Showing papers in "British Biotechnology Journal in 2013"

Statistical optimization of process variables for biodiesel production from waste cooking oil using heterogeneous base catalyst.

Abstract: In this study,the effects of methanol -to-oil molar ratio, catalyst amount and reaction time on the transesterification of waste cooking oil (WCO) to biodiesel were investigated. Methanol with calcium oxide as a heterogeneous catalyst was used for the transesterification process at a temperature of 60 o

Xylanases: An Overview

Abstract: Endo-1,4-�≤ -xylanase (Endo-�≤ -1,4-xylan, xylanohydrolase; EC. 3.2.1.8, commonly called xylanase) is an industrially important enzyme which degrades xylan randomly and produces xylooligosaccharides, xylobiose and xylose. It is mainly present in microbes and plants but not in animals. Xylanases from fungal and bacterial sources have been extensively studied and produced commercially. Its potential use in paper industries has been discussed which is directly related to reduct ion in environmental pollution. It has role in bio-bleaching paper pulp and increasing pulp brightness. Besides, it can be exploited for ethanol production and as an additive in animal feedstock to improve its nutritional value. Endo-1,4-�≤ -xylanase can also be exploited in baking and fruit j uice industries. Here, we reviewed its distribution, structural aspects and industria l/ biotechnological applications. Besides, we also discussed studies related to cloning of the gene encoding endo-1,4-�≤ -xylanase with the objectives of overproducing the enzyme and altering its properties to suit commercial applications.

Co-digestion of food waste and human excreta for biogas production.

Abstract: The ever increasing cost of fossil fuels and its attendant pollution menace has provided the pedigree to consider alternative sources of energy. An investigation was launched into the design and construction of an Anaerobic Digester system from locally available raw materials using local technology and the production of biogas from food wastes and Human excreta generated within a University campus. The experiment lasted for 60 days using a 40-liters laboratory scale anaerobic digester. The volume of gas generated from the mixture was 84,750cm and comprised of 58% CH4, 24% CO2, and 19% H2S and other impurities. The physico-chemistry of the feedstock in the digester revealed an initial drop in pH to more acidic range and a steady increase 4.52 – 6.10. The temperature remained relatively constant at mesophilic range: 22.0oC– 30.5oC throughout the study. The Carbon/Nitrogen (C/N) ratio of the feedstock before digestion was within 139:1. Population distributions of the microflora show aerobic and anaerobic bacteria to include Klebsiella spp, Bacillus spp, Escherichia coli, Clostridium spp and a methanogen of the genera Methanococcus. In most developing nations of Sub-Saharan Africa where biomass is abundant, and where biogas technology is in its infant stage, the anaerobic digestion system could be the much awaited solution. Research Article British Biotechnology Journal, 3(4): 485-499, 2013 486

Application of Box-Behnken Design for the Optimization of Citric Acid Production from Corn Starch Using Aspergillus niger

Abstract: Citric acid production from hydrolysed corn starch was optimized in this study. Response surface methodology (RSM) was employed for the analysis of the simultaneous effect of substrate concentration, broth pH and fermentation temperature on the concentration of citric acid produced during fermentation of hydrolysed corn starch. A three-variable, threelevel Box-Behnken design (BBD) comprising 15 experimental runs was used to develop a second degree statistical model for the optimisation of the fermentation conditions. The optimal fermentation conditions that resulted in the maximum citric acid concentration were substrate concentration; 50 g/L, broth pH; 2.00 and fermentation temperature; 25oC. Under these conditions, the concentration of citric acid was obtained to be 31.96 g/L. Validation of the model indicated no difference between predicted and observed values.

Antibacterial and Antiviral Activities of Essential Oils of Northern Moroccan Plants

Abstract: Aims : This study was designed to evaluate the antimicrobial activity of five essential oils (EOs) extracted from the aerial parts (leaves and flowering tops) of three species growing in the north of Morocco: Origanum elongatum , Thymus capitatus and Mentha suaveolens . Study Design: Chemical analysis of EOs, antibacterial and antiviral activities of EOs. Place and Duration of Study : Department of Biology (Faculty of Sciences), Institute of Agrochemistry and Food Technology (IATA), between September 2009 and December 2009. Methodology : The EO constituents were extracted by hydrodistillation and analysed by

