MOSP - Macao Scientific Publishers

Research Article

ISSR for comparison of cross-inoculation potential of Colletotrichum capsici causing chilli anthracnose

Kanchalika Ratanacherdchai*, Hong-Kai Wang, Fu-Cheng Lin and Kasem Soytong

Abstract: Thirty-four isolates of Colletotrichum spp. including 2 species, C. gloeosporioides and C. capsici, from anthracnose on Bell pepper, Long cayenne pepper and Bird’s eye chilli were isolated and their pathogenicity was proven via fruit inoculation. Pathogenicity tests divided pathogenic potential into low, medium and high virulence groups. It is clearly revealed that C. capsici from the three tested hosts expressed the highest virulent isolates. Cross-inoculation of three high virulent isolates of C. capsici in accordance with three chilli varieties showed that all isolates could produce anthracnose symptom in the same lesions. All tested isolates developed lesions after co-inoculation of all hosts. Inter simple sequence repeat (ISSR) analysis indicated that there are two distinct groups of C. gloeosporioides and C. capsici. Furthermore, genetic diversity was correlated with geographic distribution, while there was no clear relationship between genetic diversity and pathogenic variability. But it is clearly demonstrated that whereas C. gloeosporioides appears in the same geographic area as C. capsici, it causes lower disease incidence.[...] Read More.

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2016, 15(1), 1-6;

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Research Article

Predominant lactic acid bacteria associated with the traditional malting of sorghum grains

H. Sawadogo-Lingani*, B. Diawara, R. K. Glover, K. Tano-Debrah, A. S. Traoré and M. Jakobsen

Abstract: The traditional processes of sorghum grains malting were investigated in Tamale (Northern Ghana) from the raw materials through to the sun-dried malted sorghum including steeping and germination stages. Samples were taken at each processing stage and cultured for the isolation of LAB. The isolates were characterized by basic phenotyping, ITS-PCR / RFLP analysis and identified by partial sequencing of 16S r RNA genes. Their antimicrobial and amylolytic activities and exopolysaccharides production were also investigated. During steeping, the LAB counts in the steep water increased from 105 to 109 - 1010 cfu/ml and pH of the steep water decreased from 5.08 ± 0.22 to 4.20 ± 0.50. A total of 106 isolates were identified and the predominant isolates belonged to the species Lactobacillus fermentum (58.49%), Pediococcus acidilactici (22.64%), Weissella confusa (11.32%), Enteroccocus faecium, Pediococcus pentosaseus and Lactococcus lactis. Some isolates showed interesting antimicrobial activity (39.62%), slight amylolytic activity (37.73%) and ability to produce exopolysaccharides (90.5%). L. fermentum isolates dominated the microbiota from sorghum grains to malted sorghum. These isolates had technological properties comparable to those responsible for the acidification of sorghum beer (dolo, pito) wort produced from sorghum malt (previously studied), suggesting their potential for use as starter cultures. Suitable isolates of L. fermentum are promising candidates to be used as starter cultures from the initial step of malting, that is, the steeping and are expected to inhibit the growth and survival of pathogens and spoilage microflora, and to control the lactic fermentation of dolo and pito wort or other sorghum malt-based products like infant formulations.[...] Read More.

Keywords: Sorghum, sorghum malt, lactic acid bacteria, identification, technological properties.

2016, 15(1), 1-6;

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Research Article

Initial microbial colonization in the alimentary tract of a new â€“ born baby in different modes of parturition

I. Joseph and A. J. A. Ranjit Singh*

Abstract: The intestinal microflora is a positive health asset that crucially influences the normal structural and functional development of the mucosal immune system. In the new born, the colonization of microbes in the alimentary system starts as soon as the womb comes out. A study on the early colonized bacterial species in a new - born will help to understand the immuno protective functions in the new-born. The initial colonization of bacterial population in the alimentary tract of a new-born baby varied according to the mode of child birth. In the alimentary tract of the normally delivered baby, beneficial probiotic microbial invasion in the intestinal region is high and such microbes are mostly from the mother. In a surgically (Caesarian) delivered baby, the alimentary tract receives mostly non beneficial microbes from the environment and hence such babies are susceptible to infection.[...] Read More.

Keywords: Lactobacillus, new-born, probiotic, immune system, nosocomial infection, Caesarian, normal child birth.

2016, 15(1), 1-6;

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Research Article

Nutritional factors affecting the antifungal activity of Penicillium steckii of mangrove origin

Jyotsnamayee Sabat and Nibha Gupta*

Abstract: A fungal strain namely, Penicillium steckii was obtained from saline environment of mangrove plant Avicennia marina and evaluated for antifungal activity in PDA against dieback pathogen of rose. Amendments at the level of different carbon and nitrogen source and their concentrations, inorganic and organic salts resultant into the development of new and modified medium of basal PDA under which our fungi exhibited enhanced level of antifungal activity. Achievement of this medium composition may be useful further in process development to elaborate the antagonistic active principle against plant pathogens.[...] Read More.

Keywords: Mangrove, avicennia, antifungal, penicillium, dieback, nutrient.

2016, 15(1), 1-6;

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Research Article

Prevalence of antibodies to infectious bronchitis virus (IBV) in chickens in southwestern Nigeria

B. O. Emikpe*, O. G. Ohore, M. Olujonwo and S. O. Akpavie

Abstract: A total of 672 sera from apparently healthy commercial and indigenous chickens of different ages were screened for antibodies to infectious bronchitis (IB) virus using the enzyme-linked immunosorbent assay (ELISA). Samples obtained from breeder, layer, grower and indigenous chicken flocks in Oyo, Ogun, Ondo and Lagos states of southwestern Nigeria were screened. The total seroprevalence was 82.7% with ELISA units of 77.0 ± 8.0. Among the groups of birds, layers had the highest ELISA units of 80.0 ± 9.0, breeders had 73.0 ± 8.0 while indigenous chickens had 73.0 ± 7.0 ELISA units. These chickens, except the breeders, had no history of vaccination against infectious bronchitis. The higher prevalence observed in layers, growers and indigenous chickens may be due to field infection since maternal antibody was expected to have waned between three and four weeks of life. These findings indicate a high infectious bronchitis virus activity in southwestern Nigeria chickens hence there is an urgent need for the development of prevention and control policies against IB in Nigerian poultry farms and a national control programme for infectious bronchitis virus infection should be planned.[...] Read More.

Keywords: Chickens, infectious bronchitis virus, Nigeria, prevalence, southwest.

2016, 15(1), 1-6;

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Research Article

Bacterial contamination of crockery and cutlery within the kiosksâ€™ restaurants of the Federal University of Technology, Yola

Lynn Maori and Nandita De*

Abstract: An investigation was undertaken to determine bacteriological quality of crockery and cutlery of seven restaurants in Federal University of Technology, Yola (FUTY) kiosk. Samples (147) were collected and cultured in appropriate media and the bacterial isolates were identified according to their morphological and biochemical characteristics. The result revealed a profile of seven (7) different bacterial species including Staphylococcus aureus, Proteus vulgaris, Salmonella typhi, Escherichia coli and the species of Shigella, Klebseilla and Bacillus. The total values of bacterial count (TBC per ml) of the samples were in the range of 1.1 x 104 - 3.0 x 105 for cups, 2.2 x 104 - 1.6 x 105 for forks, 1.0 x 104 - 3.3 x 105 for knives, 1.2 x 104 – 2 .5 x 105 for plates and 1.5 x 104 - 4.7 x 105 for spoons cfu/ml. So, it may be concluded that in order to prevent food-borne infection, food inspectors should monitor the hygienic status of the restaurants within FUTY kiosk.[...] Read More.

Keywords: Salmonella, Shigella, Escherichia coli, Bacillus, Staphylococcus, Proteus vulgaris, Klebseilla.

2016, 15(1), 1-6;

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Research Article

Extracellular leucine aminopeptidase produced by Aspergillus oryzae LL1 and LL2

Shie-Jea Lin, Li-Lin Chen, Chiou-Yen Wen and Wen-Shen Chu*

Abstract: In a screen of 175 fungal strains, Aspergillus oryzae LL1 and LL2 were identified as having the highest extracellular leucine aminopeptidase (LAP) activity. LAP activity was optimal when A. oryzae LL1 was subjected to submerged fermentation with an inoculum size of 105 spores per ml and an agitation of 100 rpm at 30°C in media containing defatted soybean and rice husk with a pH of 4.5 for 72 h. Partial characterization of A. oryzae LL1 LAP revealed that it could be greatly inhibited by 2 mmol L-1 of Pb2+, Cu2+, or Fe2+. In contrast, 2 mmol L-1 Zn2+ stimulated LAP activity about one-fold compared to adding 30 mmol L-1 zinc increased DH to 33.3% for LAP LL1 during a piolot plant scale experiment hydrolysis of chopped chicken breast meat (Lin et al., 2008; Eur Food Res. Technol., 2008) . To determine the stability of A. oryzae LL1 LAP, the samples were stored at -80, -20, 4 and 25°C for eight weeks with residual activities being reduced to 84, 72, 61 and 58%, respectively. Taken together, our data suggest that A. oryzae, LL1 LAP has tremendous potential for use in the food industry.[...] Read More.

Keywords: leucine aminopeptidase, Aspergillus oryzae, culture conditions, fermentation, Zn2+.

2016, 15(1), 1-6;

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Research Article

Association of IGF-1 gene polymorphism with milk production traits and paternal genetic trends in Iranian Holstein bulls

Yousef Mehmannavaz*, Cyrus Amirinia, Mortaza Bonyadi and Rasoul Vaez Torshizi

Abstract: The effects of Insulin-Like Growth Factor-1 polymorphism were examined on milk production traits and genetic trends in the Iranian Holstein bulls. A total of 282 bulls were genotyped for SnaBI restriction site in the 5' flanking region of IGF-1 by applying PCR-RFLP method. The allele frequencies for C and T alleles were 0.562 and 0.438, respectively. The results indicated that genetic variants at the 5'-noncoding region of the bovine IGF-1 gene had a marked effect on estimated breeding values of milk (EBVM) and fat yields (EBVF). The heterozygous bulls (CT genotype) had higher EBVM and EBVF than homozygous bulls (P < 0.1) . The average substitution effects of alleles were not significant for none of studied traits (P > 0.1). The studied polymorphism of IGF-1 showed no significant association with genetic trends of milk related traits (P > 0.1). These results suggest that allelic interaction in IGF-1 polymorphism as over dominance and might be effectiveness in animal improving by crossbreeding.[...] Read More.

Keywords: IGF-1, polymorphism, Holstein, genetic trend.

2016, 15(1), 1-6;

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Research Article

Periodic changes in Prosopis cineraria associated AM population at different soil depth and its relationship with organic carbon and soil moisture

Neelam Verma, J. C. Tarafdar* and K. K. Srivastava

Abstract: Arbuscular mycorrhizal fungi (AMF) spore population was studied at eight levels of soil depth under two different sites viz., P. cineraria under agro forestry area (with moong bean) and P. cineraria as sole crop at four different quarters of the year (July, October, January, April). In agro forestry areas the spore population recorded higher as compared to P. cineraria grown as sole crop. The upper layer (0-30 cm) contains more number of AMF population in comparison to deeper layers. The trend in decrease of spore population to soil depth was as follows: 11 - 20 cm > 0 - 10 cm > 21 - 30 cm > 31 - 40 cm > 41 - 50 cm > 51 - 60 cm > 61 - 70 cm > 71 - 80 cm. AMF population was also correlated with organic carbon content at different soil depths. The per cent organic carbon content varied between sites and among the soil depths. The more organic carbon content was recorded at upper layers and decreasing towards deeper layers. The maximum moisture content was found in first quarter, that is, in month of July and minimum in fourth quarter, that is, in April. However, spore population was positively correlated with soil organic matter throughout the year but not with soil moisture. The results demonstrated that status of AMF population under P. cineraria depends on soil depth, soil organic carbon, soil moisture content and the site of plantation.[...] Read More.

Keywords: Arbuscular mycorrhiza, organic carbon content, Prosopis cineraria, soil depth, soil moisture.

2016, 15(1), 1-6;

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Research Article

A comparative study on biofilm formation of non-typeable Haemophilus influenzae and Pseudomonas aeruginosa under single culture or co-culture

Lan Liu *, Lijuan Chu , Qian Liu , Cai Wang , Yongwu Xia , Xuesong Peng

Abstract: Individual bacteria can accumulate to form biofilm which can help bacteria escape from host immune defense and many antimicrobial agents. Therefore biofilm involves a large number of bacteria which can induce chronic diseases. Here, we compared the differences of biofilm formation among Nontypeable Haemophilus influenzae (NTHi) single culture condition, Pseudomonas aeruginosa (P. aeruginosa) single culture condition and NTHi- P. aeruginosa co -culture condition. Biofilm clone formation unit (CFU) counting revealed that the number of clinical isolated P. aeruginosa was larger than clinical isolated NTHi 3 days after incubation. In co-culture condition, the CFU was significantly higher than both types of single culture condition. Consistent with this result, measurement of crystal violet staining of bacteria attached on the plate well showed that, when co-cultured, the absorbance at 570 nm enhanced 2 - 3 times compared with this two types of single culture condition. Our results were confirmed under scanning electron microscope (SEM). 3 days after inoculation, the co-cultured bacteria stacked more densely to form channels and mushroom-like structure. Whereas both single cultured groups exhibited a more or less sparse, flat-net shape. Intriguingly, both clinical isolated NTHi and P. aeruginosa showed weaker capability in the case of biofilm formation compared with their sibling strains NTHi49247 and P. aeruginosa 27853, respectively. Because all the clinical strains were isolated from children whose immuno-system had not yet developed mature or strong enough, bacteria that had less infectious capability were opportunistically invasive. Our results indicate that NTHi and P. aeruginosa can form biofilm in vitro synergistically.[...] Read More.

Keywords: Nontypeable haemophilus influenzae, Pseudomonas aeruginosa, dynamic observation, biofilm formation.

2016, 15(1), 1-6;

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Research Article

Prevalence and diagnostic aspects of sputum smear positive tuberculosis cases at a tertiary care institution in Rwanda

Claude Mambo Muvunyi, *, Florence Masaisa, Claude Bayingana, Andre Musemakweri, Leon Mutesa and Teresa Carbonell Hernandez,

Abstract: Approximately one third of the world’s population is infected with Mycobacterium tuberculosis and 9.27 million new cases of TB occurred in 2007. Developing countries disproportionately shoulder the global burden of disease with the highest estimated rates in the world, with an estimated 55% of global cases in Asia and 31% in the African region. The incidence of new sputum smear positive in Rwanda through recent national survey was an estimated 162 per 100 000 population. The aim of our study was to evaluate the prevalence of smear positive pulmonary TB among patients at the University of Butare Teaching Hospital, a tertiary health facility in South province, Rwanda. In addition, some aspects of the performance of the pulmonary TB diagnosis are discussed. The overall prevalence of sputum smear positive cases were 17.3% (63 0f 364) and most of the positive patients were within the age range 15 - 44 years. The highest percentage of TB was seen in the age group of 15 - 24 years compared with the lowest percentages in the age group below 14 years and above 45 years. A total of 63 (17.3%) suspects were found to have at least one positive. Of these, 56 (88.9% of those with one or more positive smears and 92% of those who fulfilled the case definition) were detected from the first specimen and 7 (11.1%) were positive on the second specimen but not the first. The third specimen did not have any additional diagnostic value for the detection of AFB. The prevalence of sputum smear positive cases of 17.3% increases with age up to the age 44 years. Our result show that examining two sputa smears was sufficient for the detection of AFB in our laboratory. Further research involving different laboratories from all of the regions of Rwanda is needed to reassess these findings.[...] Read More.

Keywords: Smear positive tuberculosis, prevalence, diagnostic aspects, Rwanda.

2016, 15(1), 1-6;

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Research Article

Effects of Rhizobium innoculation, lime and molybdenum on nitrogen fixation of nodulated Phaseolus vulgaris L.

Sylvie Bambara and Patrick A. Ndakidemi*

Abstract: A field and glass house experiment was conducted with the aim of evaluating Rhizobium inoculation, molybdenum (Mo) and lime supply on growth and nitrogen fixation of nodulated Phaseolus vulgaris. The experiment was laid in a split-split plot design. The experimental treatments consisted of 2 levels of Rhizobium inoculation (with rhizobia and without rhizobia) 3 levels of Mo (0, 6 and 12 g kg-1 of seeds) and 3 levels of lime (0, 2 and 3 t ha-1). Rhizobium inoculation showed significant increase in dry matter yield of different organs and decreased 15N values in all organs assessed, thus resulting to improved % nitrogen derived from atmosphere (%Ndfa) in all organs and the amount of N derived from fixation. Molybdenum and lime were significantly effective in promoting plant growth in most organs and decreased 15N values in all organs for glasshouse and field experiment. Lowest 15N values were recorded in Mo and lime supplied at the highest rates of 6 and 12 g kg-1 of seeds and 2 and 3 t lime per ha, respectively. Molybdenum and lime application also significantly improved %Ndfa in all organs and N derived from N-fixation in most organs. Significant responses in N nutrition were also reported in treatments involving the combination of Rhizobium x Mo x lime with better result being recorded in treatments involving Rhizobium inoculation and higher levels of Mo and lime.[...] Read More.

