Search results for: enterotoxigenic escherichia coli

Authors: Arun Jyothi Bheemagani, Chakrapani Pullagummi, Anupalli Roja Rani

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Exploration of the chemical constituents of the plants and pharmacological screening may provide us the basis for the development of novel agents. Plants have provided us some of the very important life saving drugs used in the modern medicine. The aim of our work was to screen the phytochemical constituents and antimicrobial and antioxidant activities of methanol extract of leaves of Annona cherimola Mill plant from Tirumala forest, Tirupathi. It was originally called Chirimuya by the Inca people who lived where it was growing in the Andes of South America, is an edible fruit-bearing species of the genus Annona from the family Annonaceae. Annona cherimola Mill is a multipurpose tree with edible fruits and is one of the sources of the medicinal products. The antibacterial activity was measured by agar well diffusion method; the diameter of the zone of bacterial growth inhibition was measured after incubation of plates. The inhibitory effect was studied against the pathogenic bacteria (Klebsiella pneumonia, Bacillus subtilis, Staphylococcus aureus and Escherichia coli (E. coli). Antioxidant assays were also performed for the same extracts by spectrophotometric methods using known standard antioxidants as reference. The studied plant extracts were found to be very effective against the pathogenic microorganisms tested. The methanolic extract of Annona cherimola Mill from showed maximum activity against Escherichia coli and Staphylococcus aureus and the least concentration required showing the activity was 0.5mg/ml. Phytochemical screening of the plants revealed the presence of flavonoids, alkaloids, steroids and carbohydrates. Good presence of antioxidants was also found in the methanolic extracts.

Keywords: annona cherimola, phytochemicals, antioxidant and antibacterial activity, methanol extract

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Authors: Sanaa Bardaweel

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Inorganic polyphosphate (poly P) is present in all living forms tested to date, from each of the three kingdoms of life. Studied mainly in prokaryotes, poly P and its associated enzymes are vital in diverse basic metabolism, in at least some structural functions and, notably, in stress responses. These plentiful and unrelated roles for poly P are probably the consequence of its presence in life-forms early in evolution. The genomes of many bacterial species, including pathogens, encode a homologue of a major poly P synthetic enzyme, poly P kinase 1 (PPK1). Genetic deletion of ppk1 results in reduced poly P levels and loss of pathogens virulence towards protozoa and animals. Thus far, no PPK1 homologue has been identified in higher-order eukaryotes and, therefore, PPK1 represents a novel target for chemotherapy. The idea of the current study is to purify the PPK1 from Escherichia coli to homogeneity in order to study the effect of active site point mutations on PPK1 catalysis via the application of site-directed mutagenesis strategy. The knowledge obtained about the active site of PPK1 will be utilized to characterize the catalytic and kinetic mechanism of PPK1 with model substrates. Comprehensive understanding of the enzyme kinetic mechanism and catalysis will be used to design and screen a library of synthetic compounds for potential discovery of selective PPK1-inhibitors.

Keywords: antimicobial, Escherichia coli, inorganic polyphosphate, PPK1-inhibitors

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Authors: G. Petruzzelli, F. Pedron, M. Grifoni, A. Pera, I. Rosellini, B. Pezzarossa

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The addition of lime as Ca(OH)2 to sewage sludge to destroy pathogens (Escherichia coli), was evaluated also in relation to heavy metal bioavailability. The obtained results show that the use of calcium hydroxide at the dose of 3% effectively destroyed pathogens ensuring the stability at high pH values over long period and the duration of the sewage sludge stabilization. In general, lime addition decreased the total extractability of heavy metals indicating a reduced bioavailability of these elements. This is particularly important for a safe utilization in agricultural soils to reduce the possible transfer of heavy metals to the food chain.

Keywords: biological sludge, Ca(OH)2, copper, pathogens, sanitation, zinc

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Authors: Ewa Brychcy, Natalia Ulbin-Figlewicz, Dominika Kulig, Żaneta Król, Andrzej Jarmoluk

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Acidic electrolyzed water (AEW) is an alternative with environmentally friendly broad spectrum microbial decontamination. It is produced by membrane electrolysis of a dilute NaCl solution in water ionizers. The aim of the study was to evaluate the effectiveness of low-concentrated AEW in reducing selected foodborne pathogens and to examine its bactericidal effect on cellular structures of Escherichia coli. E. coli and S. aureus cells were undetectable after 10 minutes of contact with electrolyzed salt solutions. Non-electrolyzed solutions did not inhibit the growth of bacteria. AE water was found to destroy the cellular structures of the E. coli. The use of more concentrated salt solutions and prolonged electrolysis time from 5 to 10 minutes resulted in a greater changes of rods shape as compared to the control and non-electrolyzed NaCl solutions. This research showed that low-concentrated acid electrolyzed water is an effective method to significantly reduce pathogenic microorganisms and indicated its potential application for decontamination of meat.