Biosorption of Lead by Pleurotus florida and Trichoderma viride

Abstract: Aims: The objective of the work is to remove Pb by Pleurotus florida and Trichoderma viride in batch studies and to study the kinetics and adsorption isotherm of Pb adsorption by fungal species and to determine the desorption performance by suitable desorbing agents. Study Design: Experimental study. Place and duration of the study: This work was carried out at Department of Biotechnology, Government College of Technology, Coimbatore, Tamil Nadu, and India for a period of five months. Methodology: The polluted sample was collected from Valankulam lake, Coimbatore. The biomass of Pleurotus florida and Trichoderma viride were used as adsorbents. Atomic Absorption Spectrophotometer was used to quantify Pb concentration. The optimum conditions of pH, adsorbent dose and contact time for biosorption were determined. Results: Maximum adsorption of Pb for Trichoderma viride and Pleurotus florida were observed at a pH of 6 and 7 respectively. The optimum quantities of adsorbent required for the removal of Pb were 0.2g for both the organisms. Adsorption of Pb was found to reach equilibrium in 1 h and 1.5 h for Pleurotus florida and Trichoderma viride respectively. Conclusion: Hence, Pleurotus florida and Tricoderma viride are suitable adsorbents for the removal of Pb from effluents. This methodology can be used for the removal of lead Research Article British Biotechnology Journal, 3(1): 66-78, 2013 67 from waste water before its disposal.

SSR- Based Genetic Diversity Assessment in Tetraploid and Hexaploid Wheat Populations

Abstract: Molecular analysis fora set of hexaploid(Triticum aestvium)and tetraploid(Triticum durum)wheat cultivars was investigated by applying 11 SSR primers set. The plant materials consisted of 45 genotypes 15 of which were Triticum aestivumand 30 ofT. durum obtained fromfour different regions Egypt, Greece, Cyprus and Italy. PCR products were separated on a 6% denaturing polyacrylamide gel electrophoresis and produced a total of 3840 DNA fragments which were used for the molecular analysis. The estimated parameters computed by POPGENE (Version 1.32) within the two population

Genetic Variability, Heritability and Genetic Advance in Pearl Millet ( Penisetum glaucum (L.) R. Br.) Genotypes

Abstract: Fifteen genotypes of pearl millet ( Pennisetum glaucum L. R. Br.)were evaluated at Sudan Universityof Science and Technology, The Demonstration Farm, College of Agricultural Studies, Shambat, during the summer season 2009 and 2010. The present study was conducted to assess the magnitude of genetic variability, heritability in broad sense and genetic advance among fifteen pearl millet genotypes for some growth and grain yield characters.A randomized complete block design with three replication swas used at each season.Highly significant differences (P "d 0.01) were observed for days to 50% flowering and days to maturity in the both seasons, for plant height, leaf area, number of grains /plant, 1000 grain weight and grain yield (t/ha) in the summer season of 2009, for panicle length in the summer season of 2010. Also highly significant differences were observed for genotypes and genotypes ◊ seasons interaction for days to 50% flowering and days to maturity. In general phenotypic coefficients of variation (PCV) estimates were higher than genotypic coefficients of variation (GCV) estimates for all the studied characters in all genotypes displaying the influence of environment effect on the studied characters. The combined results for heritability showed that the high estimates of heritability and genetic advance were scoredfor daysto 50% flowering and days to maturity indicating that these characters were under the control of additive genetic effects. The genotypes ICMV155

African Cassava: Biotechnology and Molecular Breeding to the Rescue

Abstract: SUMMARY Cassava is an important African food crop, where it is a staple to about 250 Million people It is a household name in Nigeria, the world largest producer of the root crop It is propagated from stem cuttings and well known for its adaptation to wide range of adaphoclimatic conditions and including those unfavourable for other crops However cassava production, exploitation, utilization and acceptance are limited by diseases and pests, cyanogenesis, low protein content and quality, and post-harvest physiological deterioration The breeding research activities of IITA (International Institute of Tropical Agriculture) Ibadan, Nigeria, CIAT (International Centre of Tropical Agriculture) located in Cali, Colombia and National Root Crop Research Institute (NRCRI), Umudike, Nigeria have transformed cassava to double as a food security crop as well as a cash and industrial crop Of recent, Bio Cassava Plus, an initiative sponsored by Bill and Melinda Gates, has been using experimental biotechnology approaches to address several of the main constraints to African cassava This review presents the many advantages of cassava to the small-scale farmer and its potentials for industrial applications It also describes the roles of biotic and abiotic factors hampering the production yield, root quality, nutritional adequacy, marketability and acceptance, and commercial processes

