Nafisat A. Kachalla   Ja’afaru M. Inuwa   Halima Isa   and Jude F. Chanji   

Clostridium difficile naturally inhabits intestinal tract of animals including humans. Spores of C. difficile are known contaminant in the Environment. Indiscriminate use of antibiotics has led to the emergence of a powerful resistance determinant (known as Metallo resistant genes) in many bacteria worldwide. Multidrug resistant bacteria (MRB) pose threats to health globally. This study aims to determine the distribution of Metallo resistant Clostridium difficile in hospital environment within Yola. A total of 150 surface samples from the hospital wards were collected from Federal Medical Centre, Yola (FMC) and Specialist hospital, Yola (SHY) Adamawa state, Nigeria. Clostridium difficile were recovered by inoculating samples in modified Cycloserine-cefoxitin fructose broth (CCFB) and Cycloserine-cefoxitin fructose agar (CCFA). Screening of isolates for extended spectrum β-lactamase (ESBL) and metallo-β-lactamase enzymes was done by double disc diffusion method and Imipenem-EDTA combined disc test respectively. A total of 18 C. difficile isolates were obtained and found to be extended spectrum β-lactamase (ESBL) positive. 16 (88.89 %) of the Clostridium difficile were found to be metallo-β-lactamase positive. Polymerase chain reaction showed that 11(61.1 %) of the isolates possess bla-IMP metallo resistant gene. This study suggests that multidrug resistant C. difficile with metallo resistant genes are wide spread in hospital settings and there is need for relevant authorities to improve on the sanitization of environment as well as continuous surveillance to hinder C. difficile transmission.

Moses Bali Emmanuel   Liman Mubarak Labaran   Egwim Chidi Evans   Abdulkadir Abubaka   Alex Victoria Ali   and Manya Zabe   

Lipases are enzymes that hydrolyse lipids to produce free fatty acids and glycerol. Fungi were cultured on sabouraud dextrose agar (SDA) plates and identified using microscopic techniques. Screening for lipase producers was carried out on SDA media supplemented with 3 % olive oil at ambient temperature. M. pusillus, M. canis, A. fumigatus, Yeast and T. mentagrophytes were found to produce lipases in different amounts with Yeast and M. canis being the highest producers, they were further characterised for this reason. Lipase production was carried out using submerged fermentation. Both Yeast and M. canis produced lipases maximally at 72 h. Optimum pH and temperature of activity for the lipases from Yeast were determined to be 5 and 35 ℃ respectively whereas, those from M. canis were 6 and 40 ℃ correspondingly. Yeast and M. canis lipases had preference for olive oil than vegetable and palm oils and both enzymes were activated by K⁺, Mg²⁺ and Ca²⁺ and inhibited by Fe³⁺, Hg²⁺ and Co²⁺. The lipase enzyme from Yeast had V_max of 0.0006 U/mL/Sec, K_m of 0.4242 mM and K_cat of 0.0004 S^-1 while that from M. canis had corresponding V_max, K_m and K_cat of 0.0001 U/mL/Sec, 3.2287 mM and 0.0001 S^-1.

Sanjida Sultana   Md. Maniruzzaman Sikder   Md. Sabbir Ahmmed   Aireen Sultana   and Nuhu Alam   

Fruit rot disease of strawberry caused by fungus is most economically important disease in Bangladesh, which reduces the production and quality of strawberry at both pre and post-harvest period. Present study was conducted to identify the causal organism of the fruit rot disease of strawberry through classical and molecular techniques and its eco-friendly control measures. A fungal pathogen causing fruit rot was isolated and identified as Botrytis cinerea through classical fungal taxonomy and molecular characterization based on their internal transcribed spacer (ITS) region of ribosomal DNA (rDNA). Sequence analysis showed that 5.8S of rDNA sequences were identical. The reciprocal homologies of the ITS region sequences ranged from 98 to 100%. Experimental results suggested that studied fungus (MH371474.1) was genetically similar with MF521935.1 Botrytis cinerea. Isolated fungus was evaluated on six different culture media and at five different temperature conditions. The optimum mycelial growth of B. cinerea was found on Richard agar medium at 25 ℃ temperature at 7days post inoculation (dpi), while mycelial growth was drastically reduced the temperature 30 ℃ and above. Three Trichoderma species viz., T. ressei, T. harzianum and T. asperellum were used to assess the antagonistic effect on isolated pathogenic fungus in which the mycelial growth of it was inhibited mostly by T. ressei. Experimental results revealed that Trichoderma ressei was the effective antagonistic fungus against Botrytis cinerea for the biological control.

Chetan Datta Poduri   

There is an increasing need to develop strategies for simultaneous detection of Chikungunya (ChikV), Dengue (DenV) and Zika Virus (ZikV) owing to their shared transmission ecology. Towards this aim, nine reference sequences of ChikV, DenV (1 - 4) and ZikV were aligned using ClustalW Omega Software available at the European Bioinformatics Institute (EBI) website. Internet search engines like Google (Scholar), PubMed, JSTOR, and ProQuest Central were used for literature search. Also, Conserved Domain Database (CDD) (NCBI) and ZikaVR (IMTECH, C'garh) formed constant references. One conserved region, 21 amino acids (aa) in length, common to the three viruses mapping to the Core protein of ChikV (aa 211 - 231); and the NS3 proteins of DenV (1 - 4) (aa 1608 – 1628) and ZikV (aa 1631/35 - 1651/55) has been identified by the current author in this study. This region is the part of a protease in all the three viruses studied. Thus, this conserved region can form one diagnostic tool/probe for simultaneous detection of ChikV, DenV and ZikV. Additionally, as this region is conserved, it may form one therapeutic target.

Okonkwo N. N.   Chukwura E. I.   and Mbachu A. E.   

Crude oil pollution is a global problem because of its hazardous effect to aquatic and terrestrial life, as well as having adverse effect on soil productivity. Bioremediation technology has been established as a successful alternative to physicochemical technique in cleaning up of contaminated site. This study was aimed at determining the bioremediation potentials and emulsification activity of indigenous bacteria isolated from crude oil polluted soil. Soil samples were randomly collected from Water-Smith and Base-Camp refineries in Ohaji-Egbema Local Government Area of Imo State, Nigeria. Bacteria isolation and identification were done using standard methods. The isolates were screened for crude oil biodegradation potential, and the extent of crude oil removal was determined using gravimetric method. Emulsification activity of the isolates was also determined using emulsification index (E₂₄). Data were analyzed using analysis of variance (ANOVA) at P < 0.05. Bacteria isolates such as Arthrobacter sp., Enterobacter sp., Bacillus sp. and Pseudomonas sp. were identified in this study. Pseudomonas sp. achieved the highest extent of crude oil removal within the experimental period. Moreover, highest E₂₄ of 81.80±2.8 and 84.00±4.0 was recorded in the media containing Bacillus sp. and Pseudomonas sp., respectively at 0.75mg/l crude oil concentration. Among the isolates, Pseudomonas sp. has greater potential for effective bioremediation of crude oil polluted soil as well as biosurfactant production due to its higher biodegradation efficiency and emulsification activity.

Vivian C. Onuoha   Ifeoma B. Enweani   and Ogbonnia Ekuma-Okereke   

Opportunistic infection in HIV positive individuals has a high risk of morbidity and mortality as it could be a pointer to systemic infection. Aim: This study was designed to determine the patterns of some pathogenic fungal infections isolated from HIV positive individuals from ART clinics in south eastern Nigeria. Materials and methods: A total of 132 patients (62 males and 70 females) were recruited for this study. Their sputum samples were screened for Tuberculosis (TB) using Ziehl Nelsen staining technique and were also cultured on Sabouaurd dextrose agar with and without chloramphenicol incubated at 25℃ with examination of growth every 3 days for two weeks. Identification of isolates was done using Gram staining methods, chromogenic media and slide culture. Results: 46(34.8%) of the test subjects (132) were co-infected with TB while 94(71.2%) tested positive for fungi infection; 36(60.0%) for HIV-sero positive subjects on ART and 58(80.6%) for those not on ART (P<0.05). The prevalence of fungi isolates from test subjects shows that Candida albicans has the highest frequency of 25(26.6%), followed by Penicillum marneiffei with 18(21.6%), with the least prevalent being Aspergillus flavus and Phialemonium curvatum at 2(2.1%) respectively. In HIV-sero negative indivuals, Candida albicans was of the highest prevalence with 7(53.9%), followed by Candida tropicalis with 3(23.1%) and Aspergillus fumigatus and A. flavus with the least prevalence of 1(7.7%) respectively. Furthermore, there was a positive significant correlation between TB and fungi infection in HIV-sero positive subjects (P<0.05). Conclusion: The high prevalence of fungi infection amongst TB co-infected HIV-sero positive individuals suggests a high degree of drug resistance in said patients thus, could be the mitigating factor behind the increasing morbidity and mortality among TB co-infected HIV positive subjects.

