Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

antimicrobial effect of sodium hypochlorite solutions with different pH against enterococci biofilm

Antimicrobial agents used during root canal treatment should ideally reach inaccessible sites and would selectively destroy endodontic pathogens. Different auxiliary chemical agents for root canal preparation have been proposed and the most commonly used in endodontics is sodium hypochlorite. This study aimed at evaluating of altering the pH value of sodium hypochlorite on its antibacterial effectiveness against Enterococcus faecalis [E. faecalis] biofilm. Three different sodium hypochlorite solutions with different pH values [alkaline, neutral and acidic] were prepared. One-hundred seventeen canine halves [root canal dentin] and 117 cellulose nitrate membrane filters were used. They were classified according to the test solutions into three equal groups [n=39] according to the immersion in alkaline, neutral or acidic solutions and were further subdivided into another three equal groups [n=13] according to the immersion time [5 min, 30 min and 60 min]. E. faecalis biofilm was developed onto both root canal dentin and cellulose nitrate membrane filters. The antimicrobial effect of test solutions was assessed quantitatively by bacterial count and qualitatively by Scanning Electron Microscope [SEM] examination. The antimicrobial effect of test solutions against E. faecalis biofilm developed onto root canal dentin [using bacterial count] showed that the difference in the mean bacterial count between all groups after all immersion times was statistically significant, except for the difference of acidic versus alkaline solution after 60 min immersion period. While the antimicrobial effect of test solutions against E. faecalis biofilm developed onto cellulose nitrate membrane filters provided that the mean bacterial count recorded after 5 min immersion period for the acidic sodium hypochlorite was significantly different in comparison with either the neutral or the alkaline solutions. After 30 min immersion period, the neutral sodium hypochlorite solution was significantly different in comparison with either the acidic or the alkaline solutions. Total elimination of the bacterial biofilm achieved only after the immersion for 60 min. Using [SEM] the acidic solution for both dentin and cellulose nitrate membrane filter samples showed total elimination of the bacterial biofilm only after the 60 min immersion, but neutral solution was not capable of total elimination of the bacterial biofilm at all time intervals. Finally, the alkaline solution was the only solution capable of removing the bacterial biofilm at all immersion periods. When root canals were irrigated with alkaline sodium hypochlorite the resultant radicular dentin walls were free of debris and the dentinal tubule entrances were visible

Evaluation of the sealing ability and antibacterial effect of guttaflow; a new root canal filling/sealer material using enterococci

Interest in the microbiological profile of post-treatment apical periodontitis has increased considerably during the last few years. It is generally believed that the major cause of treatment failure is the survival of microorganisms in the apical portion of the root-filled tooth. Enterococcus faecalis [E.faecalis] is the most drug resistant organism of these bacteria, and can survive in root canals for 12 months even under nutrient-deprive conditions

Aim: The study aimed to evaluate the sealing ability and the antibacterial effect of a new root canal sealer/ filling material [Guttaflow] and comparing it with other sealing materials [Topseal and gutta-percha]

Materials and Methods: To test the sealing ability of the tested materials, we used Enterococcus faecalis cultured in Brain Heart Infusion [BHI] [bacterial leakage study] ,and methylene blue dye [dye extraction study]. Sixty four caries free, extracted human anterior teeth were cut to the cementoenamel junction, prepared in a crown down technique, and divided into 2 parts [32 for each study]; one part for bacterial leakage study and the other part for dye extraction study. Then each 32 teeth were subdivided into three groups [n=10] and 2 control teeth. GroupI: obturated with a cold lateral compaction using Gutta-percha and topseal sealer. Group II: obturatd with cold lateral compaction using Gutta-percha and Guttaflow as a sealer. Group III: obturated with single cone technique using Gutta-perch and Guttaflow as filling/sealer. The antibacterial effect of the tested materials [against Enterococcus faecalis cultivated on Brain Heart Infusion Agar] were evaluated by measuring the zone of inhibition around discs made of the materials

Results: There was no statistically significant difference between the presence of coronal bacterial leakage among the three groups. For the dye study there was no statistically significant difference between Group II and Group III which showed low values of dye leakage. This was followed by Group I hasing the least absorbance [lowest dye leakage], with a significant difference between the three groups. Topseal showed the statistically significant highest mean inhibition zone [0.8 +/- 0.2]. There was no statistically significant difference between Guttapercha and GuttaFlow which showed the statistically significantly lowest mean values

Evaluation of chromagar and pastorex test in identification of staphylococcus aureus