Keywords: acidic electrolyzed water, foodborne pathogens, meat decontamination, membrane electrolysis

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Authors: Tahra El-Obeid, Reham Mousa, Amal Alzahiri

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During the past few years the traditional market of Qatar has become an attraction to many customers who eat from the numerous women street vendors selling Qatari traditional dishes. To gain an understanding on the safety of these street vended foods, we designed the study to test microbiological quality of 14 different Qatari foods sold in Souk Wagif, the main traditional market in Qatar. This study was conducted to mainly identify presence or absence of microbial pathogens. A total of 56 samples were purchased from 10 different street vendors and the samples were collected randomly on different days. The samples were tested for microbial contaminants at Central Food Laboratories, Doha, Qatar. The qualitative study was conducted using Real Time-PCR to screen for; Salmonella spp., Listeria monocytogenes, Escherichia coli and E. coli 0157:H7. Out of the 56 samples, only two samples “Biryani” and “Khabess” contained E. coli. However, both samples tested negative for E. coli O157:H7. The microbial contamination of the Qatari traditional street vended foods was 3%. This result may be attributed to the food safety training requirement set by the regulatory authorities before issuing any license to food handlers in Qatar as well as the food inspection conducted by the food health inspectors on a regular basis.

Keywords: microbiological quality, street vended food, traditional dishes, Qatar

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Authors: Samad Lotfollahzadeh

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Escherichia coli (E.coli) is the most isolated bacteria from blood circulation of septicemic calves. Given the prevalence of septicemia in animals and its economic importance in veterinary practice, better understanding of changes in clinical signs following disease, may contribute to early detection of the disorder. The present study has been carried out to detect changes of clinical signs in induced sepsis in calves with E.coli. Colisepticemia has been induced in 10 twenty-day old healthy Holstein- Frisian calves with intravenous injection of 1.5 X 109 colony forming units (cfu) of O111: H8 strain of E.coli. Clinical signs including rectal temperature, heart rate, respiratory rate, shock, appetite, sucking reflex, feces consistency, general behavior, dehydration and standing ability were recorded in experimental calves during 24 hours after induction of colisepticemia. Blood culture was also carried out from calves four times during the experiment. ANOVA with repeated measure is used to see changes of calves’ clinical signs to experimental colisepticemia, and values of P≤ 0.05 was considered statistically significant. Mean values of rectal temperature and heart rate as well as median values of respiratory rate, appetite, suckling reflex, standing ability and feces consistency of experimental calves increased significantly during the study (P<0.05). In the present study, median value of shock score was not significantly increased in experimental calves (P> 0.05). The results of present study showed that total score of clinical signs in calves with experimental colisepticemia increased significantly, although the score of some clinical signs such as shock did not change significantly.

Keywords: calves, clinical signs scoring, E. coli O111:H8, experimental colisepticemia

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Authors: Nastaran Hemmati, Farhad Nikkhahi, Amir Javadi, Sahar Eskandarion, Seyed Mahmuod Amin Marashi

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Neisseria meningitidis, Escherichia coli K, Streptococcus agalactiae, and Streptococcus pneumoniae cause 90% of bacterial meningitis. Almost all infected people die or have irreversible neurological complications. Therefore, it is essential to have a diagnostic kit with the ability to quickly detect these fatal infections. The project involved 212 patients from whom cerebrospinal fluid samples were obtained. After total genome extraction and performing multiplex quantitative polymerase chain reaction (qPCR), the presence or absence of each infectious factor was determined by comparing with standard strains. The specificity, sensitivity, positive predictive value, and negative predictive value calculated were 100%, 92.9%, 50%, and 100%, respectively. So, due to the high specificity and sensitivity of the designed primers, they can be used instead of bacterial culture that takes at least 24 to 48 hours. The remarkable benefit of this method is associated with the speed (up to 3 hours) at which the procedure could be completed. It is also worth noting that this method can reduce the personnel unintentional errors which may occur in the laboratory. On the other hand, as this method simultaneously identifies four common factors that cause bacterial meningitis, it could be used as an auxiliary method diagnostic technique in laboratories particularly in cases of emergency medicine.

Keywords: cerebrospinal fluid, meningitis, quantitative polymerase chain reaction, simultaneous detection, diagnosis testing

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Authors: kamran Bhatti

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Objective: To evaluate spectrum and resistance rates to antibacterial agents in causative pathogens of bacterial prostatitis in patients from Southern Europe, the Middle East, and Africa. Materials: 1027 isolates from cultures of urine or expressed prostatic secretion, post-massage urine or seminal fluid, or urethral samples were considered. Results: Escherichia coli (32%) and Enterococcus spp. (21%) were the most common isolates. Other Gram-negative, Gram-positive, and atypical pathogens accounted for 22%, 20%, and 5%, respectively. Resistance was <15% for piperacillin/tazobactam and carbapenems (both Gram-negative and -positive pathogens); <5% for glycopeptides against Gram-positive; 7%, 14%, and 20% for aminoglycosides, fosfomycin, and macrolides against Gram-negative pathogens, respectively; 10% for amoxicillin/clavulanate against Gram-positive pathogens; <20% for cephalosporins and fluoroquinolones against to Gram-negative pathogens (higher against Gram-positive pathogens); none for macrolides against atypical pathogens, but 20% and 27% for fluoroquinolones and tetracyclines. In West Africa, the resistance rates were generally higher, although the highest rates for ampicillin, cephalosporins, and fluoroquinolones were observed in the Gulf area. Lower rates were observed in Southeastern Europe. Conclusions: Resistance to antibiotics is a health problem requiring local health authorities to combat this phenomenon. Knowledge of the spectrum of pathogens and antibiotic resistance rates is crucial to assess local guidelines for the treatment of prostatitis.