Ndubuisi Gabriel Elemuo   Sunday Sunday Ikiensikimama   Chikwendu Ebenezer Ubani   Evans Chidi Egwim   and Justice Obinna Osuoha   

Lipases are hydrolytic enzymes responsible for the hydrolysis of triacylglycerol into glycerol and free fatty acids. The biodiversity of crude oil contaminated soil was explored for the isolation of novel potent lipase producing microorganisms. In this current study, microorganisms isolated from crude oil contaminated soils were screened for lipase activity and expression. Six fungal strains namely: Yeast, Aspergillus flavus, Aspergillus niger, Verticillus sp., Penicillum sp., and Microsporum audouini demonstrated lipase producing potentials and the best two: Verticillus sp. and Penicillum sp. were selected for enzyme production and characterization. The lipase enzyme was produced in broth medium. Optimization of selected biochemical parameters reveal that optimum temperature for lipase activity was 40℃, pH 7 with an incubation time of 168 h for lipase expressed by Verticillus sp. and 40℃, pH 8 with an incubation time of 144 h was established for lipase expressed by Penicillium sp. The kinetic parameters of the lipase enzyme revealed that the enzymes had low Km (0.19 and 0.30 respectively) for the selected substrate.

Elif Tozlu   Nasibe Tekiner   Göksel Tozlu   Recep Kotan   and Hatice Öğütçü   

Drosophila suzukii (Matsumura, 1931) (Diptera: Drosophilidae) is an invasive species originating from Southeastern Asia and spreads in a fast manner. It is among major threats in soft-shell fruit cultivation in the whole world. It was detected in 2014 in Turkey. According to international criteria, it is considered that it has the potential of threatening the fruit cultivation in Turkey where garden plants are grown widely. In this study, a total of 39 bacterial strains were isolated from 100 mature Drosophila suzukii individuals. Gram staining characteristics, catalase, oxidase and nitrate reductase activities and chitinase enzyme activities and hypersensitivity reaction of these strains were determined by using microscopical and visual inspection. The bacterial strains were identified according to their fatty acid methyl esters (FAME) analysis by using Sherlock Microbial Identification System (MIS). The identification test results of the bacterial strains were also confirmed by phylogenetic analysis and their closely related species based on the 16S rRNA sequence. The most abundant bacterial species were Paenibacillus alvei (31.57%) and Bacillus amyloliquefaciens (47.36%) according to the MIS and 16S rRNA sequence analysis results, respectively. According to the MIS results, a total of 6 strains identified as Paenibacillus alvei were identified as Bacillus amyloliquefaciens according to the 16S rRNA sequence analysis results. A total of three Paenibacillus macerans strains identified in MIS system were also identified as Bacillus amyloliquefaciens according to the 16S rRNA sequence analysis. Morphological and biochemical characteristics results of all of Bacillus amyloliquefaciens strains showed the some results. According to the 16S rRNA sequence analysis results, the other bacterial strains consist of 1 Bacillus atrophaeus (5.2%), 1 Bacillus safensis (5.2%), 1 Paenibacillus motobuensis (5.2%) and 1 Staphylococcus epidermidis (5.2%) strains. To our knowledge, this is the first study characterizing the bacterial communities of Drosophila suzukii.

Urška Rozman   Božena Kotnik Kevorkijan   Maja Rupnik   and Sonja Šostar Turk   

Hospital environment provides an important ecological niche that can serve as a reservoir for potentially pathogenic microorganisms. Hospital textiles together with moisture and heat create the right conditions for growth, dissemination and long-term survival of many microorganisms, which may results in transmission of nosocomial infections. Therefore, understanding of total microbial population on hospital textiles is an important part for assessment of public health risks. In the present work, we tested the suitability of DHPLC for estimation of total microbial population on hospital textiles. All of the identified bacteria belonged to the phyla Actinobacteria, Firmicutes and Proteobacteria and up to 63 bacterial genera/species were identified. Mostly abundant species important in perspective of HCAI were Acinetobacter spp., Corynebacterium spp., Staphylococcus spp., Sphingomonas mucosissima and Stenotrophomonas maltophilia. Microbial diversity on hospital textiles was extensive and partly coincides with the intensive care units bacterial communities and pathogens obtained from clinical samples. This is the first study describing total microbial population complexity on hospital textiles, which are important factors contributing to the hospital microbiome.

Anwuli U. Osadebe   Chinelo A. Onyiliogwu   and Gideon C. Okpokwasili   

The biodegradability of anionic surfactants in oilfield detergents was investigated using water samples from the New Calabar River and Tap water. The detergents analyzed were D.D-Y, D.G-X, SUR-500, SW-1000 and Bio-Boost. The anionic surfactant, Sodium dodecyl sulphate (SDS) was employed as a reference. The total heterotrophic counts in the water samples were determined using the spread plate technique while the detergent-utilizers were investigated using Mineral salt agar. Biodegradation was evaluated by monitoring Methylene Blue Active Substance (MBAS) using the "River die-away" method. The percentage MBAS remaining, after the 28 day study period, in increasing order was SDS < SUR-500 < SW-1000 < Bio-boost < D.D-Y < D.G-X; SDS was the most biodegraded. SUR-500, SW-1000 and Bio-boost supported the highest growth of utilizers. The detergent-utilizing bacteria isolated belonged to the genera Proteus, Acinetobacter, Enterobacter, Staphylococcus, Bacillus, Pseudomonas, Arthrobacter, Corynebacterium and Micrococcus. A steady increase in total viable count, pH and optical density was observed during biodegradation. The detergents tested could be considered biodegradable as they had an average of 91% - 97% primary biodegradation at the end of the 28 day test period which is within the biodegradability standards set by the Soap and Detergent Association (SDA) and the Organisation for Economic Co-operation and Development (OECD).

Marwa Mostafa Shalaby   Kareman Ahmed Eshra   Radwa Abd El moteleb Essa   and Ghada Mahmoud Alghazaly   

Background: Central venous catheters (CVCs) are often used as a vascular access in patients with end stage renal diseases when emergency hemodialysis is required, before maturation of arteriovenous fistula or graft, or when a permanent access becomes non-functioning. The most common complication following insertion of a CVC is infections including exit site infection, tunnel infection, or central line related blood stream infections (CLABSIs). Preventing such complications is crucial in these vulnerable patients and this can be accomplished by strict adherence to infection control guidelines. This study aimed to evaluate the efficacy of implementation of a CVC care bundle composed of best evidence based practices to see whether the rates of CLABSIs rates would decrease. Methods: the study was divided into pre and post intervention phases. The duration of each phase was 6 months during which rates of confirmed CLABSIs per 1000 catheter days as well as the causative microorganisms were recorded. The data were then compared and analyzed to evaluate the intervention. Results: CLABSIs rates decreased from 6.7 in phase 1 to 4.1 in phase 2. The relative risk reduction was 0.39. Conclusions: Implementation of bundles for insertion and maintenance of CVCs can help preventing CRBSIs which is very important to improve patient care.

Reazul Karim   Mohammad Nuruddin Mahmud   and M. A. Hakim   

Antibiotic resistance is a serious problem of present world and development of viable alternative is urgent. The research work was designed to mitigate this problem. Different types of bacterial colony were isolated from skin of 30 healthy human and their antipathogenic activity was tested against 9 pathogens. The isolate showed activity against four pathogens- Klebsiella. pneumoniae subsp. pneumoniae, Klebsiella. pneumoniae subsp. ozaenae, Staphylococcus. aureus and Pseudomonas. aeruginosa was identified as Staphylococcus. warneri. Variation was found in optimization of cultural conditions (incubation period, incubation temperature and pH) for the most potent antipathogenic metabolites production. Staphylococcus. warneri showed most potent antipathogenic activity at pH 6 to pH 9, at an incubation period of 24h to 48h and at an incubation temperature of 37℃. Antipathogenic metabolites were then detected as bacteriocin like substances. Variation was observed with bacteriocin activity against different pathogens and was found most effective against Klebsiella. pneumonia subsp. pneumonia and Klebsiella. pneumonia subsp. ozaenae Samples containing bacteriocin like substances showed heat stability up to 80℃ for 60 minutes and up to pH 8 for Klebsiella. pneumoniae subsp. pneumoniae and Klebsiella. Pneumoniae sub sp. ozaenae. Papain treated cell-free supernatant did not show any bacteriocin activity, suggesting that the substances could be antimicrobial peptides. Solvent extraction of bacteriocin was performed by using chloroform where maximum bacteriocin activity was found in interface layer rather than aqueous and organic layer.

