International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 2(11), 37-40, November (2013) Int. Res. J. Biological Sci. International Science Congress Association 37 Allozyme variation in house fly populations, Musca domestica from Allahabad, India Tripathi M.1*, Agrawal U.R.2 and Tripathi J.Department of Zoology, Iswar Saran Degree College, University of Allahabad, Allahabad-211004, INDIA Department of Zoology, CMP Degree College, University of Allahabad, Allahabad-211002, INDIAAvailable online at: www.isca.in, www.isca.me Received 5th July 2013, revised 16th August 2013, accepted 17th September 2013Abstract Allozyme variation was assessed in the four populations of the common house fly Musca domestica. Allozymes at three gene enzyme system unraveled four loci which revealed nine alleles. F statistics revealed that except XDH all the other loci show inbreeding (Fis�Fst ). Very little genetic differences have been found among the populations of M.domestica. Keywords: Musca domestica, allozyme variation, inbreeding, genetic identity, genetic distance. Introduction Enzyme polymorphism in natural population are useful tool for estimating genetic variation. The level of genetic variation within populations of a species indicate species vitality and potential for evolutionary responses to environmental changes. Musca domestica Linaeus, common house fly with cosmopolitan distribution is of great medical and sanitary importance as these are vectors of many diseases causing organism3-11. Genetic variations among house flies population have been analyzed using allozyme in different parts of the world12-19. However such studies have been sporadically carried out in Indian subcontinent20-22. In the present study allozyme variation in house fly, M. domestica from four different localities of Allahabad (U.P.) India has been analyzed to determine population diversity. Material and MethodsThe house flies Musca domestica. were collected using sweep nets from four different locations with qualitative difference in the food resources i.e., Meat shop (MS), Vegetable market (VM), Dairy farm (DF) and Solid food waste (SW). Fifty individuals were assayed for enzyme activity at three gene enzyme systems viz., malic enzyme (ME), aldehyde oxidase (AO) and xanthine dehydrogenase (XDH). Sample preparation and electrophoretic procedures were according to the method of Tripathi et al 201020. The enzyme systems studied, staining solutions, gel and electrode buffer are given in table-1. Genotype information, genetic identity and genetic distance were calculated as described earlier21. Results and DiscussionThe three gene enzyme systems analyzed in present study viz., malic enzyme (ME), aldehyde oxidase (AO) and xanthine dehydrogenase (XDH) resolved four loci among the four populations. The activity of ME and XDH were confined to single locus figure-1 and 3 while the activity of AO was confined to two loci figure-2. All the four loci analyzed were polymorphic, which revealed nine alleles. Allele frequencies and Chi-square values are presented in table-2. Three loci viz., ME, AO-1 and AO-2 showed significant departure from Hardy-Weinberg equilibrium. Flies from meat shop (MS) and dairy farm (DF) shared all the alleles while flies from vegetable market (VM) shared alleles at two locus i.e., ME and XDH with these two populations. The flies from solid food waste (SW) revealed fixed differences at ME and XDH, where it is monomorphic, however these flies shared VM at locus AO-1 and MS and DF at locus AO-2 table-2.Table1 Summary of electrophoresis and staining protocols followed in the present study Enzyme Gel/ electrode Buffer Staining buffer Substrate/Coenzyme Dyes Reference ME (E.C.1.1.1.40) 0.1M Tris-HCl (pH 8.5) 0.1M Tris-HCl (pH 7.4) Malic acid / NADP NBT PMS Tsukamoto (1989) 23 AO (E.C.1.2.3.1) 0.1M Tris-HCl (pH 8.5) 0.1M Tris-HCl (pH 7.4) Benzaldehyde NBT PMS Tsukamoto (1989) 23 XDH (E.C.1.2.1.37) 0.1M Tris-HCl (pH 8.5) 0.1M Tris-HCl (pH 7.4) Hypoxanthine / NADP NBT PMS Tsukamoto (1989) 23 International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(11), 37-40, November (2013) Int. Res. J. Biological Sci. International Science Congress Association 38 Table-2 Allele frequencies and Chi-square values in different collections of M. domestica Locus Allele MS VM DF SW ME (n=50) 0.600.430.391.00 b 0.40 0.57 0.61 - c 2 8.17* 6.72* 3.26 - AO-1 (n=50) a - 0.60 - 0.55 b - 0.40 - 0.45 c 2 - 8.17* - 7.76* AO-2 (n=50) a 0.44 - 0.35 0.48 b 0.56 - 0.65 0.52 c 2 7.45* - 0.79 6.40* XDH (n=50) a 