ISSN 2736-1756
Research Article
Advanced Journal of Microbiology Research Vol. 2012
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2012 International Scholars Journals
Full Length Research Paper
Evaluation and guidelines for use of polymerase chain reaction-computer database homology comparison (PCR-CDHC) for detection and species determination of human pathogenic microsporidia
Scot E. Dowd1* and Jeanette A. Thurston-Enriquez2
1USDA-ARS, Livestock Issues Research Unit, Lubbock, TX, Texas.
2 USDA-ARS, Soil and Water Conservation Research Unit, Lincoln, NE 68583, Texas.
Accepted 11 January, 2012
Abstract
The potential for waterborne disease and zoonotic transmission of at least two species of human pathogenic microsporidia has heightened interest in clinical and environmental detection methods for these organisms. Detection using the polymerase chain reaction (PCR) followed by computer database homology comparison (CDHC) (PCR-CDHC) was reported previously by this research group. As a result, PCR-CDHC has been employed by many research groups around the world for species determination of human pathogenic microsporidia. To validate the CDHC speciation approach, a phylogenetic tree was generated using the small subunit ribosomal DNA sequences (SSU-rDNA) of a large number of microsporidia. An index of similarity was created and used as part of an assessment of CDHCs ability to differentiate between closely related species. Polymerase chain reaction followed by dye termination PCR sequencing and subsequent CDHC of the sequences was performed on 8 species of microsporidia including four human pathogenic strains. The four non-human pathogenic microsporidia tested by this approach were those shown by the phylogenetic analyses to be very closely related to the other human pathogenic species as determined by branch length. In all cases the CDHC approach was able to correctly identify the eight species of microsporidia evaluated. To provide an example of PCR-CDHC, a “universal” and two previously published pathogen-specific microsporidia PCR protocols followed by PCR-CDHC was conducted to assess their ability to detect naturally occurring microsporidia species in swine wastewater. Only one primer set resulted in a PCR-CDHC analysis where presumptive human pathogenic microsporidia was detected. Subsequent CDHC showed these presumptive positive PCR results were actually false positives. With the appropriate primer set, PCR-CDHC proves to be a reliable method that can be used for specific species determination of human pathogenic microsporidia in samples where non-pathogenic species may be present.
Key words: Microsporidia, Sequencing, BLAST, PCR, detection.
Jeanette A. Thurston-Enriquez, Scot E. Dowd*
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2012
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2012 International Scholars Journals
Full Length Research Paper
Immune response and protection of free range chickens vaccinated orally with feeding of newcastle disease vaccine-coated cassava granules
Echeonwu G. O. N1*, Iroegbu C. U2, Echeonwu B. C3, Ngene A4, Nwosuh C. I5, Joannis T. M6 and Ndako J1.
1Virology Department, Federal College of Veterinary and Medical Laboratory Technology, National Veterinary Research Institute, Vom, Plateau State, Nigeria.
2Department of Microbiology, University of Nigeria, Nsukka, Enugu State, Nigeria.
3Poultry Department, National Veterinary Research Institute, Vom, Plateau State, Nigeria.
4Department of Veterinary Medicine, University of Nigeria, Nsukka, Enugu State, Nigeria.
5Viral Vaccines Production Department, National Veterinary Research Institute, Vom, Plateau State, Nigeria.
6Viral Research Department, National Veterinary Research Institute, Vom, Plateau State, Nigeria.
Accepted 20 April, 2012
Abstract
Cassava granules coated with Newcastle disease virus (NDV) strain V4-UPM was used to vaccinate free-range chickens in their natural habitat. Immune response, vaccine virus excretion and the efficacy of the food vaccine were assessed by standard methods. Results show that out of 218 chickens given initial food vaccine in the four locations, 138 (63.3%) produced detectable HI antibody while 202 (92.7%) had titres < 3.0. However, only 16 (7.3%) attained log2 3.0 with GMT of 3.2. This was made up of Nchara- Akanu 7(12.7%), Vandekya 0(0.0%), Fadan Karshi 1(1.7%), and Turu 8(15.1%) with GMTs of 3.3, 2.9, 3.0, and 3.6 in that order. Following the administration of a booster dose of vaccine on 194 birds in the same flocks, 170(87.6%) sero-converted with 118(60.8%) attaining log2 3.0 and GMT of 9.7. Chickens attaining HI titres up to log2 3.0 from the locations were as follows, Nchara-Akanu 26(51.0%), Vandekya 22 (51.2%), Fadan Karshi 28(53.8%), and Turu 33(68.8%) with GMTs of 12.8, 7.5, 7.0, and 12.6 respectively. Vaccinated birds excreted infective vaccine virus. Out of 55 buyback chickens challenged, 15(27.3%) died while 40(72.7%) survived. Twenty two (22) out of 24 (91.7%) unvaccinated birds challenged died and only 2(8.3%) survived. It is therefore concluded that cassava granules could be good carrier for food-borne ND vaccine delivery to village chickens in Nigeria.
Key words: Cassava, V4-UPM virus, village chickens, Newcastle disease.
Echeonwu B. C, Echeonwu G. O. N*, Joannis T. M and Ndako J, Iroegbu C. U, Ngene A, Nwosuh C. I
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2012
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2012 International Scholars Journals
Full Length Research Paper
Improvement of the quality and shelf life of concentrated yoghurt (labneh) by the addition of some essential oils
Mutlag Al.Otaibi* and Hassan El.Demerdash
Department of Food and Nutrition Sciences, College of Agricultural and Food Sciences, King Faisal University, Saudi Arabia.
