Catalog of the biting midges of Argentina (Diptera: Ceratopogonidae).

Three hundred and fiftysix species of ceratopogonids in 35 genera are listed for Argentina, with most species in Forcipomyia Meigen (67 species) Culicoides Latreille (51 species), Stilobezzia Kieffer (40 species) and Dasyhelea Kieffer (37 species). For each listed species, the following information is included: sex and/or life stage described in the original description, type locality, type status and sex, depository of type material, distribution, georeferenced localities for Argentina (provinces are underlined) and main references that include the original description and those that specify locations in the country. Specific epithets of synonyms are accompanied by type locality, type status and sex, and location of type material. New localities from Argentina for 102 species are provided, and the following 16 species are newly recorded from the country: Forcipomyia (Caloforcipomyia) glauca Macfie, Forcipomyia (C.) hatoensis Utmar & Wirth, F. (Euprojoannisia) bromeliae Saunders, F. (E.) quasiingrami Macfie, F. (Forcipomyia) zonogaster Ingram & Macfie, F. (F.) catarinensis Marino & Spinelli, F. (Lasiohelea) cornuta Saunders, F. (Metaforcipomyia) osaensis Spinelli, Marino & Borkent, F. (Phytohelea) bromelicola (Lutz), F. (Trichohelea) goniognatha Wirth & Messersmith, F. (T.) leptognatha Wirth & Messersmith, Brachypogon (Brachypogon) woodruffi Spinelli & Grogan, Physohelea turgidipes Ingram & Macfie, Bezzia gibbera (Coquillett), Bezzia schmitzorum Dippolito & Spinelli and Palpomyia versicolor Macfie. Leptoconops brasiliensis (Lutz) and Clastrieromyia dycei Spinelli & Grogan are excluded from the Argentinean fauna.

Rapid and Efficient Detection by PCR of Culicoides insignis (Diptera: Ceratopogonidae), the Main Vector of Bluetongue Virus (BTV) in the Neotropical Region.

Bluetongue virus (BTV) causes a viral, non-contagious disease that mainly affects sheep, cattle, and wild and farmed ruminants causing damage to these animals and significant economic losses. Culicoides insignis Lutz, the major BTV vector in South America, is one of the most abundant species in Argentina and commonly associated with cattle farms. The morphological identification of Culicoides spp. is routinely carried out with the aid of morphological keys, which mainly refer to the wing patterns, sensillae of palpi, and antennal flagella. Molecular tools applied to taxonomy provide a rapid and efficient method of identification of vector species. An easy protocol for the extraction of total DNA from single midges is herein described, and a forward primer for rapid and reliably test detection by polymerase chain reaction of C. insignis is developed.

Redescription of immature stages of Culicoides paraensis (Goeldi) (Diptera: Ceratopogonidae), vector of the Oropouche virus.

Oropouche fever is an emerging zoonotic disease caused by Oropouche virus (OROV). It has two distinct transmission cycles, with the anthropophilic biting midge Culicoides paraensis (Goeldi) (Diptera: Ceratopogonidae) being the primary vector in the urban cycle. Species identification of Culicoides typically has been carried out on the basis of morphological characters, but molecular tools applied to taxonomy can provide rapid and efficient methods to the identification of vector species. The aim of this work was to obtain the first DNA barcode for C. paraensis collected in Argentina and redescribe the larvae and pupae of this species. Nested PCR amplification was applied in this study to increase the DNA amplification, because the material was preserved in alcohol 70% for a long period of time. The immature stages of C. paraensis are fully described from material collected in Misiones province, Argentina. Both stages are compared with their most similar congeners. This COI sequence complements the identification based on morphological characters and the values of genetic distance between the analysed species show that this sequence is useful to discriminate between species of the Culicoides genus.

Two new species and new records of Neotropical Culicoides Latreille (Diptera: Ceratopogonidae).

The following two new species of biting midges of Culicoides Latreille are described and photographed: Culicoides carbonelli Spinelli Martínez from Uruguay, and C. dellapei Spinelli, Ronderos Díaz from Argentina. Culicoides crucifer Clastrier, 1968 and C. hoffmani Fox, 1946 are diagnosed and newly recorded from Argentina, and the studied specimens are photographed.

The immatures of three Neotropical species of Palpomyia Meigen (Diptera, Culicomorpha, Ceratopogonidae).

