Genetic Variability In Mitochondrial Dna Of The Screwworm, Cochliomyia Hominivorax (diptera: Calliphoridae), From Brazil

Restriction fragment length polymorphism (RFLP) analysis of mitochondrial DNA (mtDNA) was used to examine genetic variation and population structure of screwworm flies in four populations from Sao Paulo State, Brazil. The total DNA of 405 individuals was digested with 15 restriction endonucleases and probed with five cloned HindIII fragments representing the entire mitochondrial genome of Cochliomyia hominivorax. The survey revealed that four enzymes (HaeIII, HindIII, MspI, and PvuII) were suitable to detect mtDNA variation among all populations. Based on the fragment patterns obtained for these four enzymes, a total of 15 haplotypes in combination was detected. Heteroplasmic individuals for the PvuII pattern were obtained in one of the populations. The estimated average for nucleotide sequence divergence (δ) was 0.92%. The cladogram of the geographical distribution among the observed haplotypes suggests that the sampled screwworms probably belong to a single evolutionary lineage with populations interconnected by reduced gene flow.337-823725

Random Amplified Polymorphic Dna Of Screwworm Fly Populations (diptera: Calliphoridae) From Southeastern Brazil And Northern Argentina

The screwworm fly Cochliomyia hominivorax is one of the most important agents of traumatic myiasis throughout neotropical regions. In this work, we optimized the technique of RAPD-PCR for these species and used it to study the genetic variability among seven populations (six from southeastern Brazil and one from northern Argentina) of C. hominivorax. RAPD fingerprints showed high variation for 12 primers used, revealing 209 presumptive loci of which 198 were polymorphic. Marker pattern relationships for these different populations were used to determine genetic relatedness, as well as to examine potential patterns of gene flow. Our interpretation of Lynch and Milligan's analogue of Wright's F((ST)) was that C. hominivorax populations are genetically subdivided (F'((ST)) for pooled samples = 0.122). Our data suggested that the subdivision detected in C. hominivorax populations by RAPD can be explained by the interplay of random factors affecting allele frequency changes. These results indicate that the RAPD-PCR technique is useful for revealing genetic variation in screwworm fly populations not detected by others techniques and can represent an efficient method for understanding the genetic structure and population genetic phenomena of this important pest.424772779Apostol, B.L., Black W.C. IV, Reiter, P., Miller, B., Population genetics with RAPD-PCR markers: The breeding structure of Aedes aegypti in Puerto Rico (1996) Heredity, 76, pp. 325-334Baruffi, L., Damiani, G., Guglielmino, C.R., Bandis, C., Malacrida, A.R., Gasperi, G., Polymorphism within and between populations of Ceratitis capitata: Comparison betwee RAPD and multilocus enzyme electrophoresis data (1995) Heredity, 74, pp. 425-437Begun, D.J., Aquadro, M., Levels of naturally occurring DNA polymorphism are correlated with recombination rates in Drosophila melanogaster (1992) Nature (London), 356, pp. 519-520Bielawsky, J.P., Pumo, D.E., Randomly amplified polymorphic DNA analysis of atlantic coast striped bass (1997) Heredity, 78, pp. 32-40Black W.C. IV, Duteau, N.M., Puterka, G.J., Nichols, J.R., Pettorini, J.M., Use of the random amplified polymorphic dna polymerase chain reaction (RAPD-PCR) to detect DNA polymorphism in Aphids (Homóptera: Aphididae) (1992) Bull. Entomol. Res., 82, pp. 151-159Bush, G.L., Neck, R.W., Ecological genetics of the Screwworm fly, Cochliomyia hominivorax (Diptera: Calliphoridae) and its bearing on the quality control of mass reared insect (1976) Environ. Entomol., 5, pp. 821-826Coquerel, C., Note sur une larve d'oestridae extraite du bras d'un homme à Cayenne (1859) Rev. Mag. Zool, 2 (11), p. 356Crespi, B.J., Heterozigosity in haplodiploid thysanoptera (1991) Evolution, 45, pp. 458-464Dear, J.P., A revision of the new world Chrysomyini (Diptera:Calliphoridae) (1985) Rev. Bras. Zool., 3, pp. 109-169Dev, V., La Chance, L.E., Whitten, C.J., Polytene chromosomes, karyotype correlations