ISJ 13: 23-27, 2016 ISJ 13: 23-27, 2016 ISSN 1824-307X SHORT COMMUNICATION Cutaneous neoplasm in Phaeotabanus litigiosus (Diptera, Tabanidae) collected on the Marambaia Island, Rio de Janeiro, Brazil RR Guimarães1,2,5, RR Guimarães-Junior2,3, RS Harlan-Ronald2, R Rodrigues-Guimarães4,5, FL Carvalho6, MLC Araújo-Junior6, RW Carvalho1 1Escola Nacional de Saúde Pública Sérgio Arouca-FIOCRUZ, Rio de Janeiro, RJ, Brasil 2Centro de Educação e Pesquisas em Medicina Ambiental - CEMA, Nilópolis, RJ, Brasil 3Associação Brasileira de Ensino Universitário - ABEU, Nova Iguaçu, RJ, Brasil 4Centro Universitário de Barra Mansa - UBM, Barra Mansa, RJ, Brasil 5Universidade Estácio de Sá - UNESA, Rio de Janeiro, RJ, Brasil 6Instituto Nacional do Câncer - INCA, Rio de Janeiro, Brasil Accepted January 28, 2016 Abstract A female specimen of Phaeotabanus litigiosus (Diptera: Tabanidae) collected on Marambaia Island was found with a tumor in the abdominal integument. Histopathological examination revealed an epithelial dysplasia with anisokariosis and hyperchromasia. This is the first record of a neoplasm found in tabanid collected from natural environment. Key Words: Atlantic island; displasia; horse fly; insect disease; insect vector; neotropical region   Introduction The family Tabanidae comprises approximately 4,300 species distributed throughout the world. The tabanids are mechanical and biological vectors of many pathogens to humans and domestic and wild animals (Foil 1989; Turcatel et al., 2007). Several biotic and abiotic factors determine the behavior of tabanid populations by varying the number of individuals according to different landscapes and over years and seasons (Guimarães, 2015). Interactive biotic factors are viruses, bacteria, fungi, helminths, other arthropods, fish, reptiles, amphibians and birds, which result in predation or parasitism. Some of these factors or other intrinsic factors cause pathologies that can lead to mutations, birth defects, metabolic disorders and malignancies that affect, in some degree, the survival of individuals or populations (Philip, 1965; Nayar, 1977; Savini and Furth, 2004; Asiain and Márquez, 2009; Ferreira, 2008, 2014). Neoplasm in insects has been described from various causes, such as genetic factors, viruses infections, exposure to insecticides and experimental nerve severance. Melanotic tumors are reported in Drosophila melanogaster (Diptera: Drosophilidae) linked to genetic factors and viral ___________________________________________________________________________ Corresponding author: Ronald Rodrigues Guimarães Centro de Educação e Pesquisas em Medicina Ambiental Av. Dr. Getúlio Vargas, 1797 Centro-Olinda, Nilópolis, CEP 26525-023 Rio de Janeiro, Brasil E-mail: ronaldrguimaraes@gmail.com infections (Taylor, 1969). Insecticides can lead to tumor formation in Musca domestica (Diptera: Muscidae) (Cantwell et al., 1966). Recurrent nerve severance induce formation of tumors in digestive tract of cockroach Leucophaea maderae (Blattodea: Blaberidae) (Matz and Bergoin, 1984). The literature records no pathology related to cancer or tumors in tabanids. This study is related to a cutaneous tumor formation found in Phaeotabanus litigiosus collected on Marambaia Island, Rio de Janeiro state, Brazil. Materials and Methods During October, November and December 2013 tabanids attracted by equine bait (Equus caballus) were collected through insect net, on Marambaia Island, municipality of Mangaratiba, Rio de Janeiro state, at location known as Vacaria Velha (23°03'47" S and 43°59'16" W). This site is close to secondary tropical forest and a swamp with a small pasture area where a tied equine spent the day; these collections were made from the morning twilight and all day until around 20:00 h, at least one day in each of the three months. For