Araucnephia iberaensis n. sp., a Neotropical Black Fly with a Peculiar Distribution (Diptera-Simuliidae)

Araucnephia Wygodzinsky & Coscarón is a Neotropical black fly genus in which only one species from Central Chile is known. Another species has now been found in Corrientes province on the eastern side of the Iberá tropical swamps of Argentina, on the western border of the mountainous region of southern Brazil. This new species, A. iberaensis, is herein described and illustrated and information on its bionomics is recorded. It is an interesting species because previous to its discovery no black fly genus or subgenus from Central Chile region has been found in tropical areas, because these two regions are separated by the Monte and Pampas realms. Similarly, no Brazilian genus or subgenus has crossed the Andes mountains to Chile. A comparison with other Neotropical, Nearctic, Ethiopian (Afrotropical) and Australian Prosimuliini (sensu Crosskey & Howard) showed Araucnephia to be a valid taxon most closely related to Araucnephioides (sympatric in Chile). Araucnephia also shows great affinities with Lutzsimulium from Southeast Brazil and Argentina and Paracnephia from South Africa.

Simulium (Psilopelmia) virescens, a new black-fly species (Diptera: Simuliidae) from the southwestern region of the state of Bahia, Brazil

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.

Diversity and microdistribution of black fly (Diptera: Simuliidae) assemblages in the tropical savanna streams of the Brazilian cerrado

We describe the abiotic factors affecting the distribution of black flies at a microhabitat scale, rather than at the regional scale usually present in the literature on the Neotropics. Black fly larvae were sampled from the Tocantins River and three tributaries, located in the Brazilian savanna (state of Tocantins, Brazil) during six bi-monthly sampling periods from October 2004-August 2005. At each sampling site, 15 random quadrats (30 x 30 cm) were sampled each period and for each quadrat were determined mean water velocity, predominant substrate type (rocks, riffle litter or riparian vegetation) and depth detrended correspondence analysis (DCA) was used to determine associations with current velocity, whereas correspondence analysis (CA) was used to estimate site specific current velocity associations. Canonical correspondence analysis (CCA) was used to identify general microhabitat associations. The CCA showed that most species had a trend towards riffle litter, except for Simulium nigrimanum associated with rocky substrate and Simulium cuasiexiguum associated with riparian vegetation. The DCA showed a well defined pattern of water velocity associations. The CA revealed that the species showed different speed associations from one site to another, suggesting different competitive pressures resulting in the occurrence of different realized niches.

An Insight into the Sialotranscriptome of Simulium nigrimanum, a Black Fly Associated with Fogo Selvagem in South America

Pemphigus foliaceus is a life threatening skin disease that is associated with autoimmunity to desmoglein, a skin protein involved in the adhesion of keratinocytes. This disease is endemic in certain areas of South America, suggesting the mediation of environmental factors triggering autoimmunity. Among the possible environmental factors, exposure to bites of black flies, in particular Simulittm nigrimanum has been suggested. In this work, we describe the sialotranscriptome of adult female S. nigrimanum flies. It reveals the complexity of the salivary potion of this insect, comprised by over 70 distinct genes within over 30 protein families, including several novel families, even when compared with the previously described sialotranscriptome of the autogenous black fly, S. vitiation. The uncovering of this sialotranscriptome provides a platform for testing pemphigus patient sera against recombinant salivary proteins from S. nigrimanum and for the discovery of novel pharmacologically active compounds.

The use of bacterial larvicides in mosquito and black fly control programmes in Brazil

Bacillus spp. based larvides are increasingly replacing, with numerous advantages, chemical insecticides in programmes for controlling black fly and mosquito populations. Brazil was among the pioneers in adopting Bacillus thuringiensis israelensis (B.t.i) to control black flies. However, the major current mosquito control programme in Brazil, the Programme for Eradication of Aedes aegypti launched in 1997, only recently decided to replace temephos by B.t.i based larvicides, in the State of Rio de Janeiro. In the last decade, works developed by research groups in Brazilian institutions have generated a significant contribution to this subject through the isolation of B. sphaericus new strains, the development of new products and the implementation of field trials of Bacillus efficacy against mosquito species under different environmental conditions.

Description of Simulium damascenoi (Diptera: Simuliidae) male and the black-fly species from the State of Amapá, Brazil

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 (Chirostilbia) bifenestratum (Diptera, Simuliidae), a new black-fly species from the Atlantic forest, State of São Paulo, Brazil

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 São José do Barreiro county, state of São 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.