The first description of the fourth instar larva and pupa of Palpomyia mapuche Spinelli, Grogan & Ronderos and the pupa of P. subfuscula Ingram & Macfie are provided, as well as the redescription of the pupa of P. subaspera (Coquillett). Studied specimens were collected in lotic environments of Argentinian Patagonia, in Neuquén and Chubut Provinces. The described stages were examined and illustrated with a phase-contrast microscope. The larva was examined using a scanning electron microscope. Data on the bionomics for P. mapuche and new records for the three species are provided.

Schizonyxhelea thomsenae (Wirth), description of the pupa and first records from Argentina, Brazil and Peru (Diptera: Ceratopogonidae).

The pupa of Schizonyxhelea thomsenae (Wirth) is described and illustrated from material collected in Misiones province, Argentina. A key for the four known species of pupae of Schizonyxhelea is given, a diagnosis and photographs of the adult are also provided, and the geographic distribution of the species in the Neotropical region is enlarged including first records from Argentina, Brazil and Peru based mainly on adult specimens.

Redescription of immatures of Dasyhelea flavifrons Guérin-Méneville (Culicomorpha: Ceratopogonidae) and new contribution to the knowledge of its larval habitats.

The fourth instar larva and pupa of Dasyhelea flavifrons Guérin-Méneville are redescribed, illustrated, and photomicrographed using binocular, phase-contrast, and scanning electron microscopy. Comparisons with the American species of the grisea group were made. The immatures were collected by using a siphon bottle in tree-holes and from water collected in dead snail shells in Salta Province, Argentina, transported to the laboratory and there bred to the emergence of the adults. Details on larval habitats are given. These are the first records from Argentina and in gastrotelmata.

The Biting Midges of the Caribbean island Curaçao (Diptera: Ceratopogonidae). I. Species in the genus Dasyhelea Kieffer.

We document the first records of biting midges in the genus Dasyhelea Kieffer (Diptera: Ceratopogonidae) from Curaçao: Dasyhelea azteca Huerta & Grogan, D. bahamensis (Johnson), D. cincta (Coquillett), D. corinneae Gosseries, D. flavifrons (Guérin-Méneville) and D. grisea (Coquillett). The following six new species of Dasyhelea are described from this Caribbean island and their key features are presented in color photographs and illustrations: D. aliciae n. sp., D. recurva n. sp., D. latiala n. sp., D. rhopaloparamera n. sp., D. cyrtostyla n. sp., and D. curacaoensis n. sp. We also provide a key to males and females of all known species of Dasyhelea from Curaçao.

The immatures of Bezziachilensis Spinelli & Ronderos, 2001 (Diptera, Ceratopogonidae).

The fourth instar larva and the pupa of Bezziachilensis Spinelli & Ronderos, 2001 are described for the first time. The immature stages were collected from macrophytes and filamentous algae in streams of the Patagonian steppe, in the provinces of Neuquén and Chubut, Argentina. The described stages were photographed and illustrated with a phase-contrast microscope and scanning electron microscope. Data on the bionomics of the species, new records and tables for characters of the known larvae and pupae of Bezzia Kieffer, 1899 from the Neotropical region are provided.

The biting and predaceous midges of Guadeloupe (Diptera: Ceratopogonidae). II. Species of the subfamily Dasyheleinae.

We provide descriptions, photographs and illustrations of 13 new species and new records of eight other species of biting midges in the genus Dasyhelea Kieffer from Guadeloupe. Included are the first Guadeloupe records of D. bermudae Wirth & Williams, D. caribbeana Spinelli & Wirth, D. cincta (Coquillett), D. flavifrons (Guérin-Méneville), D. grisea (Coquillett), D. griseola Wirth, D. luteogrisea Wirth & Williams and D. pseudoincisurata Waugh & Wirth. The previously unknown male of D. caribbeana is described and illustrated, and, a key is provided for the 24 species of Dasyhelea known from Guadeloupe.

Description of the last instar larva and new contributions to the knowledge of the pupa of Dasyhelea mediomunda Minaya (Diptera, Culicomorpha, Ceratopogonidae).

The fourth instar larva of Dasyhelea mediomunda Minaya is described for the first time and a complete description of the pupa is provided, through use of phase-contrast microscope and scanning electron microscope. Studied specimens were collected in a pond connected to a small wetland "mallin" on the Patagonian steppe, Chubut province, Argentina.