and population cytology of the primary screwworm fly (1986) J. Hered., 77, pp. 427-434Gibbs, H.L., Prior, K.A., Weatherhead, P.J., Genetic analysis of populations of threatened snake species using RAPD markers (1994) Mol. Ecol., 3, pp. 329-337Graham, O.H., Symposium on eradication of the screwworm from the United States and Mexico (1985) Misc. Publ. Entomol. Soc. Am., 62, pp. 1-68Graur, D., Gene diversity in Himenoptera (1985) Evolution, 39, pp. 190-199Guimarães, J.H., Papavero, N., Pires Do Prado, A., As miíases na região Neotropical identificaçào, biologia, bibliografia (1983) Rev. Bras. Zool., 1 (4), pp. 239-416Hadrys, H., Balick, M., Schierwater, B., Applications of random amplified polymorphic DNA (RAPD) in molecular ecology (1992) Mol. Ecol., 1, pp. 55-63Hall, M., Wall, R., Myiasis of humans and domestic animals (1995) Adv. Parasitol., 35, pp. 57-334Haymer, D.S., Random amplified polymorphic DNA and microsatellites. What are they and can they tell us anything we don't already know? (1994) Ann. Entomol. Soc. Am., 87, pp. 717-722Haymer, D.S., Mei, H., McInnis, D.O., Genetic marker analysis of spatial and temporal relationships among existing populations and new infestations of the mediterranean fruit fly (Ceratitis capitata) (1997) Heredity, 79, pp. 302-309Hedrick, P., Shooting the RAPD's (1992) Nature (London), 355, pp. 679-680Hartl, D.L., (1988) A Primer of Population Genetics, , Sinauer Associates, Inc. Sunderland, MassInfante, M.E., Azeredo Espin, A.M.L., Genetic variability in mitochondrial DNA of screwworm, Cochliomyia hominivorax (Diptera: Calliphoridae), from Brazil (1995) Biochem. Genet., 33, pp. 737-756Innis, M.A., Gelfand, D.H., Optimization of PCR (1990) PCR Protocols: A Guide to Methods and Applications, pp. 3-12. , Edited by M.A. Innis, D.H. Gelfand, J.J. Sninsky, and T.J. White. Academic Press Inc., San Diego, CalifKrafsur, E.S., Whitten, C.J., Breeding structure of screwworm fly populations (Diptera:Calliphoridae) in Colima, Mexico (1993) J. Med. Entomol., 30, pp. 477-480LaChance, E.E., Whitten, C.J., Cytogenetic studies of screwworm (Diptera:Calliphoridae) populations from Southern Mexico and Jamaica. Symposium: Genetics in entomology (1986) Ann. Entomol. Am., 79, p. 729Lu, R., Rank, G.H., Use of RAPD analysis to estimate population genetic parameters in the alfalfa leaf cutting bee, Megachile rotundata (1996) Genome, 39, pp. 655-663Lynch, M., Milligan, B.G., Analysis of population genetic structure with RAPD markers (1994) Mol. Ecol., 3, pp. 91-99Nei, M., Genetic distance between populations (1972) Am. Nat., 106, pp. 283-292Nei, M., (1987) Molecular Evolutionary Genetics, , Columbia University Press. New YorkNei, M., Li, W.H., Mathemathical model for studying genetic variation in terms of restriction endonucleases (1979) Proc. Natl. Acad. Sci. U.S.A., 76, pp. 5269-5273Novo, M.T.M., De Souza, A.P., Garcia O., Jr., Ottoboni, L.M.M., RAPD genomic fingerprinting differenciates Thiobacillus ferrooxidans strains (1996) Syst. Appl. Microbiol., 19, pp. 91-95Plikaytis, B.D., Carlone, G.M., Edwards, P., Mayer, W., Robust estimation of standard curves for protein molecular weigth and linear-duplex DNA based-pair number after gel electrophoresis (1986) Anal. Biochem., 152, pp. 346-364Pomonis, J.G., Cuticular hydrocarbons of the screwworm, Cochliomyia hominivorax (Diptera:Calliphoridae). Isolation, identification and quantification as a funtion of age, sex and irradiation (1989) J. Chem. Ecol., 15, p. 2301Roehrdanz, R.L., Intraspecific genetic variability in mitochondrial DNA of screwworm fly (C.hominivorax) (1989) Biochem. Genet., 27, pp. 551-559Roehrdanz, R.L., Johnson, D.A., Mitochondrial DNA variation among geographical populations of the Screwworm fly, Cochliomyia hominivorax (1988) J. Med. Entomol., 25 (2), pp. 136-141Sambrook, J., Fritsch, E.F., Maniatis, T., (1989) Molecular Cloning. A Laboratory Manual, , Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.YStevens, J., Wall, R., The use of random amplified polymorphic DNA (RAPD) analysis for studies of genetic variation in populations of the blowfly Lucilia sericata (Diptera: Calliphoridae) in southern England (1995) Bull. Entomol. Res., 85 (4), pp. 549-555Taylor, D.B., Petterson