Search results for: congo red agar

Authors: Ashkan Zargar, Ali Taheri Mirghaed, Zein Talal Barakat, Alireza Khosravi, Hamed Paknejad

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With the increase of bacterial resistance to the chemical antibiotics, replacing it with ecofriendly herbal materials and with no adverse effects in the host body is very important. Therefore, in this study, the effect of combined essential oil (Thymus vulgaris-Origanum magorana and Ziziphora clinopodioides) on the growth behavior of Yersinia ruckeri, Aeromonas hydrophila and Lactococcus garvieae was evaluated. The compositions of the herbal essential oils used in this study were determined by gas chromatography-mass spectrometry (GC-MS) while, the investigating of antimicrobial effects was conducted by the agar-disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and bacterial growth curves determination relied on optical density (OD) at 630 nm. The main compounds were thymol (40.60 %) and limonene (15.98 %) for Thymus vulgaris while carvacrol (57.86 %) and thymol (13.54 %) were the major compounds in Origanum magorana. As regards Ziziphora clinopodiodes, α-pinene (22.6 %) and carvacrol (21.1 %) represented the major constituents. Concerning Yersinia ruckeri, disc-diffusion results showed that t.O.z (50 % Origanum majorana) combined essential oil was presented the best inhibition zone (30.66 mm) but it was exhibited no significant differences with other tested commercial antibiotics except oxytetracycline (P <0/05). The inhibitory activity and the bactericidal effect of the t.O.z, unveiled by the MIC= 0.2 μL /mL and MBC= 1.6 μL /mL values, were clearly the best between all combined oils. The growth behaviour of Yersinia ruckeri was affected by this combined essential oil and changes in temperature and pH conditions affected herbal oil performance. As regard Aeromonas hydrophila, its results were so similar to Yersinia ruckeri results and t.O.z (50 % Origanum majorana) was the best between all combined oils (inhibition zone= 26 mm, MIC= 0.4 μL /mL and MBC= 3.2 μL /mL, combined essential oil was affected bacterial growth behavior). Also for Lactococcus garvieae, t.O.z (50 % Origanum majorana) was the best between all combined oils having the best inhibition zone= 20.66 mm, MIC= 0.8 μL /mL and MBC= 1.6 μL /mL and best effect on inhibiting bacterial growth. Combined herbal essential oils have a good and noticeable effect on the growth behavior of pathogenic bacteria in the laboratory, and by continuing research in the host, they may be a suitable alternative to control, prevent and treat diseases caused by these bacteria.

Keywords: bacterial pathogen, herbal medicine, growth behavior, fish

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

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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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Authors: Amina Omer, Ikram Elsiddig

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Arctostaphylos uvaursi is a plant of the family Ericaceae, it’s used in skin care products mostly for its depigmenting action, due to the presence of hydroquinones that are well known inhibitors of tyrosinase, an enzyme involved in melanin biosynthesis in humans. The main hydroquinone found within the A. uvaursi is arbutin, which is found with varying percentage within the plant depending on the season, and area from which the plant is harvested. An in vitro experiment has shown that the arbutin found within the bearberry leaf extract inhibited the biosynthesis of melanin in human melanoma cells and in three-dimensional human skin model. Madurella mycetomatis is filamentous fungus that causes the fungal form of mycetoma known as eumycetoma, with existing anti-fungals and surgery, only 35% of people living eumycetoma are treated, M. mycetomatis has been found to shield itself against the antifungal therapy through the production of melanin decreasing the effectiveness of the therapy, therefore there is a need for a new and more effective therapy. The aim of the study was to investigate and compare the effect of arbutin powder and aqueous extract of A. uvaursi containing arbutin on the biosynthesis of melanin by M. mycetomatis. The experiment was carried out by culturing M. mycetomatis on minimal media composed of 2% agar, 15 mM glucose, 10 mM MgSO4, 29.4 mM KH2PO4, 13 mM glycin and 80mg/l gentamicin, the media was supplied with different concentration of arbutin solution (5, 25 50,and 75mg) and aqueous extract of A. uvaursi to contain arbutin with concentrations (5, 25 50,and 75mg), the plates were incubated for two month and the result was observed by the naked eye. The results revealed that the arbutin powder had an inhibitory effect on melanin synthesis by M. mycetomatis that correlated with its established inhibitory effect on melanin synthesis in humans. The inhibitory effect of arbutin on melanin synthesis by M. mycetomatis was found to be dose dependent. A. uvaursi aqueous leaf extract containing arbutin was also found to decrease melanin production by M. mycetomatis, however plates containing high concentrations of aqueous extract couldn’t be assessed for its melanin inhibitory effect due to the high content of carbohydrates in the extract that promoted the growth of fungi Asperigullus niger rendering the plates unsuitable for visual inspection. In conclusion inhibition of melanin synthesis was observed on the arbutin powder as well as the aqueous extract containing arbutin. A. uvaursi is known to exhibit anti-inflammatory activity, which can aid in wound healing that is beneficial in the chronic inflammation caused by M. mycetomatis.

Keywords: arbutin, arctostaphylos, Madurella, melanin

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Authors: T. Ewetumo, K. D. Adedayo, Festus Ben

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Knowledge of the thermal properties of foods is of fundamental importance in the food industry to establish the design of processing equipment. However, for tropical fruit juice, there is very little information in literature, seriously hampering processing procedures. This research work describes the development of an instrument for automated thermal conductivity and thermal diffusivity measurement of tropical fruit juice using a transient thermal probe technique based on line heat principle. The system consists of two thermocouple sensors, constant current source, heater, thermocouple amplifier, microcontroller, microSD card shield and intelligent liquid crystal. A fixed distance of 6.50mm was maintained between the two probes. When heat is applied, the temperature rise at the heater probe measured with time at time interval of 4s for 240s. The measuring element conforms as closely as possible to an infinite line source of heat in an infinite fluid. Under these conditions, thermal conductivity and thermal diffusivity are simultaneously measured, with thermal conductivity determined from the slope of a plot of the temperature rise of the heating element against the logarithm of time while thermal diffusivity was determined from the time it took the sample to attain a peak temperature and the time duration over a fixed diffusivity distance. A constant current source was designed to apply a power input of 16.33W/m to the probe throughout the experiment. The thermal probe was interfaced with a digital display and data logger by using an application program written in C++. Calibration of the instrument was done by determining the thermal properties of distilled water. Error due to convection was avoided by adding 1.5% agar to the water. The instrument has been used for measurement of thermal properties of banana, orange and watermelon. Thermal conductivity values of 0.593, 0.598, 0.586 W/m^o C and thermal diffusivity values of 1.053 ×〖10〗^(-7), 1.086 ×〖10〗^(-7), and 0.959 ×〖10〗^(-7) 〖m/s〗^2 were obtained for banana, orange and water melon respectively. Measured values were stored in a microSD card. The instrument performed very well as it measured the thermal conductivity and thermal diffusivity of the tropical fruit juice samples with statistical analysis (ANOVA) showing no significant difference (p>0.05) between the literature standards and estimated averages of each sample investigated with the developed instrument.