collection and transportation of biological samples the Instituto Brasileiro Chico Mendes de Biodiversidade do Ministério do Meio Ambiente issued permit nº 33382-1, SISBIO-IBAMA. The collected tabanid specimens were examined and identified according to literature (Barretto 1950; Coscarón and Papavero 2009a); 23  mailto:ronaldrguimaraes@gmail.com some specimens were mounted on entomological pins and other specimens were preserved in alcohol 70º; in one of these specimens a cutaneous injury was found on frontal side of abdomen. The specimen was examined under stereomicroscope Leica MZ 16 and the injury was photographed with Leica DFC 420 camera, at the Entomology Laboratory of Museu Nacional de História Natural do Rio de Janeiro. The injury was excised, fixed in buffered formalin and sent to the Laboratório da Seção Integrada de Tecnologia em Citopatologia do Instituto Nacional do Câncer - SITEC/INCA. The fixed material was prepared in histological sections stained with hematoxylin and eosin. The tabanid specimen and the blade with the histological sections are deposited in the Entomological Collection of Centro de Educação e Pesquisas em Medicina Ambiental - CEMA. Results P. litigiosus occurs in Brazil, in Minas Gerais, Rio de Janeiro, Sao Paulo and Parana states. During the three months of collection Vacaria Velha, Marambaia Island, 85 specimens were collected. Material examined: P. litigiosus, Brazil, Rio de Janeiro, Mangaratiba, Marambaia Island, Vacaria Velha [23°03'47 "S and 43°59'16" O], 4-5.xii.2013, Guimarães Jr. leg., Guimarães det., (1 ♂ CEMA). The observed injury was located between the first and the second left abdominal tergite (Fig. 1) about 0.5 mm in diameter. The injury showed rough and hardened surface. Histopathologic examination showed a cavity formed by a simple squamous epithelium, with an area of abnormal multiplication of integumentary cells; abnormal cells presented increased size nucleus and little cytoplasm (anisokariosis). Increased size nucleus presented granular coarsely chromatin (hyperchromasia), and evident nucleolus. There was no evidence of invasion of underlying corium or mitotic figures (Fig. 2). Discussion Neoplasic formations in insects are rarely reported in the literature: the insects are highly resistant to carcinogenesis, as most adult cells are post-mitotic, unable to multiply. However, during the larval stages, the stem cells present in the imaginal discs may change that result in neoplastic formations. Kirby and Spence (1826) refer to external wounds as precursors of tumors in insects, which does not seem to be the case of examined specimen, because any other anatomical changes on outer surface was observed. As to integument tissue formations for the account of Balazuc (1948) the meeting of a specimen of Phytodecta variabilis (Coleoptera: Chrysomelidae) presenting a large tumor formation in prothorax without histopathology. White (1929) found similar structure to fibroma, originated from connective tissue in the thorax of a honeybee (Apis mellifera (Hymenoptera: Apidae). A brain tumor, presumably formed by glial cells, was found in worker of Formica pratensis (Hymenoptera: Formicidae) without differential diagnosis of abscess (Brun, 1925). Palm (1948) described a tumor formation in ‘corpus allatum’ of a male nymph of a species of Gryllotalpa (Orthoptera: Gryllotalpidae); the tumor showed giant cells formed from the cytoplasmic and nuclear fusion of smaller cells with kariorrhexis, indistinct nuclear membrane with numerous beads of chromatin and dispersed core remains in the cytoplasm. A tumor in pharyngeal gland was described in a species of the genus Bombus (Hymenoptera: Apidae): the affected gland showed a compact structure, displaced, enlarged, with abundant connective tissue, with no connection to the common duct, with numerous secretory ducts ending blindly. The cells showed signs of hypersecretion, degeneration and kariorrhexis (Palm, 1949). Salivary gland tumor were also observed in Periplaneta americana (Blattaria: Blattidae) when the ducts were tied or removed (Sutherland, 1963, 1967). Fig. 1 Cutaneous injury in Phaeotabanus litigiosus collected on Marambaia Island, Mangaratiba, Rio de Janeiro, Brazil. The black bar measures 1 mm. Photo obtained at Entomology Laboratory of Museu Nacional de História Natural do Rio de Janeiro. 