Search results for: Sabouraud agar

Authors: Nenny Harijani, Dhandy Koesoemo Wardhana

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The aim of this study is to know the bacteriocin as antimicrobial activity produced by Lactic Acid Bacteria (LAB) as Antibacterial of Sub Clinic Mastitis on Dairy Cows. The antimicrobial is produced by LAB which isolates from cattle intestine can inhibit the growth Staphylococcus aureus, Steptocococcus agalactiae an Escherichia coli which were caused by dairy cattle subclinical mastitis. The failure of this bacteria growth was indicated by the formation of a clear zone surrounding the colonies on Brain Heart Infusion Agar plate. The bacteriocin was produced by Lactic Acid Bacteria (LAB) as antimicrobial, which could inhibit the growth of indicator bacteria Staphylococcus aureus, S.aglactiae and E.coli. This study was also developed bacteriocin to be used as a therapeutic of subclinical mastitis on dairy cows. The method used in this study was isolation, selection and identification of LAB using Mann Rogosa Sharp Medium, followed by characterization of the bacteriocin produced by LAB. The result of the study showed that bacteriocin isolated from beef cattle’s intestine could inhibit the growth Staphylococcus aureus, S. agalactiae, an Escherichia coli, which was indicated by clear zone surrounding the colonies on Brain Heart Infusion Agar plate. Characteristics of bacteriocin were heat-stable exposed to 80 0C for 30 minutes and 100 ⁰C for 15 minutes and inactivated by proteolytic enzymes such as trypsin. This approach has suggested the development of bacteriocin as a therapeutic agent for subclinical mastitis in dairy cattle.

Keywords: lactic acid bacteria, bacteriocin, staphylococcus aureus, S. agalactiae, E. coli, sub

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Authors: Alazar Nebyou, Sujata Pandit

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Lactic acid bacteria (LAB) are essential ingredients in probiotic foods, intestinal microflora, and dairy products that are capable of coping up with harsh gastrointestinal tract conditions and are available in a variety of environments. The objective of this study is to evaluate the probiotic property of LAB isolated from bovine milk. Milk samples were collected from local dairy farms. Samples were obtained using sterile test tubes and transported to a laboratory in the icebox for further biochemical characterization. Preliminary physiological and biochemical identification of LAB isolates was conducted by growing on MRS agar after ten-fold serial dilution. Seven of the best isolates were selected for the evaluation of the probiotic property. The LAB isolates were checked for resistance to antibiotics and their antimicrobial activity by disc diffusion assay and agar well diffusion assay respectively. Bile salt hydrolase activity of isolates was studied by growing isolates in a BSH medium with bile salt. Cell surface property of isolates was assayed by studying their autoaggregation and coaggregation percentage with S. aerues. All isolates were found BSH positive. In addition, BCM2 and BGM1 were susceptible to all antibiotic disks except BBM1 which was resistant to all antibiotic disks. BCM1 and BGM1 had the highest autoaggregation and coaggregation potential respectively. Since all LAB isolates showed gastrointestinal tolerance and good cell surface property they could be considered as good potential probiotic candidates for treatment and probiotic starter culture preparation.

Keywords: probiotic, aggregation, lactic acid bacteria, antimicrobial activity

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Authors: Futo Asano, Yusuke Yatsushiro, Hirokuni Miyamoto, Hiroaki Kodama

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A thermophile-fermented compost is produced using small fishes, crabs, and shrimps under a high temperature (approximately 75℃) by fermentation-associated self-heating. This compost has been used as a feed additive for pigs and hens in Japan, and the fecundity of this livestock is enhanced. Firmicutes is a dominant phylum in the microbial composition of the compost. We first reported that improvement of female larval fitness of Hercules beetle can be achieved by amendment of this compost to the humus. When the 90-d-old larvae were reared for subsequent 72 days in the humus with this compost, the growth of female larvae was significantly enhanced when compared with the growth of female larvae in the humus without the compost. In contrast, the growth of male larvae in the compost-free humus was the same as the larvae grow in the compost-amended humus. The bacterial composition of the feces of larvae was determined at 0 days and 46 days after transfer to the humus with or without the compost. The most dominant bacterium in the feces was Xylanimonas. Interestingly, the growth improvement of female larvae was associated with an increased abundance of Mollicutes in the fecal samples. These results indicate that the compost act as a probiotic material for enhancing the female larvae growth by supporting Mollicutes. Here, we tried to isolate Mollicutes from the contents of the midgut and hindgut of the 3rd instar female larvae of the Hercules beetle. These gut contents were spread onto a selective agar medium for Mollicutes (PPLO agar broth, BD Difco, NJ, USA). Although we isolated none of the Mollicutes until now, several bacteria that are closely related to Xylanimonas and Luteimicrobium were isolated. These isolates have xylanase and glucanase (CMCase) activities. We show the gut bacterial profiles of larvae and discuss how the fitness of female larvae of the Hercules beetle is improved by the compost.

Keywords: compost, beetle, mollicutes, woody biomass

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Authors: Alok Kumar, Hari Ram, Lebin Thomas, Ved Pal Singh

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Organic solvent tolerant amylases and proteases of microbial origin are in great demand for their application in transglycosylation of water-insoluble flavanoids and in peptide synthesizing reaction in organic media. Most of the amylases and proteases are unstable in presence of organic solvent. In the present work two different bacterial strains M-11 and VP-07 were isolated from the soil sample of a dairy farm in Delhi, India, for the efficient production of extracellular amylase and protease through their screening on starch agar (SA) and skimmed milk agar (SMA) plates, respectively. Both the strains (M-11 and VP-07) were identified based on morphological, biochemical and 16S rRNA gene sequencing methods. After analysis through Ez-Taxon software, the strains M-11 and VP-07 were found to have maximum pairwise similarity of 98.63% and 100% with Bacillus subtilis subsp. inaquosorum BGSC 3A28 and Bacillus anthracis ATCC 14578 and were therefore identified as Bacillus sp. UKS1 and Bacillus sp. UKS2, respectively. Time course study of enzyme activity and bacterial growth has shown that both strains exhibited typical sigmoid growth behavior and maximum production of amylase (180 U/ml) and protease (78 U/ml) by these strains (UKS1 and UKS2) was commenced during stationary phase of growth at 24 and 20 h, respectively. Thereafter, both amylase and protease were tested for their tolerance towards organic solvents and were found to be active as well stable in p-xylene (130% and 115%), chloroform (110% and 112%), isooctane (119% and 107%), benzene (121% and 104%), n-hexane (116% and 103%) and toluene (112% and 101%, respectively). Owing to such properties, these enzymes can be exploited for their potential application in industries for organic synthesis.

