., no single mortality was recorded. This research shows that plant extract from Calendula flowers is an effective antimicrobial agent against the virulent pathogenic Flavobacterium columnare disease.

Keywords: antimicrobial, Flavobacterium columnare, Clarias gariepinus, fish

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1661 Effects of Bacteria on Levels of AFM1 in Phosphate Buffer at Different Level of Energy Source

Authors: Ali M. Elgerbi, Obied A. Alwan, Al-Taher O. Alzwei, Abdurrahim A. Elouzi

Abstract:

The binding of AFM1 to bacteria in phosphate buffer solution depended on many factors such as: availability of energy, incubation period, species and strain of bacteria. Increase in concentration of sugar showed higher removal of AFM1 and faster than in phosphate buffer alone. With 1.0% glucose lactic acid bacteria and bifidobacteria showed toxin removal ranging from 7.7 to 39.7% whereas with 10.0% glucose the percentage removal was 21.8 to 45.4% at 96 hours of incubation.

Keywords: aflatoxin M1, lactic acid bacteria, bifidobacteria , binding, phosphate buffer

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1660 Application of Bacteriophages as Natural Antibiotics in Aquaculture

Authors: Chamilani Nikapitiya, Mahanama De Zoysa, Jehee Lee

Abstract:

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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1659 Investigation of the Bioactivity and Efficacy of Personal Care Products Formulated Using Extracts of Azadirachta indica A. Juss

Authors: Ade O. Oyewole, Sunday O. Okoh, Ruth O. Ishola, Adenike D. Odusote, Chima C. Igwe, Gloria N. Elemo, Anthony I. Okoh

Abstract:

Azadirachta indica (Neem tree) also referred to as an all-purpose tree is used in a wide range of medical preparations in tropical and subtropical countries for prevention and management of various livestock, crops products and human diseases. In Nigeria however, the potentials of this plant have not been fully exploited thus it causes an environmental nuisance during the fruiting season. With a rise in the demand for herbal personal care products globally extracts from different parts of the neem plant were used as the bio-active ingredients in the formulation of personal care products. In this study, formulated neem soap, body cream, lotion, toothpaste and shampoo are analyzed to determine their antibacterial, antifungal, and toxicity properties. The efficacies of these products for management of infectious diseases, both oral and dermal, were also investigated in vitro. Oil from the neem seeds obtained using a mechanical press and acetone extracts of both the neem bark and leaves obtained by the maceration method were used in the formulation and production of the neem personal care products. The antimicrobial and toxicity properties of these products were investigated by agar diffusion, and haemolytic methods respectively. The five neem products (NPs) exhibited strong antibacterial activities against four multi–drug resistant pathogenic and three none pathogenic bacterial strains (Escherichia coli (180), Listeria ivanovii, Staphylococcus aureus, Enterobacter cloacae, Vibro spp., Streptococcus uberis, Mycobacterium smegmatis), except the neem lotion with insignificant activity against E. coli and S. aureus. The minimum inhibitory concentration (MIC) range was between 0.20-0.40 mg/ mL. The 5 NPs demonstrated moderate activity against three clinical dermatophytes isolates (Tinea corporis, Tinea capitis, and Tinea cruiz) as well as one fungal strain (Candida albican) with the MIC ranging between 0.30 - 0.50 mg/ mL and 0.550 mg/mL respectively. The soap and shampoo were the most active against test bacteria and fungi. The haemolytic analysis results on the 5 NPs indicated none toxicity at 0.50 mg/ mL in sheep red blood cells (SRBC).

Keywords: antimicrobial, Azadirachta indica, multi–drug resistant pathogenic bacteria, personal care products

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1658 In vitro Biological Activity of Some Synthesized Monoazo Heterocycles Based On Thiophene and Thiazolyl-Thiophene Analogue

Authors: Mohamed E. Khalifa, Adil A. Gobouri

Abstract:

