Open Access Short communication

Mitochondrial DNA Variability among Some Saudi Arabian Goat Breeds

S. A. M. Amer

Biotechnology Journal International, Page 877-882
DOI: 10.9734/BBJ/2014/12166

Mitochondrial DNA Variability among Some Saudi Arabian Goat Breeds

Five hundred nucleotides of cytochrome b gene was sequenced for Pakistani, Tihami, Syrian, Masri and Aardi goat breeds inhabiting Saudi Arabia. Based on two computational maximum-parsimony and neighbor-joining methods, the constructed tree clustered the studied breeds into Masri and Tihami in one group and Pakistani and Aardi came basal to this group. Syrian breed came basal to all of the studied breeds. This division was based on a few mitochondrial DNA sites and few sampling size and it therefore needs to be supported in a further molecular study. The goat breeds studied herein did not show any non-synonymous genetic variability when the protein-coding gene of the mitochondrial genome was used as a molecular maker. However, the only mitogenome maker which can show base differences among these breeds was the non-coding displacement loop (d-loop). In conclusion, using the mitochondrial genes, rather than the d-loop, for studying genetic diversity among goat breeds is with no meaning and it is better to use other markers for this purpose.  



Open Access Original Research Article

Iron-Chelating Property of Phytic Acid in Thalassaemia: An In vitro Study

A. Bhowmik, D. Chattopadhyay, S. Chakravarty, T. K. Chatterjee

Biotechnology Journal International, Page 858-867
DOI: 10.9734/BBJ/2014/9781

Aims: The aim of this study is to report an oral iron chelator phytic acid (PA), on blood samples from different types of thalassaemic patients of various age groups and physical conditions.

Study Design: The In vitro iron chelating effect was evaluated by ferritin assay using ELISA.

Methodology: Blood from the iron-overloaded 30 Thalassaemic patients of different age groups, body weights and heights were collected and the serum was separated. Patients without any history of blood transfusion and chelation therapy were taken as control group. The ELISA based ferritin assay was performed with standard phytic acid (40% water solution, Fluka), using Desferrioxamine (DFO) as control (0.5g Deferoxamine mesylate USP, Novartis, USA). The serum ferritin levels were recorded in two different conditions (treated with DFO and PA) at different time intervals (10, 30 and 60 min) by measuring absorbance at 450 nm.

 Results: The Kolmogorov-Smirnov test of patient samples in two different conditions showed that the ferritin concentration, treated by DFO and PA, was significantly decrease in PA treated group compared to DFO, when used in equivalent concentrations, at intervals of 10 min (P=0.760),30 min (P=0.537) and 60 min (P=0.055).

Conclusion: The common iron chelators DFO or Deferiprone used as monotherapy may lead to transient and incomplete removal of iron, while PA showed a more complete and sustained removal of ferritin due to specific chemical binding at a wider pH range. However, further trial is required to establish its maintenance dose, comparative efficacy and mechanism of action.

Open Access Original Research Article

Enhanced Biodegradation of Spent Lubricating Oil Contaminated Soil Using Poultry Litter

E. Stephen, O. T. Temola

Biotechnology Journal International, Page 868-876
DOI: 10.9734/BBJ/2014/11567

Enhanced Biodegradation of Spent Lubricating Oil Contaminated Soil Using Poultry Litter

Enhanced biodegradation of spent lubricating oil contaminated soil using (40% w/w) poultry litter was studied for a period of 56 days. The bacterial count ranged from 1.7×106 cfu/ml – 4.0×106 cfu/ml for oil free soil (OFS), 1.7×106–9.0×105 for oil polluted soil (OPS) and 1.7×105 – 1.0×106 for Poultry Amended oil polluted soil (PAOPS) while the fungal count ranged from 1.2 x105cfu/ml to 5.0 x 105cfu/ml OFS, 4.0x10 -5.0 x105 for OPS and 1.2x105 - 5.0x 105 for PAOPS. The result revealed higher bacterial counts in poultry litter amended soil (PAOPS) compared to oil polluted soil (OPS) and oil free soil (OFS). The fungi counts were low in all treatments (OFS, OPS and PAOPS) .Ten organisms were isolated in the course of this study. The bacteria were Bacillus spp, Micrococcus spp, Pseudomonas spp, Proteus spp and Staphylococcus spp while the fungi include yeasts, Aspergillus niger, Aspergillus flavus, Mucor Spp, Penicillium spp. PAOPS had higher nitrate and phosphorus content compared to OFS and OPS. This result indicates that poultry litter could be used as biostimulating agent to enhance the biodegradation of spent lubricating oil contaminated soil.