Keywords: enterobacteriacae; escherichia coli, gram-positive pathogens, antibiotic, bacterial prostatitis, resistance

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Authors: Yu Yoshihara, Chika Tada, Moe Takada, Nyam-Osor Purevdorj, Khorolmaa Chimedtseren, Yutaka Nakai

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Water pollution from animal waste and its influence on grazing animals is a current concern regarding Mongolian grazing lands. We allocated 32 free-grazing lambs to four groups and provided each with water from a different source (upper stream, lower stream, well, and pond) for 49 days. We recorded the amount of water consumed by the lambs, as well as their body weight, behavior, white blood cell count, acute phase (haptoglobin) protein level, and fecal condition. We measured the chemical and biological qualities of the four types of water, and we detected enteropathogenic and enterohemorrhagic Escherichia coli in fecal samples by using a genetic approach. Pond water contained high levels of nitrogen and minerals, and well water contained high levels of bacteria. The odor concentration index decreased in order from pond water to upper stream, lower stream, and well. On day 15 of the experiment, the following parameters were the highest in lambs drinking water from the following sources: water intake (pond or lower stream), body weight gain (pond), WBC count (lower stream), haptoglobin concentration (well), and enteropathogenic E. coli infection rate (lower stream). Lambs that drank well water spent more time lying down and less time grazing than the others, and lambs that drank pond water spent more time standing and less time lying down. Lambs given upper or lower stream water exhibited more severe diarrhea on day 15 of the experiment than before the experiment. Mongolian sheep seemed to adapt to chemically contaminated water: their productivity benefited the most from pond water, likely owing to its rich mineral content. Lambs that drank lower stream water showed increases in enteropathogenic E. coli infection, clinical diarrhea, and WBC count. Lambs that drank well water, which was bacteriologically contaminated, had increased serum acute phase protein levels and poor physical condition; they were thus at increased risk of negative health and production effects.

Keywords: DNA, Escherichia coli, fecal sample, lower stream, well water

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Authors: S. I. Enem, S. I. Oboegbulem

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Objective: The most frequently implicated E. coli serotype causing haemorrhagic colitis and haemorrhagic uraemic syndrome (HUS) is VTEC 0157. However, non-O157 VTEC is now known to be as prevalent as VETC O157 infection (or even more) in most parts of the world. The objective of the study was to establish the occurrence of non-O157 VTEC serotypes in cattle in the Federal Capital Territory (FCT) Abuja, Nigeria. The level of significance of the infection with sex, age and season were also tested. Methods: The study was carried out in the FCT, Abuja, Nigeria which is located between latitude 8o and 90 25` North of the equator and longitude 60 45` and 7045` East of the Greenwich meridian. The cross sectional epidemiological method and multi-staged sampling technique were used in this study. Samples were collected from the freshly voided faeces of both apparently healthy and diarrhoeic cattle in selected abattoirs and cattle herds. Enriched samples were analyzed bacteriologically and biochemically after which they were characterised using commercially prepared latex agglutination test kits. Results: A total of 718 faecal samples from cattle were analyzed for the presence of VTEC non-O157. Thirty eight (5.23%) were positive for non-O157. There was no significant association (p > 0.05) between sex and infection with non-O157 VTEC in cattle. There was a significant association (P < 0.05) between age and infection with non-O157 VTEC in cattle. Calves were more associated than the adults. There was also a significant association (P < 0.05) between season and infection with non-O157 VTEC in cattle. The dry season was more associated than the wet season. Conclusion: The study established the occurrence and prevalence of non-O157 VTEC in cattle in FCT, Abuja, Nigeria. As a major food animal in Nigeria, infection in cattle provides an epidemiological causal association to the infection in humans. The result showed that warmer seasons (dry season) stimulate the presence of VTEC infection in animals and thus, as a consequence, increases the number of human cases. The prevalence was also higher in younger calves (< 6 months) probably as a result of undeveloped immune system.

Keywords: prevalence, distribution, Verocytotoxigenic escherichia coli (VTEC), non-O157 serotypes, cattle

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Authors: Henny A. Dien, Roike I. Montolalu, Feny Mentang, Jupni Keno, Reynerd S. Burdam, Siegfried Berhimpon

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Fish Sausages became popular nowadays, because of high nutritious and low in cholesterol. However, this food is also highly perishable and often contaminated by pathogen bacteria. Edible film was made from myofibril of Black Marlin (Makaira indica) waste, with addition of liquid smoke 0.8%. The aim of this study were to determine the TPC, total coliform and Escherichia coli in fish sausages coated with smoked edible film, and stored in room temperature (26-29oC), and refrigerator (5-10oC). Results shown that TPC in fish sausages coated with smoked edible film were lower than that of without coated, both for storage in room temperature and in refrigerator. Total coliform in coated with smoked edible film and stored in room temperature ranged between 7-120 MPN/g (1-4 days), while stored in refrigerator ranged between 7-93 MPN/g (1-6 days); while fish sausages coated with edible film without liquid smoke were 7-240 MPN/g (1-4 days) in room temperature, and 7-150 MPN/g in refrigerator. Total E. coli of fish sausages coated with smoked edible film and stored in room temperature ranged between 3-4 MPN/g (1-4 days), while stored in refrigerator ranged were 3 MPN/g (1-6 days); while fish sausages coated with edible film without smoked both stored in room temperature and in refrigerator, shown total E. coli 3 MPN/g during 4 days in room temperature, and 6 days in refrigerator. Total E. coli of sausages without coated stored in room temperature ranged between 7-24 MPN/g, and that of stored in refrigerator ranged between 3-4 MPN/g.