Effect of Gas Flaring on the Phytochemical and Nutritional Composition of Treculia africana and Vigna subterranean

Abstract: Aims: The effect of petroleum exploration and exploitation activities on the phytochemical and nutritional composition of African breadfruit (Treculia africana) and Bambara groundnuts (Vigna subterranean) harvested from polluted and unpolluted environment were studied. Study Design: Plant samples were collected from Izombe, an environment highly polluted by gas flaring and Eziobodo, an environment not known for gas flare exposures. Both locations are within the (Niger-Delta) rain forest region of Imo State, Nigeria. Methodology: The processed samples were subjected to proximate and phytochemical screening. The vitamins, micro and macro minerals contents were determined using standard biochemical methods and equipment. Results: Some of the phytochemicals and nutritional contents showed varying levels of significant difference (P<0.05) in samples from the two localities. Bambara groundnuts from polluted area was significantly lower (P<0.05) in moisture, ash, protein and carbohydrate when compared to samples from unpolluted environment. The concentrations of antinutrients such as oxalate, phytate, tannin and cyanogenic glycoside were significantly higher (P<0.05) in samples from polluted areas when compared to Research Article British Biotechnology Journal, 3(3): 293-304, 2013 294 samples from unpolluted areas. The results showed increased accumulation of microminerals and reduced macrominerals in samples from polluted environment. Riboflavin, vitamins A, C and E were also significantly higher (P<0.05) in samples obtained from unpolluted environment. Conclusion: Data obtained revealed that T. africana and V. subterranean are highly nutritious and their nutrient composition are responsive to pollution due to gas flaring.

Using Aedes aegypti larvae to Assess Pesticide Contamination of Soil, Groundwater and Vegetables

Abstract: In Benin, the use of synthetic pesticides in vegetable production poses a risk to the environment and human health. Vegetables, water and soil quality assessment is very important for monitoring and mitigation of these risks. The evaluation of pesticide contamination of vegetables and agricultural environment is often made using expensive methods. It is crucial for research in so-called developing countries to develop less expensive tools for pesticide risks assessment and monitoring. The aim of this study was to assess the potential of using Aedes aegypti larvae as a bio-indicator to measure the pesticide contamination of soil, groundwater and vegetables. Vegetables just before harvest, groundwater and soils samples from three production sites and vegetables samples from markets were collected from March to August 2011. Ethanol extracts of these samples were tested on first stage larvae of Aedes aegypti. The method made it possible to detect residues of chlorpyrifos-ethyl and deltamethrin in cabbage until 4 and 8 days respectively after treatment with the recommended doses for crop protection. It proved inappropriate to measure pesticides residues in market-gardening soils, since these soils contain some amounts of nitrite, nitrate and phosphate, coming from the decomposition of fertilizers which are poisonous for the first stage larvae of Aedes aegypti. Overall, the results revealed the presence of small amounts of pesticides residues in 12.5% of the vegetables collected from markets. Pesticides residues were also detected in 30.0% of vegetables collected just before harvest. Residues of pesticide were not detected in groundwater samples collected from vegetable growing areas. First stage larvae of Aedes aegypti could be used as a bio-indicator to characterize and monitor risk of pesticide contamination of vegetables in southern Benin. It could also be used for a monitoring program before running a more thorough chemical analysis to identify and quantify the pesticide molecules present in samples. (Resume d'auteur)

The Effect of Picloram on Somatic Embryogenesis of Different Explants of Strawberry (Fragaria ananassa Duch.).

Abstract: The present investigation was conducted to study the effects of different concentrations of picloram on somatic embryogenesis induction, development and maturation of three strawberry (Kurdistan, Paros and Camarosa) cultivars. For this purpose, leaf blade, nodal, petiole, stamen and flower bud calli were cultured on MS medium supplemented with picloram at 0.25, 0.5, 1 and 2 mg/L concentrations. The concentration of growth regulator, cultivar and explant type were fo und critical to somatic embryogenesis induction, development and maturation. Results obtained from the studies revealed that all explants with the exception of petiole and stamen incubated on medium formed embryonic calli. 2 mg/L picloram yielded the highe st percentage of embryonic calli and number of globular stage embryos and 1 mg/L picloram yielded the highest number of cotyledonary -stage embryos in all types of explants. The leaf explant calli and Paros cultivar were the most responsive to produce somatic embryogenesis induction, development and maturation.