Anulika Mercy Kanu   Agwu Chukwuemeka Okorie   Chidiebere Uche   and Ugochi Nwoke Awa   

The present situation of drug resistance in the world necessitates the need to intensify researches on the use of alternative strategies for effective control of infections using products obtained from plant extracts. Hence this study was undertaken to evaluate the phytochemical and antimicrobial properties of ethanoic extracts of Passiflora edulis var. flavicarpa seed by testing them against three clinically important pathogens namely: Staphylococcus aureus, Escherichia coli and Candida albicans. Agar diffusion method was used in determining the susceptibility pattern of the different seed extracts concentrations (25%, 50%, 75% and 100%) on the test organisms (Escherichia coli, Staphylococcus aureus and Candida albicans). The Passiflora edulis var. flavicarpa seed extract was subjected to phytochemical screening for presence of bioactive compounds using standard procedures. From the results, the zone of inhibition of the ethanoic extract at varying concentrations (100%, 75%, 50% and 25%) on Staphylococcus aureus ranged from 6mm to 18mm, Escherichia coli 5mm to 11mm and Candida albicans 5mm to 18mm. The growth of all the organisms was inhibited, through to varying degrees with Staphylococcus aureus and Candida albicans being more susceptible than Escherichia coli. The phytochemical screening revealed the presence of tannins, flavonoids, saponins, alkaloids, steroid, glycosides, saponin glycoside, terpenes and cyanogenic glycosides. The antimicrobial activity of this extract is possibly linked to the presence of flavonoids, alkaloids, steroid, saponins and/or tannins. The result obtained in the study is an indication that passion seed has the potential to be used as a source for new broad spectrum antimicrobials.

Anuja Sharma   Neeraj K. Aggarwal   and AnitaYadav   

Lignin is a major noncarbohydrate polyphenolic structural constituent of cell wall of all the vascular plants and is present in association with cellulose and hemicellulose. It is the second most abundant polymer after cellulose. Lignin is well known for its resistance to microbial degradation because of its high molecular weight and various biologically stable carbon-carbon and ether linkages. Seventy two fungal isolates capable of utilizing lignocellulosic biomass were isolated from different ecological samples. When subjected to primary screening by qualitative plate assay method using methyl orange, azure B, guaiacol and tannic acid as indicators for determining the lignolytic potential, 22 isolates were found positive for at least one of the indicators. Six isolates were found to exhibit all the three extracellular enzyme activities on quantitative estimation of laccase, manganese peroxidase and lignin peroxidase activities. Highest laccase activity was produced by ANF238 (3.42U/ml) while highest manganese peroxidase and lignin peroxidase activities were produced by ANF 212 (0.254U/ml) and ANF238 (0.22U/ml) respectively.

S. Saravanan   Abdul Nasar Kalanthoden   R. Karthikayan   M. Kavitha   and S. Kutti Rani   

In fermentation technology, strain improvement of baker's yeast has traditionally relied on random mutagenesis followed by screening for mutant exhibiting enhanced properties of interest. Such mutant organisms are useful in several industries. Saccharomyces cerevisiae can use sucrose as the sole source of both carbon and energy; hydrolysis of this sugar is catalyzed by the enzyme invertase. The main objective of this work is to overcome the glucose repression of invertase by invertase constitutive mutants through UV mutation. This may occur in any glucose repressible genes as a single or double mutation in repressor gene (s) which might cause constitutive synthesis of invertase. These mutated screened strains were optimized with various glucose concentration and different incubation hours for higher invertase production. The maximum synthesis of invertase was 1.066 units/ml in hour from mutant Saccharomyces cerevisiae type-2 strain.

S. Muthukrishnan   

In optimization of cellulase production by different pH, substrates, nitrogen, carbon sources were detected from penicillium citrinum micro-fungal species. The maximum cellulase activity was recorded in the range of pH- 5 to 6.5 and wheat bran (91 U/ml) and rice bran (81 U/ml) is the best raw substrate for the production of cellulase. Estimation of extracellular protein was recorded in different production medium of Penicillium citrinum (1.63 mg/ml) for maltose followed by sucrose (1.46 mg/ml), fructose (1.13 mg/ml), wheat bran (1.12 mg/ml), peptone (0.88 mg/ml) and yeast extract (0.82 mg/ml). The highest extracellular protein was recorded in pH 5.5 to 6.5 followed by substrate for wheat bran (1.12 mg/ml), rice bran (1.04 mg/ml), paddy straw (1.03 mg/ml). In nitrogen sources, ammonium sulphate (141 U/ml) and ammonium nitrate (121 U/ml) is the best nitrogen source for the production of cellulase enzyme. The maximum cellulase activity was recorded in glucose (134 U/ml) and maltose (159 U/ml) was best carbon source for the production of cellulase.

Demeke Geremew   Meseret Workineh   and Maryam Izad   

Narcolepsy is a chronic neurological disorder characterized by excessive day time sleepiness, cataplexy and hypnagogic and hypnopompic hallucination. Narcolepsy is caused by loss of hypocretine neuron. Hypocretine is an excitatory neuropeptide projecting in the central nervous system and maintains wakefulness. The mechanism of hypocretin loss in narcolepsy is elusive. However, an autoimmune mediated destruction of hypocretin neuron is suspected because of its firm association with HLA-DQB1*0602 genotype and polymorphism in the TCR-α chain. Furthermore, infection with streptococcus pyogenes, influenza A H1N1 strain and the Pandemrix vaccine itself has strong association with narcolepsy. We searched PubMed (1984 - 2015), Google Scholar (1984 - 2015), MEDLINE (1984 - 2015), and EMBASE (1984 - 2015). In the interim, bibliographies of identified articles were reviewed to find additional references. The purpose of this mini review is to summarize recent findings involving microbiological and immunological factors for narcolepsy development.

Thiruvengadam Shankar   and Thangavel Sivakumar   

Soil samples were collected from the different locations in Sathuragiri Hills (Western Ghats, Tamilnadu). The Total Fungal Counts in the soil samples of the different locations were analyzed. The percentage frequencies of occurrence of fungal isolates are identified. The fungal isolates were identified based on the morphological and cultural characteristics they were identified as Cladosporium sp, Aspergillus niger, Penicillium sp, Alternaria sp, Curvularia sp, Helminthosporium sp and Aspergillus flavus. Among the isolates maximum amount of citric acid levels was produced by Aspergillus niger (2.2 ± 0.001 g/l). The effect of different pH, temperature, carbon source, nitrogen source, incubation time and agricultural residues on citric acid production by Aspergillus niger was analysed. The maximum amount of citric acid production was recorded in pH - 6.0 (2.32 ± 0.09 g/l), temperature - 30℃ (2.85 ± 0.06 g/l), carbon source - Glucose (6.08 ± 0.10 g/l). Nitrogen source - ammonium chloride (10.22 ± 0.05g/l), incubation time - 72 hours (13.56 ± 0.04 g/l) and agricultural residue - corn ears (17.2 ± 0.14 g/l).

B. C. Behera   S. K. Singh   M. Patra   R. R. Mishra   B. K.Sethi   S. K. Dutta   and H. N. Thatoi   

As knowledge of sulphur oxidising bacterial communities in mangrove sediments is very sparse and sulphur oxidizers have wide application such as in treatment of wastewater containing high sulphate levels which causes unwanted H₂S production by sulfate reducing bacteria (SRB), oxidation of elemental sulphur to plant available sulphate for plant growth promotion, bio leaching, biocontroling agent etc. Hence, in the present study an attempt has been made to explore the diversity of sulphur oxidizing bacteria from mangroves of Mahanadi delta, Odisha to evaluate their biotechnological potential. Two sulphur oxidising bacteria (SOB-7and SOB-8) were isolated from mangrove soils of Mahanadi river delta, based on the change of colour of the thiosulphate broth medium from purple to colour less by reducing the pH. Based on morphological, biochemical and 16S rRNA gene sequencing the two strains (SOB-7) and (SOB-8) were identified as Klebsiella sp., and Micrococcus sp. respectively. The gene bank accession number of the strains are KR632644 (SOB-7) and KR632643 (SOB-8). The strain Klebsiella sp. and Micrococcus sp showed sulphate ion production ability of 243 mg/ml and 240 mg/ml respectively with decrease in pH from 7.0 to 4.0. Among these two isolates, SOB-8 showed higher sulphide oxidase production ability (126.83 U/ml) than the isolate, SOB-7 (126.0 U/ml). Maximum sulphide oxidase by both the strains were obtained at temperature 45℃, peptone as nitrogen source, thiosulphate concentration of 10 mg/ml. pH 9.0 for the isolate SOB-8 whereas pH 7.0 for the isolate, SOB-7. Partially purified enzyme of both the isolate showed higher activity at same substrate concentration of 1.5mg/ml, but varies with pH and temperature.