Keywords: Nodulated root, 15N, %Ndfa, total N.

2016, 15(1), 1-6;

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Research Article

Effects of Enterobiasis on primary school children

Ali Çeliksöz*, Mehmet Aciöz, Serpil De erli, A. Yasemin Öztop and Ahmet Alim

Abstract: The aim of the present study was to investigate the possible adverse effects of Enterobiasis vermicularis infection on primary school children in Sivas, Turkey, to know whether it affects their school success, mean weight and height or not. Seven primary schools in the central region of Sivas were included in the present study. A total number of 3659 questionnaires were completed by class teachers and parents for each student. In the questionnaire, the height and weight of children, school success and other information about children were asked. The stool and cellophane tape specimens collected from the participated students were examined by light microscopy for the diagnosis of enterobiasis. The 365 (17.0%) out of 3569 of parasitic infections were E. vermicularis along with other parasites, and the 225 (10.5%) out of 3569 had only E. vermicularis. The weights and heights were found to be lower in children with enterobiasis. When school successes of the students were compared, there were statistically significant differences between the non parasitic group and the group with E. vermicularis. Enterobiasis is still an important problem seen among primary school children causing adverse effects on school success and physical growth especially in children from the first grades of primary schools.[...] Read More.

Keywords: Enterobiasis, school success, physical growth.

2016, 15(1), 1-6;

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Research Article

Diagnostic criteria in Crimean Congo haemorrhagic fever disease and cost analysis

Sevki Hakan Eren*, lhan Korkmaz, Fatma Mutlu Kukul Güven, Can Aktas and Didem Ay

Abstract: In recent years, Crimean Congo haemorrhagic fever disease (CCHF) has created a serious health problem in our country. The disease takes place among haemorrhagic viral diseases. This study was made with CCHF diagnosed patients accepted in emergency services to determine the diagnostic criteria and cost effectivness for CCHF. CCHF patients who were accepted in our emergency service between 2004 and 2008 were analyzed retrospectively. Sensitivity, specificity, positive predictive and negative predictive value was calculated only for the patients who had 5 pathological laboratory parameters together (thromobcytopenia, high LDH, AST, ALT and CPK), and cost effectiveness analysis was made. During the five-year period, 687 patients had been evaluated. Significant pathologic results were established for alanine amino transferase (ALT), aspartate amino transferase (AST), creatine phosphokinase (CPK) enzymes, platelet counts and lactate dehydrogenase (LDH). Fatigue, fever, bleeding, gastro intestinal symptoms and diffuse pain were the symptoms seen according to their frequencies. The patients were frequently admitted in July. Mortality, costs per patient and emergency service's crowdedness can be reduced by rapid diagnosis which can easily be made with a brief history of the patient’s occupation, physical examination and mentioned laboratory results sensitivity, specificity, positive predictive and negative predictive value.[...] Read More.

Keywords: Crimean-Congo haemorrhagic fever virus, emergency service, diagnostic criteria.

2016, 15(1), 1-6;

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Research Article

Antibiotic resistance microbes in tropical mangrove sediments in east coast peninsular, Malaysia

K. C. A. Jalal*, Nur Fatin U.T, Mardiana M.A, Akbar John B, Kamaruzzaman Y. B, Shahbudin S, and Muhammad Nor Omar

Abstract: The study has been conducted at Tanjung Lumpur, mangrove swamp on January 2009 to isolate and identify the bacterial community in mangrove soil and their resistance against antibiotics. Identified bacteria were Aeromonas hydrophila group 1 and 2, Escherichia coli 1, Chryseomonas luteola, Chromobacterium violaceum, Pseudomonas aeruginosa, Serratia rubudaea, Klebsiella pnuemoniae and Enterobacter cloacae. The identified bacteria were introduced to fourteen different antibiotics to determine the bacterial susceptibility. All the isolates showed 100% resistant towards -lactam antibiotics (ampicillin, amoxicillin and penicillin), vancomycin, sulphafurazole, gentamicin, erythromycin, tetracycline, novobiocin, clindamycin and bacitracin indicates the presence of bacterial amidases and -lactamases in the bacteria which inhibit the action of - lactam antibiotics. Bacteria isolated from mangrove soil showed 66.7 and 77.8% resistance against chloramphenicol and streptomycin, respectively, suggesting that the lipid composition might play a key role in preventing the entrance or binding of antibiotic to the cell. All the isolates were susceptible to ciprofloxacin since it inhibits the enzyme topoisomerase II that cause the negative super coil in DNA and thus permits transcription or replication. All bacterial isolates showed Multi Antibiotic Resistance (MAR) index higher than 0.2 and proved high-risk sources of contamination of the environment. This study proved the presence of antibiotic resistant bacterial strains in mangrove soil that could be used for further studies.[...] Read More.

Keywords: Tanjung Lumpur, mangrove swamp, bacterial community, -lactam antibiotics, multi resistance antibiotic index.

2016, 15(1), 1-6;

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Research Article

Optimizing the conditions of - amylase by an Esturian strain of Aspergillus spp

Arun Sasi*, Manthiri Kani, A. Panneerselvam, G. Jegadeesh, K. Muthu and M Ravi Kumar

Abstract: Water samples and sediments were collected from waterways in between two back water lakes namely Vambanad and Asthamudi, in Alukadavu, Kollam along the west coast of Kerala, India. 20 fungal species were isolated and identified by lacto phenol cotton blue staining method. From this, Aspergillus flavus was selected for starch hydrolysis Agar Medium for amylase activity under submerged fermentation for amylase production. The effect of different carbon source, nitrogen source, heavy metals, amino acids, dry biomass was studied and enzyme was optimized by using different physico-chemical parameters. The purity of enzyme was analyzed by thin layer chromatography. Minimal medium with starch has produced high amylase enzyme and soluble protein dry biomass with a temperature of 30°C and pH of 6, which includes 256.2 and 108 µg/ml, respectively.[...] Read More.

Keywords: Amylase enzyme, back water fungi, Aspergillus flavus, submerged fermentation, SDS-PAGE, Ion exchange chromatography.

2016, 15(1), 1-6;

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Short Communication

Isolation and characterization of plum-seeds degrading aerobic bacteria from plum-grove soil

Haruka Kurishita, Hitomi Yoshida, Namika Yamaji, Naoko Okumura, Asuka Kirimura, Satoru Matsuda* and Hiroshi Ueno

Abstract: The aim of this study was to isolate plum-seeds degrading bacterium from plum-grove soil. Some bacteria utilizing plum-seeds were isolated from the plum-grove soil samples by biodegradation assay. Among them, five bacteria had the activity of cellulase and xylanase. Four of the strains grew in the medium containing xylan instead of glucose. Partial sequencing of 16S rRNA and genome indicated that all the isolates belong to the Psedomonas section but two bacteria are unknown. It is proposed that some Pseudomonas bacteria could be deployed as inoculant to attain the treatment of plum-seeds.[...] Read More.

Keywords: Biodegradation, plum-seeds, cellulase, xylanase, microorganism.

2016, 15(1), 1-6;

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Research Article

Uptake of textile dye Reactive Black-5 by Penicillium chrysogenum MT-6 isolated from cement-contaminated soil

Serkan Erdal and Mesut Taskin*

Abstract: The present study focused on the decolorization of textile dye Reactive Black- 5 by actively growing mycelium of fungus Penicillium chrysogenum MT-6, which was isolated from the cement-contaminated soil. Dye decolorization was probably associated with fungal growth and hyphal uptake mechanism (Biosorption/Bioadsorption) . Dye uptake was strongly depended on mycelial morphology. Small uniform pellets with 2 mm size and nutrient-poor medium were found to be better for dye uptake. Optimal conditions for dye uptake by the fungus were determined as initial pH of 5.0, shaking speed of 150 rpm, temperature of 28°C, spore concentration of 107/ml, 10 g/l sucrose and 1 g/l ammonium chloride. The maximum removal/uptake of dye by fungus was 89% (0.267 g removed-dye) with 3.83 g/l of biomass production at an initial dye concentration of 0.3 g/l in 100 h. The fungus was found to be a good bio -system for the decolorization of the medium containing Reactive Black-5. It was shown for the first time in the present study that the cement-contaminated soil was a good source of microorganisms, being capable of removing snythetic textile dye.[...] Read More.

Keywords: Penicillium chrysogenum MT-6, dye removal, optimization, reactive black-5.

2016, 15(1), 1-6;

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Research Article

Antimicrobial activity of various extracts of Centaurea cankiriense A. Duran and H. Duman

Arzu Cansaran, Nazime Mercan Do an*, Mehtap Öztekin and Gülümser Acar

Abstract: The antimicrobial activity was determined using the single disc diffusion method. The hexane, methanol and ethyl acetate extracts were assessed for antimicrobial activity against 13 bacteria and a yeast-like fungus, Candida albicans. While flower extracts of Centaurea cankiriense showed significant antibacterial activity against tested strains, the susceptibility of the test microorganisms was less pronounced in the cases of the stem extracts. Hexane extracts from both flower and stem did not show any antibacterial activity against gram-negative bacteria at test concentration, whereas ethyl acetate and methanol extract of C. cankiriense demonstrated the growth of both the gram-positive and the gram-negative bacteria. But, methanol extract inhibited the bacteria with the exception of two gram-negative bacteria namely Escherichia coli and Klebsiella pneumoniae. Minimum inhibition concentration (MIC) was determined on ethyl acetate extracts of flower and stem that showed high activity against the test bacteria. The MIC values for bacterial strains were in the range of 7.8 - 250 mg/ml. The results confirmed that E. coli (MIC = 250 mg/ml) and Morganelle morganii (MIC = 125 mg/ml) was the most resistant organisms to plant extracts. The flower extract of C. cankiriense was found to possess the strongest effect on Bacillus cereus with 7.8 mg/ml concentration.[...] Read More.

Keywords: Centaurea cankiriense, antimicrobial activity, minimum inhibition concentration, disc diffusion.

2016, 15(1), 1-6;

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Research Article

Oral colonization and boric acid susceptibility of yeast in boron mineral workers

Zafer Cetinkaya*, Semsettin Karaca , Mustafa Kulac , Ihsan Hakki Ciftci, Gulsah Asık, Osman Cenet and Nuri Kiraz

Abstract: In this study, we aimed to investigate the effects of boron on in vivo oral yeast colonization in study groups which are exposed to boron dust in different sections of the boron mine. The study was carried out in the boron mining areas of two districts (Eskisehir and Balikesir) of Turkey. We included 184 people working in open quarry and stone milling unit, 144 people working in the factory and 150 people as control group. Specimens were taken from four oral mucosal regions and cultured onto Sabouraud dextrose agar. After incubation for 3 - 7 days at 30°C, the total number of yeast colonies on the plates was considered the relative intensity of carriage, and the total number of yeast colonies on the plates was considered the relative intensity of oral carriage. The susceptibility of Candida spp. to boric acid was investigated. The frequency of Candida colonization in boron intensive area workers was found significantly higher than automatic factory workers and control groups (p = 0.012), there were no difference between automatic factory workers and control groups in point of Candida colonization (p = 0.749). We observed that oral yeast colonization had increased directly proportional with boron powder exposure in boron mine (p = 0.005) . Mean minimum inhibitory concentrations (MICs) of boric acid for Boron intensive area, 0.87 - 2.0% for automatic factory and 0.83 - 2.0% for control subjects. We observed that intensive exposure to boron mineral powders was strictly related to oral yeast colonization. Exposure to industrial boron mineral powder may cause important health problems by increasing Candida colonization in oral cavity. It may be useful to do periodical health control in boron mineral workers and population under risk.[...] Read More.

Keywords: Candida, boric acid, boron mineral, oral yeast colonization.

2016, 15(1), 1-6;

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Research Article

Antibacterial activity and mechanism of recombinant human - defensin 5 against clinical antibiotic-resistant strains

A. P. Wang, Y. P. Su, S. Wang, M. Q. Shen, F. Chen, M. Chen, X. Z. Ran, T. M. Chen and J. P. Wang*

Abstract: The increasing clinical bacterial strains resistant to conventional antibiotics have being a great challenge to the public’s health. As a novel kind of antimicrobial agent, defensins are undoubtedly worthy of exploitation for the treatment of antibiotic-resistant bacteria. To evaluate the antibacterial potency of recombinant mature human -defensin 5 (rmHD5) against clinical pathogenic strains, we examined its antibacterial kinetics and bactericidal efficacy on forty-nine bacterial strains (belonging to eleven species) with different antibiotic-resistant phenotypes, isolated from digestive and urogenital tracts of the inpatient. Meanwhile, the action mechanism of rmHD5 was analyzed by transmission electron microscopy observation and membrane permeability detection. The peptide of rmHD5 was found to possess high potency against all the tested isolates at concentrations of 6 - 12 g/ml for gram-negative (G-) bacteria and 28 - 32 g/ml for gram-positive (G+) bacteria. G- bacteria were more susceptible to the peptide than G+ bacteria. Abnormal morphological changes and increased permeabilization of the cytomembrane were observed in both G - bacteria and G+ bacteria treated with rmHD5. The antibacterial activity of rmHD5 may be tightly associated with the biomembrane permeabilization. Recombinant mHD5 is a promising candidate to be developed into therapeutic agents for bacterial infections[...] Read More.

Keywords: Antibacterial activity, mechanism, antibiotics resistant strain, human alpha defensin 5.

2016, 15(1), 1-6;

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Research Article

Fingerprinting of repetitive DNA sequences in the genus Anabaena using PCR- based techniques

Anbalagan Ezhilarasi* and Narayanaswamy Anand

Abstract: In this study, ten species of Anabaena were used to test the congruence between the traditional morphological classification system and the present molecular classification system. The electrophoretic patterns for 10 cyanobacterial strains belonging to the genera (Anabaena) were used for molecular analysis using the repeated PCR technique. Genetic diversity was assessed using the banding patterns of repetitive DNA sequences including the HIP1 sequences, enterobacterial repetitive intergenic consensus [ERIC] sequences and REP sequences that are present in the cyanobacterial genome. PCR with different sets of repetitive oligonucleotides as primers were used to evaluate the various repetitive DNA sequences. The dendrograms based on the HIP1, ERIC and REP sequences did not show a clear correlation with morphological dendrogram based on traditional classification.[...] Read More.

Keywords: Anabaena, cyanobacteria, highly iterated palindromic sequences, enterobacterial repetitive intergenic consensus sequences, repetitive extragenic palindromic sequences, polymerase chain reaction.

2016, 15(1), 1-6;

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Research Article

The effect of cultivation conditions on the mycelial growth of a dark-septate endophytic isolate

Ya-li Lv, Li-hua Sun, Fu-sheng Zhang, Yue Zhao and Shun-xing Guo*

Abstract: The EF-37 isolate, one of DSE fungi, is beneficial to the growth and development of its host plant, Saussurea involucrata Kar. et Kir. The cultivation requirements including basic culture medium, temperature, light, pH, carbon source and nitrogen compounds were studied for their effects on mycelial growth of a dark-septate endophytic (DSE) fungus EF-37 by using one-factor-at-a- time method. Potato dextrose agar (PDA) was the best medium for the growth of endophyte EF-37. Our studies showed that 20°C, 24 h dark cultivation and pH 7 significantly influenced the growth of endophyte EF-37 on PDA medium. Moreover, glucose and calcium nitrate were found to be the best nutrients for EF- 37 growth. Under the optimal cultivation conditions, DSE fungus EF-37 isolate could grow actively. This is the first study about the effect of cultivation conditions on the growth of this strain, which provides the preparatory knowledge for the biological characteristics of DSE fungus EF-37.[...] Read More.

Keywords: Cultural conditions, dark-septate endophytic (DSE) fungus EF-37, mycelial growth, optimization, Saussurea involucrata Kar. et Kir.

