Six new species of microsporidia of the genus Amblyospora (Microspora: Amblyosporidae) from blood sucking mosquitoes (Diptera: Culicidae) from the west Siberia

Microsporidia of the genus Amblyospora parasiting the adipose body of mosquito larvae of the genus Aedes and Culex has been studied with both light and electron microscopy. Six new species of microsporidia are described based on ultrastructural characteristics of spores and sporogony stages. Amblyospora flavescens sp. n. Mature spores are egg-shaped. The spore wall with three layers, about 165 nm. Exospore is two-membranous. Subexospore is absent. Endospore is electron-translucent. Polaroplast consists of three parts: lamellar, large vesicular, lamellar. The anisofilar polar filament with 10--11 coils (3 1/2 + 2 1/2 + 4-5). Fixed spores are 6.3 +/- 0.1 x 4.24 +/- 0.1 microm. Amblyospora kolarovi sp. n. Mature spores are egg-shaped. The spore wall with three layers, about 265-315 nm. Exospore shapes tucks on the surface of spore. It is two-membranous. Subexospore is quagge, structural. Endospore is electron-translucent. Polaroplast consists of two parts: lamellar and large vesicular. The anisofilar polar filament with 11-13 coils (3 + 8-10). Fixed spores are 5.4-5.6 x 3.5-4.2 microm. Amblyospora orbiculata sp. n. Mature spores are widely egg-shaped. On a back pole there is a small concavity. The spore wall with three layers, about 155 nm. Exospore is shapes tucks on a surface of spore. It is two-membranous. Subexospore is absent. Endospore is electron-translucent. Polaroplast consists of three parts: lamellar, vesicular, lamellar. Polar filament is anisofilar, with 11 1/2 coils (4 1/2 + 1 + 6). Fixed spores are 6.3 +/- 0.1 x x 4.0 +/- 0.1 microm. Amblyospora rugosa sp. n. Mature spores are egg-shaped. On a back pole there is a small concavity. The spore wall with three layers, about 225 nm. Exospore is shapes tucks on a surface of spore. It is two-membranous. Subexospore is quaggy, structural. Endospore is electron-translucent. Polaroplast lamellate. Polar filament is anisofilar, with 17 1/2 coils (3 1/2 + 1 + 13). Fixed spores are 5.3 +/- 0.1 x 3.7 +/- 0.1 microm. Amblyospora undata sp. n. Mature spores are egg-shaped. The spore wall is three-layered, about 220 nm. Exospore is shapes tucks on a surface of spore. It is two-membranous. Subexospore is quaggy, structural. Endospore is electron-translucent. Polaroplast lamellate. The anisofilar polar filament with 8 coils (3 + 5). Fixed spores are 5.0 +/- 0.1 x 3.0 +/- 0.1 microm. Amblyospora urski sp. n. Mature spores have widely oval form. The back pole is concave. The spore wall with three layers, about 280 nm. Exospore is shapes tucks on a surface of spore. It is two-membranous. Subexospore is quaggy, structural. Endospore is electron-translucent. Polaroplast lamellate. Polar filament is anisofilar, with 6 coils (2 + 4). Fixed spores are 4.4 +/- 0.1 x 2.9 +/- 0.1 microm.     

New species of Microsporidia in the genus Amblyospora from blood-sucking mosquitoes in the family Culicidae

Two microsporidians of the genus Amblyospora from mosquito larvae of Aedes and Culex were studied in Tomsk region (Siberia). Peculiarities of the sporogony and the meiospore ultrastructure were studied by light and electron microscopy. The meiospores of A. caspius sp. n. from Aedes caspius and Ae. communis measured on smears, were c. 4.8 x 3.6 microns in size. A polar filament was anisofilar with 5 wide (290 nm) and 9 narrow (190 nm) coils arranged in a single layer. A polaroplast of both species was composed of two parts: anterior lamellae and posterior fibrillae. Meiospores of A. burlaki sp. n. from Culex pipiens measured c. 4.8 x 3.5 microns. A polar filament was anisofilar with 3 wide (260 nm) and 6 narrow (110 nm) coils. The meiospore of this species had a 260 nm thick multilayered exospore.     