24  Fig. 2 Histopathological aspects of tumor found in Phaeotabanus litigiosus collected on Marambaia Island, Mangaratiba, Rio de Janeiro, Brazil; A) Cavity (cav) formed by a simple squamous epithelium (see) and epithelial displasia area (eda) (400x); B and C) Cavity (cav) and an epithelial dysplasia area (eda) with anisokariosis, hyperchromasia, coarsely granular chromatin, evident nucleolus, and little cytoplasm (ah) (1000x). 25  Among the invertebrates D. melanogaster is considered to be the most susceptible species to cancerous tumors, subject to tumors determined by variations in temperature, exposure to X-rays or by chromosomal inheritance and most carcinogenesis records on insects is made in D. melanogaster (Wautier and Wautier, 1952; Slade, 2012). Melanotic hereditary tumors in larvae of D. melanogaster tumorw strain involve encapsulation of caudal lipid bodies by hemocytes and hematopoietic organs. These hematopoietic bodies put in service a large number of blood cells, which originate lamelocytes and plasmatocytes which synthesize endogenous tissues (Perotti and Bairati, 1968; Rizki and Rizki 1974; Nappi et al., 1984; Silvers and Hanratty 1984; William and Hanratty 1984). The melanotic encapsulation performed by melanocytes is a characteristic response against aberrant tissues in Drosophila and other insects (Tascedda and Ottaviani, 2014). The Tumorosa-lethal mutation (Tum-l) is temperature-dependent and also leads to excessive proliferation of hemocytes and the formation of neoplasic melanotic tumors in the larval hematopoietic bodies (Luo et al., 1993). Rous sarcoma virus determines chromosomal abnormalities, and tumor formation in D. melanogaster that did not seem to be a really neoplasic formation, as it is known in mammals (Burdette and Yoon, 1967; Kirk et al., 1970). The polyhedrosis virus occurrs naturally in Gilpinia hercyniae (Hymenoptera: Diprionidae) and determines proliferation of the germ cells forming a tumor that surrounds the external surface of the digestive tract (Bird, 1948). Abnormal proliferation of epidermal cells larvae of Hyphantria cunea moth (Lepidoptera: erebidae) is also caused by the nuclear polyhedrosis virus and determines polyhedral formations in hypertrophied nuclei, which differs from the changes found in classical neoplasms (Watanabe, 1968). Tumors in the digestive tract caused by experimental severance of recurrent nerve in L. maderae are formed by proliferation of epithelial cells and hemocytes encapsulated by granulocytes; some authors point inflammatory processes determined by microorganisms in the digestive tract is the cause of the alteration (Scharrer, 1945a, b, 1948, 1949a, b; Matz, 1965; Taylor, 1969; Matz and Bergoin, 1984) The fact that these tumors may be transplanted and proliferate in other individuals from filtered cell or insect DNA with recurrent nerve damage, although not present tumors, indicating a likely viral etiology (Matz and Bergoin, 1984). The fluorenone derivatives induce the formation of tumor injuries in the hypodermis, intestine and lipid bodies in M. domestica (Cantwell et al., 1966). In literature, only D. melanogaster is recorded as susceptible to true tumors. Thus, the cutaneous neoplasm record in adult Tabanidae, collected in nature, is unique true record of cancer