2016, 15(1), 1-6;

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Research Article

The identification of the first isolate of influenza B virus using a duplex RT- PCR DNA sequencing in Saudi Arabia (B/Riyadh/01/2007)

Fahad N. Almajhdi

Abstract: Although Influenza virus infections threaten thousands of lives each year worldwide and bear the risk of been an epidemic, little is known about the circulating strains in Saudi Arabia. The availability of a rapid and accurate diagnostic approach is essential for effective treatment and disease control. In the current study, we optimized a duplex RT-PCR assay for the simultaneous detection of influenza A and B viruses in clinical samples. The developed assay was utilized for testing 100 nasopharyngeal aspirates collected from young children hospitalized with acute respiratory tract infection in Riyadh. Influenza B not A viruses, were identified as possible causes of the disease syndrome. Recovery of influenza B virus from clinical samples was achieved by serial passage in chicken embryos and the virus isolate was designated as Influenza B/Riyadh/01/2007. The identity of B/Riyadh/01/2007 was confirmed by sequencing the RT-PCR product. The sequenced data was submitted to the Gene Bank under the accession number GU135839. An almost complete homology was recognized with all Influenza B virus strains available on Gene Bank. Although several studies suggested the presence of influenza B virus in respiratory tract infection of Saudi children, this is the first report that describes the isolation and partial identification of the circulating virus. Further studies that fully characterize the isolated strain are ongoing.[...] Read More.

Keywords: Influenza B virus, isolation, RT-PCR, DNA sequencing, Saudi Arabia.

2016, 15(1), 1-6;

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Research Article

Development and evaluation of colloidal gold immunochromatographic strip for detection of Escherichia coli O157

Xihong Zhao, Xiaowei He*, Wenmei Li , Yao Liu, Liansheng Yang and Jihua Wang

Abstract: Escherichia coli O157:H7 is a serious and common human pathogen that can cause diarrhoea, haemorrhagic colitis, and haemolytic uraemic syndrome (HUS). In this study, the synthesis and identification of colloidal gold particles and antibody-colloidal gold conjugates probe specific to E. coli O157:H7 were performed, and the preparation of colloidal gold immunochromatographic strip based the biotin-streptavidin system was developed for detection of E. coli O157:H7.Monodispersional nanogold colloid was synthesized and preparation of nanogold-labeled polyclonal antibody probe to E. coli O157:H7 by citrate method. Combination of antibody with nanogold particles was also characterized by UV-visible (UV-vis) light absorption spectra and transmission electron microscopy (TEM). Furthermore, nanogold-labeled probe was used to develop an immunochromatographic (IC) strip for E. coli O157:H7 analysis. With this method, analysis could be completed in less than 10 min. Examination of the 65 known strains (36 E. coli O157 strains and 29 serotypes other than E. coli) showed 98.5% specifity and 100% sensitivity, only yield a false-positive reaction with Salmonella choleraesuis . The sensitivity of the IC strip was tested using 10-fold dilution E. coli O157 in foods, could be detected at a minimum of 2.3 × 103 CFU/ml without enrichment and 2.3 CFU/ml after enrichment. Application of IC strip test were performed on 265 water samples, 340 beef samples, 208 milk samples and 120 cake samples after enrichment, the specificity of the strip was 99.2, 97.9, 94.6 and 94.9%, respectively. The sensitivity of the strip was 100% agreement with tradition culture method. The established method is very useful for monitoring E. coli O157 containment in food samples.[...] Read More.

Keywords: Escherichia coli O157, colloidal gold immunochromatographic assay, rapid test.

2016, 15(1), 1-6;

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Research Article

Preliminary in-vitro antibacterial activities of ethanolic extracts of Ficus sycomorus Linn. and Ficus platyphylla Del. (Moraceae)

Adeshina Gbonjubola Olusesan*, Okeke Claire-Lorentz Ebele, Osuagwu Nonyelum Onwuegbuchulam and Ehinmidu Joseph Olorunmola

Abstract: Antibacterial therapeutic failure due to emergence of resistant bacterial strain is a worldwide phenomenon. The search for effective antibacterial substances from sources such as plants has become a necessity to overcome emergent of bacterial resistant in clinical practice. The dried leaves and stem barks of Ficus sycomorus and Ficus platyphylla were collected in Samaru-Zaria, Nigeria in July 2006 and extracted with 70% aqueous ethanol at room temperature. The antibacterial activities such as susceptibility, Minimum inhibitory concentrations (M.I.C.) and the minimum bactericidal concentrations (M.B.C.) were determined using appropriate methods. Using the same concentration of the two test plants extracts, the zones of inhibition showed by F. sycomorus ranged between 11.5 - 21.5 mm while that of F. platyphylla was from 17.0 - 22.0 mm. The values of the M.I.C and M.B.C of F. sycomorus were 1.95, 31.3 and 3.91, 250 mg/ml, respectively. Similarly, F. platyphylla displayed 1.95 and 7.81 mg/ml M.I.C. values and 3.91 to 62.5 mg/ml M.B.C. values against the test organisms. The observed antibacterial activities in this study proved that the leaves and stem bark extracts of Ficus spp. obtained in Zaria support the forcloric claims of the use of Ficus plants in the treatment of ailment such as wound dressing.[...] Read More.

Keywords: Antibacterial activity, medicinal plants, bacterial resistance, Ficus spp.

2016, 15(1), 1-6;

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Research Article

Prevalence rate and antibiotic resistant pattern of methicillin-resistant Staphylococcus aureus and methicillin-resistant Staphylococcus epidermidis

Muhammad Arfat Yameen, Hina Nasim, Naeem Akhtar, Saira Iram, Imran Javed and Abdul Hameed*

Abstract: The aim of this study was to evaluate the prevalence rate and antibiotic resistant pattern of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE). A prospective study was conducted at Holy Family Hospital Rawalpindi, Pakistan and Microbiology Research Laboratory, Quaid-i-Azam University, Islamabad, Pakistan during the period from December 2007 to August 2008. The antibiotic resistance pattern was studied for MRSA and MRSE isolated from nasal samples from patients admitted in medical and surgical intensive care units. The study was conducted on 283 isolates. The results depicted that 25% isolates of S. aureus were MRSA and 29.78% isolates of S. epidermidis were MRSE. All MRSA and MRSE were susceptible to vancomycin and quinopristin/dalfopristin while all isolates of MRSE were susceptible to teicoplanin. All the isolates of MRSA and MRSE were multidrug-resistant. The susceptibility of the isolates to the drugs varied greatly. The resistance rate of MRSA to various antibiotics was found to be as follow: cephalaxin (90%), cephalothin (58%), cephradine (86%), ciprofloxacin (80%), gentamicin (34%), imipenum (42%), levofloxacin (75%), tetracycline (49%), rifampicin (14%) and teicoplanin (3%). The resistance rate of MRSE to various antibiotics was found to be as follow: cephalaxin (64%), cephalothin (29%), cephradine (64%), ciprofloxacin (50%), gentamicin (21%), imipenum (7%), levofloxacin (21%), tetracycline (21%) and rifampicin (29%). The minimum inhibitory concentration (MIC) value for MRSA and MRSE in case of vancomycin ranged 1-4 g/ml, for tetracycline 4-128 g/ml, for rifampicin 0.5-32 g/ml and for gentamicin 0.5 – 64 g/ml. Both MRSA and MRSE showed variable susceptibility with different antibiotic groups but high susceptibility with streptogramin and glycopeptide antibiotics.[...] Read More.

Keywords: Antibiotic resistance, staphylococcus aureus, staphylococcus epidermidis, minimum inhibitory concentration.

2016, 15(1), 1-6;

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Research Article

Investigation of different infectious pathways used by Streptococcus suis type 2 and other bacteria in mice

Xingjian Cao, Zhaoming cao, Guohua Tao, Wei Xie, Xiang Chen and Shuo Wang

Abstract: The purpose of this study was to investigate the infectious pathways used by Streptococcus suis type 2 and enterohemorrhagic Escherchia coli O26 (EHEC O26) in mice. Six-week mice were intraperitoneally injected with mitomycin (0.2 mg/mouse) to decrease the protective immunity. The mice were then challenged with S. suis type 2 and EHEC O26 strains through wounded skin, stomach lavaging and intraperitoneal injection. Enterococcus faecalis ATCC 29212 and E. coli ATCC25922 were used as negative controls. Clinical and microbiological examinations were performed for all the infected mice routinely. The mortality rate of the mice infected with S. suis type 2 strain through wounded skin was 60% within 2 - 5 days. The mortality rate of the mice infected with S. suis type 2 strain through stomach lavaging was 60% within 7 - 10 days. S. suis type 2 can be isolated from the puncturing fluid of heart in the dead mice. Mortality was not observed in mice infected by S. suis type 2 and E. faecalis ATCC 29212 strains through intraperitoneal injection. However, intraperitoneal injection with EHEC O26 and E. coli ATCC25922 caused death of mice within 10 h. We demonstrated an important pathway used by S. suis type 2 strain in mouse infection model. Intraperitoneal injection with gram negative bacteria (E. coli ATCC25922 and EHEC O26) caused non-specific death in mice. These results provided fundamental bases for investigating the pathogenesis of S. suis type 2, designing vaccine and evaluating the efficiency of antimicrobial drugs.[...] Read More.

Keywords:

2016, 15(1), 1-6;

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Research Article

Antibacterial activity and mechanism of recombinant human-defensin 5 against clinical antibiotic-resistant strains

A. P. Wang, Y. P. Su, S. Wang, M. Q. Shen, F. Chen, M. Chen, X. Z. Ran, T. M. Chen and J. P. Wang*

Abstract: The increasing clinical bacterial strains resistant to conventional antibiotics have being a great challenge to the public’s health. As a novel kind of antimicrobial agent, defensins are undoubtedly worthy of exploitation for the treatment of antibiotic-resistant bacteria. To evaluate the antibacterial potency of recombinant mature human -defensin 5 (rmHD5) against clinical pathogenic strains, we examined its antibacterial kinetics and bactericidal efficacy on forty-nine bacterial strains (belonging to eleven species) with different antibiotic-resistant phenotypes, isolated from digestive and urogenital tracts of the inpatient. Meanwhile, the action mechanism of rmHD5 was analyzed by transmission electron microscopy observation and membrane permeability detection. The peptide of rmHD5 was found to possess high potency against all the tested isolates at concentrations of 6 - 12 g/ml for gram-negative (G-) bacteria and 28 - 32 g/ml for gram-positive (G+) bacteria. G- bacteria were more susceptible to the peptide than G+ bacteria. Abnormal morphological changes and increased permeabilization of the cytomembrane were observed in both G - bacteria and G+ bacteria treated with rmHD5. The antibacterial activity of rmHD5 may be tightly associated with the biomembrane permeabilization. Recombinant mHD5 is a promising candidate to be developed into therapeutic agents for bacterial infections.[...] Read More.

Keywords: Antibacterial activity, mechanism, antibiotics resistant strain, human alpha defensin 5.

2016, 15(1), 1-6;

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Research Article

Efficiency of filtration technique for isolation of leptospires from surface waters: Role of different membranes with different pore size and materials

Hami Kaboosi*, Mohammad Reza Razavi, and Ashraf al Sadat Noohi

Abstract: The objective of this research focused on evaluation of membrane filters application for isolation of Leptospira from surface water samples. The filter materials evaluated included nitrocellulose (0.22 and 0.45 µm pore diameters), polyvinylidene fluoride (Durapore 0.22 and 0.40 m pore diameters), nylon mesh (37 m), and glass fiber (1.0 µm). Millipore polyvinylidene fluoride filter (0.22 m) was examined by scanning electron microscopy to verify that leptospires were present following filtration. Our results suggest for isolate nearly 100% of Leptospira from water samples, the optimal pore diameter should be less than 0.45 µm (a standard pore size used to detect indicator microorganisms in 100 ml of water). Although filtration method can be used to isolation leptospires from surface water samples, it is unclear whether this is a useful method for detection of all leptospires exist within surface water samples. Accordingly, a large proportion of leptospires can be retained by membrane filter with a pore diameter commonly used to isolate leptospires from surface water samples (0.45 µm pore size).[...] Read More.

Keywords: Leptospira, filtration technique, leptospirosis.

2016, 15(1), 1-6;

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Research Article

The validity of the diagnostic methods in predicting pulmonary tuberculosis

Sulhattin Arslan*, Levent Özdemir, Yeltekin Demirel and Ibrahim Akkurt

Abstract: In our study, we aimed to determine the validity of diagnostic methods for tuberculosis. Eighty-one people suspected to have tuberculosis were included in the study. The validity of the applied methods for the diagnosis of tuberculosis tuberculin skin test (TST), sputum smear, and used in diagnostic chest X-ray findings (CXR), clinical features and history were evaluated as culture was considered the reference test. Included in the study of 81 people (54 males 27 females) mean age was determined as 45.04 ± 18.69. The most sensitive diagnostic methods were detected as clinical and sputum smears (89 and 86%). The sensitivity of PPD and Radiology have been identified as 0.74 and 0.73 respectively. Diagnostic method with the highest specificity value was found to be radiology. The positive predictive values of PPD, radiology, clinical and sputum smear were identified as 98, 94, 96 and 92%, respectively. PPD had the lowest negative predictive value with 26%. In our study, the validity of the diagnostic methods for the tuberculosis are compatible with the literature. These methods in the diagnosis of tuberculosis are still valid. We think our study may add to the current data in the literature about the topic.[...] Read More.

Keywords: Validity, specificity, sensitivity, PPD, TST, sputum smear, culture, tuberculosis, clinical features.

2016, 15(1), 1-6;

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Research Article

Phenotypic and molecular characterization of SHV, TEM, CTX- M and extended-spectrum -lactamase produced by Escherichia coli, Acinobacter baumannii and Klebsiella isolates in a Turkish hospital

Elif Burcu Bali, Leyla Açık* and Nedim Sultan

Abstract: A total of 94 clinical isolates were collected from Gazi University Hospital, Turkey. Presence of ESBL positivity was detected using the double disk synergy test (DDST). ESBL isolates were further typed for the blaTEM, blaSHV, blaCTX-M and blaOXA using designed primers. ESBLs were found in 65 (69.14%) isolates using DDST. Plasmid DNAs of potentially ESBL positive strains were isolated. About 7.69% of the ESBL positive isolates did not harbour plasmid DNA. According to the PCR technique, only 2 additional isolates were found to be ESBL producers. blaTEM was the commonest genotype (73.43%), followed by blaSHV (21.87%) and blaCTX-M (17.18%), either alone or in combination. ESBL positive strains of Klebsiella pneumoniae, Escherichia coli, Acinetobacter baumannii and Pseudomonas aeruginosa are increasingly found in hospital isolates. Because these strains become resistant to available antibiotics and they can pass the gene to other clinical strains, the quick detection of these strains in clinical laboratories is very important.[...] Read More.

Keywords: ESBL, double disk synergy test, plasmid, PCR.

2016, 15(1), 1-6;

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Research Article

Staphylococcal cassette chromosome mec (SCCmec) analysis and antimicrobial susceptibility profiles of methicillin- resistant Staphylococcus aureus (MRSA) isolates in a teaching hospital, Shantou, Chin

Qing Peng, Bing Hou, Shuqin Zhou, Yuanchun Huang, Dexing Hua, Fen Yao and Yuan shu Qian*

Abstract: Methicillin-resistant staphylococcus aureus (MRSA) infection has now become a major public health concern. The aim of this study is to determine the antibiotic resistance pattern and prevalence of different staphylococcal cassette chromosome mec (SCCmec) types among the MRSA isolates from a teaching hospital in Shantou, China. The minimum inhibitory concentrations (MICs) of seventeen antimicrobial agents against MRSA isolates were determined using the micro broth dilution method. SCCmec types were identified by multiplex polymerase chain reaction (PCR) strategy. The results show that all MRSA isolates were resistant to ampicillin, oxacillin, gentamycin, erythromycin and ciprofloxacin. Fewer than 6% of isolates were resistant to doxycycline, but more than 90% were resistant to cefazolin, cefuroxime, cefotaxime, cefepime, sparfloxacin and tetracycline. There was no minocycline, chloramphenicol or vancomycin resistant. S. aureus was found in this study. SCCmec type III and IIIA were predominant in our study. Our data highlighted that multidrug-resistant strains of MRSA caused severe problems in Shantou, China. However, some of the old agents, such as minocycline, doxycycline and chloramphenicol are highly effective against MRSA isolates.[...] Read More.

Keywords: Methicillin-resistant Staphylococcus aureus (MRSA), staphylococcal cassette chromosome mec (SCCmec), antimicrobial agent resistance.