Open Access Original Research Article

Assessment of Toxic Metals in Dairy Milk and Animal Feed in Peshawar, Pakistan

Bushra Iftikhar, Saatea Arif, Samina Siddiqui, Riaz Khattak

Biotechnology Journal International, Page 883-893
DOI: 10.9734/BBJ/2014/9939

Assessment of Toxic Metals in Dairy Milk and

Animal Feed in Peshawar, Pakistan

Aims: To assess the concentration of toxic heavy metals [Cadmium, Copper, Lead and Nickle] in dairy milk and animal feed of urban and rural areas and correlate them.

Study Design: It was a cross-sectional comparative study.

Place and Duration of Study: The study was conducted on the dairy milk and animal feed in milk farms of Peshawar. The samples were collected during June – July of 2010.

Methodology: The study area was divided into two zones: urban area which was located in the centre of the city and rural area comprising of peri urban villages. Fifty milk samples were collected, 30 from urban and 20 from rural areas. Feed samples were also collected. Milk and feed samples were prepared by wet digestion method using HNO3 and H2O2 as described by Association of Official Analytical Chemists (AOAC) of North America, 1990. This was followed by analysis on Atomic Absorption Spectrometer.

Results: The results showed that the mean concentration of toxic heavy metals under study namely Cadmium, Copper, Lead and Nickle were greater in dairy milk than the maximum residue limits (MRLs) of these elements as given by Joint Expert Committee on Food and Agriculture and World Health Organization. In comparison of urban and rural areas it was found that higher concentration of Cadmium, Copper, and Nickle was found in dairy milk samples from urban areas than rural areas but there was no significant difference in Lead concentration of the two areas. Their concentration in urban and rural samples were respectively Cadmium 0.69 and 0.037 with P-value <0.001, Copper 1.40 and 0.09 with P-value 0.001, Nickel 0.806 and 0.024 with P-value <0.001, Lead 2.243 and 2.082 with P -value 0.026. However in this study all these elements were within the permissible limits in animal feed except for copper in cotton khal which was 83.3mg kg-1 against the MRL of 10 mg kg-1 given by National Research Council, NRC, 2005. Also there was no significant difference in the concentration of these elements in feed of urban and rural areas. Correlation study between the concentration of these elements in milk and feed showed positive pearson’s correlation coefficient between the two particularly for copper (0.451) in sugarcane khal  and (0.341) in wheat grain, Nickle (0.342) in maize, and (0.30) in wheat bran and cotton khal.

Conclusion: The concentrations of toxic metals were higher than the maximum residue limits in milk but were within their normal limits in animal feed suggesting that milk is being contaminated by various other sources but not farm feed. Comparison between urban and rural areas showed higher concentration in urban milk samples as compared to rural except for lead probably because urban farms are located in polluted areas in the centre of the city. However no significant difference was found in animal feed of urban and rural areas because it is supplied from common source.


Open Access Original Research Article

Identification of Some Rice Genotypes Resistant to Blast Disease in Egypt

Wafaa M. Haggag, Medhat Mekhail Tawfik

Biotechnology Journal International, Page 894-903
DOI: 10.9734/BBJ/2014/10910

Blast disease caused by Pyricularia oryzae, is the most important disease of rice in Egypt and in the world. Field surveys in main production regions, revealed that the highest disease incidence was recorded in El-Behera Governorate. Seven rice cultivars were screened for resistance to blast disease.  Resistance to disease was studied under both artificially and naturally with local pathogen. A bioassay was developed based on (0-9) scale, Area Under the Disease Progress Curve (AUDPC), disease area and sporulation. Differential expression to pathogen was observed between cultivars. Among seven cultivars, Sakha 101 and Sakha 103 were highly resistant and Giza 175 and Giza 178 were highly susceptible to three strains of fungus P. oryzae. Meanwhile other lines showed moderate resistance and susceptible. At the same time there were great differences concerning the growth rate and yield between different rice cultivars.

Open Access Original Research Article

Potential of Azadirachtin and Neem (Azadirachta indica) Based Saponins as Biopesticides for In vitro Insect Pests Cellulase (Beta-1,4-Endoglucanase) Enzyme Inhibition and In vivo Repellency on Tribolium castaneum

Amtul Jamil Sami, A. R. Shakoori

Biotechnology Journal International, Page 904-917
DOI: 10.9734/BBJ/2014/6903

Aims: The work was undertaken to identify the role of Neem derived compounds (saponins and azadirachtin) on the digestive cellulose hydrolyzing enzyme activity of red flour beetle   (T. castaneum), rice grasshopper (Oxya chinensis) and red pumpkin beetle (Aulacophora foveicolis).

Place and Duration of Study: The work was carried out at the Institute of Biochemistry and Biotechnology University of the Punjab Lahore Pakistan.