0.20 0.40 0.29 1.00 b 0.42 0.60 0.45 - c 0.38 - 0.26 - c 2 0.62 1.61 2.62 - MS= Meat shop, VM = Vegetable market, DF= Dairy farm, SW= Solid food waste (in all the tables); n= number of individuals in each sample; *=Populations not in Hardy Weinberg equilibrium Figure-1 Electrophoretic phenotypes of Malic Enzyme (a) ME MS (b) ME VM (c) ME DF (d) ME SW in Musca domestica. The regions of electromorphs are indicated on the right Figure-2 Electrophoretic phenotypes of Aldehyde oxidase (a) AO MS (b) AO VM (c) AO DF (d) AO SW in Musca domestica. The regions of activity are indicated on the left and electromorphs are indicated on the right International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(11), 37-40, November (2013) Int. Res. J. Biological Sci. International Science Congress Association 39 Figure-3 Electrophoretic phenotypes of Xanthine dehydrogenase (a) XDH MS (b) XDH VM (c) XDH DF (d) XDH SW in Musca domestica.The regions of electromorphs are indicated on the right Number of alleles ranged from 1.85 to 2.07 with a mean of 1.97. The percentage of polymorphic loci ranged from 50.00% to 75.00% with a mean of 68.75% and the mean observed heterozygosity ranged from 0.250 to 0.447, with a mean of 0.356. The mean expected heterozygosity ranged from 0.483 to 0.534, with a mean of 0.510 table-3. Nei’s genetic identity values were highest between the samples collected from MS and DF (I=0.955) and the Nei’s genetic distance values were lowest between these two populations (D=0.046) table-4. As revealed by the Wright’s F statistics very little genetic variation seems to have has occurred among house fly populations analyzed in the present study. Except XDH all the other loci reveal inbreeding table-5. Conclusion It seems that the flies collected from MS and DF are genetically very similar. This may be due to the fact that the larval food substrates from the two collection sites were more or less similar as suggested by Thomas and Barker24. As well as the house fly population analyzed in the present study are characterized by a high level of inbreeding. Thus the present findings support the tenet that very little genetic differentiation has accompanied the population differentiation among house flies in Allahabad region of India22. AcknowledgementWe are grateful to Professor Raghav Ram Tewari, Cytogenetics Research laboratory, Department of Zoology, University of Allahabad for their useful suggestions, encouragement and providing laboratory facilities throughout the course of present work. Thanks are due to, Head, Department of Zoology (UGC-SAP and DST-FIST scheme sponsored), University of Allahabad, for providing all the necessary facilities. Table3 Genetic variability in different collections of house flies populations Population Sample Size Number of loci Mean Effective no. of alleles Percentage of polymorphic loci Mean observed heterozygosity (H o ) Mean expected heterozygosity (H) MS 50 4 1.85 75.00% 0.447 0.534 VM 50 4 2.07 75.00% 0.287 0.483 DF 50 4 1.95 75.00% 0.440 0.526 SW 50 4 2.01 50.00% 0.250 0.497 Mean 50 4 1.97 68.75% 0.356 0.510 = No. of heterozygotes / Total no. of individuals, H = 1- ˛ ( Nei, 1972 ), where x is the frequency of ith allele at a locus Table-4 Genetic identity (I) and genetic distance (D) among house flies in different collections (I) (D) Population MS VM DF SW MS 0.5580.9550.636 VM 0.584- 0.6070.621 DF 0.046 0.499 - 0.572 SW0.4520.4770.559 I= Jxy/Jx Jy, D= -In I, Where Jx y is the arithmetic mean of Jx y= x y over all loci, Jx is the arithmetic mean of ix = ix2 over all loci, and x (or yi) is the frequency of the ith allele in the first (or second) population. International Research Journal of Biological Sciences ________________________________________________ ISSN 2278-3202 Vol. 2(11), 37-40, November (2013) Int. Res. J. Biological Sci. International Science Congress Association 40 Table-5 Wright’s F statistics for all the variable loci. Loci F i s F st ME0.4740.243 AO-1 0.461 0.001 AO-2 0.267 0.010 XDH 0.055 0.284 Mean 0.314 0.135 References1.Murphy R.W., Sites J.W., Buth D.G. and Haufler C.H., Proteins: isozyme electrophoresis, In:Molecular systematic (eds. Hills D.M., Moritz C. and Mable B.K.), Sinauer Associates. Inc. Sunderland, Massachusetts, U.S.A., 655(1996)2.Mateus R.P. and Sene F.M., Temporal and spatial allozyme variation in the South American cactophilic Drosophila antonietae (Diptera : Drosophilidae), Biochem. Genet., 41, 219- 233 (2003)3.West L., Thehousefly. 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