Wesam Hatem Amer   and Marwa Mohamed Ezzat Abd-Elmonsef   

This study was designed to compare the activity of piperacillin/tazobactam (TZP) in combination with either netilmicin (NET) or aztreonam (ATM) against metallo-β lactamases (MβL) Pseudomonas aeruginosa. Out of 46 nosocomial Ps. aeruginosa isolates, 38 (82.6%) isolates were carbapenem-resistant by disc diffusion susceptibility testing. These were then screened by imipenem + EDTA combined disc test for MβL production. Thirty (78.9%) MβL-producing isolates detected were further tested for antibiotic synergy against NET-TZP and ATM-TZP combinations by Etest synergy and Etest/agar dilution tests. Neither combination showed antagonism. The synergistic effect of NET-TZP combination was detected in [27 (90%) and 28 (93.3%), respectively] and ATM-TZP combination in [24 (80%) and 25 (83.3%), respectively] of the tested isolates by Etest synergy and Etest/agar dilution tests, respectively. Thus, we have determined the synergistic effect of these two combinations for treating MβL-producing Ps. aeruginosa. Significance and Impact of the Study: The present study has shown valuable information on the combination treatment of Ps. aeruginosa and recommends their use for treatment of resistant Ps. aeruginosa.

Awah Nsikak Sunday   Agu Kingsley Chukwuebuka   Muokwe Juliet   Irondi Chidiebere   Okeke Chidi Benjamin   Anaukwu Chikodili Gladys   Archibong Etim Joseph   Iloanusi Confidence Ada   Ngenegbo Uche   and Umeoduagu Nnamdi   

Five brands yoghurts designated A, B, C, D and E were obtained and used for assessing their microbial quality. A ten-fold serial dilution was carried out and appropriate dilution used to inoculate duplicate plates of SDA, Eosin methylene blue, MacConkey and Nutreint agars. Discrete colonies that developed were purified and stored on agar slant at 4℃. The pH of the samples ranged from 5.6 to 6.0. In the total coliform count, sample A recorded no count while others had count from 2.0 to 5.0 x 10²cfu/ml. The total viable bacterial and fungal counts were 6.0 to 18.0 x 10⁴ and 1.0 to 6.0 x 10³cfc/ml, respectively. Five fungal genera were obtained as Aspergillus, Penicillium, Fusarium, Mucor, Geotricum, Neurospora and Absidia. Mucor species occurred in all the samples while Fusarium species occurred only in sample B. Among the samples, yoghurt sample B harbored a greater percentage of the fungal isolates. Samples C and D had the least number of fungal of three (3) each. The bacterial isolates obtained include species of Staphylococcus, Lactobacillus, Escherichia, Bacillus, Pseudomonas and Klebsiella. Sample B also recorded the highest number of bacterial isolates. The genera Bacillus occurred in all the yoghurt samples whereas Lactobacillus and E.coli occurred only in two yoghurt samples each.

Adindu Chukwuemeka Steve   Odili Lucy Chidinma   Nwagu Ezinne Emmanuella   Agu Kingsley Chukwuebuka   Awah Nsikak Sunday   Okeke Benjamin Chidi   and Orjionwe Rita Nchedo   

Phytochemical and antimicrobial analysis were carried out on the leaves, stem and bark extracts of Cola gigantea. The Phytochemicals present in the plant were Flavonoid, Tannins, Alkaloid, Cardiac glycoside, Steroids, Terpenoids and phenol. Flavonoid, Alkaloid and steroids were present in high amount in the plant than the other phytochemicals. For antimicrobial screening, the well-in-agar diffusion method was employed and the various zones of inhibition produced by the extract of the various plant parts on Escherichia coli, Bacillus subtilis, Staphylococcus aureus and Candida albicans were observed and recorded in millimeters (mm). The leaves of the plant had highest antimicrobial activities on Escherichia coli, Bacillus subtilis and Staphylococcus aureus, while the stem had highest antimicrobial activity on Candida albicans. The observed antimicrobial activity of the extract suggests its potential use in the treatment of diseases produced by the various test pathogenic microorganisms.

Chetan Datta Poduri   

This article discusses the popular Aristotelian phrase "natura non facitsaltus" using two possible approaches – a gradual evolution and a punctuated evolution. In doing so this communication shows that in whatever aspect we examine Nature, at a microscopic level we find that continuity exists. Perhaps the same occurs at macromolecular/multi-cellular levels. Nevertheless, in sexually reproducing organisms or in organisms which exhibit recombination, some amount of jumping occurs which cannot be considered random. Despite this, on a broader scale, nature seems to evolve continuously and gradually.

Jude Ogechukwu Okoye   Beatrice Omowumi Joel   and Kenneth Onyenze   

This study explored use of urine cytology as an early first-line approach in the laboratory diagnosis of dysplasia associated with urinary tract infection. Fifty (50) patients attending the obstetrics and gynaecology clinic at Madonna University Teaching Hospital and 50 apparently healthy subjects were recruited for the study. Urine samples were cultured and analyzed using the dipstick and Giemsa methods. The result of this study revealed significant higher number of nitrite positive samples, bacteria and pus cells when female test subjects were compared with apparently healthy female subjects (p<0.05). The frequency of Staphylococcus aureus, Escherichia coli, Streptococcus faecalis and C. albicans identified is 50.6, 27.3, 10.4 and 9.1 %, respectively while other isolated Coliforms amounted to 2.6%. Microscopy revealed marked cytolyses, desquamation and abnormal nuclear/cytoplasmic ratio. S. aureus and E. coli induced higher exfoliation and morphological change (72.7%) compared to other isolates. In conclusion, this study suggests that chronic bacterial infection could lead to accelerated desquamation of cells and urothelial carcinoma. More so, it revealed that females are at a higher risk of developing epithelial neoplasia following UTI when compared to the males. Thus, urine smears could be used as first line approach in the early diagnosis of urothelial malignancy.

Mbachu A.E   Chukwura E.I   and Mbachu, N.A   

Fungi capable of effectively degrading and cleaning up hydrocarbons was isolated from soil samples contaminated with used engine oil at auto-mechanic workshops (at Mgbuka-Nkpor, Nigeria) using vapour phase transfer method. The ability of the potential isolates to utilize used engine oil, diesel and petrol were assessed using gravimetric method. The ability of both the pure and consortium culture of the best potential strains to degrade the polycyclic aromatic hydrocarbons (PAHs) component of used engine oil, diesel and petrol was assessed using Gas Chromatography. A total of 8 fungal isolates were identified in this study based on their cultural and microscopic characteristics. Of these, 4 that showed high promise for hydrocarbon bioremediation potentials in screen flasks were confirmed as Candida tropicalis, Rhodosporidium toruloids, Fusarium oxysporium and Aspergillus clavatus based on their 18S rRNA gene sequencing. High biodegradation efficiency (> 70%) was recorded in the PAHs component in used engine oil, diesel and petrol with both the pure and consortium culture of the best potential strains; C. tropicalis and A. clavatus, within 16 days of incubation at 28℃. However, there was complete (100%) depletion of some PAHs such as anthracene, naphthalene, acenaphthalene, acenaphthylene, phenanthrene and benzo (k) fluoranthene) in the hydrocarbon substrates with the pure and consortium culture of the isolates within 16 days of incubation at 28℃. Both the pure and consortium culture of the isolates (Candida tropicalis and Aspergillus clavatus) could therefore be utilized in the bioremediation of used engine oil, diesel and petroleum, as well as PAHs contaminated soil.

Ester Prados   and Sergi Maicas   

The dependence of plastic materials is an increasing problem. Although plastics are very useful for humankind many disadvantages derived from the difficulties linked to recycling and disposal are well known. A feasible alternative is the production and use of bioplastics. These compounds have multiple options at the end of his life that can ensure their safety and efficacy of reuse or recovery. For example, raw materials can be returned to the manufacturer for recycling. Bioplastics synthesized through biotechnology include mainly polyhydroxyalkanoates (PHAs), common lipoidic storage materials accumulated by prokaryotes. Some processes for producing PHAs by fermentation using microorganisms have been developed at a different extent. However, biopolymers (PHA) market is under development, and therefore cannot compete with traditional plastics since manufacturing is still more expensive. In this review we have focused on the study of the production processes of bioplastics where bacteria are present, also describing the scaling to industrial level aspects and future trends.