A new species of Dasyhelea Kieffer (Diptera: Ceratopogonidae) from Brazilian Amazonia.

A new species from Manaus, Amazonas, Brazil, Dasyhelea eloyi Díaz & Ronderos, is described, illustrated and photomicrograph as larva, pupa and male and female adults based on examination with binocular and scanning electron microscopes. This new species, a typical member of the Dasyhelea grisea species group, is compared with the most similar congeners D. necrophila Spinelli & Rodriguez and the Nearctic D. pseudoincinsurata Waugh & Wirth.

New contributions to the knowledge of the immatures of Stilobezzia punctulata Lane and Stilobezzia fiebrigi Kieffer (Diptera: Ceratopogonidae).

The fourth instar larva of Stilobezzia punctulata Lane is described for the first time and the one of Stilobezzia fiebrigi Kieffer is redescribed. They are illustrated and photomicrographed from material collected in different aquatic environments of the province of Corrientes, Argentina. Both species show features typical to carnivorous-predatory larva.

Biting midges (Diptera: Ceratopogonidae) from Martín García Island, Argentina.

Nearly 230 species of biting midges have been recorded or described from Argentina; 38 of them are known from the Buenos Aires province and only one is cited from Martín García Island. This paper presents the results raised from six collecting trips which took place on the island during spring 2005, summer 2006 and autumn 2009. Diverse sampling sites including permanent and temporary aquatic environments were chosen, most of the ten sampling sites were ponds of diverse origin, some of these environments were covered with floating vegetation as Lemna gibba, Lemna minuscule, Salvinia biloba, Salvinia minima, Azolla filiculoides, Limnobium laevigatum, Pistia stratiotes, Spirodela intermedia, Wolffiella oblonga and Wolffia columbiana. Other sites were placed in urban and suburban areas. Adults were collected with sweep nets at sunrise and sunset and with light traps at intervals of four to five hours at night, depending on electricity availability on the island. Larvae and pupae were collected with different implements depending on characteristics of each surveyed aquatic habitat. In free standing water, they were captured with small sieves or hand pipettes and micropipettes, flotation techniques were utilized for sampling vegetated areas, free and rooted floating hydrophytes were extracted for removing insects among them. Thirteen species of Ceratopogonidae were collected, three of Atrichopogon Kieffer, three of Forcipomyia Meigen, two of Dasyhelea Kieffer, four of Culicoides Latreille, and one of Bezzia Kieffer, all representing new records from the island.

Biting midges (Diptera: Ceratopogonidae) from Martín García Island, Argentina

Nearly 230 species of biting midges have been recorded or described from Argentina; 38 of them are known from the Buenos Aires province and only one is cited from Martín García Island. This paper presents the results raised from six collecting trips which took place on the island during spring 2005, summer 2006 and autumn 2009. Diverse sampling sites including permanent and temporary aquatic environments were chosen, most of the ten sampling sites were ponds of diverse origin, some of these environments were covered with floating vegetation as Lemna gibba, Lemna minuscule, Salvinia biloba, Salvinia minima, Azolla filiculoides, Limnobium laevigatum, Pistia stratiotes, Spirodela intermedia, Wolffiella oblonga and Wolffia columbiana. Other sites were placed in urban and suburban areas. Adults were collected with sweep nets at sunrise and sunset and with light traps at intervals of four to five hours at night, depending on electricity availability on the island. Larvae and pupae were collected with different implements depending on characteristics of each surveyed aquatic habitat. In free standing water, they were captured with small sieves or hand pipettes and micropipettes, flotation techniques were utilized for sampling vegetated areas, free and rooted floating hydrophytes were extracted for removing insects among them. Thirteen species of Ceratopogonidae were collected, three of Atrichopogon Kieffer, three of Forcipomyia Meigen, two of Dasyhelea Kieffer, four of Culicoides Latreille, and one of Bezzia Kieffer, all representing new records from the island. Rev. Biol. Trop. 59 (3): 1183-1194. Epub 2011 September 01.