Keywords: amylase, enzyme activity, industrial applications, organic solvent tolerant, protease

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Authors: Sumeeta Khurana, Kirti Megha, Amit Gupta, Rakesh Sehgal

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Background: Amoebic keratitis is a painful vision threatening infection caused by a free living pathogenic amoeba Acanthamoeba. It can be misdiagnosed and very difficult to treat if not suspected early. The epidemiology of Acanthamoeba genotypes causing infection in our geographical area is not yet known to the best of our knowledge. Objective: To characterize Acanthamoeba isolates from amoebic keratitis patients. Methods: A total of 19 isolates obtained from patients with amoebic keratitis presenting to the Advanced Eye Centre at Postgraduate Institute of Medical Education and Research, a tertiary care centre of North India over a period of last 10 years were included. Their corneal scrapings, lens solution and lens case (in case of lens wearer) were collected for microscopic examination, culture and molecular diagnosis. All the isolates were maintained in the Non Nutrient agar culture medium overlaid with E.coli and 13 strains were axenised and maintained in modified Peptone Yeast Dextrose Agar. Identification of Acanthamoeba genotypes was based on amplification of diagnostic fragment 3 (DF3) region of the 18srRNA gene followed by sequencing. Nucleotide similarity search was performed by BLAST search of sequenced amplicons in GenBank database (http//www.ncbi.nlm.nih.gov/blast). Multiple Sequence alignments were determined by using CLUSTAL X. Results: Nine out of 19 Acanthamoeba isolates were found to belong to Genotype T4 followed by 6 isolates of genotype T11, 3 T5 and 1 T3 genotype. Conclusion: T4 is the predominant Acanthamoeba genotype in our geographical area. Further studies should focus on differences in pathogenicity of these genotypes and their clinical significance.

Keywords: Acanthamoeba, free living amoeba, keratitis, genotype, ocular

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Authors: Atin Adhikari, Sushma Kurella, Pratik Banerjee, Nabanita Mukherjee, Yamini M. Chandana Gollapudi, Bushra Shah

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Different sharp instruments, drilling machines, and high speed rotary instruments are routinely used in dental clinics during dental cleaning. Therefore, these cleaning procedures release a lot of oral microorganisms including bacteria in clinic air and may cause significant occupational bioaerosol exposure risks for dentists, dental hygienists, patients, and dental clinic employees. Two major goals of this study were to quantify volumetric airborne concentrations of bacteria and to assess overall microbial activity in this type of occupational environment. The study was conducted in several dental clinics of southern Georgia and 15 dental cleaning procedures were targeted for sampling of airborne bacteria and testing of overall microbial activity in settled dusts over clinic floors. For air sampling, a Biostage viable cascade impactor was utilized, which comprises an inlet cone, precision-drilled 400-hole impactor stage, and a base that holds an agar plate (Tryptic soy agar). A high-flow Quick-Take-30 pump connected to this impactor pulls microorganisms in air at 28.3 L/min flow rate through the holes (jets) where they are collected on the agar surface for approx. five minutes. After sampling, agar plates containing the samples were placed in an ice chest with blue ice and plates were incubated at 30±2°C for 24 to 72 h. Colonies were counted and converted to airborne concentrations (CFU/m3) followed by positive hole corrections. Most abundant bacterial colonies (selected by visual screening) were identified by PCR amplicon sequencing of 16S rRNA genes. For understanding overall microbial activity in clinic floors and estimating a general cleanliness of the clinic surfaces during or after dental cleaning procedures, ATP levels were determined in swabbed dust samples collected from 10 cm2 floor surfaces. Concentration of ATP may indicate both the cell viability and the metabolic status of settled microorganisms in this situation. An ATP measuring kit was used, which utilized standard luciferin-luciferase fluorescence reaction and a luminometer, which quantified ATP levels as relative light units (RLU). Three air and dust samples were collected during each cleaning procedure (at the beginning, during cleaning, and immediately after the procedure was completed (n = 45). Concentrations at the beginning, during, and after dental cleaning procedures were 671±525, 917±1203, and 899±823 CFU/m3, respectively for airborne bacteria and 91±101, 243±129, and 139±77 RLU/sample, respectively for ATP levels. The concentrations of bacteria were significantly higher than typical indoor residential environments. Although an increasing trend for airborne bacteria was observed during cleaning, the data collected at three different time points were not significantly different (ANOVA: p = 0.38) probably due to high standard deviations of data. The ATP levels, however, demonstrated a significant difference (ANOVA: p <0.05) in this scenario indicating significant change in microbial activity on floor surfaces during dental cleaning. The most common bacterial genera identified were: Neisseria sp., Streptococcus sp., Chryseobacterium sp., Paenisporosarcina sp., and Vibrio sp. in terms of frequencies of occurrences, respectively. The study concluded that bacterial exposure in dental clinics could be a notable occupational biohazard, and appropriate respiratory protections for the employees are urgently needed.

Keywords: bioaerosols, hospital hygiene, indoor air quality, occupational biohazards

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Authors: I. Ahmad, A. Hannan, S. Ahmad, M. Asif, M. F. Nawaz, M. A. Tanvir, M. F. Azhar

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During this research, a comprehensive survey of tree growing areas of Faisalabad district of Pakistan was conducted to observe the symptoms, spectrum, occurrence and severity of A. nilotica and E. camaldulensis decline. Objective of current research was to investigate specific fungal pathogens involved in decline of A. nilotica and E. camaldulensis. For this purpose, infected roots, bark, neck portion, stem, branches, leaves and infected soils were collected to identify associated fungi. Potato dextrose agar (PDA) and Czepak dox agar media were used for isolations. Identification of isolated fungi was done microscopically and different fungi were identified. During survey of urban locations of Faisalabad, disease incidence on Kikar and Eucalyptus was recorded as 3.9-7.9% and 2.6-7.1% respectively. Survey of Agroforest zones of Faisalabad revealed decline incidence on kikar 7.5% from Sargodha road while on Satiana and Jhang road it was not planted. In eucalyptus trees, 4%, 8% and 0% disease incidence was observed on Jhang road, Sargodha road and Satiana road respectively. The maximum fungus isolated from the kikar tree was Drechslera australiensis (5.00%) from the stem part. Aspergillus flavus also gave the maximum value of (3.05%) from the bark. Alternaria alternata gave the maximum value of (2.05%) from leaves. Rhizopus and Mucor spp. were recorded minimum as compared to the Drechslera, Alternaria and Aspergillus. The maximum fungus isolated from the Eucalyptus tree was Armillaria luteobubalina (5.00%) from the stem part. The other fungi isolated were Macrophamina phaseolina and A. niger.