Keywords: thermal conductivity, thermal diffusivity, tropical fruit juice, diffusion equation

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Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures

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Authors: Fatma A. Mohamed, Hend M. Ahmed

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This research explores the antimicrobial effects of dielectric barrier discharge (DBD) oxygen plasma treatment combined with ZnO NPs on the cotton fabric, focusing on various treatment durations (5, 10, 15, 20, and 30 minutes) and discharge powers (15.5–17.35 watts) at flow rate 0.5 l/min. After treatment with oxygen plasma and ZnO NPs, the fabric was printed with green tea (Camellia sinensis) at five different concentrations. The study evaluated the treatment's effectiveness by analyzing surface wettability, specifically through wet-out time and hydrophilicity, as well as measuring contact angles. To investigate the chemical changes on the fabric's surface, attenuated total reflectance–Fourier transform infrared (ATR-FTIR) spectroscopy was employed to identify the functional groups formed as a result of the plasma treatment. This comprehensive approach aims to understand how DBD oxygen plasma treatment and ZnO nanoparticles change cotton fabric properties and enhance its antimicrobial potential, paving the way for innovative applications in textiles. In addition to the chemical analysis, the surface morphology of the O₂ plasma/ZnO NPs-treated cotton fabric was examined using scanning electron microscopy (SEM). FTIR analysis revealed an increase in polar functional groups (-COOH, -OH, and -C≡O) on the fabric's surface, contributing to enhanced hydrophilicity and functionality. The antimicrobial properties were evaluated using qualitative and quantitative methods, including agar plate assays and modified Hoenstein tests against Staphylococcus aureus and Escherichia coli. The results indicated a significant improvement in antimicrobial effectiveness for the cotton fabric treated with plasma and coated with natural extracts, maintaining this efficacy even after four washing cycles. This research demonstrates that utilizing oxygen DBD plasma/ZnO NPs treatment, combined with the absorption of tea and tulsi leaf extracts, presents a promising strategy for developing natural antimicrobial textiles. This approach is particularly relevant given the increasing medical and healthcare demands for effective antimicrobial materials. Overall, the method not only enhances the absorption of plant extracts but also significantly boosts antimicrobial efficacy, offering valuable insights for future textile applications.

Keywords: cotton, ZnO NPs, green tea leaf, antimicrobial avtivity, DBD oxygen plasma

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Authors: Igbakura I. Luga, Alex A. Adikwu

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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 21^st 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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Authors: Ibrahim Siddig Hamid, Ikram Mohamed Eltayeb

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

Keywords: harmala, peganum, shigella, trachyspermum

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Authors: O. Antypenko, I. Vasilieva, S. Kovalenko

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Investigations for new effective and less toxic antimicrobials agents are always up-to-date. The tetrazole derivatives are quite interesting objects as for synthesis as well as for pharmacological screening. Thus, some derivatives of tetrazole demonstrated antimicrobial activity, namely 5-phenyl-tetrazolo[1,5-c]quinazoline was effective one against Staphylococcus aureus and Esherichia faecalis (MIC = 250 mg/L). Besides, investigation of the 9-bromo(chloro)-5-morpholin(piperidine)-4-yl-tetrazolo[1,5-c]quinazoline’s antimicrobial activity against Esherichia coli and Enterococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus revealed that sensitivity of Gram-positive bacteria to the compounds was higher than that of Gram-negative bacteria. So, our previously synthesized, 31 derivatives of 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas were decided to test for their in vitro antibacterial activity against Gram-positive bacteria (Staphylococcus aureus ATCC 25923, Enterobacter aerogenes, Enterococcus faecalis ATCC 29212), Gram-negative bacteria (Pseudomonas aeruginosa ATCC 9027, Escherichia coli ATCC 25922, Klebsiella pneumoniae 68) and antifungal properties against Candida albicans ATCC 885653. Agar-diffusion method was used for determination of the preliminary activity compared to well-known reference antimicrobials. All the compounds were dissolved in DMSO at a concentration of 100 μg/disk, using inhibition zone diameter (IZD, mm) as a measure for the antimicrobial activity. The most active turned to be 3 structures, that inhibited several bacterial strains: 1-ethyl-3-(5-fluoro-2-(1H-tetrazol-5-yl)phenyl)urea (1), 1-(4-bromo-2-(1H-tetrazol-5-yl)-phenyl)-3-(4-(trifluoromethyl)phenyl)urea (2) and 1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (3). IZM (mm) was 40 (Escherichia coli), 25 (Klebsiella pneumonia) for compound 1; 12 (Pseudomonas aeruginosa), 15 (Staphylococcus aureus), 10 (Enterococcus faecalis) for compound 2; 25 (Staphylococcus aureus), 15 (Enterococcus faecalis) for compound 3. The most sensitive to the activity of the substances were Gram-negative bacteria Pseudomonas aeruginosa. While none of compound effected on Candida albicans. Speaking about, reference drugs: Amikacin (30 µg/disk) showed 27 and Ceftazide (30 µg/disk) 25 against Pseudomonas aeruginosa. That is, unfortunately, higher than studied 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas. Obtained results will be used for further purposeful optimization of the leading compounds in the more effective antimicrobials because of the ever-mounting problem of microorganism’s resistance.

Keywords: antimicrobial, antifungal, compounds, 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas

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Authors: Alexis D'Agostino, Celeste Sununtunasuk, Jack Fiedler

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Background: Iron-folic acid (IFA) supplementation during antenatal care (ANC) has existed in many countries for decades. Despite this, low national coverage persists and women do not often consume appropriate amounts during pregnancy. USAID’s SPRING Project investigated pregnant women’s access to, and consumption of, IFA tablets through ANC. Cross-country analysis provided a global picture of the state of IFA-supplementation, while country-specific results noted key contextual issues, including geography, wealth, and ANC attendance. The analysis can help countries prioritize strategies for systematic performance improvements within one of the most common micronutrient supplementation programs aimed at reducing maternal anemia. Methodology: Using falter point analysis on Demographic and Health Survey (DHS) data collected from 162,958 women across 22 countries, SPRING identified four sequential falter points (ANC attendance, IFA receipt or purchase, IFA consumption, and number of tablets taken) where pregnant women fell out of the IFA distribution structure. SPRING analyzed data on IFA intake from DHS surveys with women of reproductive age. SPRING disaggregated these data by ANC participation during the most recent pregnancy, residency, and women’s socio-economic status. Results: Average sufficient IFA tablet use across all countries was only eight percent. Even in the best performing countries, only about one-third of pregnant women consumed 180 or more IFA tablets during their most recent pregnancy. ANC attendance was an important falter point for a quarter of women across all countries (with highest falter rates in Democratic Republic of the Congo, Nigeria, and Niger). Further analysis reveals patterns, with some countries having high ANC coverage but low IFA provision during ANC (DRC and Haiti), others having high ANC coverage and IFA provision but few women taking any tablets (Nigeria and Liberia), and countries that perform well in ANC, supplies, and initial consumption but where very few women consume the recommended 180 tablets (Malawi and Cambodia). Country-level analysis identifies further patterns of supplementation. In Indonesia, for example, only 62% of women in the poorest quintile took even one IFA tablet, while 86% of the wealthiest women did. This association between socioeconomic status and IFA intake held across nearly all countries where these data are available and was also visible in rural/urban comparisons. Analysis of ANC attendance data also suggests that higher numbers of ANC visits are associated with higher tablet intake. Conclusions: While it is difficult to disentangle which specific aspects of supply or demand cause the low rates of consumption, this tool allows policy-makers to identify major bottlenecks to scaling-up IFA supplementation during ANC. In turn, each falter point provides possible explanations of program performance and helps strategically identify areas for improved IFA supplementation. For example, improving the delivery of IFA supplementation in Ethiopia relies on increasing access to ANC, but also on identifying and addressing program gaps in IFA supply management and health workers’ practices in order to provide quality ANC services. While every country requires a customized approach to improving IFA supplementation, the multi-country analysis conducted by SPRING is a helpful first step in identifying country bottlenecks and prioritizing interventions.

Keywords: iron and folic acid, supplementation, antenatal care, micronutrient

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Authors: Nazlı Çetindağ, Pelin Yılmaz Çetiner, Metin Mert İlgün, Emine Birci, Gizemnur Yıldız Uysal, Özcan Hatipoğlu, Ehsan Tuzcuoğlu, Gökhan Sır

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Water scarcity is an unavoidable issue impacting an increasing number of individuals daily, representing a global crisis stemming from swift population growth, urbanization, and excessive resource exploitation. Consequently, solutions that involve the reclamation of wastewater are considered essential. In this context, household wastewater, categorized as greywater, plays a significant role in freshwater used for residential purposes and is attributed to washing. This type of wastewater comprises diverse elements, including organic substances, soaps, detergents, solvents, biological components, and inorganic elements such as certain metal ions and particles. The physical characteristics of wastewater vary depending on its source, whether commercial, domestic, or from a hospital setting. Consequently, the treatment strategy for this wastewater type necessitates comprehensive investigation and appropriate handling. The advanced oxidation process (AOP) emerges as a promising technique associated with the generation of reactive hydroxyl radicals highly effective in oxidizing organic pollutants. This method takes precedence over others like coagulation, flocculation, sedimentation, and filtration due to its avoidance of undesirable by-products. In the current study, the focus was on exploring the feasibility of the AOP for treating actual household wastewater. To achieve this, a laboratory-scale device was designed to effectively target the formed radicals toward organic pollutants, resulting in lower organic compounds in wastewater. Then, the number of microorganisms present in treated wastewater, in addition to the chemical content of the water, was analyzed to determine whether the lab-scale device eliminates microbial accumulation with AOP. This was also an important parameter since microbes can indirectly affect human health and machine hygiene. To do this, water samples were taken from treated and untreated conditions and then inoculated on general purpose agar to track down the total plate count. Analysis showed that AOP might be an option to treat household wastewater and lower microorganism growth.

