Open Access Short Research Article

Analysis of Amino Acid 530 and 549 of Hemagglutin Gene in Two Canine Distemper Virus Strains

Feng- Xue Wang, Peng Liao, Gui-Sheng Wang, Shu-Qin Zhang, Jun-Wen Liang, Fu-Lin Tian, Jian- Jun Zhao, Shi- Peng Cheng, Yong-Jun Wen

Microbiology Research Journal International, Page 367-378
DOI: 10.9734/BMRJ/2015/17349

Canine distemper virus (CDV) is prevalent among domestic dogs and causes disease in various types of carnivores worldwide. In the present study, the genotype of two CDV strains, namely, ZJJ-SD and ZJJ-LN, were investigated based on the whole hemagglutininatinine (HA) antigen gene. The CDV strains were obtained from two foxes in Shangdong province and Liaoning province of China in 2011. Before now phylogenic analysis had been carried out for only 260 CDV strains, worldwide and an analysis was performed in the amino acid substitutions at positions 530 and 549 of the HA protein. Phylogenetic analyses revealed that the two strains, ZJJ-SD and ZJJ-LN, belonged to the CDV Asia I lineage. Site 530 of HA protein was found to be relatively conserved within CDV lineages in different host species by combining the genetic sequence data with the published data from 260 CDV strains worldwide. The data analysis showed a bias toward the predicted substitution Y549H for the non-dog strains in Asia I and Europe lineages. The strain ZJJ-SD, from wild canid, has a Y549H substitution. It is one of three Y549H substitution for wild canids in Asia I lineages.

Open Access Original Research Article

Production and Characterization of a Thermostable and Ca++ Independent Amylase Enzyme from Soil Bacteria

Rachna Singh, Deepak Chanda Sharma, Vivek Kumar Shrivastav

Microbiology Research Journal International, Page 329-342
DOI: 10.9734/BMRJ/2015/13993

Aims: Present study was done with the aim of isolation of efficient amylase producing bacterial isolates from soil. Amylase enzyme production, purification, SDS-PAGE and characterization of different important parameters as pH, temperature, substrate concentration etc. was done for obtaining maximum enzyme activity.
Study Design: An experimental study.
Methodology: Isolation was done by serial dilution and spread plate method on nutrient agar at 50ºC and 11 pH. Then screening of bacterial isolates for amylase production was done using starch agar media and on addition of iodine solution bacterial colonies confirmed the amylase enzyme production by decolourization of media from purple to white due to starch hydrolysis. Bacterial isolates with highest zone of starch hydrolysis were selected for enzyme production. Amylase production was done using amylase screening broth at 50ºC, at 250 rpm for 72 h in incubator shaker. Enzyme activity was determined after centrifugation of the culture broth at 10,000 rpm for 10 min. Identification of bacterial isolate was done by performing different morphological and biochemical tests. Then enzyme purification was done using acetone and ammonium sulphate precipitation methods. Molecular weight determination was done by SDS-PAGE. For obtaining maximum enzyme activity evaluation of optimum values for important parameters such as pH, substrate concentration, temperature, reaction time, thermostability, effect of cations, effect of chelating agents and different raw starches was done.
Results: Bacterial isolate 16A showed the highest zone of starch hydrolysis and maximum amylase production (48.86 UmL-1) was obtained on 48 h of incubation at 50ºC. On the basis of morphological and biochemical characterization 16A was identified as Bacillus sp. Enzyme showed 1.6 fold purification with acetone precipitation and 1.54 fold with ammonium sulphate precipitation. Molecular weight of amylase enzyme determined was 97.4 kD approximately, by SDS-PAGE. Enzyme characterization showed that maximum enzyme activity was obtained at 15 minutes reaction time, 75% substrate concentration, 7 pH and 50ºC temperature. A good thermostability was showed by amylase enzyme for 24 h at 50ºC and 15 h at 100ºC. The enzyme activity was enhanced by Fe++ (2 X), Mn++ (3 X) and Triton X-100 (2 X) while was completely inhibited Zn++, Hg++, Cu++, Fe++, Co++, SDS, tween-20 and EDTA. On enzyme characterization maximum enzyme activity of 138 UmL-1 has been observed that is a very good activity.
Conclusion: On the basis of results during present study this is visualized that the enzyme is very suitable for many industrial applications because of its desirable qualities for its industrial applicability as Ca++independent nature and thermostability at 80-100ºC. The broad range of pH and moderate thermostability makes this amylase enzyme a useful additive to liquid detergents that can function in hard and warm water. Calcium independent amylase is suitable for manufacturing of fructose syrup, where Ca++ is an inhibitor of glucose isomerase. So this enzyme can be very useful in different industrial applications.