2016, 15(1), 1-6;

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Research Article

The potential influence of high cholesterol diet-induced oxidative stress on composition and properties of red blood cells in rabbits

Mohamed Anwar K. Abdelhalim

Abstract: Atherosclerosis and heart diseases are major causes of morbidity and mortality in adults in industrialized nations. The aim of this study was to assess the potential influence of high-cholesterol diet-induced oxidative stress on composition and properties of red blood cells (RBCs) in rabbits. Thus, percentage of hematocrit, RBCs, white blood cells (WBCs) and platelets counts, total cholesterol (TC), low density lipoprotein (LDL), triglycerides (TG) and high density lipoprotein (HDL), Thiobarbituric acid reactive substances (TBARS) serum level, antioxidant enzymes activity (Superoxide dismutase: SOD; Glutathione peroxidase: GPx), hemoglobin (Hb) and Hb derivatives (oxyhemoglobin: HbO2; carboxyhemoglobin: HbCO; sulfohemoglobin: SHb; met-hemoglobin: Met-Hb) were measured in control and high fat diet (HFD) rabbits. We found that the TC, LDL, TG and HDL (mg/dl) were significantly (p < 0.001) increased in HFD rabbits compared with control rabbits. A significant (p < 0.05) decrease in Hb (g/dl), percentage of hematocrit and RBCs count was observed in HFD rabbits compared with control rabbits while a significant increase in platelet and WBCs counts was observed. The TBARS was significantly (p < 0.05) increased in HFD rabbits compared with control rabbits while antioxidant enzymes SOD and GPx activity were significantly (p < 0.05) decreased. A significant increase in percentage of Met-Hb, HbCO and SHb was observed in HFD rabbits compared with control rabbits while a significant decrease in percentage of HbO2 was observed. This study shows that hypercholesterolemia affects the level of Hb and Hb derivatives which causes anemia and may produce reactive oxygen species (ROSs) and other free radicals increasing TBARS and decreasing SOD and GPx enzymes activities. Hypercholesterolemia may promote the conversion of HbO 2 and the fraction of unstable Hb molecules to Met-Hb, SHb and HbCO. Furthermore, increased platelet and WBCs count in HFD rabbits may be of pathophysiological importance for the progression of atherosclerosis and thromboembolic complications. This study suggests that hypercholesterolemia may produce free radicals which promote oxidation of Hb and reduce its concentration and conversion of HbO 2 to Met-Hb and the fractions of unstable Hb molecules to Met-Hb, SHb and HbCO. Furthermore, an imbalance between free radical production and antioxidant enzymes activities may lead to oxidative stress.[...] Read More.

Keywords: High cholesterol diet, oxidative stress, antioxidant enzyme activates, red blood cells, atherosclerosis.

2016, 15(1), 1-6;

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Research Article

Preliminary studies on Piliostigma thonningii Schum leaf extract: Phytochemical screening and in vitro antimalarial activity

A. Kwaji, P. U. Bassi, M. Aoill, C. M. Nneji and G. Ademowo

Abstract: Different parts of Piliostigma thonningii Scum (Caesalpinioideae) have been used medicinally. The roots and twigs have been used locally in the treatment of dysentery, fever, respiratory ailments, snake bites, hookworm and skin infections in Eastern Nigeria. The leaf extracts has been used for various ethnomedicinal purposes including the treatment of malaria all over Eastern Nigeria. In this study, we have investigated the inhibitory effects of the crude ethanol and methanol extracts for in vitro antimalarial activity against chloroquine resistant Plamodium Falciparum clone (W2-Indo-China isolates). The aqueous screening using reported methodologies for phytochemical screening and in vitro test methods, revealed the presence of alkaloids, tannins, saponnins, steroid, terponoids, flavonoids, cardiac glycosides and anthroquinoes. The crude leaf extracts obtained were tested for in vitro antimalarial activity using chloroquine resistant strain of P. Falciparum clone (W2 – Indochina isolates) . The 50% inhibitory concentrations (IC50) were evaluated after 48 - 72 h contacts between the extracts and the parasite culture. The 50% inhibitory concentration values for both the crude amide ethanolic extract and the partially purified methanolic extract ranged between 6.20 - 15.06 µg/ml. While that of chloroquine was 0.316 µg/ml. This study suggested that P. thonningii leaf extract possess a significant level of antimalarial activity.[...] Read More.

Keywords: Pilostigma thonningii leaf, phytochemical screening, antimalarial activity.

2016, 15(1), 1-6;

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Research Article

Determination of microbiological characteristics of Turkish Karin Kaymagi cheeses packaged in different materials

Salih Özdemir, Filiz Yangılar* and Cihat Ozdemir

Abstract: The aim of this study is to transport to dairy plants the making technique of Karin Kaymagi cheese and to make them hygienic cheese. Also, the effects of different packaging materials (artificial case, barrel and tripe) on microorganism counts in the cheese samples were determined. In this research, four different Karin Kaymagi cheese samples were prepared from white cheese, civil cheese, whey cheese, pasteurized cream and concentrated yoghurt mixes. These mixes were put in three different packaging materials (artificial case, barrel and tripe) and ripened at 12°C for 2, 15, 30 and 60 days. The means of total aerobic mesophilic (TAMB), lactic acid (LAB), coliforms, proteolytic, lipolytic, psychrotrophic, spore forming bacteria and yeast-moulds in the cheese samples were determined as 7.08, 6.11, 1.18, 5.13, 4.82, 5.30, 1.30 and 4.83 log cfu/g, respectively. As the LAB counts and the yeast-mould counts of samples increased, the coliform group bacteria counts decreased during ripening periods. The yeast-moulds counts of cheese samples packaged in barrel were lower than that of tripe and artificial case.[...] Read More.

Keywords: Karin Kaymagi cheese, microbiological characteristics, packaging materials.

2016, 15(1), 1-6;

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Research Article

Antibiotic sensitivity of human genital mycoplasmas

Jafar Khan*, Robeena Farzand and Pir Bux Ghumro

Abstract: The therapeutic failure of antimycoplasma drugs, subject to resistance is vital for specific microbiological investigation to overcome urogenital diseases among women. The susceptibility of 5 Mycoplasma hominis strains and 11 strains of Ureaplasma urealyticum were studied against six antibiotics namely: minocycline, lincomycin, erythromycin, tetracycline, doxycycline and rosaramicin. Both M. hominis and Ureaplasmas strains were highly resistant to tetracycline. It was observed that both rosaramicin and minocycline were active growth inhibitors for Ureaplasmas. However, erythromycin and lincomycin showed differential growth inhibitory patterns for M. hominis and Ureaplasma strains. In comparison, the minocycline and doxycycline exhibited similar antibiotic activity against Ureaplasma and M. hominis. The in vitro antibiotic sensitivity analysis for rosaramicin and erythromycin was also noted to be at the same level for all the isolates.[...] Read More.

Keywords: Mycoplasma hominis, Ureaplasma urealyticum, antibiotic sensitivity, drug resistance.

2016, 15(1), 1-6;

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Research Article

Penicillium verruculosum RS7PF: A root fungal endophyte associated with an ethno-medicinal plant of the indigenous tribes of Eastern India

R. K. Bhagobaty, S. R. Joshi* and R. Kumar

Abstract: Potentilla fulgens L., an ethno-medicinal plant of the ethnic tribes of Meghalaya in the Eastern Himalayan range of India was screened for root fungal endophytes. This plant is distributed in the wild habitats and in the traditionally preserved ‘Sacred Groves’ of Meghalaya in Eastern India, which is of late getting over-exploited for its medicinal value. Isolation of endophytes in water agar plates yielded a unique fungus that had similar morphological characteristics with the members of the genus Talaromyces. Scanning and transmission electron microscopic studies of the fungus revealed its surface morphological features, close association and extensive colonization of plant root tissues. Molecular characterization using fungal specific 18S rRNA primers showed that the fungus shared a 98% homology with Penicillium verruculosum (Genbank accession number AF510496). The isolate investigated is likely to be a novel endophytic strain of P. verruculosum which has evolved a very close symbiotic association with the host plant roots. The fungal endophyte characterized is being explored as an early colonizer in the establishment of seedlings and successful micropropagation of the plant.[...] Read More.

Keywords: Potentilla fulgens L., ethnic tribes, Sacred Grove, fungal endophyte, molecular characterization, Penicillium verruculosum.

2016, 15(1), 1-6;

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Research Article

Comparison of extract bio-activities of in-situ and in vitro grown selected bryophyte species

Aneta Sabovljevi , Marina Sokovi , Jasmina Glamo lija, Ana iri , Milorad Vuji i , Boris Pejin and Marko Sabovljevi *

Abstract: The antimicrobial activity of DMSO extracts of three bryophyte species, two mosses and a liverwort (Atrichum undulatum (Hedw.) P. Beauv., Marchantia polymorpha L. ssp. ruderalis Bischl. and Boisselier, Physcomitrella patens (Hedw.) Bruch and Schimp.) grown in nature and in axenic culture was evaluated by microdilution method against eight bacterial species ( Escherichia coli ATCC 35210, Pseudomonas aeruginosa ATCC 27853, Salmonella typhimurium ATCC 13311, Enterobacter cloacae (human isolate), Listeria monocytogenes NCTC 7973, Bacillus cereus (human isolate), Micrococcus flavus ATCC 10240 and Staphilococcus aureus ATCC 6538). All investigated bryophyte extracts are proved to be active against all bacteria tested. In general, extracts made from material grown in nature express better anti- bacterial activity comparing to those made from material grown in laboratory conditions. Some of the bacteria react the same to both extracts and some even better to the extracts made from axenically grown material.[...] Read More.

Keywords: Bryophytes, antibacterial activity, Atrichum undulatum, Marchantia polymorpha, Physcomitrella patens.

2016, 15(1), 1-6;

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Research Article

The Copepod parasite of the gills of four teleost fishes caught from the gulf of Annaba (Algeria)

Chahinez Boualleg, Hadda Ferhati, Nouha Kaouachi, Mourad Bensouilah and Sonia Ternengo

Abstract: The examination of the gills of 960 fishes belonging to four species: Diplodus annularis, Pagellus erythrinus, Lithognatus mormyrus and Mullus barbatus collected from the Gulf of Annaba, enabled us to identify 13 species of parasitic copepods: Caligus ligusticus, Caligus diaphanus, Clavellotis sp, Clavellotis sargi, Clavellotis pagri, Clavellotis strumosa, Lernaeolophus sultanus, Hatschekia pagellibogneravei, Hatschekia sp, Hatschekia mulli, Sparidicola lithognathi, Neobrachiella exigua and Alella macrotrachelus. The distribution of copepods identified, varies from one host species to another and one season to another. P. erythrinus and L. mormyrus present the important number of copepods species. However, the winter presents the highest parasitic diversity (13). Furthermore, the majority of oixenous species are counted. The rates of most values are recorded in summer (33%) and spring (25%). It is also, D. annularis which shelters more of the 2/3 population of copepods collected. The copepod H. pagellibogneravei is the most abundant. The evaluation of parasitic indices shows that, it is during the summer and spring that the maximum values are recorded.[...] Read More.

Keywords: Copepod gills, Gulf of Annaba, teleost fish, season.

2016, 15(1), 1-6;

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Research Article

Application of PCR-RFLP of gap gene method as a molecular typing tool for coagulase negative Staphylococci from bovine and human origin identified with VITEK 2

Emel Banu Buyukunal Bal*, Mehmet Ali Bal, Taner Isevi and Erkan Yula

Abstract: The aim of this study was to apply the Restriction Fragment Length Polymorphism (RFLP) of Glyceraldehyde-3-Phosphate Dehydrogenase encoding gene (gap) for testing its performance as a molecular typing tool in coagulase negative staphylococci (CNS) isolates from bovine mastitis (n = 59) and human clinical cases (n = 13) identified with VITEK 2. According to the phenotypic identification results, bovine mastitis isolates were Staphylococcus haemolyticus, Staphylococcus simulans, Staphylococcus auricularis, Staphylococcus warneri, Staphylococcus hominis, Staphylococcus capitis, Staphylococcus xylosus, Staphylococcus epidermidis and Staphylococcus cohnii. Although most of those isolates were generated PCR amplicons with gap gene specific primers, PCR amplification of gap gene failed in 29 from 72 isolates. The samples that did not produce amplicons were reamplified with Staphylococcal 16S rRNA gene specific primers. After PCR amplifications, amplicons were produced in 17 from 29 samples. Three different restriction endonucleases (AluI, MseI and RsaI) were used for PCR-RFLP analysis, among these AluI has been found the most discriminatory power for identification in species. The results of the RFLP of gap gene provide a support for the misidentification problem associated with VITEK 2 system for S. simulans, S. auricularis and S. capitis species. Moreover, more frequent failure in gap gene amplification for bovine isolates which were phenotypically identified as S. simulans, S. auricularis, S. capitis, S. xylosus and S. cohnii was not clear. In addition, the method verified the phenotypic identification for S. haemolyticus, S. warneri, S. hominis and S. epidermidis isolates with different rates at 100, 33.3, 57.1, and 66.7%, respectively.[...] Read More.

Keywords: Coagulase negative staphylococci, gap gene, PCR-RFLP.

2016, 15(1), 1-6;

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Research Article

Detection of adhesin genes and slime production among Staphylococci in orthopaedic surgical wounds

Nizami Duran*, Yunus Dogramaci, Burcin Ozer, Cemil Demir and Aydiner Kalaci

Abstract: This study was aimed at investigating: (i) three adhesin genes (clf A, fnb A and cna) in Staphylococus aureus strains, (ii) the presence of slime (ica A and ica D genes) in both Staphylococus epidermidis and S. aureus strains isolated from surgical wounds. The slime and adhesin genes were detected by multiplex PCR. The ica A/ica D positivity rates were determined as 66.2% (104/157) in a total of 157 staphylococcal strains. While the occurance rate of slime genes was 69.6% (48/69) among the S. epidermidis, this ratio was 63.6% (56/88) among the S.aureus isolates. No statistically significant difference was found between S. epidermidis and S. aureus isolates in terms of the presence of slime genes (p > 0.05). Among the 88 S. aureus strains, almost all of the strains were positive for fnb A gene (97.7%). The cna and clf A positivity rates were detected in 69 (78.4%) and 45 (51.1%) isolates, respectively. The ica A and ica D genes responsible for slime production have been found to have high prevalence. Also, the frequency of adhesin genes was determined at a high rate in S. aureus strains isolated from surgical wounds. Molecular identification of virulent staphylococcal strains may help in management in clinical decision making.[...] Read More.

Keywords: Adhesins, slime, Staphylococus aureus, Staphylococus epidermidis, wound infection.

2016, 15(1), 1-6;

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Research Article

Systematic position of Moesziomyces penicillariae among Ustilaginaceae

Gnagna Diagne-Leye*, Teresa Almaraz-Lopez, Maïmouna Sy-Ndir, Gérard Mascarell, Amadou Tidiane Ba and Christophe Roux,

Abstract: We revisited the systematic position of the fungus Moesziomyces penicillariae, the causal agent of smut of pearl millet, by using morphological characters, germination pattern of teliospores and molecular analysis of ribosomal sequences. Samples of smutted ears of pearl millet were harvested in Senegal (West Africa). Compared to the description of Moesziomyces genus sensus Vánky, our samples presented morphological differences: i) presence of a columella-like structure in sori; ii) surface ornamentations of teliospores; iii) teliospore germination similar to Ustilago and Sporisorium ones. We investigated the systematic position of our samples by aligning their Internal Transcribed Spacer (ITS) sequences of the ribosomal regions with 47 sequences from Ustilaginaceae. The resulting tree rooted with Tolyposporium junci allowed the separation of five groups among which, they are, two Ustilago and two Sporisorium. An independent clade is formed by Tranzscheliella williamsii and Tranzscheliella hypodytes species including Ustilago sparti. Moesziomyces species used in this analysis form a monophylletic group located in Ustilago 2 group, which include different Ustilago and Sporisorium species but also Pseudozyma antartica. Our results indicate the necessity to amend the Moesziomyces genus as the morphological and molecular data confirm that they are included in the Ustilago-Sporisorium complex.[...] Read More.

Keywords: Pearl millet smut, Moesziomyces penicillariae, Ustilaginaceae.

2016, 15(1), 1-6;

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Research Article

Creation of alternative energy by bio- ethanol production from pineapple waste and the usage of its properties for engine

A. B. M. S. Hossain* and A. R. Fazliny

Abstract: The world is facing problems of fossil fuels depletion. The problem of using fossil fuels can have an environmental impact due to the oxidation of CO2, SO2 and NOX emission. In order to overcome those problems, there is an alternative fuel that can replace fossil fuel such as bioethanol and biodiesel. Bioethanol can be derived from biomass with many different varieties of feedstocks such as corn, sugarcane, wood and fruits wastes that are renewable and sustainable resources, which are easily accessible and reliable and can help to clean the environment from the wastes. The experiment was conducted to prove that, bioethanol can be produced from rotten pineapples waste through the biochemical reaction called fermentation by commercial yeast, Saccharomyces cerevisiae which is suitable for running petrol engine. The influencing parameters that affect the production of bioethanol from pineapples wastes were optimized. The effects of pH, temperatures, fermentation period, substrate concentration with and without water, components of pineapples from rotten and fresh ones were investigated. From the results, the optimal yield of bioethanol in the parameters such as pH, temperatures, fermentation period, substrate concentrations was found to be 8.7% having pH 4 at 30°C using 3 g/l. The result for viscosity was found under American Society for Testing Materials (ASTM) standard in different concentration of yeast. The anhydrous ethanol was analyzed and it was found that, there was no dangerous element in it’s acceptability as a transportation fuel based on ASTM standard. The elements that were mostly contained in the samples of ethanol production from pineapples wastes were Fe, Cu, Sn, Mn, Ag, Mo, Zn, P, Ca, Mg, Si, Na, B and V, but there was no significant difference among all elements. Key word: Bioethanol, waste pineapple, yeast, temperature, viscosity.[...] Read More.