in other species of Diptera. The histopathologic diagnosis differs from any other recorded in similar injuries related to other insects. The used methodology did not allow the study of the cause of injury. Acknowledgements The authors thank Dr. M Couri, Museu Nacional de História Natural do Rio de Janeiro, for tabanid specimen photography. The authors also thank Escola Nacional de Saúde Pública Sérgio Arouca - ENSP-FIOCRUZ for financial support and CAPES (Fellowship process nº 1383383). References Asiain J, Márquez J. New teratological examples in Neotropical Staphylinidae (Insecta: Coleoptera), with a compilation of previous teratological records. Revista Mexicana de Biodiversidad 80: 129-139, 2009. Balazuc J. La teratologie des coleopteres et experiences de transplantation sur Tenebria molitor L. Mémoires du Muséum National d'Histoire Naturelle 26: 1-293, 1948. Barretto MP. Estudo sobre Tabanidas brasileiros. XIII. O gênero Phaetobanus Lutz e descrição de dois novos gêneros (Diptera, Tabanidae). Anais Faculdade de Medicina de São Paulo 25: 89-100, 1950. Bird FT. Tumours associated with a Virus Infection in an Insect. Nature 163: 777-778, 1949. Brun R. Ein Fall von Hirntumor bei der Ameise. Schweiz. Schweizer Archiv für Neurologie und Psychiatrie 16: 86-99, 1925. Burdette WJ, Yoon S. Mutations, Chromosomal Aberrations, and Tumors in Insects Treated with Oncogenic Virus. Science 155: 340-341, 1967. Cantwell GE, Shortino TJ, Robbins WE. The histopathological effects of certain carcinogenic 2-fluorenamine derivatives on larvae of the house fly. J. Invertebr. Pathol. 8: 167-174, 1966. Coscarón S, Papavero N. Catalogue of Neotropical Diptera. Tabanidae. Neotropical Diptera 16: 1- 199, 2009. Coscarón S, Papavero N. Manual of Neotropical Diptera. Tabanidae. Neotropical Diptera 6: 1- 137 2009a. Ferreira RN. A teratological specimen of Calosoma sycophanta (L.), (Coleoptera; Carabidae) from Connecticut, USA. Entomol. News 119: 307- 309, 2008. Ferreira RN. Two physical abnormalities in Coleoptera (Cerambycidae, Lucanidae) from Rhode Island, USA. Arquivos Entomoloxicos 10: 173-174, 2014. Foil LD. Tabanids as vectors of disease agents. Parasitol. Today 5: 88-96, 1989. Guimarães RR. Tabanidae (Insecta: Diptera): caracterização, ecologia e interação com a população Quilombola da Ilha da Marambaia, Rio de Janeiro, Brasil [Ph.D. thesis]. Escola Nacional de Saúde Pública-FIOCRUZ. p 187, 2015. Kirb W, Spence W. An introduction to Entomology. In: Taylor R (ed.), Shoe Lane, Londres, p 653, 1826. Kirk HD, Ewen AB, Emson HE. Melanotic lesions in two insect species: Drosophila melanogaster (Diptera) and Melanoplus sanguinipes (Orthoptera). J. Invertebr. Pathol. 15: 351-355, 1970. Luo H, Rose P, Barber D, Hanratty WP, Lee S, Roberts TM, et al. Mutation in the Jak kinase 26  http://www.sciencemag.org/search?author1=Walter+J.+Burdette&sortspec=date&submit=Submit http://www.sciencemag.org/search?author1=Jong+Sik+Yoon&sortspec=date&submit=Submit http://www.sciencedirect.com/science/article/pii/002220116690125X http://www.sciencedirect.com/science/article/pii/002220116690125X http://www.sciencedirect.com/science/article/pii/002220116690125X http://www.sciencedirect.com/science/article/pii/002220116690125X JH2 domain hyperactivates Drosophila and mammalian Jak-Statpathways. Mol. Cell Biol. 17: 1562-1571, 1997. Scharrer B. Malignant characteristics of experimentally induced tumors in the insect, Leucophaea maderae (Orthoptera). Anat. Record 100: 774-775, 1948. Matz G. Morphogenese tumorale chez les insects. J. Insect Physiol. 11: 637-638, 1965. Scharrer B. Gastric cancer experimentally induced in insects by nerve severance. Journal of Natl. Cancer Inst. 10: 375-76, 1949a Matz G, Bergoin M. Experimental tumors in Leucophaea maderae (Dictyoptera): A viral etiology. J. Invertebr. Pathol. 