Potential synthesis of a series of 3-amino-4-arylazothiophene derivatives from reaction of 2-cyano-2-phenylthiocarbamoyl acetamide and the appropriate α-halogenated reagents, followed by coupling with different aryl diazonium salts (Japp-Klingemann reaction), and another series of 5-arylazo-thiazol-2-ylcarbamoyl-thiophene derivatives from base-catalyzed intramolecular condensation of 5-arylazo-2-(N-chloroacetyl)amino-thiazole with selected B-keto compounds (Thorpe-Ziegler reaction) was performed. The biological activity of the two series was studied in vitro. Their versatility for pharmaceutical purposes was reported, where they displayed remarkable activities against selected pathogenic microorganisms; Bacillus subtilize, Staphylococcus aureus (Gram positive bacteria), Escherichia coli, Pseudomonas aeruginosa (Gram negative bacteria) and Aspergillus flavus, Candida albicans (fungi) with various degrees related to their chemical structures.

Keywords: thiophene, 2-aminothiazole, compounds, antioxidant, antitumor, antimicrobial

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1657 Microbial Quality of Raw Camel Milk Produced in South of Morocco

Authors: Maha Alaoui Ismaili, Bouchta Saidi, Mohamed Zahar, Abed Hamama

Abstract:

Thirty one samples of raw camel milk obtained from the region of Laâyoune (South of Morocco) were examined for their microbial quality and presence of some pathogenic bacteria (Staphylococcus aureus and Salmonella sp.). pH of the samples ranged from 6.31 to 6.64 and their titratable acidity had a mean value of 18.56 °Dornic. Data obtained showed a strong microbial contamination with an average total aerobic flora of 1.76 108 ufc ml-1 and a very high fecal counts: 1.82 107 ; 3.25 106 and 3.75 106 ufc.ml-1 in average for total coliforms, fecal coliforms and enterococci respectively. Yeasts and moulds were also found at average respective levels of 3.13 106 and 1.60 105 ufc.ml-1. Salmonella sp. and S. aureus was detected respectively in 13% and 30% of the milk samples. These results indicate clearly the lack of hygienic conditions of camel milk production and storage in this region. Lactic acid bacteria were found at the following average numbers: 4.25 107 ; 4.45 107 and 3.55 107 ufc.ml-1 for Lactococci, Leuconostocs and Lactobacilli respectively.

Keywords: camel milk, microbial quality, Salmonella, Staphylococcus aureus

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1656 Campylobacteriosis as a Zoonotic Disease

Authors: A. Jafarzadeh, G. R. Hashemi Tabar

Abstract:

Campylobacteriosis is caused by Campylobacter organisms. This is most commonly caused by C. jejuni, It is among the most common bacterial infections of humans, often a foodborne illness. It produces an inflammatory, sometimes bloody, diarrhea or dysentery syndrome, mostly including cramps, fever and pain. It is found in cattle, swine, and birds, where it is non-pathogenic. But the illness can also be caused by C. coli (also found in cattle, swine, and birds) C. upsaliensis (found in cats and dogs) and C. lari (present in seabirds in particular). Infection with a Campylobacter species is one of the most common causes of human bacterial gastroenteritis. For instance, an estimated 2 million cases of Campylobacter enteritis occur annually in the U.S., accounting for 5-7% of cases of gastroenteritis. Furthermore, in the United Kingdom during 2000 Campylobacter jejuni was involved in 77.3% in all cases of foodborne illness. 15 out of every 100,000 people are diagnosed with campylobacteriosis every year, and with many cases going unreported, up to 0.5% of the general population may unknowingly harbor Campylobacter in their gut annually. A large animal reservoir is present as well, with up to 100% of poultry, including chickens, turkeys, and waterfowl, having asymptomatic infections in their intestinal tracts. An infected chicken may contain up to 109 bacteria per 25 grams, and due to the installations, the bacteria is rapidly spread to other chicken. This vastly exceeds the infectious dose of 1000-10,000 bacteria for humans. In this article this disease is fully discussed in human and animals.