R.D., Jr., Population genetics and gene variation in primary and secondary screwworm (Diptera: Calliphoridae) (1994) Ann. Entomol. Soc. Am., 87, pp. 626-633Taylor, D.B., Petterson R.D., Jr., Moya-Borja, G.E., Populations genetics and gene variation in screwworms (Diptera: Calliphoridae) from Brazil (1996) Biochem. Genet., 34 (1-2), pp. 67-76Whitten, C.J., Use of the isozyme technique to asses the quality of mass-reared sterile screwworm flies (1980) Ann. Entomol. Soc. Am., 73, pp. 7-10Williams, J.G.K., Kubelik, A.R., Livak, K.J., Rafalsky, J.A., Tingey, S.V., DNA polymorphism amplified by arbitrary primers are useful as genetic markers (1990) Nucleic Acids. Res., 18, pp. 6531-6535Welsh, J., McCleland, M., Fingerprint genomes using PCR with arbitrary primers (1990) Nucleic. Acids. Res., 18, pp. 7213-7218Wright, S., (1978) Evolution and the Genetics of Populations. Vol.4. Variability within and among Natural Populations, 4. , University of Chicago Press. ChicagoYong, H.S., Low allozyme variability in Batrocera albistrigata (Insecta: Diptera: Tephritidae) from peninsular Malasya (1990) Comp. Biochem. Physiol. B Comp. Biochem. Mol. Biol., 97, pp. 119-12

Analysis Of Mitochondrial Dna Variability And Genetic Structure In Populations Of New World Screwworm Flies (diptera: Calliphoridae) From Uruguay

The New Word screwworm, Cochliomyia hominivorax (Coquerel 1858) (Diptera: Calliphoridae), is one of the most important insect pests of livestock in the Neotropical region. In this work, polymerase chain reaction-restriction fragment length polymorphism of mitochondrial DNA (mtDNA) was used to study the diversity and population structure of seven geographically distinct populations of C. hominivorax from most of the important livestock areas in Uruguay. The control region (A+T/12S) and subunits 1 and 2 of cytochrome oxidase (cox1/cox2) were amplified and digested with restriction endonucleases. Nine haplotypes were observed among the populations sampled. The mean nucleotide diversity and the haplotype diversity indicated high mtDNA variability in this species. The similarity index, average nucleotide divergence, and analysis of molecular variance results showed no evidence of subpopulation differentiation, indicating that the C. hominivorax populations of Uruguay form a single panmitic population. The distribution pattern of the genetic-variation in natural populations of C. hominivorax and the implications of these results for establishing control program are discussed. © 2005 Entomological Society of America.424589595Avise, J.C., (1994) Molecular Markers, Natural History and Evolution, , Chapman & Hall, New YorkAvise, J.C., Arnold, J., Ball, R.M., Bermingham, E., Lamb, T., Neigel, J.E., Reeb, C.A., Saunders, N.C., Intraspecific phylogeography: The mitochondrial DNA bridge between population genetics and systematics (1987) Annu. Rev. Ecol. Syst., 18, pp. 489-522Azeredo-Espin, A.M.L., Mitochondrial DNA variability in geographic populations of screwworm fly from Brazil (1993) Int. Atomic Energy Agency, 327, pp. 161-165Carballo, M., Colombo, A., Heinzen, T., Presencia de especies de dípteros Califóridos causantes de miasis cutáneas en Uruguay. Relevamiento de larvas parasitarias (instar III) en rumiantes (1990) Veterinaria, 26, pp. 4-6Dueñas, J.C.R., Panzetta-Dutari, G.M., Blanco, A., Gardenal, C.N., Restriction fragment-length polymorphism of mtDNA A+T-rich region as a genetic marker in Aedes aegypti (Diptera: Culicidae) (2002) Ann. Entomol. Soc. Am., 95, pp. 352-358Excoffier, L., Smouse, P.E., Quattro, J.M., Analysis of molecular variance inferred from metric distances among DNA haplotypes: Application to human mitochondrial DNA restriction data (1992) Genetics, 131, pp. 479-491Guimarães, J.H., Papavero, N., Prado, A.P., As miíases na região Neotropical (identificaçã o, biologia, bibliografia) (1983) Rev. Bras. Zool., 1, pp. 239-416Hall, M., Wall, R., Myiasis of human and domestic animals (1995) Adv. Parasitol., 35, pp. 256-333Heiman, M., (1997) Software Webcutter 2.0, , http://www.firstmarket.com/cutterInfante-Malachias, M.E., (1999) Estrutura Genética de Populações de Cochliomyia hominivorax (DípteraCalliphoridae) Da Região Sudeste Do Brasil: Análise Através de 3 Tipos