Keywords: usage of household water, advanced oxidation process, water reuse, modelling

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Authors: Mokgadi Miranda Hlongwane, Ntebogeng Sharon Mokgalaka, Felix Dapare Dakora

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Lessertia (L.) frutescens (syn. Sutherlandia frutescens) is a leguminous medicinal plant indigenous to South Africa. Traditionally, L. frutescens has been used to treat cancer, diabetes, epilepsy, fever, HIV, stomach problems, wounds and other ailments. This legume is endemic to the Cape fynbos, with large populations occurring wild and cultivated in the Cape Florist Region. Its widespread distribution in the Western Cape, Northern Cape, Eastern Cape and Kwazulu-Natal is linked to its increased use as a phytomedicine in the treatment of various diseases by traditional healers. The frequent harvesting of field plants for use as a medicine has made it necessary to undertake studies towards the conservation of Lessertia frutescens. As a legume, this species can form root nodules and fix atmospheric N₂ when in symbiosis with soil bacteria called rhizobia. So far, however, few studies (if any) have been done on the efficacy and diversity of native bacterial symbionts nodulating L. frutescens in South Africa. The aim of this project was to isolate and characterize L. frutescens-nodulating bacteria from five different locations in the Western Cape Province. This was done by trapping soil rhizobia using rhizosphere soil suspension to inoculate L. frutescens seedlings growing in sterilized sand and receiving sterile N-free Hoagland nutrient solution under glasshouse conditions. At 60 days after planting, root nodules were harvested from L. frutescens plants, surface-sterilized, macerated, and streaked on yeast mannitol agar (YMA) plates and incubated at 28 ˚C for observation of bacterial growth. The majority of isolates were slow-growers that took 6-14 days to appear on YMA plates. However, seven isolates were fast-growers, taking 2-4 days to appear on YMA plates. Single-colony cultures of the isolates were assessed for their ability to nodulate L. frutescens as a homologous host under glasshouse conditions. Of the 92 bacterial isolates tested, 63 elicited nodule formation on L. frutescens. Symbiotic effectiveness varied markedly between and among test isolates. There were also significant (p≤0.005) differences in nodulation, shoot biomass, photosynthetic rates, leaf transpiration and stomatal conductance of L. frutescens plants inoculated with the test isolates, which is an indication of their functional diversity.

Keywords: lessertia frutescens, nodulating, rhizobia, symbiotic effectiveness

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Authors: D. Desai, J.Dixon, N. Jain, M. Datta

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Microbial infections of scalp consist of symptomatic appearances associated with seborrhoeic dermatitis, folliculitis, furuncles, carbuncles and ringworm. The main causative organisms in these scalp-based infections are bacteria like S. aureus, P. aeruginosa and a fungus M. Furfur. Allopathic treatment of these infections is available and efficient, but occasionally, topical applications have been found to cause side effects. India is known as the botanical garden of the world and considered as the epicentre for utilization of traditional drugs. Many treatments based on herb extracts are commonly used in India. It has been observed treatment with ethnomedicines requires a higher dosage and greater time period. Additionally, repeated applications are required to obtain the full efficacy of the treatment. An attempt has been made to imbibe the traditional knowledge with nanotechnology to generate a proficient therapeutic against scalp infections. We have imbibed metallic nanoparticles with extracts from traditional medicines and propose to formulate an antimicrobial hair massager. Four commonly used herbs for treatment against scalp disorders like Zingiber officinale (ginger), Allium sativum (garlic), Azadirachta indica (neem) leaves and Citrus limon (lemon) peel was taken. 30 gms of dried homogenized powder was obtained and processed for obtaining the aqueous and ethanolic extract by soxhlet apparatus. The extract was dried and reconstituted to obtain working solution of 1mg/ml. Phytochemical analysis for the obtained extract was done. Synthesis of nanoparticles was mediated by incubating 1mM silver nitrate with extracts of various herbs to obtain silver nanoparticles. The formation of the silver nanoparticles (AgNPs) was monitored using UV-Vis spectroscopy. The AgNPs thus obtained were centrifuged and dried. The AgNPs thus formed were characterized by X Ray Diffraction, scanning electron microscopy and transmission electron microscopy. The size of the AgNPs varied from 10-20 nm and was spherical in shape. P. aeruginosa was plated on nutrient agar and comparative antibacterial activity was tested. Comparative antimicrobial potential was calculated for the extracts and the corresponding nanoconstructs. It was found AgNPs were more efficient than their aqueous and ethanolic counterparts except in the ase of C. limon. Statistical analysis was performed to validate the results obtained.

Keywords: ethnomedicine, nanoconstructs, scalp infections, Zingiber officinale

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Authors: Amal Saafan, Kareem Al Sofy, Sameh AbdelGhani, Magdy Amin

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Background: Acinetobacter spp. resistance towards β-lactam antibiotics is mediated mainly by different classes of β-lactamases production; detection of some genes responsible for production of β-lactamases is the objective of the study. Methods: One hundred fifty bacterial isolates were recovered from blood, sputum, and urine specimens from different hospitals in Egypt. Sixty-nine isolate were identified as Acinetobacter baumannii using traditional biochemical tests, CHROM agar, MicroScan and PCR amplification of blaoxa-51like gene. Acinetobacterbaumannii isolates were grouped into carbapenem resistant group (GP1), cefotaxime, ceftazidime and cefoxitin resistant group (GP2) and carbapenem and cephalosporin non-resistant group (GP3). Carbapenemase activity was screened using modified Hodge test (MHT) for GP1.Metallo-β-lactamases screening was performed for MHT positive isolates using double disk synergy test (DDST) and combined disk test (CDT). Amp C activity was screened using Amp C disk test with Tris-EDTA, DDST, and CDT for GP2. Finally, PCR amplification of blaoxa-51like, blaoxa-23like, blaIMP-like, blaVIM-like, and blaADC-like genes was performed for isolates that showed, at least, two positive results of three for both AmpC and carbapenemases phenotypic screening tests (obvious activity), in addition to GP3 (for comparison). Detection of blaoxa-51like and blaADC-like genes preceded by ISAba1 was also performed. Results: Antibiogram of 69 pure Acinetobacter baumannii isolates resulted in 57, 64, and 2 isolates enrolled into GP1, GP2, and GP3, respectively. Carbapenemase activity was shown by 49(85.9%) isolate using MHT. Metallo-β-lactamases screening revealed 32(65.3%) and 35(71.4%) using DDST and CDT, respectively.AmpC activity was shown by 43(67.2%) and 50 (78.1%) isolates using AmpC disk test with Tris-EDTA, and both DDST and CDT, respectively. Twenty-seven isolates showed obvious activity, all of them (100%) were harboring blaoxa-51like and blaADC-like genes, while blaoxa-23like, blaIMP-like andblaVIM-like genes were harbored by 23(85.2%), 9 (33.%) and no isolate respectively. Only 12 (44.4%) isolates harbored blaoxa-51like and blaADC-like genes preceded by ISAba1. GP3 isolates showed only positive blaoxa-51like and blaADC-like genes. Conclusion: It is not possible to correlate resistance with presence of blaoxa-51like and blaADC-like genes and presence of ISAba1 was immediate as transcriptional promoter. A blaoxa-23like gene played an important role in carbapenem resistance when compared with blaIMP-like and blaVIM-like gene.

