Browsing by Author "Fernando, T.S.R."

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    Determination of Morphological and Genetic Diversity of Wild Guppy (Poecilia reticulata) in Sri Lanka across the MHC Complex with Special Reference to Class IIB Region
    (Uva Wellassa University of Sri Lanka, 2013) Godagama, G.R.M.N.; Fernando, T.S.R.; Bulumulla, P.B.A.I.K.; Jayamanne, S.C.
    Wild guppies have potential in developing various strains with attractive colour patterns, tail types and tolerance to wide range of environment conditions, resistance to disease conditions due to high immunity. Application of molecular genetic markers, are important to identify diversity among wild guppies which are economically beneficial to ornamental industry and to implement conservation of these valuable genetic resources. Major histocompatibility complex (MHC) genes are highly polymorphic gene family and exon 2 of class II B gene is functionally important in immunity and disease resistance. Hence, in the present study attempts are made to assess the genetic and morphological diversity of wild guppy of Sri Lanka with special reference to immune related MCH class II B gene. A total of 238 wild guppies were collected from 10 regions to represent different agro-ecological zones of the country. The standard length in between 13-24 mm was selected (179 fishes) to collect morphological data and genomic DNA was extracted from muscle tissue using Chelex 100 DNA extraction kit. A PCR based method was used to amplify exone 2 region of candidate gene with forward (5’GTG GAT TTC AGA GAA TAT GCA 3’) and reverse (5’ TGA TTT ATC CAG AGC GGT TTG 3’) primers. Touch down PCR was followed to amplification in the temperature range of 47 to 45 . Selected fish sample consisted of 43.6% of male fishes and 56.4% female fishes. Significant association existed in tail types and colour patterns versus region. Highest variation of tail pattern types was recorded from Buttala region and 40.8% of guppies consisted round tail type. High variation of colour pattern is observed from Badulla region. 58.7% fishes had brownish gray colour pattern and 43% had golden upper body colour pattern. Variation of upper body colour in all regions was similar. PCR protocol was optimized. There was a morphological diversity between wild guppy fishes in different regions of Sri Lanka. Exon II in MHC class IIB region was amplified and optimized PCR protocol for further studies. Sequence variation based on Single Nucleotide Polymorphism (SNPs) and differences of immune response of wild guppy population is yet to be analyzed.
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    Development of Standard DNA Size Markers for Short Tandem Repeat Loci BM1818 and BM1824 of Cattle DNA Typing in Sri Lanka
    (Uva Wellassa University of Sri Lanka, 2013) Nugapola, N.W.N.P.; Bulumulla, P.B.A.I.K.; Fernando, T.S.R.; Illeperuma, R.J.
    Cattle are the most common type of livestock animal raised for meat, dairy, as draft animals and various other purposes. Sri Lankan cattle population mainly consist of indigenous cattle “Sinhala batu cattle”, European pure breeds (Jersey, Friesian, Ayrshire), Indian cattle breeds (Sahiwal, Sindhi, Tharparkar), and their crosses. Identification and traceability of cattle and their products is extremely important for proven of ownership, exploration of pedigree and animal disease control. Traditional identification methods are less effective and associated with many economic, cultural and ethical concerns and are not reliable methods of establishing maternity or paternity of cattle. Deoxyribo Nucleic Acid (DNA) based identification is based on the unique, unalterable, inherited DNA profile of an individual animal as an identifier. Testing the Short Tandem Repeats (STR) of DNA in establishing the identity and parentage is widely used for humans in Sri Lanka. However DNA based identity testing methods are needed to be developed for cattle in order to use it as an accurate method than conventional methods to address problems of cattle identity and traceability. Currently there is no proper procedure of DNA based of identification in Sri Lanka due to the unavailability of standard DNA size markers and there is an urgent need of developing standard DNA size markers for cattle STR (allelic ladders) for accurate genotyping of an individual. Therefore the present study was conducted to develop standard DNA size markers for STR loci BM1818 and BM1824. Methodology Selection of loci Two bovine specific