Keywords: decline, frequency of mycoflora, A. nilotica and E. camaldulensis, Drechslera australiensis, Armillaria luteobubalina

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Authors: Suhaida Salleh, Tee Yei Kheng

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Black pod is the most commonly destructive disease of cacao (Theobroma cacao) which cause major losses to global production of cocoa beans. The genus of Phytophthora is the important pathogen of this disease worldwide. The species of P. megakarya causes black pod disease in West Africa, whereas P. capsici and P. citrophthora cause the incident in Central and South America. In Malaysia, this disease is caused by P. palmivora which infect all stages of pod development including flower cushion, cherelle, immature and mature pods. This pathogen destroys up to 10% of trees yearly through stem cankers and causes 20 to 30% pod damages through black pod rot. Since P. palmivora has a high impact on cocoa yield, it is crucial to identify some of the abiotic factors that can constrain their growth. In an effort to evaluate the effect of different substrates and temperatures to the growth of P. palmivora, a laboratory study was done under a different range of temperatures. Different substrate for the growth of P. palmivora were used which are corn meal agar (CMA) media and detached pod of cocoa. An agar plug of seven days old of P. palmivora growth was transferred on both substrates and incubated at 24, 27, 30, 33 and 36ᵒC, respectively. The diameter of lesion on pod and the cultural growth of pathogen was recorded for 7 consecutive days. The optimum incubation temperature of P. palmivora on both substrates is at 27ᵒC. However, the growth tends to be inhibited as the temperature increases. No lesion developed on pod surface incubated at 36ᵒC and only a small lesion observed at 33ᵒC. The sporulation with the formation of white mycelial growth on pod surface was only visible at optimum temperature, 27ᵒC. On CMA, the pathogen grew over the entire range of temperatures tested. The study is, therefore, concluded that P. palmivora grow the best at temperature of 27ᵒC on both substrates and their growth begin to inhibit when the temperature rises to more than 27ᵒC. The growth pattern of this pathogen is similar on both pod surface and cultural media.

Keywords: cocoa, Phytophthora palmivora, substrate, temperature

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Authors: Sina Gharevali

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Staphylococcus aureus is one of the most important factors for nosocomial infections and infections acquired in a hospital setting role as is. Drug-resistant bacteria methicillin, which in 1961 was reported in many parts of the world, Made the role as the last drug, vancomycin, in the treatment of infections caused by the Staphylococcus aureus chain be taken into consideration. The aim of this study was to evaluate the antimicrobial effects of copper nanoparticles and compared it with antibiotics on Staphylococcus aureus resistant to vancomycin in vitro and animal model. In this study, this test was performed, and the most effective antibiotic for vancomycin-resistant Staphylococcus aureus was determined by disk diffusion method. After various concentrations of copper nanoparticles and antibiotics were prepared and vancomycin resistant Staphylococcus aureus bacteria with serial dilution method for determining antibiotic ciprofloxacin. Minimum Inhibitory Concentration and Minimum Bactericidal Concentrationcopper nanoparticles was performed. The agar dilution method for bacterial growth in different concentrations of copper nanoparticles and antibiotics ciprofloxacin was performed. The agar dilution method for bacterial growth in different concentrations of copper nanoparticles and antibiotics ciprofloxacin was performed. Then the broth dilution method for the antibiotic ciprofloxacin, nano-particles, and nano-particles of copper and copper-established antibiotic synergy MIC and MBC were obtained. MBC was obtained from the experimental animal model test method, and the results were compared. The results showed that copper nanoparticles compared with the antibiotic ciprofloxacin in vitro and animal model more effective in inhibiting the growth of Staphylococcus aureus resistant to vancomycin and ciprofloxacin and extent of the impact of the Synthetic effect of lower copper nanoparticles. Which can then be used to treat clinical research as a candidate.

Keywords: nanoparticles, copper, staphylococcus, aureus

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Authors: Sarah Almuhayya, Andrew Mcbain, Gavin Humphreys

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Background. The microbiome of the upper respiratory tract (URT) has received less research attention than other body sites. This study aims to investigate the microbial ecology of the human URT with a focus on the antagonism between the corynebacteria and staphylococci. Methods. Mucosal swabs were collected from the anterior nares and nasal turbinates of 20 healthy adult subjects. Genomic DNA amplification targeting the (V4) of the 16Sr RNA gene was conducted and analyzed using QIIME. Nasal swab isolates were cultured and identified using near full-length sequencing of the 16S rRNA gene. Isolates identified as corynebacteria or staphylococci were typed using (rep-PCR). Antagonism was determined using an agar-based inhibition assay. Results. Four major bacterial phyla (Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria) were identified from all volunteers. The typing of cultured staphylococci and corynebacteria suggested that intra-individual strain diversity was limited. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between staphylococci and corynebacteria. Despite the apparent antagonism between these genera, it was limited when investigated on agar. Of 1000 pairwise interactions, observable zones of inhibition were only reported between a single strain of C.pseudodiphtheriticum and S.aureus. Imaging under EM revealed this effect to be bactericidal with clear lytic effects on staphylococcal cell morphology. Conclusion. Nasal microbiota is complex, but culturable staphylococci and corynebacteria were limited in terms of clone type. Analysis of generated nasal microbiota profiles suggested an inverse correlation in terms of relative abundance between these genera suggesting an antagonism or competition between these taxonomic groups.