Thiruvengadam Shankar   Thangavel Sivakumar   Chandrahasan Satya   and Ponirul Ponmanickam   

Seaweed sample was collected from tide pools and rock surfaces in theKanyakumari coastal region. The bacterial isolate was identified based on the morphological and cultural characteristics. The effect of different pH on alginase production by Bacillus sp. is at pH 8.0 (10.9± 0.34 IU/ml). Among the various temperatures tested, maximum alginase production was recorded in 40°C (15.9 ± 0.39 IU/ml). The effect of metal ions tested at calcium chloride (32.41 ± 0.47 IU/ml). Sodium alginate matrix was increased gradually up to 24hrs (0.53±0.012 IU/ml). Agar-Agar matrix was better steadily up to 24 hrs (0.6± 0.017 IU/ml).Gelatin matrix was increased gradually up to 24 hrs (0.37±0.10 IU/ml)There are four variables used for the factorial analysis including pH, temperature, alginate and ammonium chloride namely X1, X2, X3 and X4 respectively. There was a considerable variation in the alginase production depending on the four chosen variables. The maximum alginase production of 36.4 IU/ml was achieved by RSM (CCD).

Mojtaba Moradi Golrokhi   Hafez Fakheri   Mohammad Reza Haghshenas   and Mohammad Ahanjan   

Background: Helicobacter pylori (H. pylori), which infects about half of the world's population, is the main factor to peptic and duodenal ulcer diseases. Resistance to antibiotics is the main reason of failure of H. pylori treatment. Objective: Since the antibiotic resistance patterns of H. pylori is varied geographically; therefore, the main purpose of this study was to determine the antibacterial effects on H. pylori infection. Material and methods: The cross-sectional study was conducted on 120 patients (51 male, 69 female) with upper Gastrointestinal symptoms which they had referred to endoscopy department. Demographic characteristics of patients were recorded prior to sampling and the situations of resistance to 6 commonly used antibiotics in treating of H. pylori infection were studied. Results: In this study, 30 isolates were obtained from the culture of gastric biopsy specimens of 120 patients. The resistance rate of H. pylori to Metronidazole, Clarithromycin, Tetracycline, Amoxicillin and Levofloxacin were 63.3%, 16.6%, 6.6%, 10% and 3.3%, respectively. There was no resistance rate of H. pylori to Furazolidone during observation. Conclusion: According to antibiotic resistance level of this study, it seems that the antibiotic susceptibility tests are always necessary in order to determine the appropriate drug regimen against to H. pylori.

Onuorah Samuel   Nwankwo Florence   and Obika Ifeanyi   

The microbial quality of commercial bottled water purchased from Eke-Awka, Ifite-Awka, Amaudo-Awka, Permanent Site-Awka, Amaenyi-Awka, Agu-Awka and Okpuno-Awka Markets, Nigeria were assessed for total bacteria, total fungi, total coliforms and faecal coliforms. The temperature and PH values were also determined. The total bacteria ranged between 100 and 350 cfu/ml, total fungi between 10 and 50 cfu/ml, total coliforms from 0 to 30 cfu/100ml, faecal coliforms from 0 to 20 cfu/100ml, temperature from 27℃ to 29℃, and PH from 5.7 to 6.3. The microbial isolates from the samples were Pseudomonas aeruginosa, Enterobacter aerogenes, Escherichia coli, Bacillus subtilis, Klebsiella pneumoniae, Penicillium digitatum and Aspergillus niger. Coliforms were not detected from the samples from Eke-Awka and Permanent Site-Awka Markets. The microorganisms were predominant in the samples from Okpuno-Awka Market (26%) while they had the least distribution in the samples from Eke-Awka Market (4%). The predominant bacteria were the gram-negative rods. Escherichia coli was the dominant bacterium (20%) while Penicillium digitatum was the dominant fungus (20%). Most of the samples did not comply with the standard set by regulatory bodies for drinking water and are therefore unsafe for drinking without adequate treatment.

Onuorah Samuel   and Obika Ifeanyi   

Used Toothbrushes serve as reservoirs for microorganisms and may play a major role in disease transmission in humans. Forty used manual toothbrushes obtained from some students of Nnamdi Azikiwe University Awka, Nigeria were analyzed bacteriologically using standard microbiological techniques. The toothbrushes were used twice per day for one month by the students. The result showed that all the toothbrushes were contaminated with bacteria while the unused ones which served as controls were not contaminated. The bacterial isolates from the toothbrushes were Streptococcus mutans, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Enterobacter aurogenes. Staphylococcus aureus was predominantly isolated (30%) from the toothbrushes, followed by Pseudomonas aeruginosa (22%), Staphylococcus epidermidis (18%), Streptococcus mutans (16%), Escherichia coli (8%) and finally Enterobacter aerogenes (6%). Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis, Streptococcus mutans, Escharichia coli and Enterobacter aerogenes were detected in 60%, 45%, 50%, 25%, 20% and 10% of the samples respectively. In view of the fact that these organisms are pathogenic and could be a potential health hazard, adequate rinsing and air-drying of the toothbrushes before storage will minimize the incidence of these bacteria and the health risk associated with them.

Kanu Anulika Mercy   Ihekwumere Ijeoma   and Kalu Emmanuel Jonah   

A study of Trichomonas vaginalis infection among 200 pregnant women attending antenatal clinics in three different public and private health facilities in Aba was carried out between August to October, 2014. High Vaginal Swabs (HVS) were randomly collected from each of the subjects and samples were analyzed using microscopy method. The presence of Trichomonas vaginalis was detected by its characteristic jerking movement in a wet preparation. Of the 200 pregnant women studied, 26 (13%) of the women were infected with T. vaginalis. Women within the age of 19 – 26 years had the highest prevalence rate of 17.95% followed by those within age group 27 – 34 years (10.64%). However, the differences were not significant (X2= 3.002, P > 0.05). Women in the third trimester had the highest prevalence of T. vaginalis infection (14.8%) and there was no significant difference between trichomoniasis and duration of pregnancy (X2= 0.434, P > 0.05). Trichomoniasis was more common in primigravidae (23.80%) than in mulitigravidae (10.13%) (X2= 5.49, P < 0.05). With respect to occupation students were the most affected (22.22%) (X2= 10. 659, P >0.05). The prevalence rate of 13% obtained in this study is high therefore clinician should routinely screen all pregnant women for the infection.

Ifediegwu, M.C.   Agu, K.C.   Awah, N.S.   Mbachu, A.E.   Okeke, C.B.   Anaukwu, C.G.   Uba, P.O.   Ngenegbo, U.C.   and Nwankwo, C.M.   

The extensive use of pesticides is one of the major causes of pollution of soil and water environments. The current method for removing such contaminants from the environment through biodegradation has been shown to be more effective than any other method. Three pesticide degrading bacteria were isolated and identified through cultural and biochemical tests as strains of Pseudomonas aeruginosa, Serretia marcescens and Klebsiella oxytoca. Their growth in mineral salt medium supplemented with 20mg/l of Chlorpyrifos was monitored at optical density of 600nm. The result showed that Pseudomonas aeruginosa had maximum growth in ten days, while Serretia marcescens and Klebsiella oxytoca recorded highest growth after six days of incubation. HPLC analysis of the residual Chlorpyrifos after 14 days incubation showed that Pseudomonas aeruginosa was able to degrade 60% of the pesticide; Klebsiella oxytoca degraded 54%, while Serretia marcescens had 53% reduction of the pesticide concentration in the mineral salt medium. The results of this research indicated that the isolated bacteria can be used for bioremediation of Chlorpyrifos contaminated soil and water ecosystems.

Okeke, B.C   Agu, K.C.   Uba, P.O.   Awah, N.S.   Anaukwu C.G.   Archibong, E.J.   Uwanta, L.I.   Ezeneche, J.N.   Ezenwa, C.U.   and Orji, M.U.   

Mixed fruit wine (pineapple and watermelon) was produced using Saccharomyces cerevisiae isolated from palm wine. Primary and secondary fermentation of the fruits lasted for 7 and 28 days respectively, during which aliquot samples analysis of pH, titrable acidity, specific gravity, alcohol content and reducing sugar were carried out using standard procedures. Specific gravity of the wine was observed to reduce drastically as the fermentation progresses. The pH of the fruit must during the period of fermentation ranged from 3.0 to 4.46. During the fermentation period, consistent increase in alcohol content was observed with time. At the end of the 28th day of fermentation, the alcohol content was observed to be 3.2%. The titrable acidity of the wine was observed to show steady trend with time throughout the period of fermentation. This study showed that acceptable wine can be produced from mixed fruits pineapple and watermelon using yeasts especially Saccharomyces cerevisiae isolated from palm wine.