Keywords: acinetobacter, beta-lactams, resistance, antimicrobial agents

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Authors: Abdul Walusansa

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In this paper, paper notes and coins were collected from general public in Kampala City where ready-to-eat food can be served, in order to survey for bacterial contamination. The total bacterial number and potentially pathogenic organisms loading on currency were tested. All isolated potential pathogens were also tested for antibiotic resistance against four most commonly prescribed antibiotics. 1. The bacterial counts on one hundred paper notes sample were ranging between 6～10918/cm cm-2,the median was 141/ cm-2, according to the data it was much higher than credit cards and Australian notes which were made of polymer. The bacterial counts on sixty coin samples were ranging between 2～380/cm-2, much less than paper notes. 2. Coliform (65.6%), E. coli (45.9%), S. aureus (41.7%), B. cereus (67.7%), Salmonella (19.8%) were isolated on one hundred paper notes. Coliform (22.4%), E. coli (5.2%), S. aureus (24.1%), B. cereus (34.5%), Salmonella (10.3%) were isolated from sixty coin samples. These results suggested a high rate of potential pathogens contamination of paper notes than coins. 3. Antibiotic resistances are commonly in most of the pathogens isolated on currency. Ampicillin resistance was found in 60%of Staphylococcus aureus isolated on currency, as well as 76.6% of E. coil and 40% of Salmonella. Erythromycin resistance was detected in 56.6% of S. aureus and in 80.0% of E. coli. All the pathogens isolated were sensitive to Norfloxacin, Salmonella and S. aureus also sensitive to Cefaclor. In this paper, we also studied the antimicrobial capability of metal coins, coins collected from different countries were tested for the ability to inhibit the growth of E. sakazakii, S. aureus, E. coli, L. monocytogenes and S. typhimurium. 1) E. sakazakii appeared very sensitive to metal coins, the second is S. aureus, but E. coli, L. monocytogenes and S. typhimurium are more resistant to these metal coin samples. 2) Coins made of Nickel-brass alloy and Copper-nickel alloy showed a better effect in anti-microbe than other metal coins, especially the ability to inhibited the growth of E. sakazakii and S. aureus, all the inhibition zones produced on nutrient agar are more than 20.6 mm. Aluminium-bronze alloy revealed weak anti-microbe activity to S. aureus and no effect to kill other pathogens. Coins made of stainless steel also can’t resist bacteria growth. 3) Surprisingly, one cent coins of USA which were made of 97.5% Zinc and 2.5% Cu showed a significant antimicrobial capability, the average inhibition zone of these five pathogens is 45.5 mm.

Keywords: antibiotic sensitivity, bacteria, currency, coins, parasites

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Authors: Waheed Anwar, Kiran Nawaz, Muhammad Saleem Haider, Ahmad Ali Shahid, Sehrish Iftikhar

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The whitefly, Bemisia tabaci (Gennadius), is a complex insect species, including many cryptic species or biotypes. Whitefly causes damage to many ornamental and horticultural crops through directly feeding on phloem sap, resulting in sooty mould and critically decreases the rate of photosynthesis of many host plants. Biological control has emerged as one of the most important methods for the management of soil-borne plant pathogens. Among the natural enemies of insects different entomopathogenic fungi are mostly used as biological control of the pest. The purpose of this research was to find indigenous insect-associated fungi and their virulence against Bemisia tabaci. A detailed survey of cotton fields in sample collection was conducted during July and August 2013 from the central mixed zone of Punjab, Pakistan. For the isolation of T. longibrachiatum, sabouraud dextrose peptone yeast extract agar (SDAY) media was used and morphological characterization of isolated T. longibrachiatum was studied using different dichotomous keys. Molecular Identification of the pathogen was confirmed by amplifying the internal transcribed spacer region. Blastn analysis showed 100% homology with already reported sequences on the database. For these bioassays, two conidial concentrations 4 × 108/mL & 4 × 104/mL of T. longibrachiatum was sprayed in clip cages for nymph and adult B. tabaci respectively under controlled environmental conditions. The pathogenicity of T. longibrachiatum was tested on nymph and adult whitefly to check mortality. Mortality of B. tabaci at nymphal and adult stages were observed after 24-hour intervals. Percentage mortality of nymphs treated with 4 x 104/mL conidia of T. longibrachiatum was 20, 24, 36 and 40% after 48, 72, 96, 72, 96, 120 and 144 hours respectively. However, no considerable difference was recorded in percentage mortality of whitefly after 120 and 144 hours. There were great variations after 24, 48, 72 and 96 hours in the rate of mortality. The efficacy of T. longibrachiatum as entomopathogenic fungi was evaluated in adult and nymphal stages of whitefly. Trichoderma longibrachiatum showed maximum activity on nymphal stages of whitefly as compared to adult stages. The percentage of conidial germination was also recorded on the outer surface of adult and nymphal stages of B. tabaci. The present findings indicated that T. longibrachiatum is an entomopathogenic fungus against B. tabaci and many species of Trichoderma were already reported as an antagonistc organism against a wide range of bacterial and fungal pathogens.

Keywords: efficacy, Trichoderma, virulence, bioassay

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Authors: Jung-En Kuan, Whei-Fen Wu

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

Keywords: enzyme, esterase, lipotic hydrolase, type IV

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Authors: Recep Kesli, Huseyin Bilgin, Yasar Unlu, Gokhan Gungor

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Aim: The aim of this study, was to investigate the presence of Helicobacter pylori (H. pylori) infection by culture, histology, and RUT (Rapid Urease Test) in gastric antrum biopsy samples taken from patients presented with dyspeptic complaints and to determine resistance rates of amoxicillin, clarithromycin, levofloxacin and metronidazole against the H. pylori strains by E-test. Material and Methods: A total of 278 patients who admitted to Konya Education and Research Hospital Department of Gastroenterology with dyspeptic complaints, between January 2011-July 2013, were included in the study. Microbiological and histopathological examinations of biopsy specimens taken from antrum and corpus regions were performed. The presence of H. pylori in biopsy samples was investigated by culture (Portagerm pylori-PORT PYL, Pylori agar-PYL, GENbox microaer, bioMerieux, France), histology (Giemsa, Hematoxylin and Eosin staining), and RUT(CLOtest, Cimberly-Clark, USA). Antimicrobial resistance of isolates against amoxicillin, clarithromycin, levofloxacin, and metronidazole was determined by E-test method (bioMerieux, France). As a gold standard in the diagnosis of H. pylori; it was accepted that the culture method alone was positive or both histology and RUT were positive together. Sensitivity and specificity for histology and RUT were calculated by taking the culture as a gold standard. Sensitivity and specificity for culture were also calculated by taking the co-positivity of both histology and RUT as a gold standard. Results: H. pylori was detected in 140 of 278 of patients with culture and 174 of 278 of patients with histology in the study. H. pylori positivity was also found in 191 patients with RUT. According to the gold standard criteria, a false negative result was found in 39 cases by culture method, 17 cases by histology, and 8 cases by RUT. Sensitivity and specificity of the culture, histology, and RUT methods of the patients were 76.5 % and 88.3 %, 87.8 % and 63 %, 94.2 % and 57.2 %, respectively. Antibiotic resistance was investigated by E-test in 140 H. pylori strains isolated from culture. The resistance rates of H. pylori strains to the amoxicillin, clarithromycin, levofloxacin, and metronidazole was detected as 9 (6.4 %), 22 (15.7 %), 17 (12.1 %), 57 (40.7 %), respectively. Conclusion: In our study, RUT was found to be the most sensitive, culture was the most specific test between culture, histology, and RUT methods. Although we detected the specificity of the culture method as high, its sensitivity was found to be quite low compared to other methods. The low sensitivity of H. pylori culture may be caused by the factors affect the chances of direct isolation such as spoild bacterium, difficult-to-breed microorganism, clinical sample retrieval, and transport conditions.