Keywords: smoke liquid, edible film, coating, sausages

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Authors: Meltem Agar, Maisem Laabei, Hannah Leese, Pedro Estrela

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Pathogenic bacteria and the diseases they cause have become a global problem. Their early detection is vital and can only be possible by detecting the bacteria causing the disease accurately and rapidly. Great progress has been made in this field with the use of biosensors. Molecularly imprinted polymers have gain broad interest because of their excellent properties over natural receptors, such as being stable in a variety of conditions, inexpensive, biocompatible and having long shelf life. These properties make molecularly imprinted polymers an attractive candidate to be used in biosensors. In this study it is aimed to produce an aptamer-molecularly imprinted polymer based electrochemical sensor by utilizing the properties of molecularly imprinted polymers coupled with the enhanced specificity offered by DNA aptamers. These ‘apta-MIP’ sensors were used for the detection of Staphylococcus aureus and Escherichia coli. The experimental parameters for the fabrication of sensor were optimized, and detection of the bacteria was evaluated via Electrochemical Impedance Spectroscopy. Sensitivity and selectivity experiments were conducted. Furthermore, molecularly imprinted polymer only and aptamer only electrochemical sensors were produced separately, and their performance were compared with the electrochemical sensor produced in this study. Aptamer-molecularly imprinted polymer based electrochemical sensor showed good sensitivity and selectivity in terms of detection of Staphylococcus aureus and Escherichia coli. The performance of the sensor was assessed in buffer solution and tap water.

Keywords: aptamer, electrochemical sensor, staphylococcus aureus, molecularly imprinted polymer

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

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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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Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

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Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

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Authors: S. M. Abd-Altwab, M. R. Noor El-Din

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This paper aims to the synthesis of new ethoxylated amide as bactericides to prevent the growth of Gram +ve and –ve bacteria of water-in-diesel fuel nanoemulsions over a long period of time as three months. To realize it, eight kinetically stable water-in-diesel fuel nanoemulsions differing in surfactant concentrations and water contents ranging from 4 to 8 and 5 to 8 wt.,wt.,% of total weight of the nanoemulsions, respectively were formed at a temperature of 20 °C. The performance of this ethoxylated amide as bactericides agents against two strains of Gram-negative bacteria, namely, Pseudomonas aeruginosa and Escherichia coli, and two strains of Gram-positive bacteria namely, Staphylococcus aureus and Bacillus subtilis, were evaluated as antimicrobial agents. The maximum and minimum antimicrobial activities were 85 and 71 % against S. aureus and E. coli, respectively, at a concentration of 5 mg/l, pH 7, and 37 °C.

Keywords: nanoemulsion, bacteriocide, diesel fuel, emulsifier

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Authors: Satwik Majumder, Dongyun Jung, Jennifer Ronholm, Saji George

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Bovine mastitis is the most common infectious disease in dairy cattle, with major economic implications for the dairy industry worldwide. Continuous monitoring for the emergence of antimicrobial resistance (AMR) among bacterial isolates from dairy farms is vital not only for animal husbandry but also for public health. In this study, the prevalence of AMR in 113 Escherichia coli isolates from cases of bovine clinical mastitis in Canada was investigated. Kirby-Bauer disk diffusion test with 18 antibiotics and microdilution method with three heavy metals (copper, zinc, and silver) was performed to determine the antibiotic and heavy-metal susceptibility. Resistant strains were assessed for efflux and ß-lactamase activities besides assessing biofilm formation and hemolysis. Whole-genome sequences for each of the isolates were examined to detect the presence of genes corresponding to the observed AMR and virulence factors. Phenotypic analysis revealed that 32 isolates were resistant to one or more antibiotics, and 107 showed resistance against at least one heavy metal. Quinolones and silver were the most efficient against the tested isolates. Among the AMR isolates, AcrAB-TolC efflux activity and ß-lactamase enzyme activities were detected in 13 and 14 isolates, respectively. All isolates produced biofilm but with different capacities, and 33 isolates showed α-hemolysin activity. A positive correlation (Pearson r = +0.89) between efflux pump activity and quantity of biofilm was observed. Genes associated with aggregation, adhesion, cyclic di-GMP, quorum sensing were detected in the AMR isolates, corroborating phenotype observations. This investigation showed the prevalence of AMR in E. coli isolates from bovine clinical mastitis. The results also suggest the inadequacy of antimicrobials with a single mode of action to curtail AMR bacteria with multiple mechanisms of resistance and virulence factors. Therefore, it calls for combinatorial therapy for the effective management of AMR infections in dairy farms and combats its potential transmission to the food supply chain through milk and dairy products.