Application of Molecular Microbiology Techniques in Bioremediation of Hydrocarbons and Other Pollutants

Abstract: Molecular microbiology techniques have revolutionized microbial ecology by paving the way for rapid, high -throughput methods for culture -independent assessment and exploitation of microbial communities present in complex ecosystems like crude oil/hydrocarbon polluted soil. The soil microbial community is relatively diversewith a high level of prokaryotic diversity. This soil species pool represents a gold mine for genes involved in the biodegradation of different classes of pollutants. Currently, less than 1% of this diversity is culturable by traditional cultivation techniques. The application of molecular microbiology techniquesinstudyingmicrobial populations in polluted sites without the need for culturing has led to the discovery of novel and unrecog nized

Efficient Micropropagation of Spilanthes acmella (L.) Murr.: A Threatened Medicinal Herb

Abstract: The present study describes an efficient and reproducible protocol for micropropagation of S. acmella . Shoot tips taken from 3 week-old aseptic seedlings were cultured on Murashige and Skoog (MS) semi-solid medium supplemented with different concentrations of TDZ. Among various concentrations, 0.25 µM TDZ was found to be optimum for shoot regeneration as it induced a maximum of 30.0 shoots per explant however with retarded growth (1.0 cm). Among different volumes of culture media, 15 ml liquid culture medium favored best response wherein a maximum of 80.2 shoots per explant with an average shoot length of 7.0 cm were induced after 6 week of subculturing. Successful in vitro rooting was induced on 2.5 µM NAA containing half-strength MS medium. Almost 96% rooted plants successfully transferred and acclimatized ex vitro under green house conditions. Morphological and physiological parameters compared with the in vivo -grown seedlings of the same age appeared to be ‘normal’ in respect to the fundamental characteristics examined.

Primary Somatic Embryos from Axillary Meristems and Immature Leaf Lobes of Selected African Cassava Varieties

Abstract: The study evaluated high-value African cassava varieties for primary somatic embryogenesis usingaxillarymeristems (AM) and immature leaf lobes (LL) on picloram based medium. The study was conducted at the Central Biotech Lab, International Institute of Tropical Agriculture (IITA), Ibadan, Nigeria between 2006 and 2009. Completely randomized design with four replicates was used for the study. Using LL explants, there were significan t (P=.05) differences in percent responding leaf lobes, percent explant with pre -embryogenic structure, PSEF and PSEE among cassava varieties. The PSEF of the only three varieties that produced mature somatic embryo were 93.6, 88.5 and 85.7% for TME 12, Ki

Changes in Heavy Metal Contents of a Waste Engine Oil Polluted Soil ExposedtoSoilpH Adjustments

Abstract: The present study investigated the role of soil pH adjustments in heavy metalsconcentrations in waste engine oil (W EO) - polluted soils. Sun-dried top soil (0-10cm) wasmeasured into buckets. WEO was added to soil and mixed thoroughly to obtain similarconcentrations of 2.5% w/w oil in soil. The polluted soil was thereafter amended with NPK(15:15:15) fertilizer to enhance microbial activity. The buckets were transferred into a wellventilated screen house with inherent constant room temperature (27oC). The entire setupwas divided into 5 sets. Each set was wetted daily with 200ml of different pH solutions (pH3, 5, 7, 9, and 11) for a period of 3 months. There were significant reductions in heavymetal concentrations particularly at pH of 5. There were significant reductions in totalhydrocarbons contents (THC) of polluted soils at 2 months after pollution from 1882.32mg/kg at pH 3 to 325 mg/kg at pH 5, compared to THC of soil at 1 week after pollution atpH 7 (3425.63mg/kg).Keywords: Contamination factor; environmental risk factor; hazard quotient; naturalattenuation; pH; soil; waste engine oil.Research Article

Constraining the Pluripotent Fate of Human Embryonic Stem Cells for Tissue Engineering and Cell Therapy – The Turning Point of Cell-Based Regenerative Medicine