Keywords: nasal, microbiota, S.aureus, microbioal interaction

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Authors: F. A. Gberindyer, M. O.Abatan, A. B. Saba

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Background: Gentamicin is an aminoglycoside antibiotic used in the treatment of infections caused by Gram-negative aerobic bacteria organisms in human and animals. In Nigeria, there are arrays of multisource generic versions of injectable gentamicin sulphate in the drug markets. There is a high prevalence of counterfeit and substandard drugs in the third world countries with consequent effect on their therapeutic efficacy and safety. Aim: The aim of this study was to investigate pharmaceutical equivalence of some of these generics used in veterinary practice in Nigeria. Methodology: About 20 generics of injectable gentamicin sulphate were sampled randomly across Nigeria but 15 were analyzed for identity and potency. Identity test was done using Fourier transform infra red spectroscopy and the spectral for each product compared with that of the USP reference standard for similarity. Microbiological assay using agar diffusion method with E. coli as a test organism on nutrient agar was employed and the respective diameters of bacterial inhibition zones obtained after 24 hour incubation at 37°C. The percent potency for each product was thereafter calculated and compared with the official specification. Result And Discussion: None of the generics is produced in any African country. About 75 % of the products are imported from China whereas 60 % of the veterinary generics are manufactured in Holland. Absorption spectra for the reference and test samples were similar. Percent potencies of all test products were within the official specification of 95-115 %. Nigeria relies solely on imported injectable gentamicin sulphate products. All sampled generic versions passed both identity and potency tests. Clinicians should ensure that drugs are used rationally since the converse could be contributing to the therapeutic failures reported for most of these generics. Bioequivalence study is recommended to ascertain their interchangeability when parenteral extra venous routes are indicated.

Keywords: generics, gentamicin, identity, multisource, potency

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Authors: Sergei Voychuk

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Introduction: Adaptive response (AR) is a manifestation of radiation hormesis, which deal with the radiation resistance that may be increased with the pretreatment with small doses of radiation. In the current study, we evaluated the potency of radiofrequency EMF to induce the AR mechanisms and to increase a resistance to UV light. Methods: Saccharomyces cerevisiae yeast strains, which were created to study induction of mutagenesis and recombination, were used in the study. The strains have mutations in rad2 and rad54 genes, responsible for DNA repair: nucleotide excision repair (PG-61), postreplication repair (PG-80) and mitotic (crossover) recombination (T2). An induction of mutation and recombination are revealed due to the formation of red colonies on agar plates. The PG-61 and T2 are UV sensitive strains, while PG-80 is sensitive to ionizing radiation. Extremely high frequency electromagnetic field (EHF-EMF) was used. The irradiation was performed in floating mode and frequency changed during exposure from 57 GHz to 62 GHz. The power of irradiation was 100 mkW, and duration of exposure was 10 and 30 min. Treatment was performed at RT and then cells were stored at 28° C during 1 h without any exposure but after that they were treated with UV light (254nm) for 20 sec (strain T2) and 120 sec (strain PG-61 and PG-80). Cell viability and quantity of red colonies were determined after 5 days of cultivation on agar plates. Results: It was determined that EHF-EMF caused 10-20% decrease of viability of T2 and PG-61 strains, while UV showed twice stronger effect (30-70%). EHF-EMF pretreatment increased T2 resistance to UV, and decreased it in PG-61. The PG-80 strain was insensitive to EHF-EMF and no AR effect was determined for this strain. It was not marked any induction of red colonies formation in T2 and PG-80 strain after EHF or UV exposure. The quantity of red colonies was 2 times more in PG-61 strain after EHF-EMF treatment and at least 300 times more after UV exposure. The pretreatment of PG-61 with EHF-EMF caused at least twice increase of viability and consequent decrease of amount of red colonies. Conclusion: EHF-EMF may induce AR in yeast cells and increase their viability under UV treatment.

Keywords: Saccharomyces cerevisiae, EHF-EMF, UV light, adaptive response

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Authors: I. Ibrahim, A. Mann

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Phytochemical screening of crude Chrysanthlum Indicum revealed the presence of carbohydrates, flavonoids, saponins, tannins, alkanoids, steroidal nucleus and cardiac glycosides. The extract was evaluated against some pathogenic organisms by agar dilution method. The minimum inhibitory concentration and minimum bacteriocidal concentration (MBC) of the active extract of Chrysanthlum Indicum shows that its extract could be a potential source of antimicrobial agents.

Keywords: extract, phytochemicals, antimicrobial, antibacterial, Chrysanthlum indicum

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Authors: Satender Saraswat, Dharmendra Prasad Singh, Rajesh Kumar Verma, Swati Sarswat

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Background and Purpose: The predominant cause of candidiasis was Candida albicans which has shifted towards non-Candida albicans Candida (NCAC) (Candida species other than the C. albicans). NCAC, earlier considered non-pathogenic or minimally virulent, are now considered a primary cause of morbidity and mortality in immunocompromised. With the NCAC spp. gaining weightage in the clinical cases, this study was conducted to determine the prevalence of NCAC spp. in different clinical specimens and to assess a few of their virulence factors. Material and Methods: Routine samples for bacterial culture and sensitivity, showing colony characteristics like Candida on Blood Agar and microscopic features resembling Candida spp. were processed further. Candida isolates were tested for chlamydospore formation, biochemical tests including sugar fermentation and sugar assimilation tests, and growth at 42oC, colony colour on HiCrome™ Candida Differential Agar, HiCandida Identification Kit and VITEK-2 Compact. Virulence factors like adherence to buccal epithelial cells (ABEC), biofilm formation, hemolytic activity, and production of coagulase enzyme were also tested. Results: Mean age of the patients was 38.46 with a male-female ratio of 1.36:1. 137 Candida isolates were recovered. 45.3% isolates were isolated from urine, 19.7% from vaginal swabs and 13.9% from oropharyngeal swabs. 55 (40.1%) isolates of C. albicans and 82 (59.9%) of NCAC spp. were identified, with C. tropicalis (23.4%) in NCAC. C. albicans (3; 50%) was the commonest species in cases of candidemia. Haemolysin production (85.5%) and ABEC (78.2%) were the major virulence factors in C. albicans. C. tropicalis (59.4%) and C. dubliniensis (50%) showed maximum ABEC. Biofilm forming capacity was higher in C. tropicalis (78.1%) than C. albicans (67%). Conclusion: This study suggests varied prevalence and virulence based on geographical locations, even within a subcontinent. It clearly demarcates the emergence of NCAC and their predominance in different body fluids. Identification of Candida to species level should become a routine in all the laboratories.