Keywords: antimicrobial resistance, culture, histology, H. pylori, RUT

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Authors: Shohei Matsuo, Saki Mikai, Hiroshi Morita

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Objectives: Shochu is a popular Japanese distilled spirits. In the production of shochu, the filamentous fungus Aspergillus kawachii has traditionally been used. A. kawachii produces two types of starch hydrolytic enzymes, α-amylase (enzymatic liquefaction) and glucoamylase (enzymatic saccharification). Liquid culture system is a relatively easy microorganism to ferment with relatively low cost of production compared for solid culture. In liquid culture system, acid-unstable α-amylase (α-A) was produced abundantly, but, acid-stable α-amylase (Aα-A) was not produced. Since there is high enzyme productivity, most in shochu brewing have been adopted by a solid culture method. In this study, therefore, we investigated production of Aα-A in liquid culture system. Materials and methods: Microorganism Aspergillus kawachii NBRC 4308 was used. The mold was cultured at 30 °C for 7~14 d to allow formation of conidiospores on slant agar medium. Liquid Culture System: A. kawachii was cultured in a 100 ml of following altered SLS medium: 1.0 g of rice flour, 0.1 g of K2HPO4, 0.1 g of KCl, 0.6 g of tryptone, 0.05 g of MgSO4・7H2O, 0.001 g of FeSO4・7H2O, 0.0003 g of ZnSO4・7H2O, 0.021 g of CaCl2, 0.33 of citric acid (pH 3.0). The pH of the medium was adjusted to the designated value with 10 % HCl solution. The cultivation was shaking at 30 °C and 200 rpm for 72 h. It was filtered to obtain a crude enzyme solution. Aα-A assay: The crude enzyme solution was analyzed. An acid-stable α-amylase activity was carried out using an α-amylase assay kit (Kikkoman Corporation, Noda, Japan). It was conducted after adding 9 ml of 100 mM acetate buffer (pH 3.0) to 1 ml of the culture product supernatant and acid treatment at 37°C for 1 h. One unit of a-amylase activity was defined as the amount of enzyme that yielded 1 mmol of 2-chloro-4-nitrophenyl 6-azide-6-deoxy-b-maltopentaoside (CNP) per minute. Results and Conclusion: We experimented with co-culture of A. kawachii and lactobacillus in order to get control of pH in altered SLS medium. However, high production of acid-stable α-amylase was not obtained. We experimented with yoghurt or milk made an addition to liquid culture. The result indicated that high production of acid-stable α-amylase (964 U/g-substrate) was obtained when milk made an addition to liquid culture. Phosphate concentration in the liquid medium was a major cause of increased acid-stable α-amylase activity. In liquid culture, acid-stable α-amylase activity was enhanced by milk, but Fats and oils in the milk were oxidized. In addition, Tryptone is not approved as a food additive in Japan. Thus, alter SLS medium added to skim milk excepting for the fats and oils in the milk instead of tryptone. The result indicated that high production of acid-stable α-amylase was obtained with the same effect as milk.

Keywords: acid-stable α-amylase, liquid culture, milk, shochu

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Authors: Assel E. Molzhigitova, Amankeldi K. Sadanov, Elvira T. Ismailova, Kulyash A. Iskandarova, Olga N. Shemshura, Ainur I. Seitbattalova

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Fire blight is a very harmful for commercial apple and pear production quarantine bacterial disease. To date, several different methods have been proposed for disease control, including the use of copperbased preparations and antibiotics, which are not always reliable or effective. The use of bacteria as biocontrol agents is one of the most promising and eco-friendly alternative methods. Bacteria with protective activity against the causal agent of fire blight are often present among the epiphytic microorganisms of the phyllosphere of host plants. Therefore, the main objective of our study was screening of local epiphytic bacteria as possible antagonists against Erwinia amylovora, the causal agent of fire blight. Samples of infected organs of apple and pear trees (shoots, leaves, fruits) were collected from the industrial horticulture areas in various agro-ecological zones of Kazakhstan. Epiphytic microorganisms were isolated by standard and modified methods on specific nutrient media. The primary screening of selected microorganisms under laboratory conditions to determine the ability to suppress the growth of Erwinia amylovora was performed by agar-diffusion-test. Among 142 bacteria isolated from the fire blight host plants, 5 isolates, belonging to the genera Bacillus, Lactobacillus, Pseudomonas, Paenibacillus and Pantoea showed higher antagonistic activity against the pathogen. The diameters of inhibition zone have been depended on the species and ranged from 10 mm to 48 mm. The maximum diameter of inhibition zone (48 mm) was exhibited by B. amyloliquefaciens. Less inhibitory effect was showed by Pantoea agglomerans PA1 (19 mm). The study of inhibitory effect of Lactobacillus species against E. amylovora showed that among 7 isolates tested only one (Lactobacillus plantarum 17M) demonstrated inhibitory zone (30 mm). In summary, this study was devoted to detect the beneficial epiphytic bacteria from plants organs of pear and apple trees due to fire blight control in Kazakhstan. Results obtained from the in vitro experiments showed that the most efficient bacterial isolates are Lactobacillus plantarum 17M, Bacillus amyloliquefaciens MB40, and Pantoea agglomerans PA1. These antagonists are suitable for development as biocontrol agents for fire blight control. Their efficacies will be evaluated additionally, in biological tests under in vitro and field conditions during our further study.

Keywords: antagonists, epiphytic bacteria, Erwinia amylovora, fire blight

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Authors: Aya Tanaka, Mariko Era, Shiho Sakai, Takayoshi Kawahara, Takahide Kanyama, Hiroshi Morita

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Objectives: Aspergillus niger and Aspergillus oryzae are used as koji fungi in the spot of the brewing. Since koji-muro (room for making koji) was a low level of airtightness, microbial contamination has long been a concern to the alcoholic beverage production. Therefore, we focused on the fatty acid salt which is the main component of soap. Fatty acid salts have been reported to show some antibacterial and antifungal activity. So this study examined antimicrobial activities against Aspergillus and Bacillus spp. This study aimed to find the effectiveness of the fatty acid salt in koji-muro as antimicrobial agents. Materials & Methods: A. niger NBRC 31628, A. oryzae NBRC 5238, A. oryzae (Akita Konno store) and Bacillus subtilis NBRC 3335 were chosen as tested. Nine fatty acid salts including potassium butyrate (C4K), caproate (C6K), caprylate (C8K), caprate (C10K), laurate (C12K), myristate (C14K), oleate (C18:1K), linoleate (C18:2K) and linolenate (C18:3K) at 350 mM and pH 10.5 were used as antimicrobial activity. FASs and spore suspension were prepared in plastic tubes. The spore suspension of each fungus (3.0×104 spores/mL) or the bacterial suspension (3.0×105 CFU/mL) was mixed with each of the fatty acid salts (final concentration of 175 mM). The mixtures were incubated at 25 ℃. Samples were counted at 0, 10, 60, and 180 min by plating (100 µL) on potato dextrose agar. Fungal and bacterial colonies were counted after incubation for 1 or 2 days at 30 ℃. The MIC (minimum inhibitory concentration) is defined as the lowest concentration of drug sufficient for inhibiting visible growth of spore after 10 min of incubation. MICs against fungi and bacteria were determined using the two-fold dilution method. Each fatty acid salt was separately inoculated with 400 µL of Aspergillus spp. or B. subtilis NBRC 3335 at 3.0 × 104 spores/mL or 3.0 × 105 CFU/mL. Results: No obvious change was observed in tested fatty acid salts against A. niger and A. oryzae. However, C12K was the antibacterial effect of 5 log-unit incubated time for 10 min against B. subtilis. Thus, C12K suppressed 99.999 % of bacterial growth. Besides, C10K was the antibacterial effect of 5 log-unit incubated time for 180 min against B. subtilis. C18:1K, C18:2K and C18:3K was the antibacterial effect of 5 log-unit incubated time for 10 min against B. subtilis. However, compared to saturated fatty acid salts to unsaturated fatty acid salts, saturated fatty acid salts are lower cost. These results suggest C12K has potential in the field of koji-muro. It is necessary to evaluate the antimicrobial activity against other fungi and bacteria, in the future.