Crepidula beklemishevi gen. et sp. n. and Dimeiospora palustris gen. et sp. n. (Microspora: Amblyosporidae)--new microsporidian genera and species from blood-sucking mosquitoes (Diptera: Culicidae) from the south of the western Siberia

Microsporidia parasitizing the adipose body of mosquito larvae of Anopheles beklemishevi and Aedes punctor has been studied. Two new genera of microsporidia are described based on lightmicroscopic and ultrastructural characteristics of spores and sporogony stages. The spore wall of Crepidula beklemishevi gen. n. et sp. n. is formed by two-membrane exospore, thick exospore, bilayer endospore and thin plasmolemma. Spores with single nucleus, polar filament anisofilar, with 6-7 coils (2+ 4-5), polaroplast consisting of three parts: macrochelicoidal, microhelicoidal and lamellar. Fixed spores 4.2 +/- 0.22 x 2 +/- 0.01 microns. The sporogony of Dimeiospora palustris gen. et. n. results in spore formation of two different types. Spores of the first type are oviform, with thick wall, single-nuclear, 6.1 x 4.9 microns. Spore wall with three layers, about 370 nm. Exospore electron-dense, subexospore moderately electrondense. Exospore and subexospore irregularly pleated on the almost spore surface and slightly thinner on anterior end only. Endospore electron-translucent. Polar filament anisofilar, with 9 coils (3 + 6). Polaroplas consists of three parts: lamellar, fine bubbled, and coarse bubbled. Spores of the second type broad-ovate, with apical pole narrower, distal pole concave, 4.6 x 3.7 microns. Spore wall with three layer, 355 nm. Exospore on the apical end irregularly pleated, consists of thin electrondense exospore, subexospore of variable electron density, endospore electron-translucent. Polar filament anisofilar, with 13 coils (3 + 10). Polaroplast has two parts: lamellar and vesicular.     

Chromosomal Evidence on the Sporogony of Amblyospora californica (Microspora: Amblyosporidae) in Culex tarsalis (Diptera: Culicidae)

ABSTRACT. Amblyospora californica is a polymorphic, eukaryotic microsporidian. Three types of sporogony producing three types of spores occur in male larvae and female adults of its mosquito host, Culex tarsalis , and an alternate copepod host, Acanthocyclops vernalis. Development of A. californica in male larvae includes merogony and sporogony. Karyogamy and meiosis was observed in sporogony in male larvae but not in the female adult or in the copepod. Chromosomal evidence showed that sporogony included two consecutive meiotic divisions and a subsequent mitosis forming an octosporont, ultimately containing eight haploid, uninucleate mature spores. In this species, the haploid number of chromosomes is nine. Macrosporoblasts and macrospores, containing 1, 2 or more nuclei, can be seen in infected male larvae. The stage of sporogony in which cytokinesis was arrested seems to determine the number of nuclei. Those with only one nucleus, we believe are due to failed nuclear division at meiosis. Although A. californica displayed a process of karyogamy and meiosis similar to that of the species from Cx. salinarius , they may not be the same species because of the difference in their chromosome numbers.     

Experimental study of the routes of transmission of Microsporidia in blood-sucking mosquitoes from southwestern Siberia

Experiments were conducted on the artificial infection of larvae of 8 species of bloodsucking mosquitoes (Aedes diantaeus, A. punctor, A. cinereus, A. flavescens, A. aegypti, Culex pipiens, Anopheles messeae, Anopheles atroparvus) with spores of Amblyospora opacita var. sibirica, Parathelohania costatus sp. n., P. obesa, P. anophelis. Negative results were obtained in all experiments.     

Amblyospora sp. (Microspora, Amblyosporidae) infecting nerve ganglia of Culex pipiens (Diptera, Culicidae) from Egypt.

A species of Amblyospora-infecting neurones of Culex pipiens is described. Diplokaryotic meronts, which divided by binary fission, were distinguished at the electron microscope level by their unthickened plasma membranes. Sporonts with an electron-dense surface coat gave rise to eight uninucleate sporoblasts within a sporophorous vesicle, cytoplasmic division occurring at the quadrinucleate or octonucleate stages. Indications that nuclear fusion and chromosome reorganization occurred in merogony and sporogony were obtained by light microscopy but meiosis was not detected at the ultrastructural level. Spores were typical of Amblyospora, being ovoid when fresh, truncate when stained, and having an exospore of two membranous layers subtended by a thick amorphous layer, an electron-lucent endospore, an anisofilar polar filament, and a polaroplast comprised of an anterior region of close-packed lamellae and a posterior region of expanded sacs. The metabolic products in the sporophorous vesicle took the form of large globules, small globules with electron-dense borders, and fine granules. These were depleted in mature sporophorous vesicles, though a surface layer of fine granules on the spores may have been derived from them. Many stages were degenerate and it is suggested that C. pipiens may be an accidental host in which the parasite could develop suboptimally in nervous tissue only. Infections in larvae hatched from eggs in the laboratory indicate that vertical transmission occurs.     