Keywords: campylobacteriosis, human, animal, zoonosis

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1655 Microbiological Analysis, Cytotoxic and Genotoxic Effects from Material Captured in PM2.5 and PM10 Filters Used in the Aburrá Valley Air Quality Monitoring Network (Colombia)

Authors: Carmen E. Zapata, Juan Bautista, Olga Montoya, Claudia Moreno, Marisol Suarez, Alejandra Betancur, Duvan Nanclares, Natalia A. Cano

Abstract:

This study aims to evaluate the diversity of microorganisms in filters PM2.5 and PM10; and determine the genotoxic and cytotoxic activity of the complex mixture present in PM2.5 filters used in the Aburrá Valley Air Quality Monitoring Network (Colombia). The research results indicate that particulate matter PM2.5 of different monitoring stations are bacteria; however, this study of detection of bacteria and their phylogenetic relationship is not complete evidence to connect the microorganisms with pathogenic or degrading activities of compounds present in the air. Additionally, it was demonstrated the damage induced by the particulate material in the cell membrane, lysosomal and endosomal membrane and in the mitochondrial metabolism; this damage was independent of the PM2.5 concentrations in almost all the cases.

Keywords: cytotoxic, genotoxic, microbiological analysis, PM10, PM2.5

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1654 Rapid Classification of Soft Rot Enterobacteriaceae Phyto-Pathogens Pectobacterium and Dickeya Spp. Using Infrared Spectroscopy and Machine Learning

Authors: George Abu-Aqil, Leah Tsror, Elad Shufan, Shaul Mordechai, Mahmoud Huleihel, Ahmad Salman

Abstract:

Pectobacterium and Dickeya spp which negatively affect a wide range of crops are the main causes of the aggressive diseases of agricultural crops. These aggressive diseases are responsible for a huge economic loss in agriculture including a severe decrease in the quality of the stored vegetables and fruits. Therefore, it is important to detect these pathogenic bacteria at their early stages of infection to control their spread and consequently reduce the economic losses. In addition, early detection is vital for producing non-infected propagative material for future generations. The currently used molecular techniques for the identification of these bacteria at the strain level are expensive and laborious. Other techniques require a long time of ~48 h for detection. Thus, there is a clear need for rapid, non-expensive, accurate and reliable techniques for early detection of these bacteria. In this study, infrared spectroscopy, which is a well-known technique with all its features, was used for rapid detection of Pectobacterium and Dickeya spp. at the strain level. The bacteria were isolated from potato plants and tubers with soft rot symptoms and measured by infrared spectroscopy. The obtained spectra were analyzed using different machine learning algorithms. The performances of our approach for taxonomic classification among the bacterial samples were evaluated in terms of success rates. The success rates for the correct classification of the genus, species and strain levels were ~100%, 95.2% and 92.6% respectively.

Keywords: soft rot enterobacteriaceae (SRE), pectobacterium, dickeya, plant infections, potato, solanum tuberosum, infrared spectroscopy, machine learning

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1653 An Acoustical Diagnosis of a Shaft-Wood Phyto-Pathogenic Damage of Sequoiadendron giganteum (Lindl.) Buccholz

Authors: Yuri V. Plugatar, Vladimir P. Koba, Vladimir V. Papelbu, Vladimir N. Gerasimchuk, Tatjana M. Sakhno

Abstract:

Using a supersonic shaft–wood tomography, the evaluation of a shaft-wood phyto-pathogenic damage level of Sequoiadendron giganteum (Lindl.) Buccholz was prosecuted. The digital bivariate reflections of the shaft tissue damage were obtained, the characteristics of comparative parameters of the wood-decay degree were given. The investigation results allowed to show up the role of some edaphic factors in their affection on a vital condition and the level of destructive processes while shaft tissue damaging of S.giganteum. It was pinned up that soil consolidation, and hydro-morphication equally make for a phyto-pathogenic damage of plants. While soil consolidation negative acting the shaft-wood damage is located in an underneath of a shaft. In the conditions of an enlarged hydro-morphication a tissue degradation runs less intensively, the destructive processes more active spread in a vertical section of a shaft. The use of a supersonic tomography method gives wide possibilities to diagnose a shaft-wood phyto-pathogenic damage.