Methodology: Total cellular proteins were isolated from the insect’s gut and salivary glands and were tested for cellulose hydrolyzing activity on substrate agar plates. Saponins and Azadirachtin were isolated from Azadirachta indica tissues and used for enzyme inhibition studies. Repellancy test was performed for T. Castaneum, using saponins and Azadirachtin. For computational studies sequence of endoglucanase gene was identified from T. castaneum genome and protein structure was deduced.

Results: Saponins were able to inhibit beta-1,4-endoglucanase  enzyme activity, present in all the three insect pests. A computational dissection of T. castaneum endoglucanase enzyme, deduced from T. castaneum genome, showed that there were five cys involved in the formation of disulphide bridges in the molecule. The disulfide bridges did not provide any protection to endoglucanase active site. Azadirachtin had no effect on cellulase activity of Oxya chinensis and Aulacophora foveicolis, while beta-1,4-endoglucanase activity of T. castaneum was inhibited. Repellency test for T. castaneum revealed that each group of compounds (Saponins and Azadirachtin) was able to repel the insect.

Conclusion: Neem derived compounds had a considerable inhibitory effect on the digestive cellulose hydrolyzing enzyme of T. castaneum.

Open Access Original Research Article

Isolation and Identification of Microorganisms from Crude oil Contaminated Soils of Dar es Salaam, Tanzania

Peter Elias Kidibule, Eva Mathias Sosovele, Anthony Manoni Mshandete

Biotechnology Journal International, Page 918-931
DOI: 10.9734/BBJ/2014/11780

Aims: The aim of this study was to isolate and identify microorganisms in crude oil polluted soils in Eastern Tanzania.

Study Design: Isolation and identification of microorganisms using conventional and molecular techniques.

Place and Duration of Study: Department of Molecular Biology & Biotechnology, University of Dar es Salaam; October 2012 to June 2013.

Methodology: Soils were collected from Mlalakua (ML), University of Dar es Salaam (UDSM) and Ubungo (Ub) local garages in Dar es Salaam, Tanzania. The selective isolation of microorganisms was done based on colony morphological appearance on agar plates and Gram reaction. Three isolates (one from each site) were subjected to molecular methods for the identification purposes. Phylogenetic analyses of bacterial 16S rRNA gene and fungal ITS region sequences were employed in this study to identify and determine the evolutionary relationships of the ML, Ub and UDSM isolates.

Results: The 16S rRNA analysis revealed that the ML isolate belonged to the genus Klebsiella, while the ITS analysis on Ub and UDSM isolates assigned them to the genus Trichosporon and Candida, respectively.

Conclusions: From these findings, further studies are recommended to assay for the biodegradative potentiality of the identified isolates for bioremediation of crude oil contaminated soils.

Open Access Original Research Article

Relation of Sirtuin 1 Gene Polymorphisms with Lipid Profile in Hemodialysis Patients

Yasser Elghobashy, Safaa Tayel, Abeer ALrefai, Said Khamis, Hany Elbarbary

Biotechnology Journal International, Page 932-945
DOI: 10.9734/BBJ/2014/12326

Relation of Sirtuin 1 Gene Polymorphisms with Lipid Profile in Hemodialysis Patients

Very little is known about the genetic variation of SIRT 1 and its effects on energy homeostasis in humans. Mammalian SIRT1deacetylates a host of target proteins that are important for apoptosis, the cell cycle, circadian rhythms, mitochondrial function and metabolism. In particular, much current research focuses on the impact of SIRT1 in glucose homeostasis, lipid metabolism and energy balance.

Objective: To study the relationship of sirtuin 1 gene polymorphisms with lipids profile in hemodialysis patients.

Subjects and Methods: This study included 70 Egyptian subjects (45 patients on hemodialysis and 25 age and gender matched healthy control group). The genotyping of SIRT1 rs7895833 in the promoter region, rs7069102 in intron 4, and rs2273773 in exon 5 was performed using polymerase chain reaction with confronting two-pair primers assay (CTPP). Serum TC, TG, HDLc, LDLc, fasting glucose, urea and creatinine were measured by standard colorimetric methods.

Results: The patients had higher diastolic and systolic BP (P<0.001), fasting blood glucose (P<0.001), TC (P<0.001), TG (P=0.006), LDLc (P=0.004), urea (P<0.001) and creatinine (P<0.001). Males and female patients differ according to cause of hemodialysis (P= 0.02) and serum creatinine (P=0.007). Control subjects of sirtuin1 rs7895833 showed significant A allele compared with patients (46% vs. 27.78% P= 0.04) while C allele of sirtuin1 rs7069102 not differ between groups (P>0.05). Sirtuin 1 rs2273773, patients showed significant lower frequency of C allele compared with control (26.67% vs.44% P= 0.04,).

Conclusion: Significant association of SIRT1 rs7895833 and rs2273773 polymorphisms with dyslipidemia and blood pressure may modulate disease course in hemodialysis.