Open Access Original Research Article

Prevalence of Dengue Virus and Malaria in Patients with Febrile Complaints in Kaduna Metropolis, Nigeria

M. O. Idoko, S. A. Ado, V. J. Umoh

Microbiology Research Journal International, Page 343-347
DOI: 10.9734/BMRJ/2015/15588

Aim: To determine the prevalence of dengue virus and malaria parasites in febrile patients in Kaduna Metropolis, Nigeria.
Study Design: Cross sectional.
Place and Duration of Study: Department of Microbiology, Ahmadu Bello University, Zaria, Kaduna State, Nigeria, during February to August of 2013.
Methodology: Blood specimens were collected from 340 consenting subjects with febrile complaints attending four hospitals in Kaduna Metropolis, Nigeria. Serological determination of dengue virus IgM was carried out using the enzyme linked immunosorbent assay. Malaria status was determined using microscopy technique. Structured questionnaire was administered to determine if the subject was on anti-malaria drug at the time of the research.
Results: A total of 6 subjects were sero-positive for dengue virus IgM giving a prevalence of 1.8%. Microscopy gave a prevalence of 46.5% for malaria. There was no statistically significant difference in the occurrence of dengue virus IgM between the subjects on anti-malaria chemotherapy and those without the therapy. About 1.3% of the malaria positive cases were also sero-positive for dengue virus IgM.
Conclusion: The occurrence of dengue virus in the studied population has been evidenced and malaria is still endemic in the studied population.

Open Access Original Research Article

Keratin Degradation by Penicillium purpurogenum Isolated from Tannery Soils in Jos, Nigeria

P. O. Nwadiaro, A. Chuku, I. A. Onyimba, I. A. Nwaukwu, D. A. Adekojo

Microbiology Research Journal International, Page 358-366
DOI: 10.9734/BMRJ/2015/16339

Background: Tannery soils are very rich in keratinous materials such as fur, wool and hair that are by-products of the tanning industry where hides and skins are processed. A keratinase producing fungus belonging to the genus Penicillium was isolated from soils containing tannery wastes in Jos, Plateau State Nigeria.
Materials and Methods: The fungus produced extra cellular protease on skim milk casein agar as an indication for keratinase production. The species had maximum proteolytic and keratinase activities in a Submerged Fermentation (SmF) using liquid basal medium supplemented with skim milk, chicken feathers and human hair as carbon and nitrogen sources.
Results: The Spectrophotometric assay of the proteolytic and keratinolytic activity of Penicillium purpurogenum, showed that it had the highest activity (13.5 U/ml) on chicken feathers. This was followed by human hair with activity of 12 U/ml. The least activity of 11.9 U/ml was observed in skim milk medium. It was observed that incubation temperature had an effect on the enzyme activity, with an optimum temperature of 37°C for both protease and keratinase.
Conclusion: This non-dermatophytic keratinolytic fungus may have potential use in biotechnological processes involving keratin hydrolysis. The results of this work reiterated that keratinolytic activity is relatively widespread among common fungi and may have an important role in keratin degradation in the natural environment.

Open Access Original Research Article

Effect of Age on Prevalence of Malaria and Oxidative Stress in Children

Olusegun Matthew Akanbi

Microbiology Research Journal International, Page 379-385
DOI: 10.9734/BMRJ/2015/16204

Objective: To determine the effect of age on prevalence of malaria infection and oxidative stress in children.
Study Design: One hundred and seventy children within the age range 0-5 years were recruited into this study from Specialist hospital, Ikare, Ondo state, Nigeria. The children were divided into three age range groups. The first group was within age range 0-1 year, the second group was within age range 2-3 years and the last group was within age range 4-5 years. The study considered those who were malaria positive only.
Methodology: Five milliliters of blood was collected by venipuncture from each child and the blood was immediately transferred into ethylene diamine tetraacetic acid (EDTA) bottle. The plasma was separated from the whole blood and was used to determine the triglyceride, total plasma protein, malondialdehyde (MDA), superoxide dismutase (SOD) and reduced glutathione (GSH) levels. The whole blood was screened for the presence of malaria parasite.
Results: The prevalence of malaria infection was significantly higher in the group within the age range 2-3 years than in other age ranges studied, but the parasite density was significantly higher (P<0.05) in the group within the age range 4-5 years than other age ranges studied. The parasite density was significantly lower in the group within the age range 0-1 year than those within the age ranges 2-3 years and 4-5 years. There was a significant increase in total protein, SOD and GSH levels in the children within the age range 0-1 year as compared with other age groups studied. The triglyceride and MDA levels were significantly higher in age group 2-3 and 4-5 years than in the children within the age range 0-1 year.
Conclusion: This study showed that the older children were more susceptible to malaria infection and oxidative stress as compared to those within the age range 0-1year, therefore, more attention should be given to these age groups.