Keywords: ABEC, NCAC, non-Candida albicans Candida, Vitek-2TM compact

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Authors: Ferhat Mohamed Amine, Boukhatem Mohamed Nadjib, Chemat Farid

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The Lamiaceae family is widely spread in Algeria. Due to the application of Thymus species growing wild in Algeria as a culinary herb and in folk medicine, the purpose of the present work was to evaluate antimicrobial activities of their essential oils and relate them with their chemical composition, for further application in food and pharmaceutical industries as natural valuable products. The extraction of the Thymus vulgaris L. essential oil (TVEO) was obtained by steam distillation. Chemical composition of the TVEO was determined by Gas Chromatography. A total of thirteen compounds were identified. Carvacrol (83.8%) was the major component, followed by cymene (8.15%) and terpinene (4.96%). Antibacterial action of the TVEO against 23 clinically isolated bacterial strains was determined by using agar disc diffusion and vapour diffusion methods at different doses. By disc diffusion method, TVEO showed potent antimicrobial activity against gram-positive bacteria more than gram-negative strains and antibiotic discs. The Diameter of Inhibition Zone (DIZ) varied from 25 to 60 mm for S. aureus, B. subtilisand E. coli. However, the results obtained by both agar diffusion and vapour diffusion methods were different. Significantly higher antibacterial effect was observed in the vapour phase at lower doses. S. aureus and B. subtilis were the most susceptible strains to the oil vapour. Therefore, smaller doses of EO in the vapour phase can be inhibitory to pathogenic bacteria. There is growing evidence that TVEO in vapour phase are effective antiseptic systems and appears worthy to be considered for practical uses in the treatment of human infections oras air decontaminants in hospital. TVEO has considerable antibacterial activity deserving further investigation for clinical applications. Also whilst the mode of action remains mainly undetermined, this experimental approach will need to continue.

Keywords: antimicrobial drugs, carvacrol, disc diffusion, Thymus vulgaris, vapour diffusion

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Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le

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Since the marine environmental conditions are extremely different from the other ones, so that marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411that are effective against wide range of pathogens: both gram-positive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the furture.

Keywords: marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis

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Authors: C. Nathanailides, P. Logothetis, G. Kanlis S. Anastasiou, L. Kokokiris, P. Mpeza

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Farmed Red porgy (Pagrus pagrus) is one of the “new candidate fish species” for the diversification of Mediterranean aquaculture which is predomintly based on the cultivation of the European sea bass, (Dicenfrarchus labrax), and the gilthead sea bream, (Sparus aurata). The quality of farmed red porgy (Pagrus pagrus) was investigated with samples obtained from the local fish market in the region of Igoumenitsa, NW Greece. Sample of the fish (ungutted and with scales) were purchased from three local fish mongers and transported to the laboratory within few minutes in foamed polystyrene boxes in ice. The average weight of whole fish ranged between 271-289g. A sample of the fish flesh taken from the upper epaxial region was transferred aseptically to a stomacher bag containing sterile Buffered Peptone Water solution (0.1%) and homogenized. After serial dilutions in 0.1% peptone water, the homogenates were spread on the surface of agar plates. Total viable counts (TVC) were determined using plate count agar after incubation at 30 oC for 3 days. The quality attributes monitored during the present work included bacterial load (total mesophilic) and the pH of the flesh. There was a marginal increase in the price of fresh red porgy sold during the summer time, with prices ranging, over a period of four seasons, from 5.85 to 7.5 per kilo. The results of the microbiological analysis indicate that with the exception of summer samples (which exhibited 5.23 (±0.13) log cfu/g), the bacterial load remained well below the legal limits and was around 3.1 log cfu/g. The pH values varied between 6.54 and 6.69. The results indicate a possible influence of season on the bacterial load of fish sold in the market. Nevertheless, the parameters investigated in the present work indicate that the bacteria load was well below the legal limit and that fish were sold within few days after harvesting. The peak of bacterial load in the summer samples may be a result of a post-harvesting contamination of the farmed fish and temperature fluctuations during handling and transportation.

Keywords: fish quality, marketing, aquaculture, Pagrus pagrus

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Authors: S. F. Afzali, W. L. Wong

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The epizootic ulcerative syndrome (EUS) causes by the oomycete fungus, Aphanomyces invadans; known to be one of the infectious fish diseases for farmed and wild fishes in fresh and brackish-water from the Asia-pacific region, America and Africa. Although, EUS had been documented by the Office International des Epizooties (OIE) since 1995, hitherto, there is neither standard chemical agents that can be used for successful treatment of this destructive infection in the time of outbreak; nor available vaccine for prevention. Plant-based remedies in controlling fish diseases are gaining much attention recently as an alternative to chemical treatments, which possess negative effects to the environment and human. In present study, Sonneratia alba, a mangrove plant belongs to the Sonneratiaceae family, was screened in vitro and in vivo for its antifungal activity against A. invadans mycelium growth and its effects on fish innate immune system and disease resistant. The in vitro tests was performed using the disc diffusion methods with measurements of minimum inhibitory concentration (MIC) and inhibition zone. For in vivo study, the S. alba extract supplemented diets were administrated at 0.0, 1.0%, 3.0%, and 5.0% on healthy goldfish, Carassius auratus, which challenged with A. invadans zoospores (100 spores/ml). To compare the significant differences in the hematological and immunological parameters obtained from the experiments, the data were analysed using the SPSS. The methanol extract of S. alba effectively inhibited the mycelial growth of A. invadans at a minimum concentration of 1000 ppm for agar and filter paper diffusion experiments. In the agar diffusion test, 500 ppm of the extract inhibited the fungus mycelial growth up to 96 hours after exposure. The mycelial growth from the edge of the pre-inoculated A. invadans agar discs treated with S. alba extracts at concentrations of 100, 500 and 1000 ppm were 15, 8 and 0 mm respectively. The results of the filter paper disc test showed that the S. alba extract at its minimal inhibitory concentration (1000 ppm) has similar qualitative inhibitory effect as malachite green at 1 ppm and formalin at 250 ppm. According to the in vivo tests findings, in the infected fish fed with 3.0% and 5.0% supplementation diet, the numbers of white blood cell and myeloperoxidase activity significantly increased after the second week of treatment. Whilst the numbers of red blood cell significantly decreased in the infected fish fed with 0.0 and 1.0% supplementation diet. After the third week of feeding, significant increases in the total protein, albumin level, lysozyme activity were recorded in the infected fish fed with 3.0% and 5.0% supplementation diet. Also, the enriched diets increased the survival rate as compared to the untreated group that suffered from 90% mortality. The present study indicated that S. alba extract may inhibit the mycelial growth of A. invadans effectively, suggesting an alternative to other chemotherapeutic agents, which brought much environmental and health concerns to the public, for EUS treatment.