Diagnosis of Staphylococcus aureus [S. aureus] is a very important to help in treatment of different clinical conditions. Rapid detection tests are the aim of many microbiological laboratories. This study aimed to compare the efficacy of CHROMagar staph aureus media [CSAM] to that of conventional media in the detection and identification of S. aureus. This study was carried out on 50 individuals attending surgical and internal medicine departments, Ain Shams University Hospitals, in the period from October 2007 to December 2008. They were suffering from different pyogenic infections. All samples were cultured on conventional media [Columbia blood agar and chocolate blood agar] and on CSAM. Isolated colonies were identified by catalase, coagulase [tube and slide] and Pastorex Staph plus agglutination test. Antibiotic sensitivity test was performed for all S. aureus isolates using the disc diffusion method. CSAM revealed better detection of S. aureus [90%] than the conventional culture medial [80%]. Pastorex Staph plus agglutination test had higher specificity and sensitivity than coagulase test. In addition, S. aureus isolates were able to resist many antibiotics. CSAM is recommended for detecting S. aureus especially in cases of mixed infections. A higher sensitivity obtained when CSAM is followed by Pastorex Staph plus

Effect of feeding some legumes traditional dishes on hypercholesterolemic rats

There is a marked increase over the past decades in the prevalence of non-infectious chronic diseases such as atherosclerosis hypercholesterolemia is one of the most prevalent risk factors for arteriosclerosis. Dietary quality has a vital role in the prevention of chronic disease. So the present work focused on the hypocholesterolemic effects of the Egyptian traditional dishes from faba bean such as [Nabet, Bissara and Medammis in a level of 10%] in hypercholesterolemic rats for 6 weeks. The chemical composition and amino acid analysis of these products were determined. The results indicated that, Nabet, Bissara and Medammis have a high nutritive value of fiber, protein, ash, carbohydrate, essential and non essential amino acids and low in fat. Also Bissara showed the highest nutritive value than other products. Results from the biological experiment showed that, feeding the hypercholesterolemic rats the basal diet supplemented with Nabet, Bissara and Medammis in a level of 10% cause significant [P > 0.05] decrease in serum total cholesterol, total lipid, triglycerides, LDL and VLDL and significantly [P> 0.05] increased the of serum HDL as compared to control group. It could be concluded that the faba bean dishes are nutritious and may improve the metabolic alterations induced by feeding a hypercholesterolaemic diet

Evaluation of mycobacterium growth indicator tube and gen-probe amplified mycobacterium tuberculosis direct test for rapid diagnosis of mycobacterium tuberculosis in sputum specimens

The Mycobacterium Growth Indicator Tube [MGIT] and Gen-Probe Amplified Mycobacterium tuberculosis Direct [AMTD] test were evaluated using 52 respiratory clinical specimens collected from suspected pulmonary tuberculosis patients. Microbiological culture on Lowenstein-Jensen medium was used as the reference method. The 52 sputum specimens, 29 [55.8%] acid fast smear positive and 23 [44.2%] acid fast smear negative, were cultivated in liquid MGIT and on solid U media. The recovary rate of MTB by MGlT was [80.8%]. The Kappa coefficient calculation showed a very good agreement [Kappa=0.83] between the 2 tests but there was no statistical difference in the sensitivity of detection of MTB between MGIT and Li methods [P>0.05]. There was a statistically significant difference [P<0.05] between the 2 methods as regards the mean time of growth detection. The mean [range] time of detection of MTB was 12.5 [5 to 30] and 19.5 [14-30] days with MGIT and Li, respectively. The recovery rate of MTB 1mm both media in combination [MGIT plus LJ] was higher [84.6%] than the recovery rate on each media separately and the difference between the 2 methods was not statistically significant [P>0.05]. Regarding the detection of MTB from smear positive and smear negative sputum, it was found that MOLT and U detected 93% and 86.2% respectively of the 29 smear positive specimens whereas they detected 65.2% and 60.8% respectively of the 23 smear negative specimens and there was no statistical difference between the 2 methods in relation to the smear type. The recovery rate of MTB by AMTD was 71%. The assay had sensitivities of 100 and 85.7% for acid fast smear positive and negative specimens, respectively. The specificity of the assay was 100% for both types of smears. Our overall results for all specimens, regardless of smear status, showed a sensitivity of 94.8%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value of 86.6%. In conclusion, Both MGIT and AMTD are reliable methods for rapid diagnosis of pulmonary tuberculosis but they should not be used instead of a solid medium; rather, they should be used in addition to it