Keywords: antimicrobial resistance, E. coli, bovine mastitis, antibiotics, heavy-metals, efflux pump, ß-lactamase enzyme, biofilm, whole-genome sequencing

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Authors: Atinderpal Kaur, Shweta Dang

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Our aim is to develop a nanoemulsion-based delivery system containing polyphenon 60 (P60) and ciprofloxacin (Cipro) for intravaginal delivery to treat urinary tract infection. In the present study Polyphenon 60 (P60) and ciprofloxacin (Cipro) were loaded in a single nano emulsion (NE) system via ultra-sonication technique and characterized for particle size, in vitro release and antibacterial efficacy against Bcl-2 level Escherichia coli bacteria. To determine in vivo pharmacokinetic parameters and intravaginal transportation of NE, gamma scintigraphy and biodistribution study was conducted by radiolabelling NE with technetium pertechnetate (99mTc). The preliminary antibacterial investigation showed synergy between these compounds with FICindex of 0.42. The developed formulation showed zeta potential +55.3 and particle size of 151.7 nm, with PDI of 0.196. The in vitro release percentage of P60 at the end of 7th hours was 94.8 ± 0.9 % whereas the release for Cipro was 75.1± 0.15 % in simulated vaginal media. MBC was identified and the findings demonstrated that in both ESBL (Extended Spectrum β- lactamase) and MBL (Metallo β- lactamase) cultures the P60+Cipro NE showed inhibition of growth of all the isolates at 2 mg/ml dilutions. The percentage per gram of radiolabelled drug was found (3.50±0.26) and (3.81±0.30) in kidney and urinary bladder, respectively at 3 h. From the findings, it was concluded that the developed P60+Cipro NE was transported efficiently throughout the target organs, had long duration of action and high biocompatibility via intravaginal administration as compared to oral administration.

Keywords: ciprofloxacin, gamma scintigraphy, intravaginal drug delivery, Polyphenon 60

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Authors: J. K. D. M. P. Madara, R. B. L. Dharmawickreme, Linu John, Ivee Boiss

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Flemingia vestita, commonly known as ‘Sohphlang’ is an important medicinal plant found in the North-Eastern region of India, which is traditionally recognized for its anthelmintic properties. This study was aimed to evaluate the phytochemical constituents and antibacterial activity of the tuber skin extracts of the plant species. Methanol, acetone, and water were used to obtain the solvent extractions of the skin peel extracts. Concentrated extracts of skin peel were tested using previously established qualitative phytochemical assays. The antibacterial efficacy of methanol tuber skin extract was tested against Gram-negative and positive microorganisms, namely, Klebsiella pneumonia, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Mycobacterium tuberculosis strains. Agar well diffusion method was employed to determine the zone of inhibition of the plant extracts. Obtained data were statistically analyzed. Methanol extracts of Flemingia vestita were found to be effective against Bacillus subtilis and Mycobacterium tuberculosis at concentrations of 0.5 mg/ml. The reported zone of inhibition for the two strains was 13.3mm ± 0.57 and 16.3mm ± 4.9, respectively. However Klebsiella pneumoniae, Pseudomonas aeruginosa and Escherichia coli were resistant to the plant extracts with no zone of inhibition. Alkaloids, glycosides, and phenols were found to be present in aqueous, methanol, and acetone extracts of the plant in qualitative phytochemical analysis.

Keywords: flemingia vestita, antibacterial activity, phytochemical screening, well diffusion method

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Authors: Ahreum Choi, Otgontuya Tsogbadrakh, Kwang-Hwan Jung

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Microbial rhodopsins are well-known seven-transmembrane proteins that have been extensively studied. These retinal-binding proteins have divided into two types. The type I is microbial rhodopsin, and type II (visual pigment) is expressed mostly in mammalian eyes. For type I rhodopsin, there are two main functions that are ion pumping activity and sensory transduction. Anabaena sensory rhodopsin (ASR) is one of the microbial rhodopsin with main function as photo-sensory transduction. Although ASR is expressed fairly well in Escherichia coli, the expression level is relatively less compare to Proteorhodopsin. In this study, full length of ASR was used to test for the expression influence by codon usage in E. coli. Eight amino acids of codon at N-terminal part of ASR were changed randomly with designed primers, which allow 8,192 nucleotide different cases. The codon changes were screened for the preferable codons of each residue, which have given higher expression yield. Among those 57 selected mutations, there are 24 color-enhanced E. coli colonies that contain ASR proteins, and it showed better expression level than the wild type ASR codon usage. This strongly suggests that high codon usage of only partial N-terminal of protein can increase the expression level of whole protein.

Keywords: 7-transmembrane, all-trans retinal, rhodopsin, codon-usage, protein expression

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Authors: Idan Chiyanzu, Maruping Mangena

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Crude glycerol, a major by-product from the transesterification of triacylglycerides with alcohol to biodiesel, is known to have a broad range of applications. For example, its bioconversion can afford a wide range of chemicals including alcohols, organic acids, hydrogen, solvents and intermediate compounds. In bacteria, the 2-oxoglutarate dioxygenase (2-OGD) enzymes are widely found among the Pseudomonas syringae species and have been recognized with an emerging importance in ethylene formation. However, the use of optimized enzyme function in recombinant systems for crude glycerol conversion to ethylene is still not been reported. The present study investigated the production of ethylene from crude glycerol using engineered E. coli MG1655 and JM109 strains. Ethylene production with an optimized expression system for 2-OGD in E. coli using a codon optimized construct of the ethylene-forming gene was studied. The codon-optimization resulted in a 20-fold increase of protein production and thus an enhanced production of the ethylene gas. For a reliable bioreactor performance, the effect of temperature, fermentation time, pH, substrate concentration, the concentration of methanol, concentration of potassium hydroxide and media supplements on ethylene yield was investigated. The results demonstrate that the recombinant enzyme can be used for future studies to exploit the conversion of low-priced crude glycerol into advanced value products like light olefins, and tools including recombineering techniques for DNA, molecular biology, and bioengineering can be used to allowing unlimited the production of ethylene directly from the fermentation of crude glycerol. It can be concluded that recombinant E.coli production systems represent significantly secure, renewable and environmentally safe alternative to thermochemical approach to ethylene production.