Keywords: Aspergillus, antimicrobial, fatty acid salts, koji-muro

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Authors: Piya Roychoudhury, Ruma Pal

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Cyanobacteria, commonly known as blue green algae were found to be an effective bioreagent for nanoparticle synthesis. Nowadays silver nanoparticles (AgNPs) are very popular due to their antimicrobial and anti-proliferative activity. To exploit these characters in different biotechnological fields, it is very essential to synthesize more stable, non-toxic nano-silver. For this reason silver-gold alloy (Ag-AuNPs) nanoparticles are of great interest as they are more stable, harder and more effective than single metal nanoparticles. In the present communication we described a simple technique for rapid synthesis of biocompatible AgNP and Ag-AuNP employing cyanobacteria, Leptolyngbya and Lyngbya respectively. For synthesis of AgNP the biomass of Leptolyngbya valderiana (200 mg Fresh weight) was exposed to 9 mM AgNO3 solution (pH 4). For synthesis of Ag-AuNP Lyngbya majuscula (200 mg Fresh weight) was exposed to equimolar solution of hydrogen tetra-auro chlorate and silver nitrate (1mM, pH 4). After 72 hrs of exposure thallus of Leptolyngyba turned brown in color and filaments of Lyngbya turned pink in color that indicated synthesis of nanoparticles. The produced particles were extracted from the cyanobacterial biomass using nano-capping agent, sodium citrate. Firstly, extracted brown and pink suspensions were taken for Energy Dispersive X-ray (EDAX) analysis to confirm the presence of silver in brown suspension and presence of both gold and silver in pink suspension. Extracted nanoparticles showed a distinct single plasmon band (AgNP at 411 nm; Ag-Au NP at 481 nm) in Uv-vis spectroscopy. It was revealed from Transmission electron microscopy (TEM) that all the synthesized particles were spherical in nature with a size range of ~2-25 nm. In X-ray powder diffraction (XRD) analysis four intense peaks appeared at 38.2°, 44.5°, 64.8°and 77.8° which confirmed the crystallographic nature of synthesized particles. Presence of different functional groups viz. N-H, C=C, C–O, C=O on the surface of nanoparticles were recorded by Fourier transform infrared spectroscopy (FTIR). Scanning Electron microscopy (SEM) images showed the surface topography of metal treated filaments of cyanobacteria. The stability of the particles was observed by Zeta potential study. Antibiotic property of synthesized particles was tested by Agar well diffusion method against gram negative bacteria Pseudomonas aeruginosa. Overall, this green-technique requires low energy, less manufacturing cost and produces rapidly eco-friendly metal nanoparticles.

Keywords: cyanobacteria, silver nanoparticles, silver-gold alloy nanoparticles, spectroscopy

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Authors: A. Mulry, C. Kealey, D. B. Brady

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Planktonic bacteria can form biofilms which are microbial aggregates embedded within a matrix of extracellular polymeric substances (EPS). They can be found attached to abiotic or biotic surfaces. Biofilms are responsible for oral diseases such as dental caries, gingivitis and the progression of periodontal disease. Biofilms can resist 500 to 1000 times the concentration of biocides and antibiotics used to kill planktonic bacteria. Biofilm development on oral surfaces involves four stages, initial attachment, early development, maturation and dispersal of planktonic cells. The Minimum Inhibitory Concentration (MIC) was determined using a range of saturated and unsaturated fatty acids using the resazurin assay, followed by serial dilution and spot plating on BHI agar plates to establish the Minimum Bactericidal Concentration (MBC). Log reduction of bacteria was also evaluated for each fatty acid. The Minimum Biofilm Inhibition Concentration (MBIC) was determined using crystal violet assay in 96 well plates on forming and pre-formed S. mutans biofilms using BHI supplemented with 1% sucrose. Saturated medium-chain fatty acids Octanoic (C8.0), Decanoic (C10.0) and Undecanoic acid (C11.0) do not display strong antibiofilm properties; however, Lauric (C12.0) and Myristic (C14.0) display moderate antibiofilm properties with 97.83% and 97.5% biofilm inhibition with 1000 µM respectively. Monounsaturated, Oleic acid (C18.1) and polyunsaturated large chain fatty acids, Linoleic acid (C18.2) display potent antibiofilm properties with biofilm inhibition of 99.73% at 125 µM and 100% at 65.5 µM, respectively. Long-chain polyunsaturated Omega-3 fatty acids α-Linoleic (C18.3), Eicosapentaenoic Acid (EPA) (C20.5), Docosahexaenoic Acid (DHA) (C22.6) have displayed strong antibiofilm efficacy from concentrations ranging from 31.25-250µg/ml. DHA is the most promising antibiofilm agent with an MBIC of 99.73% with 15.625µg/ml. This may be due to the presence of six double bonds and the structural orientation of the fatty acid. To conclude, fatty acids displaying the most antimicrobial activity appear to be medium or long-chain unsaturated fatty acids containing one or more double bonds. Most promising agents include Omega-3-fatty acids Linoleic, α-Linoleic, EPA and DHA, as well as Omega-9 fatty acid Oleic acid. These results indicate that fatty acids have the potential to be used as antimicrobials and antibiofilm agents against S. mutans. Future work involves further screening of the most potent fatty acids against a range of bacteria, including Gram-positive and Gram-negative oral pathogens. Future work will involve incorporating the most effective fatty acids onto dental implant devices to prevent biofilm formation.

Keywords: antibiofilm, biofilm, fatty acids, S. mutans

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Authors: Agumah N. B. Orji, M. U., Oru C. M., Ugbo, E. N., Onwuliri E. A Nwakaeze, E. A.,

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ERG11 gene has been reported to be one of the genes whose expression is responsible for resistance of Candida to various triazole drugs, which are first line treatment for candidiasis. This study was carried out to determine the prevalence of Triazole (Fluconazole and voriconazole) resistant Candida albicans expressing ERG11 gene from adult females in Abakaliki. Urine and vaginal swab samples were randomly collected from volunteers after obtaining their consent to participate in the study. A total of 565 adult females participated in the study. A total of 340 urine specimens and 288 vaginal swab specimens were collected. Direct wet mount technique, as well as culture in Trichomonas broth, were used to examine the urine and vaginal swab specimens for the presence of motile Trichomonads. The Trichomonas broth used was selective for both T. vaginalis and C. albicans. Broths that yielded budding yeast cells after microscopy were subcultured on to Sabouraud dextrose agar, after which Germ tube test was carried out to confirm the presence of C. albicans. Biochemical tests, including carbohydrate fermentation and urease utilization, were also performed. Antibiogram of C. albicans isolates obtained from this study was carried out using commercially available azole drugs. Fluconazole and voriconazole were selected as Triazole drugs used for this study. Nystatin was used as a tangential control. An MIC test was carried out with E-strips on some of the resistant C. albicans isolates A total of 6 isolates that resisted all the azole drugs were selected and screened for the presence of ERG11 gene using Reverse transcriptase polymerase chain reaction technique. The total prevalence recorded for C. albicans was 13.0%. Frequency was statistically higher in Pregnant (7.96%) than non pregnant (5.09%) volunteers (X2=0.94 at P=0.05). With respect to clinical samples, frequency was higher in vaginal swabs samples (7.96%) than Urine samples (5.09%) (X2=9.05 at P=0.05). Volunteers within the age group 26-30 years recorded the highest prevalence (4.46%), while those within the age group 36-40 years recorded the lowest at 1.27%(X2=4.34 at P=0.05). In pregnant female participants, the highest frequency was recorded with those in their 3rd trimester (4.14%), while lowest incidence was recorded for those in their first trimester (0.80%). Antibiogram results from this study showed that C. albicans isolates obtained from this study resisted Fluconazole (72%) more than Voriconazole (57%). Only one out of the six selected isolates yielded resistance in the MIC test. Results obtained from the RT-PCR showed that there was no expression of ERG11 gene among the fluconazole resistant isolates of C. albicans. Observed resistance may be due to other factors other than expression of ERG11 gene.