Species composition of mosquitoes (Diptera, Culicidae) and possibility of the West Nile virus natural foci formation in the South of Western Siberia

In 2004 June-July collections of mosquito adults and small mammals were carried out in two areas of Novosibirsk Region (forest-steppe and steppe zones), where the West Nile virus (WNV) was for the first time recorded in birds with different migration status in 2002-2004. Seventeen species of mosquitoes were found; significant changes in their species composition and abundance, as compared with latest faunistic studies made in the sixties-seventies of the last century, are revealed. WNV markers (antigen, RNA) are found in small mammals; highly sensitive to the WNV replication mosquito species are also found. These facts allow supposing a possibility of the formation of stable West Nile virus natural foci in the South of Western Siberia, under conditions of forest-steppe and steppe zones.     

Molecular phylogeny and evolution of mosquito parasitic Microsporidia (Microsporidia: Amblyosporidae)

Amblyospora species and other aquatic Microsporidia were isolated from mosquitoes, black flies, and copepods and the small subunit ribosomal RNA gene was sequenced. Comparative phylogenetic analysis showed a correspondence between the mosquito host genera and their Amblyspora parasite species. There is a clade of Amblyospora species that infect the Culex host group and a clade of Amblyospora that infect the Aedes/Ochlerotatus group of mosquitoes. Parathelohania species, which infect Anopheles mosquitoes, may be the sister group to the Amblyospora in the same way that the Anopheles mosquitoes are thought to be the sister group to the Culex and Aedes mosquitoes. In addition, by sequence analysis of small subunit rDNA from spores, we identified the alternate copepod host for four species of Amblyospora. Amblyospora species are specific for their primary (mosquito) host and each of these mosquito species serves as host for only one Amblyospora species. On the other hand, a single species of copepod can serve as an intermediate host to several Amblyospora species and some Amblyospora species may be found in more than one copepod host. Intrapredatorus barri, a species within a monotypic genus with Amblyospora-like characteristics, falls well within the Amblyospora clade. The genera Edhazardia and Culicospora, which do not have functional meiospores and do not require an intermediate host, but which do have a lanceolate spore type which is ultrastructurally very similar to the Amblyospora spore type found in the copepod, cluster among the Amblyospora species. In the future, the genus Amblyospora may be redefined to include species without obligate intermediate hosts. Hazardia, Berwaldia, Larssonia, Trichotuzetia, and Gurleya are members of a sister group to the Amblyospora clades infecting mosquitoes, and may be representatives of a large group of aquatic parasites.     

On the fauna and ecology of mosquitoes (Diptera: Culicidae) in forest-steppe and steppe regions of Western Siberia

Fauna and ecology of the mosquitoes from the southern forest-steppe (Chany Lake) and the northern steppe (Karasuksky lakes) have been investigated during 2004-2006. Species composition, dominant species, biotopical and seasonal distribution are examined. Detailed data on the biotopical and seasonal distribution, sex ratio and terms of development for the dominant species Coquillettidia richiardii (Ficalbi, 1889) are reported.     

Quantitative assessment of emergence of blood-sucking mosquitoes (Diptera,Culicidae) by the hydrobiological method and by cone-shaped traps

Two methods of estimating the emergence and transfer of biomass of blood-sucking mosquitoes (Diptera and Culicidae) are compared. The phonological and quantitative data obtained by the hydrobiological method (using an aquatic net and a tray) and using cone-shaped traps are different. Nevertheless, the general characteristics of the studied processes are similar. Results of the hydrobiological method showed that the total value of the annual transfer of biomass by Culicidae accounts for 3.157 g wet weight m^-2 per year; that of cone-shaped traps is 3.457 g wet weight m^-2 per year.     