Keywords: crude glycerol, bioethylene, recombinant E. coli, optimization

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Authors: Yong-Cin Chen, Meng-Jiy Wang

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This study reports to prepare cuprous oxide (Cu2O) particles with different dimension and shape for evaluating the antibacterial applications. In the preparation of Cu2O, the surfactant, cetyltrimethylammonium bromide (CTAB), was used as templates to modulate the size of the prepared Cu2O particles. Furthermore, ammonia water was used for adjusting the pH environment that four different shapes of particles including cubic, spherical, octahedral, and star-like Cu2O were synthesized. The physical characteristics of Cu2O particles were evaluated by scanning electron microscope (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), UV/VIS spectrophotometer, and zeta potential meter/particle size analyzer (ZetaPALS). The resistance to bacteria was investigated against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) by applying the synthesized Cu2O particles that the qualitative analyses were facilitated by measuring the inhibition zone on Agar plate.

Keywords: copper oxide, cupric oxide, nanoparticles, antibacetrial

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Authors: M. T. Amina, A. A. Alazba, U. Manzoor

Abstract:

Solar disinfection (SODIS) is a viable method for household water treatment and is recommended by the World Health Organization as cost effective approach that can be used without special skills. The efficiency of both SODIS and solar collector disinfection (SOCODIS) system was evaluated using four different sources of water including stored rainwater, storm water, ground water and treated sewage. Samples with naturally occurring microorganisms were exposed to sunlight for about 8-9 hours in 2-L polyethylene terephthalate bottles under similar experimental conditions. Total coliform (TC), Escherichia coli (E. coli) and heterotrophic plate counts (HPC) were used as microbial water quality indicators for evaluating the disinfection efficiency at different sunlight intensities categorized as weak, mild and strong weathers. Heterotrophic bacteria showed lower inactivation rates compared to E. coli and TC in both SODIS and SOCODIS system. The SOCODIS system at strong weather was the strongest disinfection system in this study and the complete inactivation of HPC was observed after 8-9 hours of exposure with SODIS being ineffective for HPC. At moderate weathers, however, the SOCODIS system did not show complete inactivation of HPC due to very high concentrations (up to 5x10^7 CFU/ml) in both storm water and treated sewage. SODIS even remained ineffective for the complete inactivation of E. coli due to its high concentrations of about 2.5x10^5 in treated sewage compared with other waters even after 8-9 hours of exposure. At weak weather, SODIS was not effective at all while SOCODIS system, though incomplete, showed good disinfection efficiency except for HPC and to some extent for high E. coli concentrations in storm water. Largest reduction of >5 log occurred for TC when used stored rainwater even after 6 hours of exposure in the case of SOCODIS system at strong weather. The lowest E. coli and HPC reduction of ~2 log was observed in SODIS system at weak weather. Further tests with varying pH and turbidity are required to understand the effects of reaction parameters that could be a step forward towards maximizing the disinfection efficiency of such systems for the complete inactivation of naturally occurring E. coli or HPC at moderate or even at weak weathers.

Keywords: efficiency, microbial, SODIS, SOCODIS, weathers

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Authors: Nunyong Fuengkajornfung, Pattama Hirunyophat, Tidarat Sanphom

Abstract:

The objectives of this research were: To study of the recipe standard of Thai steamed dumpling, to study the ratio of modified starch in Thai steamed dumpling, to study chemical elements analyzing and Escherichia coli in Thai steamed dumpling. The experimental processes were designed in two stages as follows: To study the recipe standard of Thai steamed dumpling and to study the ratio of rice flour: modify starch by three levels 90:10, 73:30, and 50:50. The evaluation test used 9 Points Hedonic Scale method by the sensory evaluation test such as color, smell, taste, texture and overall liking. An experimental by Randomized Complete Block Design (RCBD). The statistics used in data analyses were means, standard deviation, one-way ANOVA and Duncan’s New Multiple Range Test. Regression equation, at a statistically significant level of .05. The results showed that the recipe standard was studied from three recipes by the sensory evaluation test such as color, odor, taste, spicy, texture and total acceptance. The result showed that the recipe standard of second was suitably to development. The ratio of rice flour: modified starch had 3 levels 90:10, 73:30, and 50:50 which the process condition of 50:50 had well scores (like moderately to like very much; used 9 Points Hedonic Scale method for the sensory test). Chemical elements analyzing, it showed that moisture 58.63%, fat 5.45%, protein 4.35%, carbohydrate 30.45%, and Ash 1.12%. The Escherichia coli is not found in lab testing.