Alrededor de 230 especies de ceratopogónidos han sido registradas o descritas en Argentina, 38 de ellas son conocidas para la provincia de Buenos Aires y sólo una ha sido previamente citada para la Isla Martín García. En este trabajo se presentan los resultados obtenidos a partir de muestreos realizados en seis viajes a la isla durante la primavera de 2005, verano de 2006 y otoño de 2009; se seleccionaron diez sitios de muestreo con ambientes acuáticos permanentes y temporales. La mayoría de los muestreos se llevaron a cabo en cuerpos de agua de diverso origen; algunos de éstos se hallaban cubiertos con vegetación flotante compuesta por Lemna gibba, Lemna minuscule, Salvinia biloba, Salvinia minima, Azolla filiculoides, Limnobium laevigatum, Pistia stratiotes, Spirodela intermedia, Wolffiella oblonga y Wolffia columbiana; mientras que los restantes se realizaron en zonas urbanas y suburbanas. Los adultos se capturaron con redes entomológicas a la salida y puesta del sol y con trampas de luz a intervalos de cuatro a cinco horas por la noche, dependiendo de la disponibilidad de electricidad en la isla. Las larvas y pupas fueron recolectadas con diferentes metodologías según las características de los ambientes estudiados; en el agua libre se capturaron con tamices pequeños o pipetas y micropipetas, y en cuerpos de agua con vegetación se utilizó la técnica de flotación para separar los insectos de las hidrófitas flotantes y arraigadas. Se identificaron trece especies de Ceratopogonidae, 3 de Atrichopogon Kieffer, 3 de Forcipomyia Meigen, 2 de Dasyhelea Kieffer, 4 de Culicoides Latreille, y uno de Bezzia Kieffer, todos ellos representan nuevos registros para la isla.

Reaction of Müller cells in an experimental rat model of increased intraocular pressure following timolol, latanoprost and brimonidine.

The aim of this study was to evaluate the reaction of Müller cells in an experimental rat model of intraocular pressure (IOP) and their response to treatment with ocular hypotensive drugs. Episcleral vein cauterization in unilateral eyes of Wistar rats was performed to produce elevated IOP. The animals were divided into five groups: control, experimental, and experimental treated with timolol, latanoprost or brimonidine. Histological sections of retina were studied by immunochemistry with antibodies to glial fibrillary acidic protein (GFAP), and the percentage of labeled area was measured to evaluate the degree of reactive gliosis. In the experimental group, the Müller cells showed hypertrophy and a significant increase in GFAP (4.39+/-0.32%) in relation to retinas of the control group (2.05+/-0.14%). Gliosis was detected in all three treated groups, with a varying increase in GFAP intensity. The timolol-treated group showed the most intense and persistent glial reactivity after 3 months of treatment (13.89+/-0.63%). Treatment with brimonidine, however, resulted in a decrease in the level of GFAP immunoreactivity (8.37+/-0.4%). The group treated with latanoprost showed the lowest glial reactivity (4.8+/-0.36%). Given that all three drugs are effective hypotensive agents, their neuroprotective effect could be related with other factors, such as gliosis, which, over long periods may have noxious effects on the neurons. Thus, hypotensives like brimonidine, and specially latanoprost, may afford greater neuroprotection to the ganglion cells by attenuating the retinal glial reaction.

Experimental model of ocular hypertension in the rat: study of the optic nerve capillaries and action of hypotensive drugs.

PURPOSE: To investigate quantitatively the effect of elevated intraocular pressure (IOP) on the microvasculature of the optic nerve with and without topical treatment with two hypotensive drugs, timolol and latanoprost. METHODS: Three groups of rats underwent cauterization of three episcleral veins to produce elevated IOP in the right eye. Two of these groups were treated with timolol or latanoprost for 3 months. Eyeballs were incubated with anti-GLUT-1 polyclonal antibody. GLUT-1-positive capillaries of the optic nerve head (ONH) and optic nerve exit (ON) were examined and analyzed for their number per square millimeter, volume fraction, length per unit volume, surface area per unit volume, and mean diameter. RESULTS: An increase in IOP resulted in a significant decrease in microvessel density in the laminar region (LR) and postlaminar region (PR) and ON compared with the control group. The other parameters fell significantly in all regions of the optic nerve. Topical treatment with timolol or latanoprost did not modify the density of the capillaries, although the other parameters increased significantly compared with the untreated experimental group. Additionally, the mean diameter of the capillaries in the LR and the PR recovered after treatment. CONCLUSIONS: The results indicate that the capillaries of the LR and the PR of the ONH are the most susceptible to IOP elevation. The authors suggest that timolol and latanoprost have a certain vascular action by increasing the available blood volume, surface area per unit volume, length per unit volume, and diameter of the capillaries of the ONH in these two regions.