43: 424-428, 1984. Scharrer B. Tumor mortality and sex mortality in Licophaea maderae (Orthoptera). Anat. Record 105: 624, 1949b. Matz G, Weil JH, Joly P, Ebel JP. Transmission of tumors in Locusta migratoria Linnaeus by nucleic acid extracted from the tumors. J. Invertebr. Pathol. 8: 8-13, 1966. Silvers M, Hanratty WP. Alterations in the production of hemocytes due to a neoplastic mutation of Drosophila melanogaster. J. Invertebr. Pathol. 44: 324-328, 1984. Nappi AJ, Kmiecik J, Silvers M. Cellular immune competence of a Drosophila mutant with neoplastic hematopoietic organs. J. Invertebr. Pathol. 44: 220-227, 1984. Slade H. How can we learn about human neoplasms by studying animals, plants, fish and insects? In: Neoplasms: principles of development and diversity, Jones and Bartlett Publishers, USA, pp 67-84, 2012. Nayar JL. Abdominal Teratology in Eristalis tenax (Linnaeus) from Libya and India (Diptera: Syrphidae). Oriental 11: 639-664, 1977. Sutherland DJ. Experimentally induced tumours in Periplaneta americana L. J. Insect Physiol. 9: 131-135, 1963. Palm NB. Notes on the structure of the corpora allata in Gryllotalpa. Kungl. Fysiografiska Siillskapets i Lund Forhandlingar 17: 1-11, 1948. Sutherland DJ. The development of salivary tumours in Periplaneta americana (L.) as induced by duct ligation. J. Insect Physiol. 13: 137-152, 1967. Palm NB. The pharyngeal gland in Bombus Latr. and Psithyrus Lep., with a description of a case of pathological development of the pharyngeal gland. Opuscula Entomologica 14: 27-47, 1949. Tascedda F, Ottaviani E. Tumors in invertebrates. Inv. Surv. J. 11: 197-203, 2014. Taylor RL. Formation of tumorlike lesions in the cockroach Leucophaea maderae after nerve severance. J. Invertebr. Pathol. 13: 167-187, 1969. Perotti ME, Bairati JrA. Ultrastructure of the melanotic masses in two tumorous strains of Drosophila melanogaster (tuB3 and Freckled). J. Invertebr. Pathol. 10: 122-138, 1868. Turcatel M, Carvalho CJB, Rafael JA. Mutucas (Diptera: Tabanidae) do estado do Paraná, Brasil: chave de identificação pictórica para subfamílias, tribos e gêneros. Biota Neotropica 7: 265-278, 2007. Philip CB. The Identity and relantionships of Tabanus (Neotabanus) vittiger and notes on two cases of teratology in Tabanidae (Diptera). Ann. Entomol. Soc. Am. 58: 876-80, 1965. Watanabe H. Abnormal cell proliferation in the epidermis of the fall webworm, Hyphantria cunea, induced by the infection of a nuclear- polyhedrosis virus. J. Invertebr. Pathol. 12: 310- 315, 1968. Rizki TM, Rizki RM. Topology of the caudal fat body of the tumorw mutant of Drosophila melanogaster. J. Invertebr. Pathol. 24: 37-40, 1974. Savini V, Furth, DG. Teratología en Coleoptera: un caso en Gioia bicolor (Blake, 1969) (Chrysomelidae, Alticinae) de Jamaica. Entomotropica 19: 165-167, 2004. Wautier V, Wautier J. Le Cancer et les Invertébrés. Bulletin Mensuel de la Société Linnéenne de Lyon 3: 66-96, 1962. White PB. Note on a case of fibroma in a honey- bee. J. Pathol. Bacteriol. 24: 138-39, 1921. Scharrer B. Experimental tumors in an insect. Science 102: 102, 1945a. William MS, Hanratty P. Alterations in the production of hemocytes due to a neoplastic mutation of Drosophila melanogaster. J. Invertebr. Pathol. 44: 324-328, 1984. Scharrer B. Experimental tumors after nerve section in an insect. Proc. Soc. Exp. Biol. Med. 60: 184- 89, 1945b.   27  Ferreira RN. Two physical abnormalities in Coleoptera (Cerambycidae, Lucanidae) from Rhode Island, USA. Arquivos Entomoloxicos 10: 173-174, 2014. Foil LD. Tabanids as vectors of disease agents. Parasitol. Today 5: 88-96, 1989. Guimarães RR. Tabanidae (Insecta: Diptera): caracterização, ecologia e interação com a população Quilombola da Ilha da Marambaia, Rio de Janeiro, Brasil [Ph.D. thesis]. Escola Nacional de Saúde Pública-FIOCRUZ. p 187, 2015.