Keywords: Bioethanol, waste pineapple, yeast, temperature, viscosity.

2016, 15(1), 1-6;

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Research Article

Emulsifying bacteria in produce water from Niger-Delta, Nigeria

S. A. Balogun* and O. E. Fagade

Abstract: The study reports the diversity of emulsifying bacteria in produce water obtained from Bonny Oil Terminal, Niger Delta, Nigeria. Physico–chemical analysis of produce water showed a high chloride ion content of 650.0 mg/L. Total aerobic mesophilic bacterial count was 8.6 x 106 CFU/mL while the oil utilizing bacterial count was 1.32 x 106 CFU/mL. The predominant bacteria genera were Bacillus, Pseudomonas, Serratia and Klebsiella. The ratio of the surface active agent producers in the mesophilic bacteria was 0.34%, while 2.2% was recorded among the oil utilizing bacteria. The highest emulsification and de-emulsification indices of 65 and 50% recorded respectively were for Pseudomonas mallei. The result obtained was discussed in relation to the use of the emulsifying bacteria in microbial enhanced oil recovery (MEOR) in the Niger Delta, Nigeria.[...] Read More.

Keywords: Emulsifying bacteria, emulsification-index, surface-active agents, biosurfactants, MEOR, crude oil, Niger-Delta.

2016, 15(1), 1-6;

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Research Article

A survey on epidemiology of urinary tract infections and resistance pattern of uropathogens in an Iranian 1000-bed tertiary care hospital

Ava Behrooozi, Mohammad Rahbar * and Jalil Vand Yousefi

Abstract: Urinary tract infections (UTIs) are one of the most common infectious diseases diagnosed in communities and hospitalized patients. The aim of this study was to determine frequency of occurrence and antimicrobial susceptibility patterns of uropathogens in Milad hospital of Tehran, Iran. In a prospective study from March to June 2009, a total of 11308 urine sample from patients admitted in Milad hospital of Tehran were analyzed. All specimens were inoculated on routine culture media. Bacterial isolates were identified by conventional bacteriological methods. Susceptibility testing was performed by standard methods as recommended by clinical laboratory standard institute. 11308 urine samples were cultured and 1020 pathogen were isolated. Escherchia coli with 620 (60.78%) isolates was the most common causative agent of UTI followed by Klebsiella pneumoniae with 115 (11.27%) isolates. Among gram positive Cocci Enterococcus spp with 110 (10.78%) isolates and Staphylococcus aureus with 81 (7.94%) isolates were predominant organisms. Of 1020 patients, 227 (22.25%) were male and 793 (77.74%) were female. Of 1020 patients, 224 (21.96%) of patients were hospitalized and 796 (78.03%) were outpatients. Of 224 hospitalized patients, 85% of isolates of E. coli were resistant to ampicillin, while this figure was 90% for K. pneumoniae. Resistant to other antibiotics were also prevalent. Nitrofurantoin was the most effective antibiotics against E. coli and Enterococcus spp. In conclusion, our study revealed that bacterial resistance in uropathogens in our hospital continues to be a great problem and needs drug resistance surveillance periodically.[...] Read More.

Keywords: Urinary tract infections, drug resistance.

2016, 15(1), 1-6;

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Research Article

Mitochondrion activity and dispersal of Aspergillus fumigatus and Rhizopus oryzae

Desmond M. Ncango, Chantel W. Swart, Carolina H. Pohl, Pieter W. J. Van Wyk and Johan L. F. Kock*

Abstract: We investigated the effects of anti-inflammatory and anti-mitochondrial compounds on spore dispersal in the pathogens, Aspergillus fumigatus and Rhizopus oryzae. When acetylsalicylic acid (ASA) and other non steroidal anti- inflammatory drugs (NSAIDs) were added to bio-assays of A. fumigatus and R. oryzae, spore-releasing structures were targeted first at lower concentrations. Similar results were obtained when oxygen was limited. These spore-releasing structures contained increased levels of mitochondrion activity compared to hyphae. We concluded that increased mitochondrion activity is necessary for dispersal of A. fumigatus and R. oryzae.[...] Read More.

Keywords: Aspergillus fumigatus, bio-assay, mitochondrion activity, non steroidal anti-inflammatory drugs, Rhizopus oryzae, spore-releasing structure.

2016, 15(1), 1-6;

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Research Article

Isolation and molecular characterization of phosphate solubilizing Enterobacter and Exiguobacterium species from paddy fields of Eastern Uttar Pradesh, India

Arvind Kumar, Poonam Bhargava and Lal Chand Rai*

Abstract: Six phosphate solubilizing bacteria (PSB) were isolated from paddy fields of Eastern Uttar Pradesh, India harboring low available phosphorus. Taxonomic delineation employing morphological, biochemical, 16S rRNA gene sequences and phylogenetic affiliations suggests that they are members of Enterobacter and Exiguobacterium genera. Of the six isolates, Enterobacter sp. LCR1 and LCR2 exhibited high level (568 - 642 g/ml) of phosphate solubilization in NBRIP liquid medium. Exiguobacterium sp. LCR4 and LCR5 showed increased phosphate solubilization efficiency under alkaline pH while Enterobacter sp. LCR3 remained unaffected. At high salt and temperature, Enterobacter sp. LCR1 and LCR2 produced 1.6 fold soluble phosphorus in comparison with earlier studies. Thus, these isolates may be useful for the development of potential bio-inoculants for soils having alkaline pH, high salt, temperature and insoluble phosphorus.[...] Read More.

Keywords: Phosphate solubilizing bacteria, phylogenetic analysis, Enterobacter spp., Exiguobacterium spp., 16S rRNA gene sequencing.

2016, 15(1), 1-6;

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Research Article

Screening, identification and statistic optimization of a novel exopolysaccharide producing Lactobacillus paracasei HCT

Rihua Xu, Shimin Ma, Yang Wang, Lisha Liu and Pinglan Li*

Abstract: The exopolysaccharides (EPS) producing ability was evaluated by phenol -sulphuric acid method and the Lactobacillus HCT was the highest yield EPS-producing strain, which was isolated from feces of Bama centenarian in Guangxi of China. It was identified as Lactobacillus paracasei HCT with carbohydrate assimilation profiling and 16S rRNA gene sequencing. L. paracasei HCT was found to be a novel EPS producing strain. Preliminary one factor tests were employed to obtain the favorable conditions for EPS yield and microorganism growth in a chemically defined medium and found that carbon nitrogen ratio (C/N ratio), cultivation time and temperature had the most significant influences. Box–Behnken experimental design and response surface methodology (RSM) were adopted to further study the interactive effects of these three variables on EPS yield and cells growth. The optimal culture conditions for EPS production were: C/N ratio 9.090, cultivation time 60.67 h and temperature 29.2°C. In these conditions, the maximum EPS yield was 39.0736 mg/ml which was 4 times more than the original yield. But the optimal conditions for cell growth were: C/N ratio 8.643, 58.75 h and 31.9°C, logarithm value of viable cell count reached 7.921. These predicted values were also verified by validation experiments.[...] Read More.

Keywords: Lactobacillus paracasei, exopolysaccharides, chemically defined medium, Box-Behnken design.

2016, 15(1), 1-6;

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Research Article

Observation on the occurrence and transmission pattern of Salmonella gallinarum in commercial poultry farms in Ogun State, South Western Nigeria

Agbaje M.*, Davies R., Oyekunle M. A., Ojo O. E., Fasina F. O. and Akinduti P. A.

Abstract: Reports of heavy mortality in three commercial layer poultry farms and two meat-type turkey farms in Ogun State were received separately between March and December, 2008 at the Veterinary Teaching Hospital (VTH), University of Agriculture, Abeokuta, Nigeria. We investigated the cause of mortality, isolated and serotyped the aetiological agent, and carried out antimicrobial susceptibility testing. All five isolates serotyped were Salmonella Gallinarum and showed identical patterns of resistance and susceptibility to all antimicrobials used. The occurrence of ciprofloxacin resistance in all five isolates is of importance since fluoroquinolone resistance has implications for both veterinary and human therapy and the abuse of such medications in poultry could result in the emergence of resistant zoonotic organisms.[...] Read More.

Keywords: Occurrence, transmission pattern, Salmonella gallinarum poultry farms.

2016, 15(1), 1-6;

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Research Article

Evaluation of salivary gland scintigraphy, magnetic resonance and diffusion-weighted imaging in clinical diagnosis of SjÃ¶grenâ€™s Syndrome

Mei Li, Hao-Jie Dai, Zhen-Chang Wang*, Zhen-Gang Wang, Li Cui, Qing-Gang Xu and Jing Li

Abstract: To assess diagnostic effectiveness of Sjögren’s syndrome (SS), salivary gland scintigraphy (SGS), magnetic resonance (MR) and diffusion-weighted imaging (DWI) were performed in 54 patients (primary SS, secondary SS and connective tissue disease (CTD) but without SS). Results showed that SGS and diffusion-weighted magnetic resonance imaging (MRI) provided high-resolution images of the parotid glands that enabled apparent diffusion coefficient (ADC) measurements. After overall analysis, all of the oral activity indices, the maximum accumulation (MA) and uptake rate (UR) of the parotid gland clearly decreased as clinical severity progressed, and statistically significant differences were observed. Our technique may prove helpful in diagnosing the progression of SS beyond its early stages.[...] Read More.

Keywords: Diffusion-weighted imaging, salivary gland scintigraphy, Sjögren’s syndrome, receiver operating characteristic curve.

2016, 15(1), 1-6;

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Research Article

Differentiation of Salmonella enterica based on PCR detection of selected somatic and flagellar antigens

Mohd Elbagir Elhassan Nori and Kwai Lin Thong*

Abstract: Serotyping is the basis of Salmonella surveillance. However, the limitations of the traditional serotyping have stimulated rapid research and development in DNA-based serotyping. The aim of this study was to apply a combination of sequential multiplex PCRs targeting the O, H and Vi antigens to serotype a panel of 122 recently isolated human- and foodborne- Salmonella strains. The mPCR that targets the o (genes wzxC2, rfbJ, prt, tyv, wzxE, wzxC1, prt) and Vi (viaB) antigens successfully subtyped the strains into serogroups C2 (n = 35, 28.7%), B (n = 33, 27.1%), D9 (n = 29, 22.9%), E (n = 21, 17.2%), C1 (n = 2, 64%) and A (n = 2, 1.64%). Eight of the Salmonella strains from serogroup D were positive for Vi antigen. Two multiplex PCRs were optimized for detection of H1 antigens (Ha, Hb, Hd, r, z10, z6, g and m) and H2 antigens (1.5, 1.2, 1.6 and enx). Overall, the multiplex PCRs of O, H and Vi antigens results correctly serotyped 94 of 122 strains (77%). The most frequent serovars encountered were Salmonella weltevrerden, Salmonella enteritidis, Salmonella typhimurium, Salmonella hadar and Salmonella typhi. Application of DNA based technique for serogrouping and serotyping of the selected Salmonella enterica was found to be robust, quick, specific and reliable for the specific antigenic targets and is useful in the study area which lack complete serotyping facilities.[...] Read More.

Keywords: Salmonella, somatic antigens (O), flagella antigens (H), serotyping, multiplex PCR.

2016, 15(1), 1-6;

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Research Article

Characterisation of non human origin probiotic Lactobacillus plantarum with cholesterol-lowering property

Sasithorn Sirilun, Chaiyavat Chaiyasut*, Duangporn Kantachote and Plearnpis Luxananil

Abstract: This study evaluated the probiotic and cholesterol-lowering property of Lactobacillus spp. isolated from non human origin. Four strains, TGCM 15, TGCM 26, TGCM 33 and TGCM 128 were selected and identified as Lactobacillus plantarum. These strains tolerated to 0.15% and 0.30% (w/v) bile salt and resisted to pH values 2 - 8 with survival rate more than 50% during 2 h of growth. In addition, all strains exhibited strong antimicrobial activities against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 25923, Bacillus cereus ATCC 11778, Pseudomonas aeruginosa ATCC 27853, Salmonella typhi, Shigella sonnei and Candida albicans ATCC 90028. TGCM 15 and TGCM 33 exhibited the bile salt hydrolase (BSH) activity and cholesterol-lowering properties with the reduction rate more than 50% by active cells. In particular, TGCM 15 exhibited the significantly highest cholesterol-lowering activity at 81.46%. Conversely, TGCM 26 was determined to have the significantly lowest activity at 25.41%. The percentages of cholesterol-lowering by resting and dead cells of TGCM 15 and TGCM 33 were not significantly different. The resting and dead cells of all strains exhibited the cholesterol-lowering activities in the range of 13.11 - 23.28 and 11.44 - 19.53%, respectively. According to these results, the 4 strains of L. plantarum have revealed the probiotic’s potential in cholesterol-lowering property.[...] Read More.

Keywords: Probiotic, Lactobacillus plantarum, bile salt hydrolase, cholesterol-lowering property.

2016, 15(1), 1-6;

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Research Article

Culture-dependent and culture-independent approaches to study the bacterial and archaeal diversity from Jordanian hot springs

Hanan I. Malkawi* and Manal N. Al-Omari

Abstract: The microbial diversity was investigated at five major hot springs in Jordan (Ashounah, Waggas, Zara, Zarqa Ma'in and Afra springs) using both microbiological culture-based and molecular culture-independent approaches. One hundred and thirty two (132) bacterial isolates were obtained and characterized morphologically. Out of 132 isolates, 125 isolates were gram positive rods, while the other seven isolates were gram positive coccobacilli. The bacterial growth at different high temperatures was determined, and revealed that 19 out of 132 isolates were able to grow at high temperatures of up to 75°C. All bacterial isolates grew at pH 7.0, except one single isolate from Zarqa Ma'in spring which was able to grow at pH 3.5. Culture-independent approaches using polymerase chain reaction (PCR) was used for the amplification of specific 16S rDNA sequences of Bacteria, Archaea, Green sulfur bacteria, Green nonsulfur bacteria, Heliobacteria, and methanogenic Archaea from metagenomic DNA extracted directly from water and mat samples from each thermal spring. Nine water samples and 9 mat samples from all spring sites revealed the presence of bacteria yielding the amplicon size of (1500 bp), while 4 water samples from Zarqa Ma'in springs and 5 mat samples from Zarqa Ma'in springs and Waggas well revealed the presence of archaea with an amplicon size of (650 bp). When the primer pair targeting purple phototrophic bacteria was used, positive PCR amplification product (229 bp) appeared in 6 mat samples from Zarqa Ma'in, Afra, Zara, Ashounah and Waggas springs. Also PCR identification of bacterial isolates using primer pair specific to the 16S rDNA gene sequences of the genus Bacillus indicated that 96.97 % (128 out of 132) of bacterial isolates have the size of PCR amplicon (320 bp) for the genus Bacillus.[...] Read More.

Keywords: Bacteria, archaea, diversity, hot springs, Jordan.

2016, 15(1), 1-6;

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Research Article

Early detection of ovarian carcinoma by proteome profiling based on magnetic bead separation and matrix-assisted laser desorption/ionization time of flight mass spectrometry

Dong-Mei Fan, Hui-Rong Shi *, Zhi-Min Chen, Qing-Hua Wu, Hui-Na Liu and Rui-Tao Zhang

Abstract: In this study, affinity purification with magnetic beads and matrix-assisted laser desorption/ionization time of flight mass spectrometry analysis was used to screen candidate biomarkers of ovarian carcinoma. Mass spectrometry compiled from serum of 30 cases of ovarian carcinomas was compared with that from serum of 10 cases of benign group and13 cases of normal group. Interesting peaks were further sieved from ascites and cystic fluid. Peptides were deduced by searching in http://us.expasy.org/tools/tagident.html and were detected by immunohistochemistry and enzyme-linked immunosorbent assay. Results showed that two peaks of 2881 and 2897Da elevated in the serum, malignant ascite and cyst fluid of cancer group when compared with normal and benign groups were deduced as gastrin -releasing peptide which was higher in the malignant tissues and serum than other two groups. It was concluded that gastrin-releasing peptide in serum might be a potential marker for ovarian carcinoma.[...] Read More.

Keywords: Gastrin-releasing peptide, matrix-assisted laser desorption/ionization time of flight mass spectrometry, magnetic bead, and ovarian carcinoma.