Abstract: To date, the lack of a clinically-suitable source of engraftable human stem/progenitor cells with adequate neurogenic potential has been the major setback in developing safe and effective cell-based therapies for regenerating the damaged or lost CNS structure and circuitry in a wide range of neurological disorders. Similarly, the lack of a clinically-suitable human cardiomyocyte source with adequate myocardium regenerative potential has been the major setback in regenerating the damaged human heart. Given the limited capacity of the CNS and heart for self-repair, there is a large unmet healthcare need to develop stem cell therapies to provide optimal regeneration and reconstruction treatment options to restore normal tissues and function. Derivation of human embryonic stem cells (hESCs) provides a powerful in vitro model system to investigate molecular controls in human embryogenesis as well as an unlimited source to generate the diversity of human somatic cell types for regenerative medicine. However, realizing the developmental and therapeutic potential of hESC derivatives has been hindered by the inefficiency and instability of generating clinically-relevant functional cells from pluripotent cells through conventional uncontrollable and incomplete multi-lineage differentiation. Recent advances and breakthroughs in hESC research have overcome some major obstacles in bringing hESC therapy derivatives towards clinical applications, including establishing defined culture systems for de novo derivation and maintenance of clinical-grade pluripotent hESCs and lineage-specific differentiation of pluripotent hESCs by small molecule induction. Retinoic acid was identified as sufficient to induce the specification of neuroectoderm direct from the pluripotent state of hESCs and trigger a cascade of neuronal lineage-specific progression to human neuronal progenitors and neurons of the developing CNS in high efficiency, purity, and neuronal lineage specificity by promoting nuclear translocation of the neuronal specific transcription factor Nurr-1. Similarly, nicotinamide was rendered sufficient to induce the specification of cardiomesoderm direct from the pluripotent state of hESCs by promoting the expression of the earliest cardiac-specific transcription factor Csx/Nkx2.5 and triggering progression to cardiac precursors and beating cardiomyocytes with high efficiency. This technology breakthrough enables direct conversion of pluripotent hESCs into a large supply of high purity neuronal cells or heart muscle cells with adequate capacity to regenerate CNS neurons and contractile heart muscles for developing safe and effective stem cell therapies. Transforming pluripotent hESCs into fate-restricted therapy derivatives dramatically increases the clinical efficacy of graft-dependent repair and safety of hESC-derived cellular products. Such milestone advances and medical innovations in hESC research allow generation of a large supply of clinical-grade hESC therapy derivatives targeting for major health problems, bringing cell-based regenerative medicine to a turning point.

Optimization of Sweet Potato Starch Hydrolyzate Production and Its Potential Utilization as Substrate for Citric Acid Production

Abstract: Aims: The aims of this work was optimization of two -step enzymatic hydrolysis of sweet potato starch using statistical approach and subsequent utilization of the hydrolyzate obtained for citric acid production. Methodology: Box Behnken design was used in this s tudy to generate a total of 17 individual experiments for each step of the hydrolysis (liquefaction and saccharification steps). These were designed to study the effect of temperature, time and pH on the sweet potato starch hydrolyzate (SPSH) concentration . The optimization was carried out using response surface methodology (RSM). The SPSH obtained was used to culture Aspergillus nigerfor citric acid production. Results:A statistically significant quadratic regression model (P<

Characteristics of Penicillin G Acylase Immobilized onto Iron Oxide Nanoparticles

Abstract: Penicillin G acylase was immobilized onto iron oxide nanoparticles coated with polyethyleneimine and then cross linked with glutaraldehyde solution. The FTIR spectrum of immobilized enzyme showed peak at 1648cm -1 which can be attributed to the C=N bonds of Schiffs base linkage formed between glutaraldehyde and amino group of penicillin G acylase. By considering the FTIR spectrum of nano particle coated with polyethyleneimine, adsorption of penicillin G acylase has taken place and then glutaraldehyde cross linked enzyme onto activated support. Catalytic properties of nano penicillin G acylase were improved upon immobilization as compared to its free counterpart. The optimal pH and temperature were determined to be 7.0, 10.0, 50 and 75∫C for free and immobilized penicillin G acylase, respectively. Thermal stabilities of both nano and free penicillin G acylase were studied .The Km value of immobilized nanozyme was calculated from Lineweaver Burck plot to be 0.23 �oM while that of free penicillin G acylase was 0.28�oM. In this way nano penicillin G acylase with improved catalytic properties was developed as compared to its soluble counterpart.