Keywords: fungal pathogen, goldfish, organic extract, treatment

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Authors: Okolie Pius Ifeanyi, Emerenini Emilymary Chima

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Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits using Molecular Nested PCR analysis and the efficacy of cell free supernatants from Lactic Acid Bacteria (LAB) isolated from fresh fruits for in vitro control of some tomato pathogens. Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples. The inhibitory potentials of supernatant obtained from LAB isolates of fruits origin that were molecularly characterized were investigated against some tomato phytopathogens using agar-well method with the view to develop biological agents for some tomato disease causing organisms. Methodology: Gram positive, catalase negative strains of LAB were isolated from fresh fruits on Man Rogosa and Sharpe agar (Lab M) using streaking method. Isolates obtained were molecularly characterized by means of genomic DNA extraction kit (Norgen Biotek, Canada) method. Standard methods were used for Nested Polymerase Chain Reaction (PCR) amplification targeting the 16S rRNA gene using universal 16S rRNA gene and LAB specific primers, agarose gel electrophoresis, purification and sequencing of generated Nested PCR products (Macrogen Inc., USA). The partial sequences obtained were identified by blasting in the non-redundant nucleotide database of National Center for Biotechnology Information (NCBI). The antimicrobial activities of characterized LAB against some tomato phytopathogenic bacteria which include (Xanthomonas campestries, Erwinia caratovora, and Pseudomonas syringae) were obtained by using the agar well diffusion method. Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%). The cell free supernatants of LAB from fresh fruits origin (Weissella cibaria, Weissella confusa, Leuconostoc paramensenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Lactobacillus pentosus) can inhibits these bacteria by creating clear zones of inhibition around the wells containing cell free supernatants of the above mentioned strains of lactic acid bacteria. Conclusion: This study shows that potentially LAB can be quickly characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer. Tomato disease causing organisms can be most likely biologically controlled by using extracts from LAB. This finding will reduce the potential hazard from the use of chemical herbicides on plant.

Keywords: nested pcr, molecular characterization, 16s rRNA gene, lactic acid bacteria

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Authors: Nadia Chougui, Nawal Makhloufi, Farouk Rezgui, Elias Benramdane, Carmen S. R. Freire, Carla Vilela, Armando J. D. Silvestre

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Native to Mexico, Opuntia ficus-indica was introduced in southern Spain, and thereafter, it was spread throughout the Mediterranean Basin by the Spanish conquerors in the 16th and 17th centuries. O. ficus-indica is a tropical and subtropical plant able to grow in arid and semi-arid regions, such as the Mediterranean and Central America regions. The culture of Opuntia covers about 200,000 ha in North Africa. This tree is used against soil erosion and desertification for fruit production and is encouraged to promote the livestock sector. It has recently received ever-increasing attention from researchers worldwide for the multivalent pharmaceutical and cosmetical potential of its different compartments (fruits, seeds, cladodes). The present study investigated the elaboration by casting method and characterization of new biodegradable films composed of cladodes powder (CP) of the plant raw material mentioned above, and a marine seaweed derivative, namely agar (A). The effect of glycerol concentration on the properties of the films was evaluated at four different contents (30, 40, 50 and 60 wt.%). The films present UV-blocking properties, thermal stability as well as moderate mechanical performance and water vapor transmission rate (WVTR). The results point to an increase in thickness, elongation at break, moisture content, water solubility, and WVTR with increasing glycerol content. On the contrary, Young’s modulus, tensile strength and contact angle decreased as glycerol concentration increased. The best combination is obtained for the film with 30% glycerol, based on an intermediate compromise between physical, mechanical, thermal and barrier properties. All these outcomes express the potentiality of the powder obtained from grinding the OFI cladodes as raw material to produce low-cost films for the development of sustainable packaging materials.

Keywords: Opuntia ficus-indica cladodes powder, agar, biobased films, effect of plasticizer, sustainable packaging

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Authors: Yasir Bashawri, Vincent N. Chigor James McDonald, Merfyn Williams, Davey Jones, A. Prysor Williams

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Resistance to antibiotics has become a threat to public health. As a result of their misuse and overuse, bacteria have become resistant to many common antibiotics. Βeta lactam (β-lactam) antibiotics are one of the most significant classes of antimicrobials in providing therapeutic benefits for the treatment of bacterial infections in both human and veterinary medicine, for approximately 60% of all antibiotics are used. In particular, some Enterobacteriaceae produce Extend Spectrum Beta Lactamases (ESBLs) that enable them to some break down multi-groups of antibiotics. CTX-M enzymes have rapidly become the most important ESBLs, with increases in mainly CTX-M 15 in many countries during the last decade. Global travel by intercontinental medical ‘tourists’, migrant employees and overseas students could theoretically be a risk factor for spreading antibiotic resistance genes in different parts of the world. Bangor city, North Wales, is subject to sudden demographic changes due to a large proportion (>25%) of the population being students, most of which arrive over a space of days. This makes it a suitable location to study the impacts of large demographic change on the presence of ESBLs. The aim of this study is to monitor the presence of ESBLs in Escherichia coli and faecal coliform bacteria isolated from Bangor wastewater treatment plant, before, during and after the arrival week of students to Bangor University. Over a five-week period, water samples were collected twice a week, from the influent, primary sedimentation tank, aeration tank and the final effluent. Isolation and counts for Escherichia coli and other faecal coliforms were done on selective agar (primary UTI agar). ESBL presence will be confirmed by phenotypic and genotypic methods. Sampling at all points of the tertiary treatment stages will indicate the effectiveness of wastewater treatment in reducing the spread of ESBLs genes. The study will yield valuable information to help tackle a problem which many regard to be the one of the biggest threats to modern-day society.