2016, 15(1), 1-6;

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Research Article

Demineralized bone matrix gelatin as scaffold for tissue engineering

Yong Pan, Shiwu Dong, Yong Hao, Tongwei Chu, Changqing Li, Zhengfeng Zhang and Yue Zhou*

Abstract: The aim of the present study was to do a comparative light microscopical and histochemical analysis of bone regeneration processes in rabbits, with the use of demineralized bone matrix gelatin scaffolds. Our histological results showed that no gelatin scaffold was extruded, indicating that neither body fluid nor blood swept the gelatin scaffold from the implantation site, nor was the gelatin scaffold biodegraded. Additionally, no abscess or inflammation of the peripheral osseous tissues at the implantation site was observed, suggesting that the implantation of the gelatin scaffold in the vertebra bone defect did not cause histopathology or exhibit malbiocompatibility with the peripheral osseous tissues. Key word: Demineralized bone matrix gelatin scaffolds, tissue engineering.[...] Read More.

Keywords: Demineralized bone matrix gelatin scaffolds, tissue engineering.

2016, 15(1), 1-6;

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Research Article

Isolation of Escherichia coli O157:H7 from manure fertilized farms and raw vegetables grown on it, in Tabriz city in Iran

Jalil Khandaghi*, Vadood Razavilar and Abolfazl Barzgari

Abstract: Concern for pathogens in vegetables has risen because of increasing number of outbreaks of foodborne illnesses caused by consumption of fresh and minimally processed vegetables. Using bovine feces as fertilizer, which is the common organic fertilizer on farmlands in Iran, can pose risk of spreading and transmission of enteric pathogens such as Escherichia coli O157:H7 both by direct contact and consumption of produces grown on such farms. We obtained a total of 282 samples of soil and vegetables (lettuce, cabbage, carrot and radish sprout), from manure fertilized agriculture farms in Tabriz city. Phenotypic/biochemical characteristics followed by biochemical tests were performed. Multiplex PCR technique was used to confirm E. coli O157:H7 isolates. The data indicated that 5 samples of soils (1.77 %) and one sample of vegetable (0.35 %) of total samples were contaminated with E. coli O157:H7. Organic fertilizer used in farm lands in this study poses a supposed low risk in vegetables.[...] Read More.

Keywords: Foodborne, bovine feces, Escherichia coli O157:H7, raw vegetables, multiplex PCR.

2016, 15(1), 1-6;

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Research Article

Frequency of extended spectrum beta-lactamase (ESBLs) producing Escherichia coli and klebseilla pneumonia isolated from urine in an Iranian 1000-bed tertiary care hospital

Ava Behrooozi, Mohammad Rahbar* and Jalil Vand Yousefi

Abstract: Betal-lactam antibiotics are the most frequently prescribed antibiotics. Emerging resistance to these antibiotics among gram-negative bacilli limited their utility. This study was done to determine the frequency of extended spectrum beta-lactmase (ESBLs) producing Escherichia coli and Klebsiella pneumoniae isolated from urine in Milad Hospital of Tehran. This study was done on 735 urine gram-negative bacilli isolates including 620 strains of E. coli and 115 strains of K. pneumoniae in microbiology laboratory of Milad Hospital in Tehran, Iran. ESBls resistance was detected in 132 (21%) of isolates of E. coli and 18 (12%) in those with K. pneumoniae. Of 150 patients which had positive ESBLs isolates, 104 were outpatients and 46 others were hospitalized. Nearly 80% ESBls isolates were resistant to co-trimoxazole and nitrofuantion was the most effective antibiotic against ESBLs producing isolates. Our study revealed that there is a high frequency isolates of ESBls producing strains of E. coli and K. pneupmiae in both community and hospital. This has a significant implication for patients’ management. Further drug resistance surveillance in our hospitals and molecular characteristics of ESBls isolates in our country is necessary.[...] Read More.

Keywords: Extended spectrum beta-lactamase (ESBLs), Escherichia coli, Klebseilla pneumonia, urinary tract infections.

2016, 15(1), 1-6;

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Research Article

Monogenean parasites in Sparid fish (Pagellus genus) in eastern Algeria coastline

Nouha Kaouachi*, Chahinez Boualleg, Mourad Bensouilah and Bernard Marchand

Abstract: A spatial distribution study of parasite populations of gills from 540 specimens belonging to Pagellus genus: Pagellus acarne, Pagellus erythrinus and Pagellus bogaraveo (Sparidae) was realized from the eastern Algeria coastline. A preliminary description allows the distinguishing of eight parasite species belonging to two subclasses: Monopisthocotylea and Polyopisthocotylea. Rates of parasite species and parasitic indices of host species were calculated. The study findings show that the parasitic populations vary depending on the site and the host species within the same genus.[...] Read More.

Keywords: Parasitism, monogenean, Pagellus erythrinus, Pagellus acarne, Pagellus bogaraveo, Algerian coastline.

2016, 15(1), 1-6;

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Research Article

Occurrence of tetracycline-resistant Aeromonas hydrophila infection in Korean cyprinid loach (Misgurnus anguillicaudatus)

Jin Woo Jun, Ji Hyung Kim, Dennis K. Gomez, Casiano H. Choresca Jr, Jee Eun Han, Sang Phil Shin and Se Chang Park*

Abstract: In this paper, we described a mass mortality of cyprinid loaches, Misgurnus anguillicaudatus from a private fish farm in Korea. Diffuse bleeding was observed on the skin around the anal orifice. Bacterial pathogens from kidneys samples of moribund fish were cultured, identified and confirmed to be Aeromonas hydrophila using morphological, biochemical test and genetical analysis. The isolated A. hydrophila strains were resistant to commercial antibiotics and the presence of the tetracycline resistance gene (tet E) was detected by genetical analysis. Pathogenecity test was performed using healthy 4-week-old cyprinid loaches by intraperitoneal (IP) injection and the LD50 concentration of the pathogen was determined to be 6.0 × 107 CFU fish-1. In this paper, we confirmed the infection of farm-raised cyprinid loaches with tetracycline-resistant A. hydrophila.[...] Read More.

Keywords: Cyprinid loach (Misgurnus anguillicaudatus), Aeromonas hydrophila, tetracycline resistance gene, pathogenecity test.

2016, 15(1), 1-6;

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Research Article

Bacteria population of some commercially prepared yoghurt sold in Enugu State, Eastern Nigeria

Nwagu Tochukwu Nwamaka* and Amadi Emmanuel Chike

Abstract: The demand for fermented dairies is on the increase in developing countries following consumer awareness of some of the health benefits attributed to prolonged intake. Eight samples of yoghurt produced from Enugu, Eastern Nigeria was collected from various locations. Samples were incubated using various media for the isolation, identification and enumeration of the bacteria population within the yoghurt. Reports indicated that total viable count of bacteria was in the range of 1.4 x 106 – 2.2 x 107 cfu /ml. Not all bacterial isolates were lactose fermenters. Members of the genus Staphylococcus, Aeromonas, Klebsiella, Pseudomonas, Bacillus, Streptococcus and Lactobacillus were isolated with 100, 25, 50, 33.3, 25, 12.5 and 25% frequency of occurrence, respectively. Viable counts of lactic acid bacteria were low indicating that probiotics effect following consumption may be poor. Occurrence of pathogenic organisms indicates improper handling and inadequate sanitary measures. More care should be taken during yoghurt fermentation.[...] Read More.

Keywords: Lactobacillus, fermentation, yoghurt, bacteria, contaminant.

2016, 15(1), 1-6;

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Research Article

Influences of light and temperature on membrane potential and respiratory viability of an aerobic anoxygenic phototrophic bacterium Erythrobacter sp. JL475

Zaiqing Peng and Nianzhi Jiao*

Abstract: Flow cytometry’s (FCM) measurement of membrane potential (MP) and cell respiration viability based on continuous culture was used to investigate the responses of aerobic anoxygenic phototrophic bacteria (AAPB) in the heterotrophic growth and regulation mechanism of photosynthesis to environmental changes. An AAPB strain Erythrobacter sp. JL475 and a non-AAPB strain Erythrobacter sp. JL316 were used as the experimental bacteria, both of which were isolated from the South China Sea. The results showed that light -cultured AAPB showed higher MP and biomass at 10°C, suggesting an obvious stimulation of light on AAPB growth. By contrast, dark-cultivated JL475 had higher MP and biomass at higher temperature (20, 30 and 40°C). The rate of heterotrophic respiration at different temperature environment ranked as follows: dark-cultivated JL316 > dark-cultivated JL475 > light/dark cycling cultivated JL475. Light undoubtedly increased the cell viability of AAPB, especially of apoptosis cells. The CTC+% at different carbon concentration ranked as follows: light/dark cycling cultivated JL475 > dark-cultivated JL316 > dark-cultivated JL475. It was concluded that the heterotrophic respiration would played a key role in energy metabolism of AAPB, photosynthesis may provide an advantage for AAPB to survive in a variety of diverse environments.[...] Read More.

Keywords: Flow cytometry, aerobic anoxygenic phototrophic bacteria, membrane potential, respiration viability, biomass.

2016, 15(1), 1-6;

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Research Article

Isolation, recombinant expression and characterization of the dprA gene product in Streptomyces rimosus NRRL 2455

Khaled Mohamed Anwar Aboshanab

Abstract: Dihydrodipicolinate synthase (DHDPS) catalyses the first enzymatic step in the lysine-biosynthetic pathway in bacteria, plants and some fungi. In this study, cloning and submission of 3639 bp from Streptomyces rimosus NRRL 2455 to the EMBL database under the accession code EU617017 were carried out. The entire dprA gene was cloned and heterologously expressed in Escherichia coli BL21 (DE3) and the gene product was purified and analysed spectrophotometrically. A dprA- knock-out mutant was created and showed a reduction of mycelia growth and spore formation by about 43 and 37%, respectively, as compared to the wild strain. Fed with meso-diaminopimelic acid, the dprA- mutant regained its capability of mycelial growth, however no significant effect on spore formation was observed confirming that dprA gene product was involved in the biosynthesis of dihydrodipicolinate in S. rimosus NRRL 2455. This is the first report about identification of dihydrodipicolinate synthase of the order streptomycetes.[...] Read More.

Keywords: Dihydrodipicolinate synthase, lysine biosynthesis, Streptomyces rimosus NRRL 2455, dprA knock-out mutants

2016, 15(1), 1-6;

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Research Article

Research advance and superiority of microdiesel production with biowastes

Chuan-chao Dai*, Jie Tao, Yu Wang and Yu-min Hao

Abstract: Lignocellulosic biowastes can be utilized to produce microdiesel, a promising alternative energy source for limited crude oil and food based biofuels. As an alternative fuel, microdiesel produced from biowastes it has superior environmental benefits, economically competitive, producible in sufficient quantities and the energy conversion efficiency is also estimated in the article in theory. Microdiesel derived from biowastes is a potential renewable and carbon neutral alternative to petroleum fuels. Unfortunately, microdiesel from biowastes cannot realistically satisfy even a small fraction of the existing demand for transport fuels. As demonstrated here, microdiesel appears to be the only source of renewable biodiesel that is capable of meeting the global demand for transport fuels. Approaches for making microdiesel economically competitive with petrodiesel are also discussed.[...] Read More.

Keywords: Microdiesel, lignocellulosic biowastes, environmental benefits, potential supply.

2016, 15(1), 1-6;

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Research Article

Molecular identification and codon optimization analysis of major virulence encoding genes of Aeromonas hydrophila

Vijai Singh,*, Dharmendra Kumar Chaudhary, Indra Mani , Pallavi Somvanshi, Gaurav Rathore and Neeraj Sood

Abstract: Aeromonas hydrophila is an opportunistic pathogen of aquatic and terrestrial animals and human beings. The aim of this study is to investigate 25 isolates of A. hydrophila from 40 fish muscles and 12 water samples. All isolates are screened for the presence of virulence factors such as aerolysin, hemolysin and lipase both phenotypically as well as genotypically. All the isolates produced lipase, whereas only 60% of isolates produced b-hemolysis with RBCs. The presence of 3 virulence genes aerolysin, hemolysin and lipase were confirmed by PCR assay, which gave positive PCR amplification for conserved fragment. These were also codon optimized for over expression in Escherichia coli and obtained the high codon adaptation index in comparison to wild type of DNA sequences. These finding may help to better expression of gene for scale up production of vaccine candidates. Keywords: Aeromonas hydrophila, virulence factor, PCR, codon optimization, vaccine.[...] Read More.

Keywords: Aeromonas hydrophila, virulence factor, PCR, codon optimization, vaccine.

2016, 15(1), 1-6;

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Research Article

Isolation and characterization of Streptomyces species with antifungal activity from selected national parks in Kenya

James Oluoch Nonoh, Wilber Lwande, Daniel Masiga, Rafi Herrmann, James K. Presnail, Eric Schepers, Matilda Angela Okech, Richard Bagine, Paul Mungai, Aggrey Bernard Nyende and Hamadi Iddi Boga*

Abstract: Microorganisms and their natural products are potentially important for the biological control of crop diseases without detrimental effects to the environment. In this study, acetonitrile-methanol extracts of 361 actinobacterial isolates obtained from Aberdares, Arabuko Sokoke, Lake Bogoria, Mt Kenya, Kakamega, Ruma, Shimba Hills and Imenti forest national parks in Kenya were screened for antagonism against Fusarium oxysporum, Fusarium spp and Colletotrichum kahawae, which are important crop pathogens. Twenty-three isolates showed antagonistic activity to one or all of the test fungi. Five isolates that were antagonistic against all test fungi were investigated further and were also found to have antibacterial activity against Staphylococcus aureus and Escherichia coli. Morphological and physiological studies show that the isolates belong to streptomycetes. Phylogenetic analysis of amplified actinobacterial 16S rRNA gene confirmed that all the five antagonistic isolates formed close phylogenetic clusters with known members of Streptomyces species with a (97 - 100%) sequence identity. The results suggest that protected areas may be ideal habitats for isolation of antagonistic actinobacterial species which may have the potential for beneficial application in biological control of fungal pathogens. However, further investigation by characterization of the antifungal and antibacterial compounds produced will be necessary.[...] Read More.

Keywords: Protected areas, soil streptomyces, bio-prospecting, antimicrobial, phytopathogens.

2016, 15(1), 1-6;

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Research Article

Function analysis of the predicted protein-tyrosine phosphatases gene (DR2161) in Deinococcus radiodurans

Shu Haiyan* and Tian Baoming

Abstract: Protein phosphorylation on tyrosine has been demonstrated to occur in a wide array of bacterial species and appears to be ubiquitous among prokaryotes. In Deinococcus radiodurans, after the predicted protein-tyrosine phosphatase (PTP) gene DR2161 was deleted, the radiation resistance of the bacterium changed a little. The natural resistance-associated macrophage protein gene DR1709 was the possible target of PTP. But the radio resistance of the double mutant (DR1709 and DR2161 that were deleted) was almost the same as that of M1709. When the strain protease secretion was measured, the clearing area of M2161 was smaller than that of the wild type. But the protease secretion of the double mutant was similar to that of M1709. The influence on the bacterium DR2161, that was deleted seemed to have been covered by DR1709 being disrupted. The four strains (the wild type, M1709, M2161 and the double mutant) sensitivity to high concentrate of Mn2+ and Fe2+ were very similar, showing that they had similar resistance to high concentrate of Mn2+ or Fe2+. In liquid defined minimal medium (DMM) with 200 nM Mn, M1709 and the double mutant almost can not grow. But in DMM with 200 nM Fe, the two strains grew as quickly as the wild type. M1709 reaction to low concentrate of Mn2+ and Fe2+ was not affected by DR2161. The expression of DR1709 was not regulated by DR2161.[...] Read More.

Keywords: Deinococcus radiodurans, protein-tyrosine phosphatase, natural resistance-associated macrophage protein, DR1709, DR2161.

2016, 15(1), 1-6;

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Research Article

Escherichia coli phoA gene inactivation, insertion and restoration based on the red system

J. M. Wambale, Ni zhaohui , J. N. Kimatu and Fan Li*

Abstract: The red recombinase system of bacteriophage lambda has been used to inactivate chromosomal genes in Escherichia coli K-12 through homologous recombination using linear PCR products. Here, E. coli BW 25113 mutants were created by changing the localization of genes on the genome using a method which contains both inactivation and reinsertion steps. The phoA gene, the structural gene for alkaline phosphatase, was inactivated by having the ORF and its regulator region replaced with a kanamycin cassette flanked by FLP recognition target sites. The site for insertion was selected based on the intergenic distance and non-coding regions, thus the new neighboring genes sequences were not affected. The Insertion was carried out by designing primers armed with homologous sequences to the insertion site; the primers contained the enzyme restriction sites in order to ligate the gene PCR product with the FRT-kan -FRT marker prior to the electroporation. The PCR reactions, cultures in the media supplemented with appropriate antibiotics and comparative gene expression study were carried out to confirm the knockout, the insertion and the expression of the gene in its new location. This procedure and the created E. coli mutants can be efficiently used for the study of bacterial genomics, especially in systems biology to understand the relation between the genes loci on the genome and their expression alongside new neighboring genes and others with the same functional group.[...] Read More.