Keywords: Sequoiadendron giganteum (Lindl.) Buccholz, supersonic tomography, diagnosis, phyto-pathogenic damage, a vital condition

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1652 Evaluation of Bacterial Composition of the Aerosol of Selected Abattoirs in Akure, South Western Nigeria

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

Abstract:

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

Keywords: abattoir, aerosol, bacterial composition, environment

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1651 Studies on the Bioactivity of Different Solvents Extracts of Selected Marine Macroalgae against Fish Pathogens

Authors: Mary Ghobrial, Sahar Wefky

Abstract:

Marine macroalgae have proven to be rich source of bioactive compounds with biomedical potential, not only for human but also for veterinary medicine. Emergence of microbial disease in aquaculture industries implies serious loses. Usage of commercial antibiotics for fish disease treatment produces undesirable side effects. Marine organisms are a rich source of structurally novel biologically active metabolites. Competition for space and nutrients led to the evolution of antimicrobial defense strategies in the aquatic environment. The interest in marine organisms as a potential and promising source of pharmaceutical agents has increased in the last years. Many bioactive and pharmacologically active substances have been isolated from microalgae. Compounds with antibacterial, antifungal and antiviral activities have been also detected in green, brown and red algae. Selected species of marine benthic algae belonging to the Phaeophyta and Rhodophyta, collected from different coastal areas of Alexandria (Egypt), were investigated for their antibacterial and antifungal, activities. Macroalgae samples were collected during low tide from the Alexandria Mediterranean coast. Samples were air dried under shade at room temperature. The dry algae were ground, using electric mixer grinder. They were soaked in 10 ml of each of the solvents acetone, ethanol, methanol and hexane. Antimicrobial activity was evaluated using well-cut diffusion technique In vitro screening of organic solvent extracts from the marine macroalgae Laurencia pinnatifida, Pterocladia capillaceae, Stepopodium zonale, Halopteris scoparia and Sargassum hystrix, showed specific activity in inhibiting the growth of five virulent strains of bacteria pathogenic to fish Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio anguillarum, V. tandara, Escherichia coli and two fungi Aspergillus flavus and A. niger. Results showed that, acetone and ethanol extracts of all test macroalgae exhibited antibacterial activity, while acetone extract of the brown Sargassum hystrix displayed the highest antifungal activity. The extracts of seaweeds inhibited bacteria more strongly than fungi and species of the Rhodophyta showed the greatest activity against the bacteria rather than fungi tested. The gas liquid chromatography coupled with mass spectrometry detection technique allows good qualitative and quantitative analysis of the fractionated extracts with high sensitivity to the smaller amounts of components. Results indicated that, the main common component in the acetone extracts of L. pinnatifida and P. capillacea is 4-hydroxy-4-methyl2-pentanone representing 64.38 and 58.60%. Thus, the extracts derived from the red macroalgae were more efficient than those obtained from the brown macroalgae in combating bacterial pathogens rather than pathogenic fungi. The most preferred species over all was the red Laurencia pinnatifida. In conclusion, the present study provides the potential of red and brown macroalgae extracts for development of anti-pathogenic agents for use in fish aquaculture.

Keywords: bacteria, fungi, extracts, solvents

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1650 In Vivo Evaluation of the Therapeutic Effect on Intestinal Disorders by Thermophilic Streptococcus Isolated from Camel Milk

Authors: A. T. Laiche, M. L. Tlil, Benine B., S. Bechoua

Abstract:

The aim of this study is to isolate and select, from camel milk from El-Oued region in Algeria, a strains of lactic acid bacteria and possessing probiotic properties ; and to evaluate their potential effect on intestinal disorders in Wistar ratsmThe results relating to the selection of probiotic strains confirms that the Thermophilic streptococcus exhibits the best probiotic activity performance, with a resistance important to different degrees of pH and to bile salts, and a remarkable antibacterial activity and resistance to antibiotics compared to the other four isolated strains. In the in vivo study, diseases are induced in rats at the level of the digestive system, it was reported that the administration of Escherichia coli and castor oil caused an intestinal disorders. The microscopic observation of the histological section of the intestine showed a damaged intestinal structure and some symptoms of its irritation, including a decrease in the height of the villi and the presence of others destroyed cells, and after treatment with Streptococcus thermophilus, the microscopic observation of the cut of the histological section of the intestine showed almost complete disappearance of the mentioned symptoms, The dosage of the hematological parameters by complete blood count (CBC) is in agreement with the results of the histological sections.