Keywords: Thai snack in Rattanakosin era, Thai steamed dumpling, modify starch, recipe standard

Procedia PDF Downloads 322

Authors: Davidson E. Egirani, Nanfe R. Poyi, Napoleon Wessey

Abstract:

This paper would report on the effect of solid waste on water resource quality in the Gbarain catchment of the Niger Delta Region of Nigeria. The Gbarain catchment presently hosts two waste-dump sites located along the flanks of a seasonal flow stream and perennially waterlogged terrain. The anthropogenic activity has significantly affected the quality of surface and groundwater in the Gbarain catchment. These wastes have made the water resource environment toxic leading to the poisoning of aquatic life. The contaminated water resources could lead to serious environmental and human health challenges such as low agricultural yields to loss of vital human organs. The contamination is via geological processes such as seepage and direct infiltration of contaminants into watercourses. The results obtained from field and experimental investigations followed by modeling, and graphical interpretation indicate heavy metal load and fecal pollution in some of the groundwater. The metal load, Escherichia coli, and total coliforms counts exceed the international and regional recommended limits. The contaminate values include Lead (> 0.01 mg/L), Mercury (> 0.006 mg/L), Manganese (> 0.4 mg/L and Escherichia coli (> 0 per 100ml) of the samples. Land use planning, enactment, and implementation of environmental laws are necessary for this region, for effective surface water and groundwater resource management.

Keywords: aquatic life, solid waste, environmental health, human health, waste-dump site, water-resource environment

Procedia PDF Downloads 143

Authors: A. V. Chandewar, Sanjay Bais

Abstract:

Backgound: In India, drugs of herbal origin have been used in traditional systems of medicines such as Unani and Ayurveda since ancient times. WHO limit for Escherichia coli is 101/gm cfu, for Staphylococus aureus 105/gm cfu, and for Pseudomonas aeruginosa 103/gm cfu and for Salmonella species nil cfu. WHO mentions maximum permissible limits in raw materials only for arsenic, cadmium, and lead, which amount to 1.0, 0.3, and 10 ppm, respectively. Aim: The main purpose of the investigation was to document evidence for the users, and practitioners of marketed haematinic herbal formulations. In the present study haematinic herbal formulations marketed in Yavatmal India were determined for the presence of microbial and heavy metal content. Method: The investigations were performed by using specific medias and atomic absorption spectrometry. Result: The present work indicates the presence of heavy metal contents in herbal formulations selected for study. It was found that arsenic content in formulations was below the permissible limit in all formulations. The cadmium and lead content in six formulations were above the permissible limits. Such formulations are injurious to health of patient if consumed regularly. The specific medias were used to determining the presence of Escherichia coli 4 samples, Staphylococcus aureus 3 samples, and P. aeruginosa 4 samples. The data indicated suggest that there is requirement of in process improvement to provide better quality for consumer health in order to be competitive in international markets. Summary/Conclusion: The presence of microbial and heavy metal content above WHO limits indicates that the GMP was not followed during manufacturing of herbal formulations marketed in India.

Keywords: toxicological standardization, heavy metals, microbial contamination, haematinic herbal formulations

Procedia PDF Downloads 448

Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

Abstract:

For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

Procedia PDF Downloads 448

Authors: R. Gounina-Allouane, N. Ouali, F. Z. Berrabah, A. Bentaleb

Abstract:

For a long time, the Gram-positive spore-forming bacteria Bacillus thuringiensis (Bt) has been widely used in biological control against devastating and disease vectors insects. This is due to the insecticidal activity of its crystalline parasporal inclusion (crystals) predominantly comprised of one or more proteins (Cry and Cyt proteins) also called δ-endotoxins, produced during sporulation. The shape and composition of Bt crystals vary among strains and crystalline proteins are extremely varied (more than 475 cry gene were discovered). The insecticidal activity of Bt crystals is very well studied, thus their insecticidal mode of action is well established, however, their antimicrobial effect is largely unknown. The lack of data on the antimicrobial effect of crystalline proteins of Bt and the need for searching new antimicrobial molecules encouraged us to carried out this study. The antibacterial effect of δ-endotoxines produced by two Bt stains; a strain isolated from soil at northern of Algeria (Bt 7.2.B), and a strain isolated from a bioinsecticide (Bacillus thuringiensis var aizawai), activated by proteolysis, was assayed on clinical bacterial strains and ATCC collection ones respectively. Gram positive and negative clinical bacterial strains (Escherichia coli, Klebsiella pneumonaie, Pseudomonas aeruginosa, Staphylococcus aureus) were sensitive to activated Bt 72B endotoxins. Similarly, bacterial strains from ATCC collection (Escherichia coli ATCC 25922, Pseudomonas aerugenosa ATCC 27853, Staphylococcus aureus ATCC 25923) were sensitive to activated B. thuringiensis var aizawai δ-endotoxines. The activated δ-endotoxins were separated by SDS-PAGE.