A new black fly isolate of Bacillus thuringiensis autoagglutinating strain highly toxic to Simulium pertinax (Kollar) (Diptera, Simuliidae) larvae

Formulations containing the entomopathogenic Bacillus thuringiensis serovar israelensis strain IPS-82 has been widely applied for mosquito control around the world. Strain IPS-82 is highly active against Aedes aegypti but less active against other well-known vectors such as Culex quinquefasciatus and Simulium spp. larvae. Eighteen strains of B. thuringiensis were isolated from Simulium pertinax larvae naturally occurring in rivers of Southeast Brazil with one demonstrating special toxic effects. Simulated field tests against S. pertinax larvae showed that the native Brazilian autoagglutinanting B. thuringiensis (LFB-FIOCRUZ 1035) has an LC50 at least 25 times lower than the standard IPS-82 strain. The same bacterial preparation was also tested against Ae. aegypti larvae in laboratory trials and the LC50 values obtained with LFB-FIOCRUZ 1035 were at least three times lower than the one for the IPS 82 strain. The results indicate that this strain is more toxic than the standard B. thuringiensis serovar israelensis (H14) in the two Dipteran species tested. It is noteworthy that differences between LC50 values were more pronounced in S. pertinax larvae, the source of the original isolation.

Single and concomitant experimental infectionsby Endotrypanum spp. and Leishmania (Viannia) guyanensis (Kinetoplastida: Trypanosomatidae) in the Neotropical sand fly Lutzomyia longipalpis (Diptera: Psychodidae)

Lutzomyia longipalpis females received single and mixed infections with Endotrypanum and Leishmania. Two biological parameters were analyzed: the percentage of infected females and the distribution of flagellates in the gut of the females. The principal comparisons were performed between (1) two strains of Endotrypanum, (2) cloned versus primary sample of one strain of Endotrypanum, (3) Endotrypanum versus Leishmania guyanensis, and (4) the pattern of flagellates behaviour by optical microscopy in females with single or mixed infection versus the identification of parasites isolated from digestive tracts by isoenzyme electrophoresis. Flagellates of Endotrypanum showed distinct patterns of infection suggesting that there is variation between and within strains. The distribution of Endotrypanum and L. guyanensis differed significantly in relation to the colonization of the stomodeal valve. In co-infection with L. guyanensis, a large number of flagellates were seen to be plentifully infecting the stomodeal valve in significantly more specimens than in females infected by Endotrypanum only. However, the electrophoretic profiles of isoenzymes of parasites recovered from all co-infected specimens corresponded to Endotrypanum. This suggests that the mere correlation sand fly infection-biochemical analysis of isolates may induce parasitological incorrect consideration.

Simulium (Inaequalium) marins, a new species of black fly (Diptera: Simuliidae) from inselbergs in Brazil

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 São Paulo, and in a small stream in the Serra dos Órgãos National Park, state of Rio de Janeiro, Brazil.

An investigative study on the effects of black fly (diptera: simuliidae) sugar meals on reproductive success and parasite transmission /

Black flies are opportunistic sugar-feeders. They take sugar meals from Homopteran honeydew secretions or plant nectars, depending on availability. Homopteran honeydew secretions contain both simple and complex carbohydrates while plant nectars contain primarily simple carbohydrates. In order to determine whether honeydew secretions offer more energy than plant nectars to their insect visitors a study of wild-caught black flies was undertaken in Algonquin Provincial Park, Canada during the spring of 1 998 and 1 999. It was hypothesized that female black flies maintained on honeydew sugars will survive longer, produce more eggs and have a greater parasite vectoring potential than those maintained on artificial nectar or distilled water. Results demonstrated that: (1) host-seeking female Prosimulimfuscum/mixtum and Simulium venustum maintained on artificial honeydew did not survive longer than those maintained on artificial nectar when fed ad libitum; (2) fiiUy engorged S. venustum and Simulium rugglesi maintained on artificial honeydew did not produce more eggs than those maintained on artificial nectar when fed ad libitum; and (3) S. rugglesi did not have a greater vectoring potential of Leucocytozoon simondi when maintained on artificial honeydew as opposed to artificial nectar when fed ad libitum. However, all flies maintained on the two sugars (artificial honeydew and artificial nectar) survived longer, produce more eggs and had greater vectoring potential than those maintained on distilled water alone.

Behavioural characteristics in phylogenetics : a case study using black fly (Diptera: Simuliidae) cocoon spinning behaviour