Keywords: camel milk, probiotic, pathogenic bacteria, intestinal disorders, lactic acid bacteria

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1649 Isolation and Characterization of Indigenous Rhizosphere Bacteria Producing Gibberellin Acid from Local Soybeans in Three Different Areas of South Sulawesi

Authors: Asmiaty Sahur, Ambo Ala, Baharuddin Patanjengi, Elkawakib Syam'un

Abstract:

This study aimed to isolate and characterize the indigenous Rhizosphere bacteria producing Gibberellin Acid as plant growth isolated from local soybean of three different areas in South Sulawesi, Indonesia. Several soil samples of soybean plants were collected from the Rhizosphere of local soybeans in three different areas of South Sulawesi such as Soppeng, Bone and Takalar. There were 56 isolates of bacteria were isolated and grouped into gram-positive bacteria and gram negative bacteria .There are 35 isolates produce a thick slime or slimy when cultured on media Natrium Broth and the remaining of those produced spores. The results showed that of potential bacterial isolated produced Gibberellin Acid in high concentration. The best isolate of Rhizosphere bacteria for the production of Gibberellin Acid is with concentration 2%. There are 4 isolates that had higher concentration are AKB 19 (4.67 mg/ml) followed by RKS 17 (3.80 mg/ml), RKS 25 (3.70 mg / ml) and RKS 24 (3.29 mg/ml) respectively.

Keywords: rhizosphere, bacteria, gibberellin acid, soybeans

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1648 The Effect of Bacteria on Mercury's Biological Removal

Authors: Nastaran Soltani

Abstract:

Heavy metals such as Mercury are toxic elements that enter the environment through different ways and endanger the environment, plants, animals, and humans’ health. Microbial activities reduce the amount of heavy metals. Therefore, an effective mechanism to eliminate heavy metals in the nature and factory slops, is using bacteria living in polluted areas. Karun River in Khuzestan Province in Iran has been always polluted by heavy metals as it is located among different industries in the region. This study was performed based on the data from sampling water and sediments of four stations across the river during the four seasons of a year. The isolation of resistant bacteria was performed through enrichment and direct cultivation in a solid medium containing mercury. Various bacteria such as Pseudomonas sp., Serratia Marcescens, and E.coli were identified as mercury-resistant bacteria. The power of these bacteria to remove mercury varied from 28% to 86%, with strongest power belonging to Pseudomonas sp. isolated in spring making a good candidate to be used for mercury biological removal from factory slops.

Keywords: bacteria, Karun River, mercury, biological removal, mercury-resistant

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1647 Rapid Identification and Diagnosis of the Pathogenic Leptospiras through Comparison among Culture, PCR and Real Time PCR Techniques from Samples of Human and Mouse Feces

Authors: S. Rostampour Yasouri, M. Ghane, M. Doudi

Abstract:

Leptospirosis is one of the most significant infectious and zoonotic diseases along with global spreading. This disease is causative agent of economoic losses and human fatalities in various countries, including Northern provinces of Iran. The aim of this research is to identify and compare the rapid diagnostic techniques of pathogenic leptospiras, considering the multifacetedness of the disease from a clinical manifestation and premature death of patients. In the spring and summer of 2020-2022, 25 fecal samples were collected from suspected leptospirosis patients and 25 Fecal samples from mice residing in the rice fields and factories in Tonekabon city. Samples were prepared by centrifugation and passing through membrane filters. Culture technique was used in liquid and solid EMJH media during one month of incubation at 30°C. Then, the media were examined microscopically. DNA extraction was conducted by extraction Kit. Diagnosis of leptospiras was enforced by PCR and Real time PCR (SYBR Green) techniques using lipL32 specific primer. Out of the patients, 11 samples (44%) and 8 samples (32%) were determined to be pathogenic Leptospira by Real time PCR and PCR technique, respectively. Out of the mice, 9 Samples (36%) and 3 samples (12%) were determined to be pathogenic Leptospira by the mentioned techniques, respectively. Although the culture technique is considered to be the gold standard technique, but due to the slow growth of pathogenic Leptospira and lack of colony formation of some species, it is not a fast technique. Real time PCR allowed rapid diagnosis with much higher accuracy compared to PCR because PCR could not completely identify samples with lower microbial load.

Keywords: culture, pathogenic leptospiras, PCR, real time PCR

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