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Simukunin from the Salivary Glands of the Black Fly Simulium vittatum Inhibits Enzymes That Regulate Clotting and Inflammatory Responses  [PDF]
Hitoshi Tsujimoto, Michail Kotsyfakis, Ivo M. B. Francischetti, Jai Hoon Eum, Michael R. Strand, Donald E. Champagne
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0029964
Abstract: Background Black flies (Diptera: Simuliidae) feed on blood, and are important vectors of Onchocerca volvulus, the etiolytic agent of River Blindness. Blood feeding depends on pharmacological properties of saliva, including anticoagulation, but the molecules responsible for this activity have not been well characterized. Methodology/Principal Findings Two Kunitz family proteins, SV-66 and SV-170, were identified in the sialome of the black fly Simulium vittatum. As Kunitz proteins are inhibitors of serine proteases, we hypothesized that SV-66 and/or ?170 were involved in the anticoagulant activity of black fly saliva. Our results indicated that recombinant (r) SV-66 but not rSV-170 inhibited plasma coagulation. Mutational analysis suggested that SV-66 is a canonical BPTI-like inhibitor. Functional assays indicated that rSV66 reduced the activity of ten serine proteases, including several involved in mammalian coagulation. rSV-66 most strongly inhibited the activity of Factor Xa, elastase, and cathepsin G, exhibited lesser inhibitory activity against Factor IXa, Factor XIa, and plasmin, and exhibited no activity against Factor XIIa and thrombin. Surface plasmon resonance studies indicated that rSV-66 bound with highest affinity to elastase (KD = 0.4 nM) and to the active site of FXa (KD = 3.07 nM). We propose the name “Simukunin” for this novel protein. Conclusions We conclude that Simukunin preferentially inhibits Factor Xa. The inhibition of elastase and cathepsin G further suggests this protein may modulate inflammation, which could potentially affect pathogen transmission.
Larval description of Simulium (Notolepria) cuasiexiguum and Simulium (Chirostilbia) obesum and new records of black fly species (Diptera: Simuliidae) in the States of S?o Paulo and Minas Gerais, Brazil
Pepinelli, Mateus;Hamada, Neusa;Trivinho-Strixino, Susana;
Neotropical Entomology , 2006, DOI: 10.1590/S1519-566X2006000500019
Abstract: we describe the larval stage of simulium cuasiexiguum shelley, luna dias, maia-herzog & lowry and simulium obesum vulcano and provide new distribution records of four black fly species in the state of s?o paulo (simulium cuasiexiguum, simulium metallicum s.l. bellardi, simulium stellatum gil azevedo, figueró & maia-herzog and simulium oyapockense s.l. floch & abonnenc) and three species in the state of minas gerais (simulium duodenicornium pepinelli, hamada & trivinho-strixino, simulium obesum and lutzsimulium pernigrum lutz).
Description of Simulium damascenoi (Diptera: Simuliidae) male and the black-fly species from the State of Amapá, Brazil
Hamada, Neusa;Ale-Rocha, Rosaly;Luz, Sérgio Luíz Bessa;
Memórias do Instituto Oswaldo Cruz , 2003, DOI: 10.1590/S0074-02762003000300010
Abstract: five species are included in the simulium siolii group, which is placed in the subgenus psaroniocompsa (diptera: simuliidae). of these five species, only two (simulium siolii py-daniel and simulium tergospinosum hamada) have been described in all their life stages, except eggs. knowledge of the taxonomic characters of all life stages of a species is important in order to clarify interspecific and higher-level taxonomic relationships. the objectives of the present study are to describe the male of simulium damascenoi py-daniel, to provide a list of black-fly species their bionomics and distributions in the state of amapá, brazil, and to provide an identification key for larvae and pupae for these species.