de Marcadores Genéticos, , Ph.D. dissertation, State University of Campinas (UNICAMP), Campinas, SP, BrazilInfante-Malachias, M.E., Yotoko, K.S.C., Azeredo-Espin, A.M.L., Random amplified polymorphic DNA of screwworm fly populations (Diptera: Calliphoridae) from southeastern Brazil and northern Argentina (1999) Genome, 42, pp. 772-779Infante-Vargas, M.E., Azeredo-Espin, A.M.L., Genetic variability in mitochondrial DNA of screwworm, Cochliomyia hominivorax (Diptera: Calliphoridae), from Brazil (1995) Biochem. Genet., 33, pp. 237-256(2000) Genetic Sexing and Population Genetics of Screwworms, , IAEA-314-D4-00CT2176. International Atomic Energy Agency, Vienna, AustriaKrafsur, E.S., Whitten, C.J., Breeding structure of screwworm fly populations (Diptera-Calliphoridae) in Colima, Mexico (1993) J. Med. Entomol., 30, pp. 477-480Litjens, P., Lessinger, A.C., Azeredo-Espin, A.M.L., Characterization of screwworm flies Cochliomyia hominivorax and Cochliomyia macellaria by PCR-RFLP of mitochondrial DNA (2001) Med. Vet. Entomol., 15, pp. 183-188Lessinger, A.C., Azeredo-Espin, A.M.L., Evolution and structural organization of mitochondrial DNA control region of myiasis-causing flies (2000) Med. Vet. Entomol., 14, pp. 71-80Lessinger, A.C., Junqueira, A.C.M., Lemos, T.A., Kemper, E.L., Da Silva, F.R., Vettore, A.L., Arruda, P., Azeredo-Espin, A.M.L., The mitochondrial genome of the primary screwworm fly Cochliomyia hominivorax (Diptera: Calliphoridae) (2000) Insect Mol. Biol., 9, pp. 521-529McElroy, D., Moran, P., Bermingham, E., Kornfield, I., (1992) Restriction Analysis Package (REAP), Version 4.0, , University of Maine, Orono, ME(2002), http://www.mgap.gub.uy/Nei, M., (1987) Molecular Evolutionary Genetics, , Columbia University, New YorkNei, M., Li, W.H., Mathematical model for studying genetic variation in terms of restriction endonucleases (1979) Proc. Natl. Acad. Sci. U.S.A., 76, pp. 5269-5273Nei, M., Tajima, F., DNA polymorphism detectable by restriction endonucleases (1981) Genetics, 97, pp. 145-163Roehrdanz, R.L., Intraspecific genetic variability in mitochondrial DNA of the screwworm fly (Cochliomyia hominivorax) (1989) Biochem. Genet., 27, pp. 551-569Roehrdanz, R.L., Johnson, D.A., Mitochondrial DNA variation among geographical populations of the screwworm fly, Cochliomyia hominivorax (1988) J. Med. Entomol., 25, pp. 136-141Rokas, A., Ladoukakis, E., Zouros, E., Animal mitochondrial DNA recombination revisited (2003) Trends Ecol. Evol., 18, pp. 411-417Ross, K.G., Krieger, M.J.B., Shoemaker, D.D., Vargo, E.L., Keller, L., Hierarchical analysis of genetic structure in native fire ant populations: Results from three classes of molecular markers (1997) Genetics, 147, pp. 643-655Schneider, Roessli, S.D., Excoffier, L., (2001) Arlequin Ver. 2.01. Software for Population Genetics Data Analyses, , Genetics and Biometry Laboratory. Department of Anthropology and Ecology, University of Geneva, SwitzerlandSimon, C., Frati, F., Beckenbach, A., Crespi, B., Liu, H., Flook, P., Evolution, weighting and phylogenetic utility of mitochondrial gene sequences and compilation of conserved polymerase chain reaction primers (1994) Annu. Entomol. Soc. Am., 87, pp. 651-701Taylor, D.B., Peterson II, R.D., Population genetics and gene variation in primary and secondary screwworm (Diptera: Calliphoridae) (1994) Ann. Entomol. Soc. Am., 87, pp. 626-633Taylor, D.B., Szalanski, A.L., Peterson II, R.D., Mitochondrial DNA variation in screwworm (1996) Med. Vet. Entomol., 10, pp. 161-169Thomas, D.B., Mangan, R.L., Oviposition and wound visiting behavior of the screwworm fly Cochliomyia hominivorax (Coquerel) (1989) Ann. Entomol. Soc. Am., 82, pp. 526-534Torres, T.T., Brondani, R.P.V., Garcia, J.E., Azeredo-Espin, A.M.L., Isolation and characterization of microsatellite markers in the new world screw-worm Cochliomyia hominivorax (Diptera: Calliphoridae) (2004) Mol. Ecol. Notes, 4, pp. 182-184Vargas-Teran, M., Hursey, B.S., Cunningham, E.P., Eradication of the screwworm from Libya using the sterile insect technique (1994) Parasitol. Today, 10, pp. 119-122Wyss, J.H., Galvin, T.J., Central America regional screwworm eradication program (benefit/cost study) (1996) Ann. N.Y. Acad. Sci., 791, pp. 241-24