Keywords: candida, ERG11, triazole, nigeria

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Authors: M. S. Raza, A. Kapil, Sonu Tyagi, H. Gautam, S. Mohapatra, R. Chaudhry, S. Sood, V. Goyal, R. Lodha, V. Sreenivas, B. K. Das

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Background: Acute bacterial meningitis remains the major cause of mortality and morbidity. More than half of the survivors develop the significant lifelong neurological abnormalities. Diagnosis of the hospital acquired acute bacterial meningitis (HAABM) is challenging as it appears either in the post operative patients or patients acquire the organisms from the hospital environment. In both the situations, pathogens are exposed to high dose of antibiotics. Chances of getting multidrug resistance organism are very high. We have performed this experiment to find out the etiological agents of HAABM and its antibiotics susceptibility pattern. Methodology: A perspective study was conducted at the Department of Microbiology, All India Institute of Medical Sciences, New Delhi. From March 2015 to April 2018 total 400 Cerebro spinal fluid samples were collected aseptically. Samples were processed for cell count, Gram staining, and culture. Culture plates were incubated at 37°C for 18-24 hours. Organism grown on blood and MacConkey agar were identified by MALDI-TOF Vitek MS (BioMerieux, France) and antibiotic susceptibility tests were performed by Kirby Bauer disc diffusion method as per CLSI 2015 guideline. Results: Of the 400 CSF samples processed, 43 (10.75%) were culture positive for different bacteria. Out of 43 isolates, the most prevalent Gram-positive organisms were S. aureus 4 (9.30%) followed by E. faecium 3 (6.97%) & CONS 2 (4.65%). Similarly, E. coli 13 (30.23%) was the commonest Gram-negative isolates followed by A. baumannii 12 (27.90%), K. pneumonia 5 (11.62%) and P. aeruginosa 4(9.30%). Most of the antibiotics tested against the Gram-negative isolates were resistance to them. Colistin was most effective followed by Meropenem and Imepenim for all Gram-negative HAABM isolates. Similarly, most of antibiotics tested were susceptible to S. aureus and CONS. However, E. faecium (100%) were only susceptible to vancomycin and teicoplanin. Conclusion: Hospital acquired acute bacterial meningitis (HAABM) is becoming the emerging challenge as most of isolates are showing resistance to commonly used antibiotics. Gram-negative organisms are emerging as the major player of HAABM. Great care needs to be taken especially in tertiary care hospitals. Similarly, antibiotic stewardship should be followed and antibiotic susceptibility test (AST) should be performed regularly to update the antibiotic patter and to prevent from the emergence of resistance. Updated information of the AST will be helpful for the better management of the meningitis patient.

Keywords: CSF, MALDI-TOF, hospital acquired acute bacterial meningitis, AST

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Authors: R. A. Abdel Rahman, A. E. Abdel Wahab, E. A. Goghneimy, H. F. Mohamed, E. M. Salama

Abstract:

Infectious diseases are a major cause of death worldwide. The treatment of infections continues to be problematic in modern time because of the severe side effects of some drugs and the growing resistance to antimicrobial agents. Hence, the search for newer, safer and more potent antimicrobials is a pressing need. Herbal medicines have received much attention as a source of new antibacterial drugs since they are considered time-tested and comparatively safe both for human use and the environment. In the present study, the antimicrobial activity of marjoram (Origanum majorana L.) essential oil on some gram positive and gram negative reference bacteria, as well as some hospital resistant microbes, was tested. Marjoram oil was extracted and the oil chemical constituents were identified using GC/MS analysis. Staphylococcus aureas ATCC 6923, Pseudomonus auregonosa ATCC 9027, Bacillus subtilis ATCC 6633, E. coli ATCC 8736 and two hospital resistant microbes isolates 16 and 21 were used. The two isolates were identified by biochemical tests and 16s rRNA as proteus spp. and Enterococcus facielus. The effect of different concentrations of essential oils on bacterial growth was tested using agar disk diffusion assay method to determine the minimum inhibitory concentrations and using micro dilution method to determine the minimum bactericidal concentrations. Marjoram oil was found to be effective against both reference and hospital resistance strains. Hospital strains were more resistant to marjoram oil than reference strains. P. auregonosa growth was completely inhibited at a low concentration of oil (4µl/ml). The other reference strains showed sensitivity to marjoram oil at concentrations ranged from 5 to 7µl/ml. The two hospital strains showed sensitivity at media containing 10 and 15µl/ml oil. The major components of oil were terpineol, cis-beta (23.5%), 1,6 – octadien –3-ol,3,7-dimethyl, 2 aminobenzoate (10.9%), alpha terpieol (8.6%) and linalool (6.3%). Scanning electron microscope (SEM) and transmission electron microscope (TEM) analysis were used to determine the difference between treated and untreated hospital strains. SEM results showed that treated cells were smaller in size than control cells. TEM data showed that cell lysis has occurred to treated cells. Treated cells have ruptured cell wall and appeared empty of cytoplasm compared to control cells which shown to be intact with normal volume of cytoplasm. The results indicated that marjoram oil has a positive antimicrobial effect on hospital resistance microbes. Natural crude extracts can be perfect resources for new antimicrobial drugs.

Keywords: antimicrobial activity, essential oil, hospital resistance microbes, marjoram

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Authors: Recep Kesli, Huseyin Bilgin, Yasar Unlu, Gokhan Gungor

Abstract:

Aim: The aim of this study was to compare diagnostic values of two different molecular-based tests (GenoType® HelicoDR ve Seeplex® H. pylori-ClaR- ACE Detection) in detection presence of the H. pylori from gastric biopsy specimens. In addition to this also was aimed to determine resistance ratios of H. pylori strains against to clarytromycine and quinolone isolated from gastric biopsy material cultures by using both the genotypic (GenoType® HelicoDR, Seeplex ® H. pylori -ClaR- ACE Detection) and phenotypic (gradient strip, E-test) methods. Material and methods: A total of 266 patients who admitted to Konya Education and Research Hospital Department of Gastroenterology with dyspeptic complaints, between January 2011-June 2013, were included in the study. Microbiological and histopathological examinations of biopsy specimens taken from antrum and corpus regions were performed. The presence of H. pylori in all the biopsy samples was investigated by five differnt dignostic methods together: culture (C) (Portagerm pylori-PORT PYL, Pylori agar-PYL, GENbox microaer, bioMerieux, France), histology (H) (Giemsa, Hematoxylin and Eosin staining), rapid urease test (RUT) (CLOtest, Cimberly-Clark, USA), and two different molecular tests; GenoType® HelicoDR, Hain, Germany, based on DNA strip assay, and Seeplex ® H. pylori -ClaR- ACE Detection, Seegene, South Korea, based on multiplex PCR. Antimicrobial resistance of H. pylori isolates against clarithromycin and levofloxacin was determined by GenoType® HelicoDR, Seeplex ® H. pylori -ClaR- ACE Detection, and gradient strip (E-test, bioMerieux, France) methods. Culture positivity alone or positivities of both histology and RUT together was accepted as the gold standard for H. pylori positivity. Sensitivity and specificity rates of two molecular methods used in the study were calculated by taking the two gold standards previously mentioned. Results: A total of 266 patients between 16-83 years old who 144 (54.1 %) were female, 122 (45.9 %) were male were included in the study. 144 patients were found as culture positive, and 157 were H and RUT were positive together. 179 patients were found as positive with GenoType® HelicoDR and Seeplex ® H. pylori -ClaR- ACE Detection together. Sensitivity and specificity rates of studied five different methods were found as follows: C were 80.9 % and 84.4 %, H + RUT were 88.2 % and 75.4 %, GenoType® HelicoDR were 100 % and 71.3 %, and Seeplex ® H. pylori -ClaR- ACE Detection were, 100 % and 71.3 %. A strong correlation was found between C and H+RUT, C and GenoType® HelicoDR, and C and Seeplex ® H. pylori -ClaR- ACE Detection (r:0.644 and p:0.000, r:0.757 and p:0.000, r:0.757 and p:0.000, respectively). Of all the isolated 144 H. pylori strains 24 (16.6 %) were detected as resistant to claritromycine, and 18 (12.5 %) were levofloxacin. Genotypic claritromycine resistance was detected only in 15 cases with GenoType® HelicoDR, and 6 cases with Seeplex ® H. pylori -ClaR- ACE Detection. Conclusion: In our study, it was concluded that; GenoType® HelicoDR and Seeplex ® H. pylori -ClaR- ACE Detection was found as the most sensitive diagnostic methods when comparing all the investigated other ones (C, H, and RUT).