Abdul Matin, W Islam   and O Ali Mandal   

The anitbacterial potency screening of the crude extracts of leaves, seed and stem wood of Adenanthera pavonina L. were tasted against 15 Gram-positive and Gram-negative bacteria along with a standard antibiotic, Ciprofloxacin. The in vitro screening for antimicrobial potency was carried out by following the agar diffusion and micro broth dilution techniques. Among the fifteen bacterial strains tested the chloroform fraction of the seed extract showed promising antibacterial efficacy against Salmonella-β-haemolyticus, Serratia dysenteriae and Escherarichia coli. The susceptibility order of extracts were seed>leaf>stem. The MIC values of the chloroform seed extract were determined 256µg/ml againstB. cerus; 128 µg/ml against S.-β–haemolyticus, S. dysenteriaeand 64 µg/ml against Klebsiella sp. The MIC values of the chloroform extracts of stem wood were128µg/ml S.-β–haemolyticus; 64 µg/ml against B. megaterium, S. sonnei and 32 µg/mlagainst S. typhi.

Onuorah Samuel   Obika Ifeanyi   Orji Michael   and Odibo Frederick   

The primary function of aviation fuel is to provide propulsive energy to the aircraft. Microorganisms may contribute to aging instability in the fuel, induce corrosion of the storage tanks and pipeworks, form mats which can block filters and pipelines and increase wear in fuel pumps eventually leading to engine failure, therefore, in this study, the microbial contaminants in the commercial aviation fuel obtained from Benin City Airport, Nigeria were isolated, characterized, identified and their hydrocarbon-utilizing potentials determined using standard methods. The microbial contaminants were Micrococcus luteus, Serratia marcescens, Pseudomonas aeruginosa, Bacillus subtilis, Citrobacter freundii, Klebsiella aerogenes, Acinetobacter calcoaceticus, Fusarium oxysporum, Penicillium italicum, Rhizopus stolonifer, Aspergillus fumigatus and Candida tropicalis. Serratia marcescens had the highest percentage distribution (16%) in the fuel. Serratia marcescens, Pseudomonas aeruginosa, Acinetobacter calcoaceticus, Rhizopus stolonifer and Aspergillus fumigatus were the best utilizers of the fuel among the organisms isolated. There was a drop in the initial pH of the mineral salts oil medium inoculated with the isolates indicating hydrocarbon utilization. The result of this work showed that the fuel examined was contaminated with bacteria and fungi, therefore the incorporation of effective biocides, frequent inspection and drainage of water from fuel tanks and pipeworks are recommended.

Onuorah Samuel  Obika Ifeanyi  and Orji Michael  

Tuberculosis (TB) and human immunodeficiency virus (HIV) co-infection is a major challenge in tuberculosis prevention and control. The prevalence of HIV in tuberculosis patients aged between 10 and 59 years receiving medical care at General Hospital Onitsha, Nigeria was investigated. Seventy-five of the patients were males while eight-five were females. Their sputum and blood samples were screened for tuberculosis and HIV. The sputum samples were examined by microscopy and culture. Patients between 20 and 29 years of age had the highest number of tuberculosis cases (30.7%) while those aged between 50 and 59 years had the least number of cases (9.4%). Forty-five males (28.1%) and fifty females (31.2%) tested positive to HIV respectively. The prevalence of HIV among the tuberculosis patients was 59.3%. Twenty-eight patients (29.5%) composed of twelve males (12.6%) and sixteen females (16.9%) between 20 and 29 years had TB/HIV while eleven patients (11.6%) comprising six males (6.3%) and five females (5.3%) between 50 and 59 years were co-infected. Acid-fast bacilli were detected by culture in 92 (57.5%) of the tuberculosis patients while they were detected by microscopy in 68 (42.3%) of the patients indicating the higher sensitivity of culture for tuberculosis case detection.

Sergi Maicas   and Jose Juan Mateo   

The fermentation of grape must to produce wine is a biologically complex process, carried on by yeasts and malolactic bacteria. The yeasts present in spontaneous fermentation may be divided into two groups, the Saccharomyces yeasts, particularly S. cerevisiae, and the non-Saccharomyces yeasts which include members of the genera Rhodotorula, Pichia, Candida, Debaryomyces, Metschtnikowia, Hansenula and Hanseniaspora. S. cerevisiae yeasts are able to convert sugar into ethanol and CO₂ via fermentation. They have been used for thousands of years by mankind for the production of fermented beverages and foods, including wine. Their enzymes provide interesting wine organoleptic characteristics. β-Glucosidase activity is involved in the release of terpenes to wine, thus contributing to varietal aroma. β-Xylosidase enzyme is also interesting in industry due to its involvement in the degradation of hemicellulose by hydrolyzing its main heteroglycan (xylan). The ability of yeasts to release proteases has been observed by many researchers because of their potential to degrade haze proteins in wine and to generate nutrient sources for microorganisms. Moreover, these enzymes are interesting in biotechnology, for use in food processing such as cheese, pickles or sausage.

Onuorah Samuel   and Orji M. U.   

Tomato contains large amount of water which makes it more susceptible to spoilage by fungi. These fungi that spoil tomatoes produce mycotoxins that are detrimental to human health. This study was therefore carried out to isolate, characterize and identify the fungi associated with the spoilage of tomato fruits sold in Okpuno, Amaudo, Eke- Awka, Nodu and Amaikwo Markets in Awka, Nigeria. The average fungal counts ranged between 1.3 x 10³ and 2.0 x 10³ cfu/ml, while the fungal isolates were Aspergillus niger, Rhizopus stolonifer, Fusarium oxysporum, Saccharomyces cerevisiae, Alternaria alternata, Penicillium digitatum and Geotrichum candidum. The percentage occurrence of the isolates in the fruits from Eke Awka was the highest (32.73%) while that of the fruits from Nodu market was the least (12.73%). Aspergillus niger had the highest percentage occurrence (47.27%) in the fruits studied, with Saccharomyces cerevisiae and Geotrichum candidum having the lowest percentage occurrence (3.64%). Pathogenicity test result showed that Aspergillus niger also had the highest decay diameter of 30mm in the healthy tomato fruits while Geotrichum candidum had the lowest decay diameter. Proper handling and adequate storage facilities must therefore be employed to prolong the shelf life of tomato fruits.

Tarun Kumar   Anuradha Sourirajan   and Kamal Dev   

Cold adapted and extracellular β-D-galactosidase (EC 3.2.1.23) with high specific activity has potential in food industry. Two psychrophlic bacterial isolates (A5-2 and B8) were screened from soil collected from permanent glaciers of Himachal Pradesh, India. Both A5-2 and B8 isolates showed growth between 4-25℃, but not at temperature higher than 30℃, hence classified as psychrotolerants. Biochemical characteristics and 16S rDNA sequencing identified the isolates as Serratia quinivorans A5-2 and Serratia quinivorans B8, and deposited in NCBI GenBank under accesion numbers KJ 176660 and KJ 176661, respectively. The cold active, extracellular β-D-galactosidase activity of A5-2 isolate was three fold higher compared to its intracellular activity. Comparatively, the B8 bacterial showed negligible intracellular activity, and its extracellular activity was two folds higher as compared to that of A5-2 isolate. Interestingly, growth and β-D-galactosidase activity of A5-2 was enhanced in lactose supplemented medium;whereas, growth of B8 isolate was unaffected and its β-D-galactosidase activity was enhanced when grown in lactose supplemented medium. β-D-galactosidase activity was also increased, when the isolates were grown in galactose supplementation medium, but decreased when grown in the presence of glucose. The presence of milk sugars like lactose, glucose, or galactose, or the milk metal ions, namely Ca²⁺ and Na⁺ ions did not inhibit the activity of β-D-galactosidase.

Abdel-Naser A. Zohri   Nanis H. Gomah   and Maysa A. Ali   

This study was aimed to investigate the feasibility of bio- ethanol production by batch fermentation of kareish cheese whey. Two forms of whey; untreated (crude) whey containing 5% lactose and treated whey (deproteinized and concentrated to 14% lactose) were utilized. Fermentation processes were performed by two strains of Kluyveromyces marxianus and four strains of Saccharomyces cerevisiae which were previously recognized as ethanol- producing strains. Effects of different initial pH values, as well as, external supplementation of treated whey by four different nitrogen sources on the rate of ethanol production by two of the highest producing strains were also investigated. All the studied yeast strains were able to grow and produce ethanol from both crude and treated whey. Levels of ethanol production ranged between 3.4- 18.5g/l and 24.11-57.66 g/l from crude and treated whey, respectively. The most suitable initial pH maximizing ethanol yield was 5.5 and the best added nitrogen source was yeast extract. Maximum ethanol levels produced by K. marxianus ZMS3GU133329 and S.cerevisiae EC1118 from treated whey adjusted to pH 5.5 and supplemented by 0.3% yeast extract reached to 69.85 and 65.36 g/L, corresponding to 97.8 and 91.4% of the theoretical values, respectively. The kinetic parameters and productivity were calculated and discussed for all experiments.