STR markers (BM1818, BM1824) recommended by the Food and Agriculture Organization (FAO) and the International Society for Animal Genetics (ISAG) (FAO, 2011) and previous studies done by Goor and Van de (2011) were selected for this study. Sample collection and DNA extraction Blood samples (n=38) and tissue samples (n=12) were randomly collected from unrelated animals from different areas of the country according to the recommendations provided by FAO 100 extraction protocol described by Phillips (2009) with minor modifications. DNA from tissue samples was extracted by ChargeSwitch Forensic DNA Purification Kit (Invitrogen™, Life Technologies Corporation) according to the kit’s manual. PCR amplification PCR amplification of extracted DNA samples was carried out with 5 µL of genomic DNA in a 50 µL reaction volume, consisted of 5 µL of 10X STR buffer (DreamTaq Green Buffer - Thermo Scientific), 5 µL of each dNTP (Guangzhou Geneshun Biotech Ltd.), 5 µL of primer (FAO, 2011 and Goor and Van de 2011) (Integrated DNA Technology), 0.30 µL of Taq DNA polymerase (DreamTaq-Thermo scientific) and 30 µL of sterilized distilled water. The PCR reaction was carried out in a Veriti 96 well thermal cycler (Applied Biosystem, USA) under the following PCR cycling conditions (Sodhi et al., 2006): 2 min at 94 °C, followed by 30 cycles of 1 min at 94 °C, 1 min at annealing temperature (56 to 62 °C) of each primer, 1 min at 72 °C, and final extension of 10 min at 72 °C. Subsequent to PCR, the products were subjected to 3 % w/v agarose gel electrophoresis to verify the success of the PCR reaction. Then the PCR products were subjected to 7 % denaturing polyacrylamide gel electrophoresis. Amplified products were detected using DNA Silver Staining procedure described by Bassam et al. (1991) and Promega technical manual. Allelic ladder construction Allelic ladders were constructed by two methods as follows (1) DNA samples that can contribute to maximum number of polymorphic alleles to each locus were selected, pooled and amplified by PCR reaction (100 µL reaction volume), (2) PCR products that can contribute to maximum number of polymorphic alleles to each locus were selected and pooled to create the allelic ladders. Results and Discussion According to the Table 1, allelic range of the BM1818 locus was from allele 11 to 21. The highest frequency was shown by allele No.18 at a percentage of 35.0 percent. Allele 11 had the lowest frequency with 1.3 percent. When considering BM1824 locus, allelic range was from allele 11 to 19. The highest frequency was shown by allele No.13 at a percentage of 34.1 percent. Two alleles (11 and 18) showed the lowest allelic frequency of 1.2 percent.
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    Effect of Enrobing on Quality Traits of Chicken Nuggets Produced Using Steam Cooking and Without Steam Cooking
    (Uva Wellassa University of Sri Lanka, 2016) Samudini, W.M.S.; Jayasena, D.K.D.D.; Fernando, T.S.R.; Wijesundera, W.M.N.M.; Athula, D.L.K.; Pemasire, P.F.S.
    The aim of this study was to examine the effect of enrobing on quality traits of chicken nuggets produced using steam cooking and without steam cooking. Nuggets were manufactured, based on nuggets preparation procedure of Ceylon Agro Industries (pvt) Ltd, with four treatments Tr 1 (initial product), Tr2 (steam cooked nugget without enrobed), Tr3 (enrobed fried nugget produced without steam cooking) and Tr4 (enrobed, fried steam cooked nugget). Nuggets were evaluated for microbiological counts, pH, colour, breading pickup and sensory properties during storage. Higher initial aerobic plate counts were recorded in uncoated nuggets same as Staphylococcus aureus. After 21 days, uncoated nuggets were resulted highest aerobic plate counts and Staphylococcus aureus. Tr4 recorded zero initial Staphylococcus aureus counts during storage time. The pH of the control samples in respect to the enrobed, diminished during first fourteen days of storage and then in 21 day of storage it showed an increment. Same pattern was exhibited in enrobed samples but the increment was less than the control samples. The L*, a*and b* values changed significantly (p
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    Evaluation of Escherichia coli, Salmonella serovars and Staphylococcus aureus Contamination of Retail Chicken Meat in Badulla District, Sri Lanka
    (Uva Wellassa University of Sri Lanka, 2013) Madurangi, G.H.P.; Chandrasena, G.; Fernando, T.S.R.; Wjesundara, W.M.N.M.; Bulumulla, P.B.A.I.K.