Keywords: Helicobacter pylori, GenoType® HelicoDR, Seeplex ® H. pylori -ClaR- ACE Detection, antimicrobial resistance

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Authors: S. H. Gasa, L. Singh, B. Pillay, A. O. Olaniran

Abstract:

Wastewater treatment plants (WWTPs) have been implicated as the leading reservoir for antibiotic resistant bacteria (ARB), including Enterococci spp. and antibiotic resistance genes (ARGs), worldwide. Enterococci are a group of clinically significant bacteria that have gained much attention as a result of their antibiotic resistance. They play a significant role as the principal cause of nosocomial infections and dissemination of antimicrobial resistance genes in the environment. The main objective of this study was to ascertain the role of WWTPs in Durban, South Africa as potential reservoirs for antibiotic resistant Enterococci (ARE) and their related ARGs. Furthermore, the antibiogram and resistance gene profile of Enterococci species recovered from treated wastewater effluent and receiving surface water in Durban were also investigated. Using membrane filtration technique, Enterococcus selective agar and selected antibiotics, ARE were enumerated in samples (influent, activated sludge, before chlorination and final effluent) collected from two WWTPs, as well as from upstream and downstream of the receiving surface water. Two hundred Enterococcus isolates recovered from the treated effluent and receiving surface water were identified by biochemical and PCR-based methods, and their antibiotic resistance profiles determined by the Kirby-Bauer disc diffusion assay, while PCR-based assays were used to detect the presence of resistance and virulence genes. High prevalence of ARE was obtained at both WWTPs, with values reaching a maximum of 40%. The influent and activated sludge samples contained the greatest prevalence of ARE with lower values observed in the before and after chlorination samples. Of the 44 vancomycin and high-level gentamicin-resistant isolates, 11 were identified as E. faecium, 18 as E. faecalis, 4 as E. hirae while 11 are classified as “other” Enterococci species. High-level aminoglycoside resistance for gentamicin (39%) and vancomycin (61%) was recorded in species tested. The most commonly detected virulence gene was the gelE (44%), followed by asa1 (40%), while cylA and esp were detected in only 2% of the isolates. The most prevalent aminoglycoside resistance genes were aac(6')-Ie-aph(2''), aph(3')-IIIa, and ant(6')-Ia detected in 43%, 45% and 41% of the isolates, respectively. Positive correlation was observed between resistant phenotypes to high levels of aminoglycosides and presence of all aminoglycoside resistance genes. Resistance genes for glycopeptide: vanB (37%) and vanC-1 (25%), and macrolide: ermB (11%) and ermC (54%) were detected in the isolates. These results show the need for more efficient wastewater treatment and disposal in order to prevent the release of virulent and antibiotic resistant Enterococci species and safeguard public health.

Keywords: antibiogram, enterococci, gentamicin, vancomycin, virulence signatures

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Authors: M. I. Ribah, M. Jibir, Y. A. Bashar, S. S. Manga

Abstract:

Food safety is a significant and growing public health problem in the world and Nigeria as a developing country, since food-borne diseases are important contributors to the huge burden of sickness and death of humans. In Nigeria, traditional ready-to-eat meat products (RTE-MPs) like balangu, tsire, guru and dried meat products like kilishi, dambun nama, banda, were reported to be highly appreciated because of their eating qualities. The consumption of these products was considered as safe due to the treatments that are usually involved during their production process. However, during processing and handling, the products could be contaminated by pathogens that could cause food poisoning. Therefore, a hazard identification for pathogenic bacteria on some traditional RTE-MPs was conducted in Kebbi and Sokoto States, Nigeria. A total of 116 RTE-MPs (balangu-38, kilishi-39 and tsire-39) samples were obtained from retail outlets and analyzed using standard cultural microbiological procedures in general and selective enrichment media to isolate the target pathogens. A six-fold serial dilution was prepared and using the pour plating method, colonies were counted. Serial dilutions were selected based on the prepared pre-labeled Petri dishes for each sample. A volume of 10-12 ml of molten Nutrient agar cooled to 42-45°C was poured into each Petri dish and 1 ml each from dilutions of 102, 104 and 106 for every sample was respectively poured on a pre-labeled Petri plate after which colonies were counted. The isolated pathogens were identified and confirmed after series of biochemical tests. Frequencies and percentages were used to describe the presence of pathogens. The General Linear Model was used to analyze data on pathogen presence according to RTE-MPs and means were separated using the Tukey test at 0.05 confidence level. Of the 116 RTE-MPs samples collected, 35 (30.17%) samples were found to be contaminated with some tested pathogens. Prevalence results showed that Escherichia coli, salmonella and Staphylococcus aureus were present in the samples. Mean total bacterial count was 23.82×106 cfu/g. The frequency of individual pathogens isolated was; Staphylococcus aureus 18 (15.51%), Escherichia coli 12 (10.34%) and Salmonella 5 (4.31%). Also, among the RTE-MPs tested, the total bacterial counts were found to differ significantly (P < 0.05), with 1.81, 2.41 and 2.9×104 cfu/g for tsire, kilishi, and balangu, respectively. The study concluded that the presence of pathogenic bacteria in balangu could pose grave health risks to consumers, and hence, recommended good manufacturing practices in the production of balangu to improve the products’ safety.

Keywords: ready-to-eat meat products, retail outlets, public health, safety assessment

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Authors: Tahisa M. Pedroso, Hérida R. N. Salgado

Abstract:

The microbiological turbidimetric assay allows the determination of potency of the drug, by measuring the turbidity (absorbance), caused by inhibition of microorganisms by ertapenem sodium. Ertapenem sodium (ERTM), a synthetic antimicrobial agent of the class of carbapenems, shows action against Gram-negative, Gram-positive, aerobic and anaerobic microorganisms. Turbidimetric assays are described in the literature for some antibiotics, but this method is not described for ertapenem. The objective of the present study was to develop and validate a simple, sensitive, precise and accurate microbiological assay by turbidimetry to quantify ertapenem sodium injectable as an alternative to the physicochemical methods described in the literature. Several preliminary tests were performed to choose the following parameters: Staphylococcus aureus ATCC 25923, IAL 1851, 8 % of inoculum, BHI culture medium, and aqueous solution of ertapenem sodium. 10.0 mL of sterile BHI culture medium were distributed in 20 tubes. 0.2 mL of solutions (standard and test), were added in tube, respectively S1, S2 and S3, and T1, T2 and T3, 0.8 mL of culture medium inoculated were transferred to each tube, according parallel lines 3 x 3 test. The tubes were incubated in shaker Marconi MA 420 at a temperature of 35.0 °C ± 2.0 °C for 4 hours. After this period, the growth of microorganisms was inhibited by addition of 0.5 mL of 12% formaldehyde solution in each tube. The absorbance was determined in Quimis Q-798DRM spectrophotometer at a wavelength of 530 nm. An analytical curve was constructed to obtain the equation of the line by the least-squares method and the linearity and parallelism was detected by ANOVA. The specificity of the method was proven by comparing the response obtained for the standard and the finished product. The precision was checked by testing the determination of ertapenem sodium in three days. The accuracy was determined by recovery test. The robustness was determined by comparing the results obtained by varying wavelength, brand of culture medium and volume of culture medium in the tubes. Statistical analysis showed that there is no deviation from linearity in the analytical curves of standard and test samples. The correlation coefficients were 0.9996 and 0.9998 for the standard and test samples, respectively. The specificity was confirmed by comparing the absorbance of the reference substance and test samples. The values obtained for intraday, interday and between analyst precision were 1.25%; 0.26%, 0.15% respectively. The amount of ertapenem sodium present in the samples analyzed, 99.87%, is consistent. The accuracy was proven by the recovery test, with value of 98.20%. The parameters varied did not affect the analysis of ertapenem sodium, confirming the robustness of this method. The turbidimetric assay is more versatile, faster and easier to apply than agar diffusion assay. The method is simple, rapid and accurate and can be used in routine analysis of quality control of formulations containing ertapenem sodium.

Keywords: ertapenem sodium, turbidimetric assay, quality control, validation

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