Improved Sample Preparation for PCR-Based Assays in the Detection of Xanthomonads Causing Bacterial Leaf Spot of Tomato

Abstract: Aims: To develop a sampling procedure for PCR-based screening of bacterial leaf spot (BLS)-causing xanthomonads without DNA extraction from infected tomato plants. Place and Duration of Study: University of Copenhagen, Denmark and Sokoine University of Agriculture, Morogoro, Tanzania between July 2008 and November 2010. Methodology: Flinders Technology Associates (FTA) plant cards and Chromatography paper or Whatman paper strips (WPS) were spotted with bacterial suspensions from 24h-old cultures from reference strains of BLS-causing xanthomonads, or sap obtained by grinding or hand maceration of plant tissue, were used as templates in PCR reactions or isolation of live bacterial cells on Nutrient agar (NA) media. Samples were tested by PCR with Xan 7 genus/-specific Xanthomonas primers or in multiplex with 26S rRNA primers. Isolation of bacteria was done by streaking aliquots of 75 μl of a suspension from a disc Research Article British Biotechnology Journal, 3(4): 556-574, 2013 557 (2-mm-punch by Harris Micro Punch®) in triplicate, removed from each of the FTA plant card and WPS onto NA media. Results: The FTA plant card spotted with pure cultures of reference strains of xanthomonads and sap from grinding or direct maceration of plant tissue resulted in more clear PCR bands (402 bp) and (594 bp of rRNA gene in multiplex) than the WPS samples. Sensitivity of detection by the FTA paper-based PCR was ≈ 5.0 x 10, while that of the WPS was > 1.0 x 10 CFU/ml. The WPS (but not the FTA) was proved to be useful for saving living bacteria cells for up to one week of storage at ambient temperatures. Conclusion: Both FTA plant card and WPS can be used for PCR detection of BLScausing xanthomonads in tomato. However, the FTA plant card is recommended as it produced clearer PCR products than WPS. WPS is recommended for experiments requiring isolation of live bacterial cells on NA media.

Characterization of Dairy Effluents by Physicochemical Parameters

Abstract: Aims: The dairy industry faces growing scrutiny of its environmental stewardship. The potential impact of an individual operation on the environment varies with animal concentration, weather, terrain, soils, and numerous other conditions. It is hoped that management practices found on dairy industry will benefit by the management practices. So properly applied the Management Practices, the factual study of dairy effluent by variousphysico-chemical characters concern for environmental health and safety. Study Design:Effluent samples were collected from dairy industries. The samples were characterized by physical parameters like pH, temperature, TS, etc. and chemical parametersBOD, COD, DO etc. Place and Duration of Study:Effluent samples were collected from dairy industry of district Kolhapur Maharashtra (India).Physico -chemical characteristics of the effluent during the months between March to August 2011. Methodology: Total 4 samples of dairy effluents were collected by composite sampling at

Kinetic study of neem biodiesel production.

Abstract: Aims: The kinetic features of neem biodiesel production were studied to establish theconversion parameters that govern optimal product yield in minimum reaction span. The mechanistic features of the dual acid/base catalytic conversion were investigated to gain an insight into the influence of methoxide nucleophilic attack and catalytic restoration on reaction yield in the final stage. Study Design: The reaction kinetics was investigated by UV -Vis absorption

Genetic Characterization of Insulin Growth Factor-1 and Its Receptor Genes in Egyptian Buffalo (Bubalus bubalis L.)

Abstract: Aim: The somatotropic axis (SA) comprises genes associated with economically important quantitative traits in livestock like mammary and muscle growth as well as carcass traits. Insulin growth factor -1 (IGF-1) and its receptor ( IGF-1R) are two important genes belonging to the SA. The aim of this study was to evaluate the genetic polymorphism ofIGF1/SnaBI andIGF-1R/TaqIrestriction sites in Egyptian buffalo. Methodology :Genomic DNA was extracted from blood samples of 100 healthy buffaloes maintained at the Mahlet Mussa and ElGmeasa herds from 2010 to 2012. PCR was performed using primers flanking a 250 -bp fragment of the regulatory region of the buffalo IGF-1 gene and a616-bp fragment of theIGF-1R gene encompassing 51-bp from exon 12, 479-bp from intron 12 and 86bp from exon 13. The PCR-amplified fragments were digested withSnaBI (IGF-1) and TaqI (IGF-1R), electrophoresed and analyzed on agarose gels stained with ethi dium bromide.The two amplified fragments were also sequenced and aligned with published sequences. Results:All buffaloes investigated in this study were genotyped BB (i.e., negative for the SnaBI restriction site at position 224 ^225 (TAC^GTA) of theIGF-1regulatory region) and Research Article