    Food safety is a global challenge for most developing countries. Food borne diseases are mainly caused by E.coli, Salmonella and S. aureus. And foods originated from animal are more susceptible for spoiling. Thus, the present study aimed at evaluating the Escherichia coli, Salmonella serovars and Staphylococcus aureus contamination in retail chicken meat from Badulla district to analyze the microbiological quality of the retail chicken meat in Badulla District. Twenty retail shops were randomly selected from seven secretary divisions in Badulla district. Two whole chicken samples were collected from each retail shop and transferred to the laboratory under refrigerated condition. 25 g of chicken meat samples from different cuts (breast, back, thigh, wings and whole) were taken. Each meat sample was pre enriched with 225 ml of buffered peptone water and placed in incubator at 37 for 24 hours. Loops full of pre enriched samples were streaked on Eosin Methylene Blue agar, Brilliant Green agar and Manitol Salt agar to isolate of E.coli, Salmonella and S.aureus respectively. Inoculated plates were incubated at 37 for 24 hours. Presumptive colonies on each agar plate, sub cultured on nutrient agar plates and incubated at 37 for another 24 hours. Presumptively positive colonies of E.coli, Salmonella on nutrient agar plates were bio-chemically confirmed with Simmons Citrate agar and S.aureus by catalase test. Prevalence of Salmonella in thigh, breast, back and wing cuts were 28.92 %, 20.48 %, 19.28% and 13.25 % respectively. Prevalence of Salmonella in whole chicken sample was 18.07%. No significance association was observed for the prevalence of Salmonella with different chicken meat cuts (P >0.05). Prevalence of Escherichia coli in thigh, breast, back and wing cuts were 18.99%, 26.58%, 26.58% and 11.39% respectively. Prevalence of Escherichia coli in whole chicken sample was 16.46%. There was a significance association between chicken part and the prevalence of Escherichia coli in retail chicken meat in Badulla District. Contamination rates of S .aureus in different cuts of retail chicken meat were thigh (20.99%), breast (25.93%), back (24.69%) and wing (11.11%). A significant association was observed in prevalence of S. aureus in different cuts of chicken carcass taken from the retail outlets of Badulla district (P<0.05). The highest occurrence of Salmonella was reported in Badulla division (19.28%). Incidences of Escherichia coli (24.05%) and S. aureus (18.52%) were significantly high in Bandarawela division. The findings of this study are vital to the public health risk of the country and emphasis the need of developed programme to assure the quality and safety of poultry meat at retail market
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    A Preliminary Study on Developing a Feed Ration with Poultry Offal Meal for Young Fattening Pigs
    (Uva Wellassa University of Sri Lanka, 2013) Perera, D.S.R.S.P.S.; Tharangani, R.M.H.; Fernando, T.S.R.; Nambapana, M.N.M.; Jeewanthi, G.
    There are several important factors to be concerned when conducting a commercial piggery including feeding and housing. Feeding is one of the most important tasks based on meeting the animal’s requirements for energy and protein to optimize the growth performance and carcass lean content (Pettigrew and Esnaola, 2001). In general feed cost accounts for 55 % - 75 % of the total cost in any commercial pig farm (Ologhobo et al., 2012). As an alternative for high cost ingredients, some local feed that are inexpensive can be used in feed formulation for the fatteners which provide complete nutrition. Poultry Offal meal (POM) is one of the highly important feed stuffs available for the animal feed formulation (Hansen et al., 2006). Recycling of poultry offal waste helps to reduce the high cost of dietary ingredients and feeding that are associated with intensive animal production systems as well as to reduce the environmental and health hazards to human (Ologhobo et al., 2012). The objective of this experiment was to develop a ration with POM for feeding fattening pigs to reduce the cost of production and supply required level of nutrients. Methodology The current study was carried out at the Maxies & Company (Pvt) Ltd, Wennappuwa. One month old twelve fatteners were divided into four groups and fed with three experimental feed rations (Feed A, 23 % POM; Feed B, 20 % POM and Feed C, 18 % POM) and the commercial pig starter ration was given as the control feed for the fourth group. All three experimental feed rations were formulated with different proportions of POM and other ingredients as listed in Table 1., according to the swine nutritional recommendations of National Research Council (NRC, 1995).