Keywords: extended spectrum β-lactamase, enterobacteriaceae, international travel, wastewater treatment plant

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Authors: Jesryl B. Paulite, Irene Alcantara-Papa, Teofila O. Zulaybar, Jocelyn T. Zarate, Virgie Ugay

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Banana is one of the major fruits in the Philippines in terms of volume of production and export earnings. The Philippines export of fresh Cavendish banana ranked No.1 with 22% share. One major threat to the industry is Fusarium wilt caused by Fusarium oxysporum f. sp. cubense. It tops as a major concern today affecting the Philippine banana industry since 2002 up to the present in Mindanao. Because of environmental and health issues concerning the use of chemical pesticides in the control of diseases, utilization of microorganisms has been significant in recent years as a promising alternative. This study aims to evaluate the potential of actinomycetes to control Fusarium wilt in Cavendish banana. The in-vitro experiments was carried out in Complete Randomized Design (CRD) while field experiment was laid out in a Randomized Complete Block Design (RCBD) with three treatments and three replications. Actinomycetes were isolated from mangrove soils in areas in Quezon and Bataan, Philippines. A total of 199 actinomycetes were isolated and 82 actinomycetes showed activity against the local Fusarium oxysporum (Foc) by agar plug assay. The test for antagonisms (AQ6, AQ30, and AQ121) of three best isolates Foc to were selected inhibiting Foc by 21.0mm, 22.0mm and 20.5mm, respectively. The same actinomycetes inhibited well Foc Tropical Race 4 showing 24.6 mm, 20.2mm and 19.0 mm zones of inhibition by agar plug assay, respectively. Combinations of the three isolates yielded an inhibition of 13.5 mm by cup cylinder assay. These findings led to the formulation of the mixed actinomycetes as biocontrol agents against Foc. A field experiment to evaluate the formulated mixed actinomycetes against Foc in a Foc infested field in Kinamayan, Sto Tomas, Davao Del Norte, Philippines. was conducted. Results showed that preventive method of application of the mixed actinomycetes against Foc showed promising results. A 56.66% mortality was observed in control set-up (no biocontrol agent added) compared to 33.33% mortality in preventive method. Further validation of the effectiveness of the mixed actinomycetes as biocontrol agent is presently being conducted in Asuncion, Davao Del Norte, Philippines.

Keywords: actinomycetes, biocontrol agents, cavendish banana, Fusarium oxysporum f. sp. cubense

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Authors: Messaouda Belaid, Salima Kebbouche-Gana, Djamila Benaziza

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Our study focuses on determining the antibacterial activity of some honeys from northern Algeria. To test this activity, the agar well diffusion methods was employed. The bacterial strains tested were Staphylococcus aureus, Bacillus subtilis, Streptococcus faecalis, Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeroginosae. The results showed that all the microbes tested were inhibited by all honey used in this study but Those bacteria that appear to be more sensitive to all honey tested are Staphylococcus aureus and Pseudomonas aeroginosae.

Keywords: honey, antibacterial activity, Northern Algeria, Staphylococcus aureus

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Authors: Hyvee Gean Cabuso, Arvin Taruc, Danielle Villanueva, Channela Anais Hipolito, Jia Bianca Alfonso

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Introduction: Denture adhesive aids to provide additional retention, support and comfort for patients with loose dentures, as well as for patients who seek to achieve optimal denture adhesion. But due to its growing popularity, arising oral health issues should be considered, including its possible impact that may alter the microbiological condition of the denture. Changes as such may further resolve to denture-related oral diseases that can affect the day-to-day lives of patients. Purpose: The study aims to assess and compare the microbiological status of dentures without adhesives versus dentures when adhesives were applied. The study also intends to identify the presence of specific microorganisms, their colony concentration and their possible effects on the oral microflora. This study also aims to educate subjects by introducing an alternative denture cleaning method as well as denture and oral health care. Methodology: Edentulous subjects age 50-80 years old, both physically and medically fit, were selected to participate. Before obtaining samples for the study, the alternative cleaning method was introduced by demonstrating a step-by-step cleaning process. Samples were obtained by swabbing the intaglio surface of their upper and lower prosthesis. These swabs were placed in a thioglycollate broth, which served as a transport and enrichment medium. The swabs were then processed through bacterial culture. The colony-forming units (CFUs) were calculated on MacConkey Agar Plate (MAP) and Blood Agar Plate (BAP) in order to identify and assess the microbiological status, including species identification and microbial counting. Result: Upon evaluation and analysis of collected data, the microbiological assessment of the upper dentures with adhesives showed little to no difference compared to dentures without adhesives, but for the lower dentures, (P=0.005), which is less than α = 0.05; therefore, the researchers reject (Ho) and that there is a significant difference between the mean ranks of the lower denture without adhesive to those with, implying that there is a significant decrease in the bacterial count. Conclusion: These results findings may implicate the possibility that the addition of denture adhesives may contribute to the significant decrease of microbial colonization on the dentures.

Keywords: denture, denture adhesive, denture-related, microbiological assessment

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Authors: Admire Isaac Tichafa Shayanowako, Mark Laing, Hussein Shimelis

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Striga asiatica is among the leading abiotic constraints to maize production under small-holder farming communities in southern African. However, confirmed sources of resistance to the parasitic weed are still limited. Conventional breeding programmes have been progressing slowly due to the complex nature of the inheritance of Striga resistance, hence there is a need for more innovative approaches. This study aimed to achieve partial resistance as well as to breed for compatibility with Fusarium oxysporum fsp strigae, a soil fungus that is highly specific in its pathogenicity. The agar gel and paper roll assays in conjunction with a glass house pot trial were done to select genotypes based on their potential to stimulate germination of Striga and to test the efficacy of Fusarium oxysporum as a biocontrol agent. Results from agar gel assays showed a moderate to high potential in the release of Strigalactones among the 33 OPVs. Maximum Striga germination distances from the host root of 1.38 cm and up to 46% germination were observed in most of the populations. Considerable resistance was observed in a landrace ‘8lines’ which had the least Striga germination percentage (19%) with a maximum distance of 0.93 cm compared to the resistant check Z-DPLO-DTC1 that had 23% germination at a distance of 1.4cm. The number of fusarium colony forming units significantly deferred (P < 0.05) amongst the genotypes growing between germination papers. The number of crown roots, length of primary root and fresh weight of shoot and roots were highly correlated with concentration of fusarium macrospore counts. Pot trials showed significant differences between the fusarium coated and the uncoated treatments in terms of plant height, leaf counts, anthesis-silks intervals, Striga counts, Striga damage rating and Striga vigour. Striga emergence counts and Striga flowers were low in fusarium treated pots. Plants in fusarium treated pots had non-significant differences in height with the control treatment. This suggests that foxy 2 reduces the impact of Striga damage severity. Variability within fusarium treated genotypes with respect to traits under evaluation indicates the varying degree of compatibility with the biocontrol.