Keywords: Gene, operon, knockout, insertion, systems biology, electroporation.

2016, 15(1), 1-6;

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Research Article

Antimicrobial activity of the lichen Lecanora frustulosa and Parmeliopsis hyperopta and their divaricatic acid and zeorin constituents

Kosani Marijana*, Rankovi Branislav and Sukdolak Slobodan

Abstract: Antibacterial and antifungal activity of the acetone, methanol and aqueous extracts of the lichen Lecanora frustulosa and Parmeliopsis hyperopta and their divaricatic acid and zeorin constituents has been screened in vitro against the following species of microorganisms: Bacillus mycoides, Bacillus subtilis, Staphylococcus aureus, Enterobacter cloaceae, Escerichia coli, Klebsiella pneumoniae, Aspergillus flavus, Aspergillus fumigatus, Botrytis cinerea, Candida albicans, Fusarium oxysporum, Mucor mucedo, Paecilomyces variotii, Penicillium purpurescens, Penicillium verrucosum and Trichoderma harsianum. The antimicrobal activity was estimated by the disc-difusion method and determination of the minimal inhibitory concentration (MIC) by the Broth tube Dilution method. The bacteria were more sensitive related to the tested fungi. The smallest MIC values were 0.39 mg/mL against bacteria and 1.56 mg/mL against fungi. Acetone and methanol extracts of the investigated lichens showed relatively strong antimicrobial activity, whereas aqueous extracts were inactive. Divaricatic acid and zeorin also showed strong activity against bacteria and fungi. There was no antimicrobial activity against E. coli species. The lichen extracts maximum activity showed in the concentrations of 0.78 mg/mL and the lichen components demonstrated maximum activity in the concetrations of 0.39 mg/mL. Generally, the tested lichen extracts and lichen compounds demostrated a strong antimicrobial effect against the tested microorganisms. That suggest a possibility of their use in the treatment of various diseases caused by these and similar microorganisms.[...] Read More.

Keywords: Antimicrobial activity, lichen extracts, lichen components.

2016, 15(1), 1-6;

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Research Article

Rationalizing some medicinal plants used in treatment of skin diseases

Adebayo-Tayo B.C, Adegoke A.A, *Okoh, A.I and Ajibesin K.K

Abstract: The hydroethanolic extracts obtained from ten plant species being used as components of skin disease remedy in Southwest Nigeria were subjected to phytochemical analysis and screened for antimicrobial activity. The antimicrobial activity was determined against Escherichia coli NCIB 86, Staphylococcus aureus NCIB 8588, Klebsiella pneumoniae NCIB 418, Pseudomonas aeruginosa NCIB 950, Proteus vulgaris NCIB 67, Bacillus subtilis NCIB 3610, Candida albicans and Aspergillus flavus by agar diffusion method. Terpenes, flavonoids, tannins and saponin were detected in the active plants through phytochemical screening and confirmed by thin layer chromatography (TLC) . The plant extracts demonstrated antimicrobial effect against bacteria and the fungal cultures used in this study at different levels. The plant species Funtumia elastica, Raphyostylis beninensis, Butyrospermum paradoxum, Serataria caudula, Parkia biglobosa and Curculigo pilosa showed significant antimicrobial activities against the test organisms. Curculigo pilosa, F. elastic and P. biglobosa gave the highest zone of inhibition of 30 mm at 80 mg/ml against Aspergillus flavus, Bacilus subtilis and Proteus vulgaris while Curculigo pilosa also gave the least zone of inhibition of 2 mm at 80 mg/ml against Klebsiella pneumoniae. This relative high active index explains the antimicrobial effects of the remedy for skin diseases.[...] Read More.

Keywords: Phytochemical analysis, antimicrobial activity, TLC, skin disease, Funtumia elastica.

2016, 15(1), 1-6;

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Research Article

Effect of blood group and demographic characteristics on malaria infection, oxidative stress and haemoglobin levels in South Western Nigeria

O. M. Akanbi, J. A. Badaki, O. Y. Adeniran and O. O. Olotu

Abstract: Malaria infection generates oxidative stress which has serious effect on the haemoglobin (Hb) level of the infected individuals. This work studied the effect of blood group, age and gender on prevalence of malaria, oxidative stress and Hb level. 5 ml of blood samples were collected into EDTA bottle from 120 volunteered adult males and non-pregnant females. The plasma was separated and used to quantify oxidative stress by measuring malonaldehyde (MDA) and superoxide dismutase (SOD) levels using colorimetric method, while haematological parameter and malaria parasite screening was done using a whole blood. The parasite density and MDA levels were significantly higher (P < 0.05) in individuals with blood group A than those with blood group B and O. SOD and Hb levels were lower in those with blood group A than those with blood group B and O. The parasite density and MDA levels were significantly higher in age group 18 - 21 years than other age groups but SOD was higher (P < 0.05) in age group 22 - 25 and 26 - 30 than age group 18 - 21. The parasite density and MDA were higher (P < 0.05) in males than females in this study. The findings indicate that gender, sex and blood group have impact on malaria infection and oxidative stress.[...] Read More.

Keywords: Malaria, blood grouping, gender, oxidative stress, haemoglobin, age.

2016, 15(1), 1-6;

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Research Article

Genetic characterization of osmotolerant fermentative saccharomyces yeasts from Tanzania suitable for industrial very high gravity fermentation

D. Sumari ,*, K. M. M. Hosea and F. S. S. Magingo

Abstract: Naturally occurring yeasts were sought from diverse Tanzanian environments and screened for industrial application. The yeasts were isolated from environments such as traditional brews and wines from various parts of Tanzania. In this regard, a total of thirty yeast isolates were screened for their suitability in Industrial Very High Gravity Fermentation (VHGF). Five of these isolates were able to grow and ferment medium with 40% initial sucrose concentration. Whereby, three were able to grow and ferment medium with 700 g/Litre (70% w/w) initial sucrose concentration. One of the three isolates coded LB2 isolated from a traditional Makonde sorghum brew was able to ferment a medium with initial sucrose concentration of 1000 g/Litre (100% w/w) at 30°C. On the basis of PCR-RFLP of the internal transcribed spacer (ITS) of the ribosomal DNA (rDNA), all the three most osmotolerant isolates were identified to belong to the Saccharomyces sensu stricto clade. Phylogenetic analysis of the 650 bp D1D2 domain of the large subunit 26S rDNA of the isolate LB2 clustered this isolate away from the so far known typical osmotolerant yeasts. The fermentation by LB2 isolate of 100% gravity substrate observed in this work is higher than any other encountered in the literatures reviewed.[...] Read More.

Keywords: Osmotolerant, PCR-RFLP, Saccharomyces, traditional brews, very high gravity fermentation (VHGF).

2016, 15(1), 1-6;

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Research Article

Susceptibility pattern of pseudomonas aeruginosa against various antibiotics

Farida Anjum and Asif Mir*

Abstract: To determine current trends of antibiotic resistance among clinically significant pseudomonas aeruginosa strains causing various nosocomial and community acquired infections. A total of 100 clinical isolates of P. aeruginosa from inpatient and outpatient were studied from June 2008 to May 2009 for its prevalence and susceptibility profiles. Most of the isolates were from pus followed by urine, sputum, blood, ear swab and catheter tip. Uropathogenic P. aeruginosa infections were higher in females than males, ratio was found more among young and elderly debilitated patients. Ciprofloxacin, Piperacillin, Imipenem were found more effective for treatment of infections in outpatients but for inpatients, parental therapy with newer aminoglycosides and third and forth generation cephalosporins need to be advocated as the P. aeruginosa causing nosocomial infections exhibits a high degree of drug resistance. Ninety nine percent of the clinical isolates were resistant to six commonly used antibiotics with highest resistance to ampicillin (100%) and cefuroxime (100%) followed by amoxycillin (99%), co-trimoxazole (99%), Tetracycline (99%), Cefazoline (99%). The in vitro sensitivity pattern of 100 isolates of P. aeruginosa showed highest sensitivity to imipenem (97%) followed by amikacin (79%), tobramycin (70%), ceftazidime (62%), ciprofloxacin (73%), cefoperazone (60%), piperacillin (65%), Gentamycin (34%) and Cefotaxime (14%). ESBLs producing strains (33%) were also less in number but were much more resistant to -lactam and other antibiotics. The results indicate that P. aeruginosa is the most common gram -negative bacterium responsible for the nosocomial as well as community acquired infections. The excessive use of antibiotics has not only led to treat the P. aeruginosa infections but also the emergence of antibiotic resistance. The development of multidrug resistant P. aeruginosa is currently one of the greatest challenges to the effective management of infections. This suggests that in addition to curative measures promptly preventive measures such as hygienic as well as better hospital and postoperative care in administration should be adopted.[...] Read More.

Keywords: P. aeruginosa, antibiotic resistance, nosocomial infections.

2016, 15(1), 1-6;

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Research Article

Protective effects of sucralfate on anti-H. pylori VacA IgY in vivo and in vitro

Zhou Xun, Guo Li-yuan, Yang Zhibang* and Chen Xiaoping

Abstract: This study aimed to evaluate the protective effects of sucralfate on immunoglobulin yolk (IgY) and to provide a theory basis for preparing IgY oral preparations which can tolerate acid and pepsin. Engineering bacteria DH5 -vacA-pQE30 were largely induced to express recombinant vacuolate cytotoxin antigen (VacA). VacA antigen was inoculated in Lohman hen and IgY was purified. Different concentrations of sucralfate were added into IgY solution including different concentrations of pepsin at different pH values. IgY solution containing different concentrations of sucralfate underwent seven times of repeated freeze thawing, and then placed at room temperature for 1 d, 4 w. The IgY residual antibody activity (AR/AC) of each experiment group was detected by ELISA. Helicobacter pylori infected mice were intragastrically administrated with 30% sucralfate and different concentrations of IgY solution and then chronic inflammatory reaction of gastric mucosa was observed to evaluate the protective effects of sucralfate on IgY. At pH = 1.5, 60% sucralfate could maintain 48.9% IgY residual antibody activity. At pH = 2.0, 30% sucralfate could enhance the IgY residual antibody activity to 86.5%, at 50% sucralfate could fully maintain the IgY antibody activity. At pH = 3.0, 30% sucralfate could maintain 87.6% IgY antibody activity, while over 40% sucralfate could almost fully maintain the IgY antibody activity. Under pH 1.5, 3.0 and 0.02 mg/ml pepsin conditions, the residual antibody activities of IgY solution containing 10, 30 and 50% sucralfate were 63.2, 72.4 and 82.2%, respectively. Over 30% sucralfate could enhance the anti-freeze thawing ability of IgY. After one month at room temperature, over 80% antibody activity could still be maintained. Intragastric administration with 0.5 mg IgY plus 30% sucralfate/day could effectively prevent and cure gastric mucosal injure caused by H. pylori infection, the effect of which increased eight folds as compared with the group without sucralfate. Over 30% sucralfate can enhance the tolerance of VacA IgY to low pH value and pepsin and enhance the anti-freeze thawing ability, therefore, it is an ideal IgY protective agent.[...] Read More.

Keywords: Helicobacter pylori, vacuolate cytotoxin antigen, immunoglobulin yolk, Sucralfate, antibody activity.

2016, 15(1), 1-6;

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Research Article

Biofilm formation potential of oral streptococci in related to some carbohydrate substrates

Arezoo Tahmourespour*, Rooha Kasra Kermanshahi, Rasoul Salehi and Nafiseh Ghasemi Pero

Abstract: In this study, the effect of sugars type (glucose, sucrose, fructose and the mixture of glucose and fructose) and best substrate concentration are investigated on biofilm accumulation. Then, the role of this substrate examined on gtfB expression level by real time RT PCR. A total of 40 Streptococci strains isolated from dental caries and plaque of patients. The ability of them to form biofilm measured in the presence of some sugars on micro titer plates. The mean of resulted optical density of formed biofilm in the presence of sucrose was higher than other carbohydrates significantly (p < 0.05). In the next step biofilm formation of best Streptococcus mutans isolates monitored at time intervals and revealed that the number of attached bacteria increased with the increase of sucrose concentration. The results corresponded to a non linear increase of attached bacteria. The line of best fit for the calibration of the attached bacteria versus sucrose concentration required a 3rd -order polynomial (R2=1). The comparison between the mRNA level of gtfB in planktonic, biofilm and unattached cells of S. mutans by real time RT PCR showed that, the level of gtfB gene expression in the biofilm condition was significantly higher than the planktonic condition.[...] Read More.

Keywords: Biofilm formation, carbohydrate, gene expression, oral streptococci.

2016, 15(1), 1-6;

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Research Article

Nasal colonization of symptomatic pneumonia patients in University of Benin Teaching Hospital, Benin City, Nigeria by multiple antibiotic resistant Staphylococcus aureus

E. E. Akortha* and M. J. Ikenebomeh

Abstract: A study of the antibiotic resistance pattern of Staphylococcus aureus from the nasal cavities of patients with pneumonia attending the University of Benin Teaching Hospital, Benin City, Nigeria was conducted. The survey revealed S. aureus to be the most predominant cause of Staphylococcal pneumonia with an occurrence rate of 38.5%. Other microorganisms encountered included Staphylococcus epididermis, Streptococcus viridans and Brahamella catarrhalis. Patients in age group 1 - 5 years had the highest incidence rate of 60 and 53.8% in males and females, respectively, while patients within ages 11 - 30 years had the least incidence. There was no significant difference between the rate of occurrence of staphylococcal preumonia in males and females at p > 0.05. High resistance levels was recorded towards tetracycline (100%), chloramphenicol (95%), erythromycin (85%) and ampicillin (80%) while moderate resistance levels was recorded towards saprfloxacin (60%), ciprofloxacin (45%), gentamycin (40%) and ofloxacin (20%). The observation in the study, calls for need for effective surveillance of antibiotic resistance.[...] Read More.

Keywords: Antibiotic resistance, Staphylococcus aureus, nasal cavities.

2016, 15(1), 1-6;

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Research Article

Growth control of listeria monocytogenes in experimental cheese samples by lactobacillus casei RN 78 and its bacteriocin

Naheed Mojgani¹, Mansoureh Ameli, Narges Vaseji , Mohammad A. Hejazi, Mohammad A K Torshizi and Cyrus Amirinia*

Abstract: Anti-listerial effect of freeze dried fractions of partially purified bacteriocin (Fd-PPL) and the producer strain Lactobacillus casei RN 78 strain was evaluated in experimental cheese samples during storage at two different temperatures (4 and 35˚C). With the addition of 6400 AU/g of Fd- PPL fractions to the cheese samples of the initial concentration of 4.81 +/- 0.06 log CFU/ml of Listeria monocytogenes was reduced up to 0.91 +/- 0.01 log CFU/ml. Whilst, in the presence of 107 CFU/ml live bacterial cells of L. casei RN 78 of the viability of the sensitive cells decreased sharply (0.71 log CFU/ml). The effect was more pronounced after 24 h of incubation and high levels of antibacterial activity (12800 AU/ml) was seen in these samples. An enhanced reduction in the pH was recorded (4.8 - 5.0) in the cheese samples inoculated with live producer cells compared to control samples without any enrichment. A synergistic bactericidal effect of Lactocin RN 78 in combination with 3% Sodium chloride in cheese samples was observed, and L. monocytogenes population were reduced to 0.69 log CFU/ml within 90 days of incubation, at 35˚C. In contrast to the producer strain, the antibacterial effect of Lactocin was more pronounced in cheese samples stored at 4˚C. The texture of the experimental cheese samples including odour, colour and consistency in different batches were also recorded through out the study.[...] Read More.

Keywords: Lactobacillus casei, anti-listerial, bacteriocin, cheese, biological preservative.

2016, 15(1), 1-6;

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Research Article

Cloning and phylogenetic analysis of gD gene of human herpes simplex virus type 2 in Iranian isolates

Hassan Momtaz, Payam Ghasemi Dehkordi *, Abbas Ali Rezaeian, Ramin Yaghobi, Mohammad Kargar and Manochehr Momeni

Abstract: Herpes simplex virus type 2 (HSV-2) is a member of herpesviridae family and subfamily of alphaherpesvirinae. HSV-2 infection often causes genital herpes in women and men, abortion, infant herpes and non-infectious meningitides. Glycoprotein D and B, respectively, are coded by gD and gB gene. They are the adhesion of HSV-2 attached to the surface of the epithelial cells and are used for making vaccine. To determine the phylogenetic analysis of HSV-2 gD gene and cloning of it in E. coli, infected samples from Esfahan and Chaharmahal Va Bakhtiari provinces in Iran, which were multiplied by PCR and then the segments having 1013bp lengths related to 3 infected samples for gD gene were selected in cloning pCR 4-TOPO plasmid and sequencing. After determining the nucleotide sequences of HSV-2 gD gene, they were compared with samples reported in Iran and other countries. The results showed 2.8 - 10% genetic differentiation which enjoyed more affinity with nucleotide sequenced in USA (K02373) with 97.2% and the biggest difference is in Sweden sample (EU018093) with 90%. The current research showed high homology in the sequence of gD gene of HSV-2. Then, with presence of these affinities in the research samples and other species of Iran and other countries, a good vaccine can be made with high efficiency against all HSV-2 species in the world.[...] Read More.