Accepted 09 March, 2012
Abstract
Three essential oils, namely thyme, marjoram and sage, were added to concentrated yoghurt (labneh) at concentrations of 0.2, 0.5 and 1.0 parts per million (ppm). Subsequently, the chemical, microbiological and organoleptic properties of freshly prepared labneh and of the labneh stored at 5˚C ± 1 for up to 21 days were determined. Addition of essential oils affected the pH, soluble nitrogen -to-total nitrogen, total volatile fatty acid and acetaldehyde values of the prepared labneh. On the other hand, total solids and fat-to-dry matter values were only slightly affected. Total viable counts, as well as counts of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus in the treated labneh increased and reached a maximum after 7 days of storage where after it decreased until the end of the storage period. Yeasts and moulds, coliform bacteria and spore-forming bacteria were not detected in the treated labneh. Of the different treated labneh, labneh containing 0.2 ppm thyme, marjoram or sage oils were organoleptically the most acceptable, and it had a good body and texture that was similar to that of the untreated control. From the results of this study, it can be concluded that 0.2 ppm of thyme, marjoram or sage can be used in order to increase the shelf life of labneh for up to 21days.
Key words: Labneh, essential oils, shelf life, chemical properties, microbiological properties
Hassan El.Demerdash, Mutlag Al.Otaibi*
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Volume 2012
Available online at http://internationalscholarsjournals.org/journal/ajmr
© Advanced Research Journals
Full Length Research Paper
Chemical analysis and antimicrobial activity of the essential oil of Syzigium aromaticum (clove)
G. A. Ayoola1*, F. M. Lawore1, T. Adelowotan2, I. E. Aibinu2, E. Adenipekun2, H. A. B. Coker1 and T. O. Odugbemi2
1Department of Pharmaceutical Chemistry, Faculty of Pharmacy, University of Lagos, CMUL campus, Lagos, Nigeria. 2Depatment of Medical Microbiology and Parasitology, College of Medicine, University of Lagos, Lagos, Nigeria.
Accepted 30 June, 2011
Abstract
Steam distillation of the dry flower buds of Syzigium aromaticum (clove) yielded 7% (w/w) of the pure light yellow oil. Gas chromatography-mass spectrometry (GC-MS) analysis of the oil revealed that the components were eugenol, caryophyllene, eugenol acetate and alpha-humelene, with eugenol being the main component. The antimicrobial sensitivity of the volatile oil against some Gram-negative bacteria (Escherichia coli ATCC 35218, Escherichia coli, Klebsiella pneumoniae, Salmonella paratyphi, Citrobacter spp. and Enterobacter cloacae), a Gram-positive bacterium (Staphylococcus aureus ATCC 25923), and a fungus (Candida albicans) showed a broad spectrum of activity. The minimum inhibitory concentration (MIC) was determined for each organism as 2.4, 1.6, 0.27, 0.016, 0.23, 1.63, 0.73 and 0.067 mg/ml for S. aureus ATTC 25923, E. cloacae, S. paratyphi, K. pneumoniae, E. coli ATTC 35218, E. coli, Citrobacter spp. and C. albicans, respectively. Antioxidant screening of clove oil with 2,2-diphenyl-picryl-hydrazyl radical (DPPH) was positive, indicating the presence of free radical scavenging molecules which can be attributed to the presence of eugenol, a phenolic compound.
Key words: Syzigium aromaticum, clove oil, antimicrobial, antifungal, Gram-negative bacteria, Gram-positive bacteria, Gas chromatography, mass spectrometry.
F. M. Lawore, I. E. Aibinu, H. A. B. Coker and T. O. Odugbemi, G. A. Ayoola*, T. Adelowotan, E. Adenipekun
Page: 1 - 10
Research Article
Advanced Journal of Microbiology Research Vol. 2012
Available online at http://internationalscholarsjournals.org/journal/ajmr
© 2012 International Scholars Journals
Full Length Research Paper
An improved method for the extraction of nematodes using iodixanol (OptiPrep Ô)
Dewang Deng1, Allan Zipf1, Y. Tilahun1, G.C.Sharma1, J. Jenkins2 and K. Lawrence3
1Department of Natural Resources and Environmental Science, Alabama A&M University, Normal, AL.
2USDA-ARS-CSRL, Mississippi State, MS
3Entomology and Plant Pathology, Auburn University, Auburn, AL
Accepted 16 April, 2012
Abstract
A new nematode extraction technique was established, which is based on an iso-osmotic density- gradient medium (OptiPrepTM). This technique resulted in significantly higher numbers of clean eggs and vermiform nematodes that retain higher viability (48.6%) than samples processed with the sucrose method (28.7%). Nematodes survived exposure to OptiPrepTM for 22 hours without significant mortality whereas all nematodes died in the sucrose medium. OptiPrepTM provided a suitable, non-toxic alternative to the traditional density gradient material for the isolation of nematodes. This technique is convenient and relatively simple, with the added benefit of yielding cleaner samples compared to traditional isolation techniques.
Key words: nematode extraction, density gradient, nematology.
Dewang Deng, Allan Zipf, Y. Tilahun, G.C.Sharma , J. Jenkins and K. Lawrence
Page: 1 - 10
Research Article
E. I Eziashi and A. A. Adekunle, I. B. Omamor
Page: 1 - 10