Nevzat Ünal   Adil Karadağ   Keramettin Yanık   Kemal Bilgin   Murat Günaydın   and Asuman Birinci   

Fungi are the factors causing nosocomial infections but they are not considered in the first hand still, their importance is gradually increased as they are frequently detected microorganisms. We aimed to determine in vitro efficiency of electrolyzed water on different molds and yeasts. Materials and Methods-In School of Medicine Hospital of Ondokuz Mayıs University, Samsun, Turkey, the effect of electrolyzed water produced in Envirolyte (Envirolyte Industries International Ltd., Estonia) device on the yeast fungi and mold fungi in different concentrations and contact period isolated from clinical samples with qualitative suspension test method. As media Sabouraud dextrose agar, and as Dey-Engley Neutralizing broth (Sigma-Aldrich, USA) were used. Results-Envirolyte electrolyzed water was detected to be effective against all yeasts used in the study as well as Candida albicans, Candida tropicalis, Candida parapsilosis, Candida glabrata, Candida krusei, Candida lusitaniae,Trichosporon spp. and mold; Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger isolates in the concentrations of 1/1, 1/2, 1/5, 1/10 in 1 minute and other testing durations.Based upon the results we acquired, we believe that Envirolyte electrolyzed water is a cheap and easy to obtain and natural disinfectant for controlling the nosocomial infections.

Sara Marchi   Ilaria Giovanelli   Raffaele Gargiulo   Daniela Nozzi   Agostino Barozzi   Mario Sarti   Ramona Iseppi   Immacolata Anacarso   and Carla Sabia   

The dissemination of carbapenemase-producing Klebsiella pneumoniae (KPC) strains is an increasing problem worldwide. KPC ß-lactamases are Ambler class A enzymes mostly plasmid-encoded; their global spread represents a threat to clinical patients care and public health. Between September-December 2012, have been isolated 60 Klebsiella pneumoniae strains that showed a decrease of susceptibility to carbapenems, collected from patients at the Provincial Laboratory of Clinical Microbiology ‘S. Agostino-Estense' Hospital (Modena, Italy). Species identification and antimicrobial susceptibility testing were carried out using the Vitek 2 and studied for carbapenemases production using phenotypic confirmatory tests. The presence of carbapenemase genes were confirmed by PCR and sequencing. The localization of beta-lactamase genes was established by conjugation experiments. We reported the emergence of KPC-2-producing Klebsiella pneumoniae isolates PCR and sequencing experiments revealed the presence of blaKPC-2 gene in 56/60 isolates. The KPC-encoding plasmid was analysed by transconjugation experiments and DNA sequencing. The findings indicate the need to supplement surveillance systems based on susceptibility data with the surveillance of resistance mechanisms.

Fatthy Mohamed Morsy   Nivien Allam Nafady   Mohamed Hemida Abd-Alla   and Deia Abd Elhady   

Studying of nanoparticle structures is gaining momentum because of their great potential in improving several fields of science such as agriculture. The water soluble fraction of the extracellular polysaccharides (EPS)/matrix of the highly EPS producing cyanobacterium Nostoc commune have been used as a potent reducing and capping agent for green synthesis of silver nanoparticles. The size of these nanoparticles with the EPS coat was found to be in the range of 15–54 nm as analyzed using transmission electron micrographs. Interestingly, after washing the EPS coated silver nanoparticles by ethanol, the size of nanoparticles reduced to less than 15 nm due to the formation of silver oxide nanoparticles and removal of the EPS coat. Silver nanoparticles showed antibacterial properties against Escherichia coli. The minimum inhibitory concentration (MIC) was 0.012 mg/ml while the minimum bactericidal concentration (MBC) was 0.016 mg/ml. The slight difference between the MIC and MBC suggests that such silver nanoparticles act as a potent bactericidal agent against E. coli. Presoaking seeds of crop plants (Sorghum and broad bean) in five-fold MBC of silver nanoparticles (0.08 mg/ml) did not adversely affect the germination of Vicia faba L. and Sorghum bicolor plants. Concomitantly, such fivefold MBC concentration of silver nanoparticles was powerful sterilizing agent for seeds and grains against seed/grain-borne microorganisms. The results showed gradual depletion of the total colony forming units (CFU) in seeds and grains sterilized with silver nanoparticles than those sterilized with chlorine. These results suggest that the water soluble fraction of the extracellular polysaccharides (EPS)/matrix of Nostoc commune can be used as a potent reducing and capping agent for green synthesis of silver nanoparticles and that silver nanoparticles can be used as a potent surface sterilizing agent of seed crops against phytopathogenic fungi.

Tripti Yadav   

Bacterial diseases are contagious and can result in many serious or life-threatening complications, such as blood poisoning (bacteremia), kidney failure, and toxic shock syndrome. In diagnosing bacterial infections, the rapid identification of bacteremia at an early stage of the disease is critical for a favorable outcome. Furthermore, it is important that exact information be obtained on the stage of the disease rapidly in order to choose and initiate the appropriate therapy. In recent years many new techniques have been added in the diagnostic tools. The observations were based on a survey carried out in population of Bassi village near Jaipur. It was found that many people were prone to bacterial infections due to unhygienic conditions and improper sanitation. The patients affected which died were basically unconcerned, careless and ignorant. Many patients got hospitalization and some were prescribed oral antibacterials at home. The data showed that maximum number of patients had Salmonellosis, followed by Botulism. 6 Bacterial diseases were common in the human population of Bassi village, near Jaipur. Knowledge of the disease outbreaks further leads to treatment techniques and prevention of deaths in the population. Different types of antibiotics were effective for treating specific types of bacteria. Antibiotics may be given orally, intravenously, or by intramuscular injection, depending on the type and severity of bacterial disease in patients. General types of antibiotics included-Aminoglycosides, Cephalosporins, Macrolides, Quinolones, Penicillins and tetracyclines.

N.S. Vardanyan   and A.K. Vardanyan   

Sulphur-oxidizing bacteria strain SO-1 and strain SO-2 have been isolated respectively from leaching pulp of zinc and copper concentrates. The cells of isolated bacteria are rods with the size of 0.3-0.5 x 0.7-2.0 mkm. They are capable of oxidizing elemental sulphur and tetrathionate with optimal growth temperature of 35℃ and pH 2.7-3.0. The strains are considered to be facultative chemolithoautotrophic. Analysis of 16S rRNA gene sequences has shown that the isolated strain SO-1 forms a single cluster with corresponding sequences of a type strain of Acidithiobacillus thiooxidans ATCC 19 377 having 97.5% similarity with them. Strain SO-2 of sulfur-oxidizing bacteria isolated from leaching pulp of copper concentrate shows a high level of similarity with corresponding sequences of the type strain of Acidithiobacillus albertensis DSM-14366. We come to the conclusion that the strain SO-1 participating in the oxidation of zinc concentrate can represent a new species of the genus Acidithiobacillus distinct from A.thiooxidans and the strain SO-2 is a new strain of A.albertens.

Yakovleva L. M.   and Ogorodnyk L.E.   

After 15 or more years after accident at the Chernobyl nuclear power plant there was isolated a number of new for Ukraine phytopathogenic bacteria. All of them are characterized by a very high aggressiveness, which is preserved at the storage of bacteria in the laboratory. Among these bacteria are called: Erwinia amylovora, Curtobacterium flaccumfaciens and Curtobacterium sp. – previously missing in Ukraine, strongly aggressive Pantoea agglomerans – previously deemed to epiphyte, Pantoea agglomerans - not described in literature and a new form or, perhaps, a new species with some similarity of properties Clavibacter michiganensis subsp. michiganensis. In article there were presented some features of symptoms of bacterial diseases, caused by these agents.

Luis J. del Valle   Michael L. Jaramillo   Miguel Talledo   Maria J. Pons   Lidia Flores   Ruth L. Quispe   Pablo Ramírez   Ruth García-de-la-Guarda   Débora Alvarado   Abraham Espinoza-Culupú   Juana del Valle   Martha Vargas   and Joaquim Ruíz   

We designed a 16S rRNA gene PCR-RFLP scheme to identify all currently described Bartonella spp. The 16S rRNA genes of all Bartonella spp. were in-silico analyzed in order to design a RFLP technique able to discriminate among different species. The restriction enzymes selected were MaeIII, MseI, Sau96I, BsaAI, DrdI, FokI, BssHII, BstUI, AluI, TspDTI and HphI which, according to a decision-making tree, facilitated the differentiation of all the currently described species of Bartonella.The technique was experimentally tested in different species of Bartonella, including human pathogenic B. bacilliformis and B. henselae with a 100% of concordance with the in-silico predicted patterns.This novel RFLP assay could be used to identify both human and non-human pathogenic Bartonella in diagnostic, phylogenetic and epidemiologic studies.