Description of Simulium damascenoi (Diptera: Simuliidae) male and the black-fly species from the State of Amapá, Brazil  [cached]
Hamada Neusa,Ale-Rocha Rosaly,Luz Sérgio Luíz Bessa
Memórias do Instituto Oswaldo Cruz , 2003,
Abstract: Five species are included in the Simulium siolii group, which is placed in the subgenus Psaroniocompsa (Diptera: Simuliidae). Of these five species, only two (Simulium siolii Py-Daniel and Simulium tergospinosum Hamada) have been described in all their life stages, except eggs. Knowledge of the taxonomic characters of all life stages of a species is important in order to clarify interspecific and higher-level taxonomic relationships. The objectives of the present study are to describe the male of Simulium damascenoi Py-Daniel, to provide a list of black-fly species their bionomics and distributions in the state of Amapá, Brazil, and to provide an identification key for larvae and pupae for these species.
Simulium (Inaequalium) marins, a new species of black fly (Diptera: Simuliidae) from inselbergs in Brazil
Pepinelli, Mateus;Hamada, Neusa;Currie, Douglas C;
Memórias do Instituto Oswaldo Cruz , 2009, DOI: 10.1590/S0074-02762009000500012
Abstract: a new species of black fly, simulium (inaequalium) marins (diptera: simuliidae), is described based on the male, female, pupa and larva. this new species was collected from two localities: a small stream on the pico dos marins, a high mountain with granite outcrops in piquete county, state of s?o paulo, and in a small stream in the serra dos órg?os national park, state of rio de janeiro, brazil.
Macrogenomic Evidence for the Origin of the Black Fly Simulium suzukii (Diptera: Simuliidae) on Okinawa Island, Japan  [PDF]
Peter H. Adler, Yao-Te Huang, Will K. Reeves, Sam Kyu Kim, Yasushi Otsuka, Hiroyuki Takaoka
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0070765
Abstract: To determine the geographic origin of the black fly Simulium suzukii on Okinawa Island, Japan, macrogenomic profiles derived from its polytene chromosomes were compared with those of mainland and other insular populations of S. suzukii and of the isomorphic Simulium tani species complex. The Okinawan population is a chromosomally unique cytoform, designated ‘D,’ which is essentially monomorphic and differs by about 27 fixed rearrangements from the chromosomal standard sequence for the subgenus Simulium and by two fixed differences from its nearest known relative, representing the type of S. suzukii, on the main islands of Japan. Chromosomal band sequences revealed two additional, sympatric cytoforms of S. suzukii, designated ‘A’ and ‘B,’ each with species status, in Korea, and a third cytoform, designated ‘C,’ on Hokkaido, Japan. A new cytoform, ‘K,’ of S. tani from Malaysia, representing the type of S. tani, is more closely related to cytoforms in Thailand, as are populations from Taiwan previously treated as S. suzukii but more closely aligned with S. tani and newly recognized as cytoform ‘L’ of the latter nominal species. Rooting of chromosomal band sequences by outgroup comparisons allowed directionality of chromosomal rearrangements to be established, enabling phylogenetic inference of cytoforms. Of 41 macrogenomic rearrangements discovered in the five new cytoforms, four provide evidence for a stepwise origin of the Okinawan population from populations characteristic of the main islands of Japan. The macrogenomic approach applied to black flies on Okinawa Island illustrates its potential utility in defining source areas for other species of flies including those that might pose medical and veterinary risks.
Simulium (Chirostilbia) bifenestratum (Diptera, Simuliidae), a new black-fly species from the Atlantic forest, State of S?o Paulo, Brazil
Hamada, Neusa;Pepinelli, Mateus;
Memórias do Instituto Oswaldo Cruz , 2004, DOI: 10.1590/S0074-02762004000100008
Abstract: the larva, pupa, male, and female of simulium bifenestratum n. sp. are described and illustrated. the pupae of the new species have 10 gill filaments, thick at their base and arranged in a three-dimensional way, surrounding the head and thorax. its pupal cocoon is peculiar, not found in any of the known brazilian black-fly species; it is very thick and hard with two openings in the anterior region. s. bifenestratum n. sp. was collected in one stream in the bocaina mountain chain, atlantic forest, in s?o josé do barreiro county, state of s?o paulo, in a high (1500 m) natural grassland. larvae and pupae were collected on the edges of small waterfalls and in places with-high speed laminar water flow, attached to the bedrock.