Keywords: Bacillus thuringiensis, crystals, cry proteins, δ-endotoxins, antibacterial activity

Procedia PDF Downloads 430

Authors: Omneya M. Helmy, Mona T. Kashef

Abstract:

Aminoglycosides are used in treating a wide range of infections caused by both Gram-negative and Gram positive bacteria. The presence of 16S rRNA methyl transferases (16S-RMTase) is among the newly discovered resistance mechanisms that confer high resistance to clinically useful aminoglycosides. Cephalosporins are the most commonly used antimicrobials in Egypt; therefore, this study was conducted to determine the isolation frequency of 16S rRNA methyl transferases among third generation cephalosporin-resistant clinical isolates in Egypt. One hundred and twenty three cephalosporin resistant Gram-negative clinical isolates were screened for aminoglycoside resistance by the Kirby Bauer disk diffusion method and tested for possible production of 16S-RMTase. PCR testing and sequencing were used to confirm the presence of 16S-RMTase and the associated antimicrobial resistance determinants, as well as the genetic region surrounding the armA gene. Out of 123 isolates, 66 (53.66%) were resistant to at least one aminoglycoside antibiotic. Only one Escherichia coli isolate (E9ECMO) which was totally resistant to all tested aminoglycosides, was confirmed to have the armA gene in association with blaTEM-1, blaCTX-M-15, blaCTX-M-14 and aac(6)-Ib genes. The armA gene was found to be carried on a large A/C plasmid. Genetic mapping of the armA surrounding region revealed, for the first time, the association of armA with aac(6)-Ib on the same transposon. In Conclusion, the isolation frequency of 16S-RMTase was low among the tested cephalosporin-resistant clinical samples. However, a novel composite transposon has been detected conferring high-level aminoglycosides resistance.

Keywords: aminoglcosides, armA gene, β lactmases, 16S rRNA methyl transferases

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Authors: M. E. Abalaka, O. A. Falusi, M. Galadima, D. Damisa

Abstract:

The antimicrobial activity of aqueous, ethanolic and methanolic extracts of Chromolaena odorata (Siam weed) was evaluated against four wound isolates: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Klebsiella pneumoniae at the concentrations of 200mg/ml, 100mg/ml, 50mg/ml and 25mg/ml respectively. S. aureus and E. coli showed high susceptibility to the various extracts than the other test isolates. The aqueous extract showed activity against Staphylococcus aureus with a mean diameter of zone of inhibition of 16 ± 3.00 at concentration of 200mg/ml and as low as 8 ± 0.00 at concentration of 25mg/ml; E. coli showed susceptibility with a mean diameter of zone of inhibition of 18 ± 2.00 and 10 ± 0.00 at a concentration of 200mg/ml and 25mg/ml respectively. Pseudomonas aeruginosa and Klebsiella pneumoniae were resistant to the aqueous extract. Methanol extract showed activity against Staphylococcus aureus with a mean diameter of zone of inhibition at 28 ± 4.00 and 12 ± 2.30 at a concentration of 200mg/ml and 25mg/ml respectively; while E. coli was susceptible with mean diameter of zone of inhibition of 18 ± 2.00 and as low as 12 ± 0.00 at a concentration of 200mg/ml and 50mg/ml respectively, Pseudomonas aeruginosa showed considerable susceptibility with mean diameter of zone of inhibition of 13 ± 1.00 and 12 ± 0.00 at a concentration of 200mg/ml and 100mg/ml respectively. The ethanol extract showed activity against S. aureus with a mean diameter zone of inhibition of 15 ± 2.00 and 9 ± 0.00 at a concentration of 200mg/ml and 25mg/ml respectively: E. coli showed susceptibility with a mean diameter zone of inhibition of 20 ± 4.00 and 13 ± 2.00 at a concentration of 200mg/ml and 25mg/ml respectively. Pseudomonas aeruginosa showed considerable susceptibility with a mean diameter zone of inhibition of 13 ± 1.00 and 9 ± 0.00 at a concentration of 200mg/ml and 100mg/ml respectively. The results above indicate the efficacy and potency of the crude extracts of Chromolaena odorata leaf on the tested wound isolates.

Keywords: antibacterial, Chromolaena odorata, leaf extracts, test isolates

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Authors: Afzan Mahmad, Santibuana Abd Rahman, Gouri Kumar Dash, Mohd. Syafiq Bin Abdullah

Abstract:

Antibacterial activity of the methanol extract of fruit peel of Punica granatum Linn (Family: Punicaceae) was evaluated against two Gram positive and two Gram negative bacteria. The Gram positive bacteria included Staphylococcus aureus, Streptococcus pneumoniae and the Gram negative organisms included Escherichia coli and Pseudomonas aeruginosa respectively. The culture media used for antibacterial assay was Mueller Hinton agar for the growth of S. aureus, E. coli, and P. aeruginosa. The media used for the growth of S. pneumoniae was Mueller Hinton blood agar. The antibacterial assay was performed through Disc diffusion technique. The methanol extract was tested at three different concentrations (50, 100 and 200 mg/ml). Standard antibiotic discs containing vancomycin (30 μg) for S. pneumoniae, penicillin (10 units) for S. aureus, ceftriaxone (30 μg) for E. coli and ciprofloxacin (5 μg) for P. aeruginosa were used for the activity comparison. The results of the study revealed that the extract possesses antibacterial activity against S. aureus, S. pneumoniae and P. aeruginosa at all tested concentrations. The maximum zone of inhibition of 19 mm of the extract at 200 mg/ml was observed against S. pneumoniae. However, no zone of inhibition was observed against E. coli at the tested concentrations of the extract. Based on the results obtained in this study, it may be concluded that the fruit peel of P. granatum possess broad spectrum of antibacterial activity against a number bacteria.

Keywords: Punica granatum Linn., methanol extract, antibacterial, zone of inhibition

Procedia PDF Downloads 394