Keywords: Herpes simplex virus type 2 (HSV-2), PCR, gD gene, phylogenenetic analysis, Iran, Chaharmahal Va Bakhtiari province, Esfahan province.

2016, 15(1), 1-6;

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Research Article

Prevalence of bacterial infection responsible for bovine mastitis

DaRong Cheng*, ShanYuan Zhu, ZhaoHua Yin, WenWei Ding, ZhiXia Mu, ZhiRui Su and HuaiChang Sun

Abstract: Mastitis continues to be the most economically important disease of dairy cattle, and current data on mastitis is even less readily available. To investigate into the prevalence of the pathogens responsible for bovine mastitis, 100 raw milk samples were obtained from the dairy cows with clinical or sub-clinical bovine mastitis in 5 farms in Jiangsu Province, China. All the samples were submitted to bacterial isolation and identification by morphologic examination and biotyping. The data revealed that Escherichia coli was the commonest organism in mastitis cases, being implicated in 82% cases, and Streptococcus uberis (53%) continues to be a prevalent pathogen closely followed by Staphylococcus aureus (41%), Streptococcus dysgalactiae (29%) and Streptococcus agalactiae (27%). In addition, Str. uberis and S. aureus were more frequently associated with clinical mastitis than sub-clinical case, while the infection rates of other bacteria were similar. Further more, Staphylococcus epidermids (15%) and Staphylococcus saprophyticus (10%), previously considered as naught pathogenic bacteria, were also detected in the diseased mammary gland of the problem cows.[...] Read More.

Keywords: Mastitis, bovine, bacteria, infection, prevalence.

2016, 15(1), 1-6;

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Research Article

Evaluation of serum copper level during Giardia intestinalis infection

Eser Kilic, Recep Saraymen, Ozlem Miman and Suleyman Yazar*

Abstract: Alterations of serum copper concentrations are commonly found in patients suffering from gastrointestinal infections and with hepatic, renal, cardiovascular and malignant diseases. In this study, we aimed to investigate the changes of total content of the essential element of copper level in patients infected with Giardia intestinalis. Copper concentration was measured in 64 patients who were positive for intestinal parasite of G. intestinalis. Scores obtained for the positive samples, their age and sex, matched 60 G. intestinalis negative healthy controls. The mean concentration of copper in serum was significantly higher in G. intestinalis positive subjects than in their controls both in females (p < 0.05) and males (p < 0.05). No correlation could be demonstrated between age and mean value of copper in G. intestinalis positive in both females and males control (p > 0.05). Copper level was found to be clearly increased in the patients infected with G. intestinalis compared to controls. Although, our findings suggest altered element metabolism due to the pathophysiologic changes of G. intestinalis, a detailed biochemical and molecular studies on large sample size are needed to put forward a relationship between G. intestinalis (may be other parasitic diseases) and serum element level changes.[...] Read More.

Keywords: Giardia Intestinalis, copper, atomic absorption.

2016, 15(1), 1-6;

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Research Article

Seroprevalence of toxoplasmosis in high school girls in Bushehr city, South-west of Iran, 2009

Moradali Fouladvand*, Afshin Barazesh, Behrouz Naeimi, Keivan Zandi and Saeed Tajbakhsh

Abstract: Toxoplasmosis is a parasitic disease prevalent all over the world and it’s causal agent is Toxoplasma gondii. Toxoplasmosis is very important in pregnant women. Seroepidemiological survey of toxoplasmosis in young girls before marriage for identifying non immune girls could be used to prevent congenital toxoplasmosis. A seroepidemiological study of toxoplasma IgG-antibody in 516 high school girls was conducted. Sample cases were chosen randomly from public high school girls. Blood samples were collected and the obtained sera were examined for anti toxoplasma IgG and IgM antibodies by using ELISA method. Prevalence of toxoplasmosis in high school girls in Bushehr city was 22.1%. There was significant correlation between seropositivity of toxoplasma antibodies and factors such as, contact with cat, raw milk consumption, raw vegetables consumption and food consumption habits, but there was no significant correlation between seropositivity and availability of drinking water, keeping pet and information about toxoplasmosis. About 78% of high school girls in Bushehr city were seronegative in toxoplasma IgG antibody. It is recommended to the health managers to design the educational measures and develop the serological tests for identifying the non immune girls before marriage to prevent congenital toxoplasmosis.[...] Read More.

Keywords: Elisa, toxoplasmosis, high school girls, seroprevalence.

2016, 15(1), 1-6;

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Research Article

Isolating fungal endophyte from Paris polyphylla Smith var. yunnanensis and identifying their antibacterial ability

Q. Xuan, F. K. Bao, H. M. Pan and A. H. Liu*

Abstract: Eighteen fungal endophytes strains parasitizing the famous Chinese medicinal plant Paris polyphylla Smith var. yunnanensis (Franch.) Hand. Mazz were isolated from surface-sterilized plant tissues, such as rhizome, root, stem, leaf and flower. The effect of endophytes on the growth of human pathogenic microbes was evaluated in vitro, using disc diffusion assay. According to the characteristics of cultures and DNA sequences, the WRF7 and WRF7’ belonged to the species Penicillium chrysogenum. The extract of WRF7’ inhibited a broad range of human -pathogenic bacteria, but that of WRF7 almost had no similar effect. And the metabolites of WRF7’ could not stop the growth of test fungus Candida albicans. The study indicates that P. chrysogenum becomes fungal endophyte of Paris polyphylla Smith var. yunnanensis and its isolate WRF7’ makes metabolites that are inhibitory to all test bacteria in vitro. Therefore, the host plant suppressing pathogens owing to its endophytes and inhibitory mechanisms of endophytes to test bacteria and fungi are different.[...] Read More.

Keywords: Fungal endophyte, Paris polyphylla Smith var. yunnanensis (Franch.) Hand. Mazz, isolating, identifying, antibacterial ability.

2016, 15(1), 1-6;

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Research Article

Beneficial effects of Bacillus subtilis on field-grown tomato in Burundi: Reduction of local Fusarium disease and growth promotion

Venant Nihorimbere, Marc Ongena*, Hélène Cawoy, Yves Brostaux, Pascal Kakana, Emmanuel Jourdan and Philippe Thonart

Abstract: The biocontrol potential of Bacillus subtilis S499 was evaluated on tomato in open field sites in low altitude area of the plain of Imbo in Burundi. This strain was tested in order to reduce the impact of an important fungal disease giving rise to large losses in local plantings. The causing pathogen was isolated from diseased leaves at different locations in the fields and identified as Fusarium most probably related to the semitectum species according to the fermentation profile, morphology and gene homology. Results of assays performed in two successive years on the same site indicated that bacterial treatment on seeds significantly increased growth and fruit yield of tomato plants and also provided a high level of protection against the disease caused by this Fusarium pathogen. This is the first reported study on this disease and based on the data collected, B. subtilis S499 may represent an effective solution as biocontrol agent where other chemical options have failed.[...] Read More.

Keywords: Bacillus subtilis, biocontrol, Fusarium disease, tomato plants.

2016, 15(1), 1-6;

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Research Article

Production and characterization of protease enzyme from Bacillus laterosporus

B. Usharani and M. Muthuraj*

Abstract: Production and partial purification of protease enzyme by Bacillus laterosporus was the aim of this study. B. laterosporus was allowed to grow in shake flask broth culture for purpose of inducing protease enzyme. The protease enzyme was purified by ammonium sulfate precipitation followed by dialysis and further concentrated by Amicon tubes. After concentration, the protein was subjected to 12% Zymogram gel with gelatin and the molecular weight of the protease enzyme was 15 kDa. The protease activity increased as there was increase in enzyme concentration; optimum substrate concentration (starch) was 1.0% (w/v); an optimum incubation temperature was 40°C. Purified protease enzyme had a maximum activity at pH 7.0 of phosphate buffer and the optimum incubation time was 24 h. The protease isolated from B. latrosporus is a mesophilic protease. It is stable at pH 7, at 40°C temperature, and this enzyme can be exploited commercially.[...] Read More.

Keywords: Bacillus laterosporus, PMSF, -mercaptoethanol, protease, fermentation.

2016, 15(1), 1-6;

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Research Article

Effect of Mentha longifolia L. ssp longifolia essential oil on the morphology of four pathogenic bacteria visualized by atomic force microscopy

Hajlaoui Hafedh*, Ben Abdallah Fethi, Snoussi Mejdi, Noumi Emira and Bakhrouf Amina

Abstract: The chemical composition and the antibacterial activity of the essential oil of Mentha longifolia L. ssp longifolia are being investigated in the present work. The minimal inhibitory concentration (MIC) of this oil against four gram+ and gram- reference bacteria including Salmonella typhimurium LT2, Escherichia coli ATCC 35218, Microccus luteus NCIMB 8166 and Staphylococcus aureus ATCC 25923 was used to study the morphological alteration of bacterial wall cell visualized by atomic force microscopy (AFM). The chemical analysis of the essential oil showed the presence of 34 compounds. The most important ones were: menthol (32.51%), menthone (20.71%), pulegone (17.76%), 1,8-cineole (5.61%), terpineol-4 (4.87%) and piperitone (2.16%). The MIC for bacteria ranged from 0.19 to 1.56 mg/ml. We found that M. longifolia (Menthol chemotype) has a high antibacterial effect. Cell wall of the tested bacteria was damaged at MIC concentrations. This susceptibility is more accentuated in S. typhimurium and E. coli (rod bacteria), whereas the damage is less important in coccoid bacteria (S. aureus and M. luteus).[...] Read More.

Keywords: Mentha longifolia, essential oil, Staphylococcus aureus, Microccus luteus, Escherichia coli, Salmonella typhimurium, cell morphology, atomic force microscope.

2016, 15(1), 1-6;

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Research Article

Comparison of cell growth and ethanol productivity on different pretreatment of rice straw hemicellulose hydrolysate by using Candida shehatae CICC 1766

Wan-Li Sun, and Wei-Yi Tao*

Abstract: In this work, the cell growth and ethanol productivity was tested for lignocellulosic biomass sugars of overliming-detoxified and NaOH-neutralized rice straw acid hydrolysate by using Candida shehatae CICC 1766. When the acetic acid reached 3 g/L in defined xylose medium, the final ethanol concentration was 33% of that in the medium without acetic acid addition. C. shehatae CICC 1766 could bear lower pH (ethanol yield 13.5 g/L at pH 4.0) in overliming-detoxified hydrolysate. At the optimal pH 5.0 the ethanol yield attended to 16.1 g/L. Ethanol yield in NaOH-neutralized hydrolysate was 13.7 g/L which is better than in overliming-detoxified hydrolysate (7.0 g/L) . It suggested that NaOH-neutralized hydrolysate could be directly fermented by using C shehatae CICC 1766.[...] Read More.

Keywords: Rice straw, Candida shehatae, hemicellulose hydrolysate, ethanol fermentation.

2016, 15(1), 1-6;

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Research Article

In vitro activity of Thaumatococcus daniellii and Megaphrynium macrostachyum against spoilage fungi of white bread and â€˜Ebaâ€™, an indigenous staple food in Southern Nigeria

Grillo Joseph Adebayo* and Lawal Adekunle Kolawole

Abstract: Fresh white bread and ‘eba’ (a carbohydrate; > 80% starch, indigenous staple food in southern Nigeria) were purchased from traders in Lagos, Nigeria and stored under aseptic conditions in the laboratory at room temperature (27 ± 2°C) (11 h of darkness and 15 h of daylight) for duration of 144 h, during which samples were analyzed at 48 h intervals for fungal population using the plate count technique. The sensitivity of the spoilage fungi to aqueous and methanol extracts of leaves of Thaumatococcus danielli and Megaphrynium macrostachyum was determined using the broth dilution method. Fungal populations increased with hour of storage. For bread, Penicillium sp was dominant throughout with average populations of 2.0 x 101 and 8.01 x 106 cfu/g at 0 and 144 h, respectively, while Mucor sp was dominant in eba with average populations of 1.0 x 101 and 9.5 x 106 cfu/g at 0 and 96 h, respectively. For T. danielli, while the methanol extract had greater activity against the spoilage fungi than the aqueous extract, the reverse was observed for M. macrostachyum. The lowest minimum inhibitory concentration (MIC) of T. danielli was 25 mg/ml of the methanol extract against Saccharomyces cerevisiae and Saccharomyces chevalieri and its lowest minimum cidal concentration (MCC) was 50 mg/ml of the methanol extract against S. cerevisiae and Penicillium sp. For M. macrostachyum, the lowest MIC and MCC were 25 and 50 mg/ml of the aqueous extract respectively against S. chevalieri. The results highlight the potential of extracts of T. daniellii and M. macrostachyum as sources of alternative natural preservatives of bread and ‘eba’. [...] Read More.

Keywords: Thaumatococcus danielli, Megaphrynium macrostachyum, minimum inhibitory concentration, minimum cidal concentration.

2016, 15(1), 1-6;

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Research Article

Genetic diversity of Fusarium oxysporum f. sp. cubense isolates from Malaysia

S. K. Leong, Z. Latiffah* and S. Baharuddin

Abstract: Genetic diversity of Fusarium oxysporum f. sp. cubense (FOC) isolates were analyzed using enterobacterial repetitive intergenic consensus PCR (ERIC-PCR), random amplified microsatellites (RAMS) and restriction fragment length polymorphism of Intergenic Spacer (RFLP-IGS) . The three molecular techniques showed intraspecific variations and the banding patterns generated using each technique were highly variable. UPGMA cluster analysis of the combined data of the three techniques showed 35.7 - 100% genetic similarity among the FOC isolates. Based on the dendrogram, most of the isolates including four isolates from Indonesia were clustered together in two main clusters. Sequence analysis of TEF-1 gene of some of the isolates of FOC using parsimony and distance methods also showed intraspecific variations. The highly variable banding patterns shown by using molecular methods supported the hypothesis that the isolates of FOC co-evolved with the edible bananas and dissemination of the pathogen was probably through infected rhizomes.[...] Read More.

Keywords: Fusarium oxysporum f.sp. cubense, enterobacterial repetitive intergenic consensus PCR, random amplified microsatellites, restriction fragment length polymorphism of Intergenic Spacer.

2016, 15(1), 1-6;

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Research Article

Isolation and characterisation of new spore-forming lactic acid bacteria with prospects of use in food fermentations and probiotic preparations

Ali Bayane*, Bréhima Diawara, Robin Dauphin Dubois, Jacqueline Destain, Dominique Roblain and Philippe Thonart

Abstract: Five spore-forming bacteria producer of lactic acid were isolated from soils sampled in the vicinity of poultry farms in Burkina Faso. All isolates were Gram-positive, motile, mesophilic, facultative anaerobic, catalase positive rods, and with L(+) lactic acid production. The isolates have been characterized and identified by a polyphasic approach, combining various phenotypic and genetic characteristics. The 16S-rDNA-sequence analyses revealed the membership of two isolates to the genus Bacillus and the three other to the genus Paenibacillus. The physiological and biochemical analyses showed that the isolates were quite different from known spore forming lactic acid bacteria. Several relevant technological properties were observed, particularly the resistance of the isolates to bile salts and acidic conditions, even the productions of amylolytic and proteolytic enzymes, which could make them good candidates for certain technological applications such as food fermentations and probiotic formulations. Furthermore, the isolation of these microorganisms in the vicinity of farms reinforces the feasibility of their involvement in animal feedstuffs preparations. In conclusion, this work shows an important diversity within the spore-forming lactic acid bacteria and confirms the conclusions of previous works, which have already shown that the SFLAB (Spore Forming Lactic Acid Bacteria) were good candidates for food fermentation and the probiotic formulations.[...] Read More.

Keywords: Lactic acid, spore-forming bacteria, biodiversity, bacillus, 16S rDNA.

2016, 15(1), 1-6;

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Research Article

Antifungal activities of neem (Azadirachta indica) seed kernel extracts on postharvest diseases in fruits

Jingfa Wang, Jian Li, Jiankang Cao and Weibo Jiang*

Abstract: To learn the antifungal effects of neem seed kernel extract (NE) on the post harvest diseases, pathogens of Monilinia fructicola, Penicillium expansum, Trichothecium roseum and Alternaria alternate isolated from the infected fruit were treated with NE in vitro. Results showed that growth of the four pathogens could be significantly (P < 0.05) reduced by NE. The diseases in fruit of plum (Prunus salicina) or Yali pear (Pyrus bertschneideri) inoculated with the pathogens could be prevented remarkably by treated fruit with NE.[...] Read More.

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2016, 15(1), 1-6;

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