1-1 is torically, the predominan t method of reconstructing phylogenies has been through the use of morphological characters. There are new techniques now gaining acceptance, including molecular techniques al1d chromosomal information. Altl10ugh the study of behaviour has been used in a comparative framework, these analyses have, historically, been based on intuition. Hennig (1966) devised a neV\' method of reconstructing phylogenies which provided a 110ncircular method for formulating, testing and refining phylogenies. Subsequent s)Tstematists had virtually abandoned ecological and beha\lioural data as primary indicators of phylogenetic relationships (Brooks and McLennan 1991). Therefore, in a modern cladistic framework (sensu Hennig) the analysis of behavioural traits remains underrepresented as a method of reconstructing phylogenies. This thesis will reconstruct the phylogeny for species of black flies (Diptera: Simuliidae), using two steps. The first step is to thoroughl)' understand and explain the cocoon spinning in black fly larvae. There have bee115 previous descriptions of cocoon spinning, but all were incomplete or erroneous. The advances in technology, including video recorders and VCRs, have allowed this behaviour to be analyzed in great detail in 20 different species. A complete description of the cocoon spinning of Simulium \littatum is given. This description will be used as a template for the other species observed. The description and understanding of cococ)n spinning was the first step in undertaking a phylogenetic analysis using this behaviour. The behaviour was then broken down and analyzed, revealing 23 characters, 3 either qualitative and quantitative in nature. These characters were assessed in a cladistic framework (sensu Hennig) and a phylogenetic tree was reconstructed with a e.I of 0.91 and an R.I. of 0.96. This phylogenetic tree closely resembles a previously established pllylogenetic tree produced from morphological and cytological information. The importance of this result is the indication that, contrary to some authors, behavioural characters, if used properly, can add very informative characters to a data set.

The effect of feeding regime on larval black fly (Diptera: simuliidae) primary head fan ray number

Identification of larval simuliids has always been difficult due to the morphological similarity many species bear to one another. For this reason all characters available have been drawn upon to aid in species identification, including head fan ray number. Even in light of an increasing body of anecdotal reports that head fan ray number is not fixed, it has continued to be used to aid species identification. In the current experiment simuliid larvae were reared under controlled laboratory conditions to last instar in one of three feeding regimes. Out of nine trials, the results of six showed a significant inverse relationship between feeding regime and head fan ray number. In addition to the laboratory experiments, larvae were also collected from the field over the course of the spring and summer, 1994. From these samples significant interspecific and intraspecific variations in head fan ray number were found both spatially and temporally within Algonquin Park. From these data it is concluded that head fan ray number for the species analysed is a developmentally plastic character, which varies in response to food availability. Furthermore, given the extreme variations in head fan ray number found in some species, I recommend that head fan ray number not be used as an aid to identification unless it can be shown to be a fixed character for the species in question.

An investigation into the effects of homopteran honeydew sugars versus floral nectar sugars on black fly longevity, flight performance and digestion /

Floral nectar is thought to be the primary carbohydrate source for most dipteran species. However, it has been shown that black flies (Burgin & Hunter 1997 a,b,c), mosquitoes (Foster 1995; Burkett et al. 1999; Russell & Hunter 2002), deer flies (Magnarelli & Burger 1984; Janzen & Hunter 1998; Ossowski & Hunter 2000), horse flies (Schutz & Gaugler 1989; Hunter & Ossowski 1999) and sand flies (MacVicker et al. 1990; Wallbanks et al. 1990; Cameron et al. 1992, 1995; Schlein & Jacobson 1994, 1999; Hamilton & EI Naiem 2000) feed on homopteran honeydew as well as floral nectar. Prior to 1997 floral nectar was thought to be the main source of carbohydrates for black flies. However, Burgin & Hunter (1 997a) demonstrated that up to 35% of black flies had recently consumed meals of homo pte ran honeydew. This information has necessitated a re-assessment of many life history aspects of black flies. Attempts are being made to examine the effects of nectar versus honeydew on black fly fecundity and parasite transmission (Hazzard 2003). Recently, Stanfield and Hunter (unpublished data) have shown that in female black flies, honeydew sugars produce flights of longer distance and duration than do nectar sugars. This thesis examines two aspects of black fly biology as it relates to sugar meal consumption. First, the effects of honeydew and nectar on black fly longevity are examined. Second, the proximate causation behind longer flight performances in honeydew-fed flies will be examined. The comparison between these two sources is important because nectar is composed of mainly simple sugars (monosaccharides and disaccharides) whereas honeydew is composed of both simple and complex sugars (including trisaccharides and tetrasaccharides ).

Characterization of black fly (Diptera: Simuliidae) silk proteins

Black fly (Simuliidae) silk is produced by the larvae and pharate pupae and is used for anchorage and cocoon production. There exists limited information on simuliid silks, including protein composition and genetic sequences encoding such proteins. The present study aimed to expand what is known about simuliid silks by examining the silks of several simuliid species and by making comparisons to the silk of non-biting midges (Chironomidae). Silk glands were dissected out of larval and pupal simuliids, and protein contents were separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and visualized with silver stain. Protein contents were compared by mass in kilodaltons (kDa) between life stages and among species. Polymerase chain reaction (PCR) was used to expand upon known gene sequence information, and to determine the presence of genes homologous to chironomid silk. SDS-PAGE of cocoons revealed the presence of a 56 kDa and a 67 kDa protein. Silk gland contained as many as 28 different proteins ranging from 319 kDa to 8 kDa. Protein profiles vary among species, and group into large (>200), intermediate(>100), and small (<100) protein classes as is found in chironomids. It is likely that silk evolved in a common ancestor of simuliids and chironomids