Simulium (Chirostilbia) bifenestratum (Diptera, Simuliidae), a new black-fly species from the Atlantic forest, State of S o Paulo, Brazil  [cached]
Hamada Neusa,Pepinelli Mateus
Memórias do Instituto Oswaldo Cruz , 2004,
Abstract: The larva, pupa, male, and female of Simulium bifenestratum n. sp. are described and illustrated. The pupae of the new species have 10 gill filaments, thick at their base and arranged in a three-dimensional way, surrounding the head and thorax. Its pupal cocoon is peculiar, not found in any of the known Brazilian black-fly species; it is very thick and hard with two openings in the anterior region. S. bifenestratum n. sp. was collected in one stream in the Bocaina mountain chain, Atlantic forest, in S o José do Barreiro county, state of S o Paulo, in a high (1500 m) natural grassland. Larvae and pupae were collected on the edges of small waterfalls and in places with-high speed laminar water flow, attached to the bedrock.
Polytene chromosomes of an Indian Himalayan black fly Simulium (Nevermannia) praelargum (Diptera: Simuliidae)  [cached]
Willie HENRY, Subrata Kumar DEY, Rakesh VARMA, Sachin THAPA,William S PROCUNIER
Current Zoology , 2010,
Abstract: High quality polytene chromosome maps (n=3) of a Himalayan Simuliid Simulium praelargum Datta, 1973 are presented and represent the first cytological description of a taxon found in the feuerborni group, subgenus Nevermannia. Polytene chromosomes one (I) and two (II) are metacentric, chromosome three (III) is submetacentric with the length of each chromosome occupying 37.25 %, 31.36 % and 31.34 % of the total complement length, respectively. Typical simuliid diagnostic intergeneric chromosomal markers are found within the polytene complement of this species. The nucleolar organizer (N.O.) is found at the base of the short arm of chromosome one (IS), the Ring of Balbiani (R.B.), double bubble (D.B.) and triad occur in the short arm of chromosome two (IIS), the Parabalbiani Ring (P.B.) and grey band (gb) occur in the long arm of chromosome two (IIL) and the Blister (BL) and Capsule (Ca) occur in the short arm of chromosome three (IIIS).Terminal bands at the end of IIIS are heterochromatinized and present atypically with respect to other simuliid fauna. Populations studied so far are unique among the Simuliidae in that they exhibit chromosome structural monomorphism. These high resolution polytene chromosome maps will form the basis for future cytological characterization and phylogenetic comparisons amongst members of the feuerborni group [Current Zoology 56 (4): 437–444, 2010].
Simulium (Psilopelmia) virescens, a new black-fly species (Diptera: Simuliidae) from the southwestern region of the state of Bahia, Brazil
Hamada, Neusa;Silva, Nayra Gomes da;Pereira, Eleny da Silva;
Memórias do Instituto Oswaldo Cruz , 2012, DOI: 10.1590/S0074-02762012000100015
Abstract: the last-instar larva, pupa, male and female of simulium virescens sp. nov. are described and illustrated. this species has a peculiar larva, which has an elongated head capsule and light-green colour. the first thoracic segment has tubercle on its dorsal region and the third thoracic segment has one pair of tubercles; the first to the fourth abdominal segments have one pair of tubercles on each segment. until now this new species had only been collected at the type locality, which is on the middle stretch of the correntina river in the southwestern portion of the state of bahia, brazil. females were voraciously biting humans during the field work. this new species represents the second species of simulium(psilopelmia) in brazil and the first registered outside of the brazilian amazon region.
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