Autogeny in populations of blood-sucking mosquitoes (Culicidae) on the southern Yamal

After three-year studies a degree of autogeny in populations of Aedes communis. A. pullatus and A. hexodontus was defined. In various seasons a share of autogenous females in A. communis was stable enough amounting to 30-37%. At the same time considerable variations in the degree of autogeny in some micropopulations of this species were observed. Variations in the ratio between autogenous and non-autogenous individuals reflect a dissimilar effect of varied combinations of ecological factors upon larvae of mosquitoes.     

Updated checklist of the mosquitoes (Diptera: Culicidae) of Croatia

Improvement of morphological and molecular identification methods allows the detection of new species of mosquitoes. The mosquito fauna of Croatia currently includes 52 species, belonging to eight genera, including Anopheles (12 species), Aedes (24 species), Coquillettidia (one species), Culex (seven species), Culiseta (six species), Orthopodomyia (one species), and Uranotaenia (one species). This is an updated checklist, which includes five new species found in Croatian mosquito fauna. Two of these are invasive mosquito species, Aedes albopictus (Skuse, 1895) and Aedes japonicus (Theobald 1901), which are spreading across Europe and Croatia. The other three species, Culex laticinctus (Edwards 1913), Culex torrentium (Martini 1925), and Anopheles daciae (Linton, Nicolescu & Harbach 2004) are autochthonous species which haven't been recorded so far. Since there are several more invasive species spreading across Europe, we assume that this is not the final list.     

Updated checklist of the mosquitoes (Diptera: Culicidae) of Belgium

Most information about the systematics and bioecology of Belgian mosquitoes dates back from before 1950, and only scattered information was produced during the last decades. In this paper we review and update the list of mosquito species recorded in Belgium, from first report (1908) to 2015. Six genera and 31 species were recorded so far, including 28 autochthonous species and three invasive alien species recently recorded in Belgium: Aedes albopictus (Skuse 1894), Ae. japonicus japonicus (Theobald 1901), and Ae. koreicus (Edwards 1917). The six genera are Anopheles (five species), Aedes (sixteen species), Coquillettidia (one species), Culex (four species), Culiseta (four species), and Orthopodomyia (one species).     

The mosquitoes (Diptera: Culicidae) of Tabasco,Mexico

To document and update the mosquito species of Tabasco, Mexico, field collection trips were conducted in the two physiographic regions of Tabasco: the coastal plain of the southern gulf and the mountains of Chiapas and Guatemala. Mosquitoes were collected as immature and adult stages during the dry and rainy seasons from 2014 through 2015. Additionally, the Reference Collection of Arthropods of Medical Importance (CAIM‐InDRE) containing mosquitoes of Tabasco was re‐examined. In total, 4,913 specimens were collected and examined, which are divided into seven tribes, 18 genera, 27 subgenera, and 104 species. Of these, one genus (Shannoniana Lane and Cerqueira), two subgenera (Georgecraigius Reinert, Harbach and Kitching, and Carrollia Lutz), and 21 species are new records for the mosquito fauna of Tabasco. Culex metempsytus Dyar is a new record for Mexico and Wyeomyia jocosa (Dyar and Knab) is removed from the Mexican mosquito fauna. Seventeen species historically reported were not found in the field collections conducted here. Taxonomic notes, new distribution limits, and comments about the medical importance of species of mosquitoes of Tabasco are discussed. Tabasco is the second state in Mexico with the largest mosquito richness (104 species), followed by Veracruz with 139 species.     

Boric acid bait kills adult mosquitoes (Diptera: Culicidae)

The toxicity of boric acid solutions to adult Anopheles quadrimaculatus Say, Culex nigripalpus Theobald, and Aedes albopictus Skuse was evaluated in the laboratory. Median lethal concentrations (LC50 in %) at 24-h exposure for male and female An. quadrimaculatus were 0.317 and 0.885, respectively; for Cx. nigripalpus, 0.273 and 0.560, respectively; and for Ae. albopictus, 0.174 and 0.527, respectively. The LC50 values at 48-h exposure for male and female An. quadrimaculatus were 0.101 and 0.395, respectively; for Cx. nigripalpus, 0.098 and 0.255, respectively; and for Ae. albopictus, 0.078 and 0.244, respectively. In laboratory tests, access for 48 h to sucrose (10%) water containing 1% boric acid (boric acid bait) resulted in 98% mortality in blood fed, gravid, and parous Ae. albopictus. When offered a choice between boric acid bait and sucrose water, 52% of male and 33% of female Ae. albopictus ingested sufficient boric acid bait in 24 h to cause death; after 48 h, respective percent mortalities were 88 and 58%. In outdoor tests, in a walk-in screened cage (156 m3) containing 1,250 female Ae. albopictus, mosquito biting rates on the exposed forearm of a human subject in 3-min exposure were reduced > or = 78% for the boric acid bait treatment, compared with a sucrose water control.     