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    Prevalence of Gastrointestinal Parasites in Cattle in Badulla District (with special reference to Cryptosporidium spp)
    (Uva Wellassa University of Sri Lanka, 2010) Fernando, T.S.R.
    Due to the recognition of subclinical infections with negative impacts on production as diseases, control of gastrointestinal (GI) parasites in cattle is becoming established. Cryptosporidiosis is a gastrointestinal zoonotic disease and there is no effective therapy for this disease. This study examines prevalence of gastrointestinal parasites in Badulla district including Cryptosporidium spp and identifies the significance' of age, water source and feeding pattern for the prevalence of Cryptosporidium in cattle. The prevalence of gastrointestinal parasites including Cryptosporidium in faeces of 250 cattle in three age categories was examined. The eggs of gastrointestinal parasites were identified using the Mc Master method. Larval culture was done to identify the Genera of parasites. Oocysts of Cryptosporidium were demonstrated using the Shearther's sucrose flotation method followed by staining with modified Ziehl Neelsen technique. Prevalence of gastrointestinal parasites and. Cryptosporidium in Badulla district was 57.20% and 15.20% respectively. Trichostrongyl spp, Haemonchus spp, Strongyloid spp, Toxocara spp, Trichuris spp, Moneiza spp,. Eimeria spp were the common gastrointestinal parasites in cattle in Badulla district. Prevalence, of Cryptosporidium was significantly higher in cattle of <6 months (57.89%) compared with 7-12 months and >12 months of age (P value<0.05). Cryptosporidium oocysts were detected in cattle using surface and well water with the highest prevalence of infection (81.58%) were with surface water. There was no significant association of prevalence of Cryptosporidium oocysts with feeding pattern (P value>0.05). These animals are likely to play an important role in the epidemiology of cryptosporidiosis in cattle and humans. These findings clearly demonstrate that cattle farmers and the people living in villages amidst cattle in Badulla district are more exposed to the infection. Key Words: Cryptosporidium spp, Gastrointestinal parasites, Cattle
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    Prevalence of Gastrointestinal Parasites in Cattle in Badulla District(With special Reference to Cryptosporidium spp)
    (Uva Wellassa University of Sri Lanka, 2010) Fernando, T.S.R.; Rajapakshe, R.P.V.J.; Bullumulla, P.B.A.I.K.; Wijesundara, K.
    Due to the recognition of subclinical infections with negative impacts on production as diseases, control of gastrointestinal (GI) parasites in cattle is becoming established. Cryptosporidiosis is a gastrointestinal zoonotic disease and there is no effective therapy for this disease. This study examines prevalence of gastrointestinal parasites in Badulla district including Cryptosporidium spp and identifies the significance of age, water source and feeding pattern for the prevalence of Cryptosporidium in cattle. The prevalence of gastrointestinal parasites including Cryptosporidium in faeces of 250 cattle in three age categories was examined. The eggs of gastrointestinal parasites were identified using the Mc Master method. Larval culture was done to identify the Genera of parasites. Qa Oocysts of Cryptosporidium were demonstrated using the Shearther's sucrose flotation method followed by staining with modified Ziehl Neelsen echnique. Prevalence of gastrointestinal parasites and Cryptosporidium in Badulla district was 57.20% and 15.20% respectively. Trichostrongyl spp, Haemonchus spp, Strongyloid spp, Toxocara spp, Trichuris spp, Moneiza spp, Eimeria spp were the common gastrointestinal parasites in cattle in Badulla district. Prevalence of Cryptosporidium is significantly higher in cattle of <6 months (57.89%) compared with 7-12 months and >12 months of age (P<0.05). Cryptosporidium oocysts were detected in cattle using surface and well water with the highest prevalence of infection (81.58%) were with surface water. There was no significant association of prevalence of Cryptosporidium oocysts with feeding pattern (P>0.05). These animals are likely to play an important role in the epidemiology of cryptosporidiosis in cattle and humans. These findings clearly demonstrate that cattle farmers and the people living in villages amidst cattle in Badulla district are more exposed to the infection. Key Words: Cryptosporidium spp, Gastrointestinal parasites, Cattle