MOSP - Macao Scientific Publishers

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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.

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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.

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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.

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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.

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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.

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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.

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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).

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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.

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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.

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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.

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