Epizootiological studies of Amblyospora camposi (Microsporidia: Amblyosporidae) in Culex renatoi (Diptera: Culicidae) and Paracyclops fimbriatus fimbriatus (Copepoda: Cyclopidae) in a bromeliad habitat

The epizootiology of Amblyospora camposi was studied in a natural population of Culex renatoi, a bromeliad-inhabiting mosquito, and its intermediate host, Paracyclops fimbriatus fimbriatus, over a 2-year period. Twenty Eryngium cabrerae plants were sampled monthly from January 2003 to January 2005 and the prevalence of A. camposi in P.f. fimbriatus and Cx. renatoi populations was determined. The monthly prevalence rates of meiospore infections in Cx. renatoi larvae never exceeded 5.5% and was detected in 50% of the monthly samples. Meiospores were available in plants over the course of the study at a mean concentration of 2 x 10(4) meiospores/ml. Within each plant the parasite was maintained by horizontal transmission. P.f. fimbriatus with vegetative stages and mature spores were found regularly in bromeliads suggesting efficient meiospore infectivity to field copepod populations. The mean concentration of spores from copepods found in plants was 8 x 10(2) spores/ml. Infections in copepods were detected in 54% of the monthly samples with a prevalence rate ranging from 0.55 to 17.4% and an overall average of 5.1%. Vegetative stages in fourth instar mosquito larvae (probably derived from the horizontal pathway via spores formed in copepods) were detected in 12.5% of the monthly samples with an overall prevalence rate of 1.1%. Infections in female and male adults were detected in 20.8% of the monthly samples with an overall average of 4.1% and 6.8%, respectively.     

Checklist of Iranian mosquitoes (Diptera: Culicidae).

The mosquito fauna of Iran includes seven genera, 64 species, and three subspecies. The records of 12 other species should be verified. There are 24 species in the most recent checklist of Iranian Anopheles. Two species, An. peditaeniatus and An. fluviatilis species V, have been reported since. An. atroparvus, An. labranchiae, and An. martinius of the Maculipennis Group, and An. cinereus, An. nigerrimus, and An. rhodesiensis rupicola were recorded previously but are not included in the checklist. The checklist of Iranian Culicinae includes ten species of the tribe Aedini, but there are some records of four other species: Aedes aegypti, Ochlerotatus berlandi, Oc. chelli, and Oc. dorsalis. The genus Culex includes 19 species, excluding Cx. impudicus, which has not been collected recently, and some doubtful records of Cx. univittatus, Cx. vishnui, and Cx. vagans. The genus Culiseta includes five species and the genera Coquillettidia and Uranotaenia each include one species in Iran. No information is available for the An. subpictus, Oc. caspius, Oc. detritus, and Oc. pulcritaris species complexes in Iran. The An. claviger and Cx. pipiens complexes and the An. hyrcanus group require review.     

An annotated list of the mosquitoes (Diptera: Culicidae) of Mississippi

There has been no previous systematic statewide study of mosquitoes in Mississippi. This survey, resulting in the collection of over 400,000 specimens, was conducted by the authors from 2003 to 2007 throughout much of the state using CO₂‐baited CDC light traps and larval dipping. In addition, a health department contract mosquito surveillance technician collected several thousand specimens from the state from 2001 to 2003. Lastly, specimens housed at the Mississippi State University Entomological Museum, obtained from previous surveys, were included as vouchers for species occurring in the state. The collection records and literature show 60 species as occurring or having occurred in Mississippi. Voucher specimens representing 57 of the 60 species discussed are deposited in the Mississippi Entomological Museum or in the U.S. National Museum of Natural History (USNM), Washington, D.C.