Obiukwu C.E.   and Otokunefor T.V.   

The microbiological quality of refinery effluent, and water and sediments of the effluent receiving Okrika sector of bonny estuary was investigated. Treated waste water (TWW) recorded the highest total heterotrophic bacteria (THB) of 1.6 X 109 cfu/ml, while raw waste water had the highest bacterial diversity of 24.82%. The THB population of the water body was lowest [8.6X103cfu/ml] at the effluent outfall site (SS1) in January compared to the highest [1.10X108cfu/ml] recorded at the control station (SS6). The sediment samples were richer in microbial population density with highest value of 8.6X109cfu/g and least value of 7.8X104cfu/g recorded at SS6 and SS1 respectively. While fungi population showed significant upstream - downstream variation in sediment, the population of fungi in the water samples was relatively stable (3 logs) throughout the study period. The most predominant bacterial genera in the refinery effluent include Bacillus sp (23.37%), Pseudomonas sp (12.41%), and Staphylococcus sp. (10.59%). Similarly, Bacillus sp (15.73% & 16.77%) and Pseudomonas sp (13.72% & 11.44%) were the most prevalent in the river water and sediment respectively. Penicillum sp, Fusarium sp dominated the fungi population. The effluent outfall site (SS1) recorded the least generic diversity of 13.16% and 13.92% both for water and sediment samples respectively. Pollution of the Bonny estuary by refinery effluent resulted in changes in the microbial population density and disappearance of organisms like Chromobacter species, Vibro species, E. coli and Serratia species from the affected sites.

HasinaKarki, Ahmed Mustafa   and ArlisLaMaster, Shree Dhawale   

Experiments were conducted to investigate the presence of antibiotic resistant bacteria in hatchery-reared fish that are important in commercial aquaculture. Two fish species, tilapia (Oreochromisniloticus) andcohosalmon(Oncorhynchuskisutch)were obtained from certified hatcheries to investigate ifantibiotic resistant bacteria are present in their guts. The antibiotic resistant bacteria were isolated on the basis of their resistance to ampicillin. Six out of 8 tilapia and 3 out of 8 salmon yielded antibiotic resistant isolates. These results indicated prevalence of antibiotic resistant bacteria in guts of both fish species. Five bacterial isolates were taken from each of these nine fish that yielded ampicillin resistant bacteria (45 bacterial isolates in total) to test antibiotic sensitivity using six additional antibiotics: penicillin, vancomycin, chloramphenicol, tetracycline, streptomycin, and gentamicin. In addition to ampicillin, all isolates showed resistance to penicillin and sensitivity to gentamicin.Overall, ten different resistance phenotypes were observed and many isolates displayed multidrug resistance.

Maddela Naga Raju   and Kadiyala Venkateswarlu   

A thiadiazine compound, buprofezin, and an organophosphorous insecticide, acephate, singly, were assessed for their nontarget effects on activities of proteases, urease and acid phosphatase in NPK (nitrogen, phosphorous and potassium)-fertilizer amended and unamended cotton soils. The studied enzyme activities were adversely affected by the insecticides above 5 or 7.5 µg g⁻¹ concentrations. Additionally, activities were declined after three applications of insecticides. Therefore, the results of the present study clearly indicate that the soil application of buprofezin or acephate at higher rates and repeated applications greatly affected the activities of studied enzymes in unamended or NPK-amended soils.

Mohamed Hemida. Abd-Alla   El-Sayed. A. El-Sayed   and Abdel-Hamied. M. Rasmey   

Out of 210 actinomycetes isolates belonged to the genus Streptomyces recovered from rhizospheric soil of Triticum vulgaris, Zea mays and Vicia faba cultivated in Assuit and New Valley Governorates of Egypt, ten isolates were capable of producing high amounts of L-glutaminase enzyme. The most potent L-glutaminase producer isolate was Streptomyces sp. ASU319 recovered from rhizosphere of Triticum vulgaris. The higher producer actinomycete isolate was identified by sequencing of 16S rRNA as Streptomyces variabilis ASU319 and was deposited in the GenBank nucleotide sequence database under accession number KC145278. Factors affecting L-glutaminase production by the Streptomyces variabilis ASU319(KC145278) were examined and the results revealed that the maximum L-glutaminase value was obtained when the isolate cultivated in the production broth medium supplemented by glutamine 8 mg/ml, adjusted at pH 4 and incubated at 35℃. These results suggest that L-glutaminase-producing Streptomyces variabilis ASU319 could be used as a plant growth promoting rhizobacteria by increasing the ammonia content in the rhizosphere soil across degradation of the agricultural wastes. Also this isolate can be used in both pharmaceutical and food industrial application for L-glutaminase production on commercial scale.

A.K. Vardanyan Vardanyan N.S. and Markosyan L.M. 

Leptospirillum sp. str. ZC isolated from bioleaching pulp of zinc concentrate is capable of oxidizing Fe (II) with the optimal temperature of 37℃. In pure culture bacteria is unable to oxidize pyrite, however, its growth together with sulfur oxidizing A.albertensis str.SO^-2 significantly enhances the efficiency of pyrite bioleaching. The correlation between pyrite biooxidation and adhesion of Leptospirillum spp. bacteria on mineral surface has been found. It has been revealed that in case of mixed culture the cells of Leptospirillum sp. str. ZC are adhered on pyrite surface only after its initial colonization by sulfur oxidizing bacteria. It is supposed that bioleaching of pyrite by Leptospirillum spp. bacteria occurs by means of indirect contact leaching.

Tanushree Barua Malini Shariff and S.S. Thukral 

The aim of the study was to determine the prevalence of AmpC producing E. coli, Klebsiellapneumoniae and Klebsiellaoxytoca and toevaluate and compare four different phenotypic methods viz; Modified Three Dimensional Test, AmpC Disk Test, AmpC Disk Test with Tris-EDTA and Inhibitor based detection test for the detection of AmpCβ-lactamases in these isolates, collected during a period of 2003 to 2009 from various hospitals of Delhi, India. Characterization of these enzymes was carried out by polymerase chain reaction (PCR) as well as by nucleotide sequencing. Phenotypic results showed that Inhibitor based detection test, Modified three dimensional test and AmpC disk test detected 40.8% of E. coli and 34% of Klebsiella spp. to be AmpC producers. However, Inhibitor based detection test was found to be simple, highly specific and sensitive in detecting these enzyme producers. PCR and nucleotide sequencing showed the presence of blaCMY-2 and blaACT-1 genes in all the positive isolates.

Monika Bhardwaj Bhoj Raj Singh Sandeep Kumar and Abhijit M Pawde 

In the study nasal and throat swabs were collected from 78 apparently healthy and 69 cases of kennel cough from dogs of different age, sex, breed and places and analysed for presence of Bordetella bronchiseptica through bacterial isolation and multiplex PCR (mPCR). The pathogen could be isolated from an apparently healthy Labrador bitch while with mPCR, 7 (4.76%, 4 from healthy dogs and 3 from sick dogs) samples were positive for genus specific amplicon and three (2.04%, one from healthy and two from sick Labrador bitches) were positive for B. bronchiseptica species specific amplicons. The kennel cough had no significant association (p, 0.49) with the detection of B. bronchiseptica but sex (p, 0.03) appeared to be the significant predisposing factors associated with the infection.

Immacolata Anacarso Carla Condò Carla Sabia Patrizia Messi Simona de Niederhausern Moreno Bondi and Ramona Iseppi 

We investigated the presence of antibiotic-resistances and others important virulence factors in coagulase negative and positive staphylococci, isolated from several sources (food, environmental and clinical). Thirty-four staphylococci were isolated and identified (28 coagulase-positive were identified as Staphylococcus aureus, 6 remaining Coagulase Negative Staphylococci were identified as 3 S. epidermidis, 2 S. warneri, 1 S. hycus). All the strains were analyzed both with phenotypic and genotypic assays in order to evaluate the virulence factors and the susceptibility against antibiotics and to correlate them to the sources of isolation. Results showed that isn't a direct correlation between features researched and isolation, and the different virulence factors are as widespread as antibiotic-resistance. The isolation from food of a strain of Staphylococcus aureus with a high methicillin resistance was very interesting. 15 of the 34 strains were found to possess multi-drug resistance and in 7 cases with Minimum Inhibitory Concentration(MIC) really very high; also in this case there is no distinction between the coagulase-positive and coagulase-negative staphylococci and their origin of isolation.