Keywords: maize, Striga asiaitca, resistance, compatibility, F. oxysporum

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Authors: Kiran Nawaz, Ahmad Ali Shahid, Sehrish Iftikhar, Waheed Anwar, Muhammad Nasir Subhani

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Chili (Solanum annum L.) attacks by many fungal pathogens, including members of Oomycetes which are responsible for root rot in different chili growing areas of the world. Oomycetes pathogens cause economic losses in different regions of the Pakistan. Most of the plant tissues, including roots, crowns, fruit, and leaves, are vulnerable to Phytophthora capsici. It is very difficult to manage the Phytophthora root rot of chili as many commercial varieties are tremendously vulnerable to P. capsici. The causal agent of the disease was isolated on corn meal agar (CMA) and identified on a morphological basis by using available taxonomic keys. The pathogen was also confirmed on the molecular basis through internal transcribed spacer region and with other molecular markers.The Blastn results showed 100% homology with already reported sequences of P. capsici in NCBI database. Most of the farmers have conventionally relied on foliar fungicide applications to control Phytophthora root rot in spite of their incomplete effectiveness. In this study, in vitro plate assay, seed soaking and foliar applications of 6 fungicides were evaluated against root rot of chili. In vitro assay revealed that significant inhibition of linear growth was obtained with Triflumizole at 7.0%, followed by Thiophanate methyl (8.9%), Etridiazole (6.0%), Propamocarb (5.9%) and 7.5% with Mefenoxam and Iprodione for P. capsici. The promising treatments of in vitro plate bioassay were evaluated in pot experiments under controlled conditions in the greenhouse. All fungicides were applied after at 6-day intervals. Results of pot experiment showed that all treatments considerably inhibited the percentage of P. capsici root rot incidence. In addition, application of seed soaking with all six fungicides combined with the foliar spray of the same components showed the significant reduction in root rot incidence. The combine treatments of all fungicides as in vitro bioassay, seed soaking followed by foliar spray is considered non-harmful control methods which have advantages and limitation. Hence, these applications proved effective and harmless for the management of soil-borne plant pathogens.

Keywords: blastn, bioassay, corn meal agar(CMA), oomycetes

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Authors: Isma Benmazouz, Bálint Joszef Nagy, Bence Bálacs, Gábor Kardos, László Kővér

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Vancomycin-resistant enterococci (VRE) are among the major nosocomial threats, which have a potential for zoonotic transmission due to the ubiquity of enterococci in the environment and in animal microbiota, e.g., wild birds. . In order to assess the prevalence in an urbanized bird species, 221 fecal samples were collected from Hooded crows (Corvus cornix) in 2020. Fecal samples were screened using VRE agar plates. None of the samples yielded VRE. The absence of VRE isolates in sampled urban hooded crows indicates that crows residing in the city do not necessarily constitute a reservoir of VREs.

Keywords: resistance, crows, Enterococci, wild birds

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Authors: Shradha S. Binani, Pravin S. Bodke, Ravi V. Joat

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An efficient method has been described for the synthesis of 6-(substituted aryl)-4-(2'- hydroxy-5'-chlorophenyl)-1, 6-dihydropyrimidine-2-thiol, as a beneficial antibacterial and antifungal agents. The diketones of title compounds were synthesized in four steps and subsequently these diketones were further reacted with thiourea in the presence of DMF, which led to the formation of dihydropyrimidine derivatives 5 (a-f). Compounds 5 (a-f) were screened for their in vitro antibacterial and antifungal activity by agar well method. Compounds 5b, 5c, 5e, and 5f were exhibited significant antimicrobial potential against tested strains at 50ug/ml and 100ug/ml concentrations. Six novel dihydropyrimidine analogues have been synthesized, characterized and found to be promising antibacterial and antifungal agents.

Keywords: diketones, dihyropyrimidine, antimicrobial activity, thiol group

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Authors: Fatma Acheuk, Akila Hamichi, Siham Semmar

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The crude ethanolic extract from Solenostemma argel was obtained by maceration of leaves and stems of the plant. Phytochimical study revealed the richness of the species on flavonoids, alkaloids, tannins and glycosides. Antimicrobial activity of the growth of clinical isolates of Eschirichia coli, Pseudomonas aeriginosa, Staphylococus aureus and Bacillus Subtilis was carried out using agar disc diffusion. The results of the study revealed that the test compound has antimicrobial activity against gram-positive bacteria which are resistant to commonly antimicrobial agents used. However, no effect was observed on other species tested.

Keywords: Solenostemma argel, crude extract, phytochemical screening, antimicrobial activity

Procedia PDF Downloads 348

Authors: Hanouf A. S. Al Nuwaysir, Nadine Moubayed, Abir Ben Bacha, Islem Abid

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Consumption of waste water continues to cause significant problems for human health in both developed and developing countries. Many regulations have been implied by different world authorities controlling water quality for the presence of coliforms used as standard indicators of water quality deterioration and historically leading health protection concept. In this study, the European directive for the detection of Escherichia coli, ISO 9308-1, was applied to examine and monitor coliforms in water samples collected from Wadi Hanifa and neighboring wells, Riyadh governorate, kingdom of Saudi Arabia, which is used for irrigation and industrial purposes. Samples were taken from different locations for 8 months consecutively, chlorine concentration ranging from 0.1- 0.4 mg/l, was determined using the DPD FREE CHLORINE HACH kit. Water samples were then analyzed following the ISO protocol which relies on the membrane filtration technique (0.45µm, pore size membrane filter) and a chromogenic medium TTC, a lactose based medium used for the detection and enumeration of total coliforms and E.coli. Data showed that the number of bacterial isolates ranged from 60 to 300 colonies/100ml for well and surface water samples respectively; where higher numbers were attributed to the surface samples. Organisms which apparently ferment lactose on TTC agar plates, appearing as orange colonies, were selected and additionally cultured on EMB and MacConkey agar for a further differentiation among E.coli and coliform bacteria. Two additional biochemical tests (Cytochrome oxidase and indole from tryptophan) were also investigated to detect and differentiate the presence of E.coli from other coliforms, E. coli was identified in an average of 5 to 7colonies among 25 selected colonies.On the other hand, a more rapid, specific and sensitive analytical molecular detection namely single colony PCR was also performed targeting hha gene to sensitively detect E.coli, giving more accurate and time consuming identification of colonies considered presumptively as E.coli. Comparative methodologies, such as ultrafiltration and direct DNA extraction from membrane filters (MoBio, Grermany) were also applied; however, results were not as accurate as the membrane filtration, making it a technique of choice for the detection and enumeration of water coliforms, followed by sufficiently specific enzymatic confirmatory stage.

Keywords: coliform, cytochrome oxidase, hha primer, membrane filtration, single colony PCR

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