Study of the effects of ocular hypotensive drugs on number of neurons in the retinal ganglion layer in a rat experimental glaucoma.

PURPOSE: We investigated the effects of antiglaucomatous drugs on neurons in the retinal ganglion layer (RGL) in an experimental model of elevated intraocular pressure (IOP). METHODS: Three episcleral veins of rats with normal IOP were cauterized. Three months later, we examined the effects on the number of neurons in the RGL as well as in rats submitted to treatment with timolol, latanoprost, or brimonidine. The IOP was measured using a calibrated Tono-Pen XL tonometer before and immediately after cauterization and every 2 weeks for the following 3 months as well as immediately before perfusion. RESULTS: The IOP was 14.85+/-0.65 mmHg in the control group, whereas it was 1.25-fold higher (33.5+/-1.06 mmHg) in the experimental group. After treatment, the IOP returned to baseline levels. The mean number of neurons per mm2 in the RGL was 33% lower in the experimental group (283+/-10 cells/mm2) compared with the control group (423+/-11 cells/mm2). In the groups treated with timolol, latanoprost, or brimonidine, the neuronal loss was less (331+/-10, 360+/-15, and 333+/-3 cells/mm2, respectively), although values did not return to baseline levels. CONCLUSIONS: This experimental model provokes an immediate, constant, and prolonged increase in IOP and the application of hypotensive agents affords a certain degree of protection to neurons in the RGL.

Nitric oxide synthase in retina and optic nerve head of rat with increased intraocular pressure and effect of timolol.

We investigated the expression of nitric oxide synthase (NOS) isoforms -1, -2 and -3 in the retina and optic nerve head (ONH) in an experimental rat model of elevated intraocular pressure (IOP) before and after treatment with timolol, to assess whether its neuroprotective action is associated with the activity of these enzymes. Episcleral vein cauterization in unilateral eyes of Wistar rats was performed to produce elevated IOP. Histological sections of retina and ONH from animals with normal IOP, with elevated IOP, and elevated IOP treated with timolol, were studied by immunohistochemistry with antibodies to NOS-1, NOS-2, and NOS-3. In the control rats, NOS-1 was localized to photoreceptor inner segments, amacrine cells and bipolar cells in the retina, and in astrocytes, pericytes and vascular nitrergic terminals in the ONH. NOS-3 immunostaining localized to the endothelial cells. The rats with elevated IOP showed increased expression of NOS-1 in the plexiform layers of the retina and reactive astrocytes in the ONH. These cells also showed NOS-2 positivity. The rats treated with timolol showed reduced expression of NOS-1 in the retina and ONH. NOS-2 was only detected in a few groups of astrocytes in the ONH. NOS-3 was unchanged in both elevated IOP and timolol-treated groups. These results show that excessive levels of NO synthesized by the NOS-1 and -2 isoforms, considered neurotoxic, might contribute to the progressive lesions of retinal ganglion cell axons. Their reduction after treatment suggests a possible neuroprotective effect of timolol in neurons exposed to excessive amounts of NO.

NADPH-diaphorase activity in the superficial layers of the superior colliculus of rats during aging.

Neurons in the superficial layers of the superior colliculus are key elements in the visual system of rodents since they receive extensive afferent projections from retinal ganglion cells. The NADPH-diaphorase histochemical technique was used to detect differences in neuronal nitric oxide synthase (nNOS) in the superficial layers of the superior colliculus (sSC) of young adult (3 months) and aged (24 and 26 months) rats. The orientation of the dendritic processes of NADPH-diaphorase-positive neurons, cross-sectional area, and number of neurons per mm2 were analyzed. NADPH-d histochemistry revealed a high number of NADPH-d-positive cells in the stratum zonale and stratum griseum superficiale in adult and aged animals. NADPH-d-positive neurons were classified into the following morphological types: marginal, horizontal, pyriform, narrow-field vertical, wide-field vertical, and stellate. During aging, narrow field vertical and wide field vertical neurons present somatic atrophy and an increase in dendritic processes with dorsoventral orientation, whereas wide field vertical neurons show a decrease in those with lateromedial orientation. Marginal neurons undergo somatic hypertrophy at 26 months when compared with those at 3 months. The remaining types of neurons do not undergo size changes. Finally, the number of NADPH-d-positive neurons per mm2 in the various types of morphology does not significantly change with age. It is suggested to be likely that the aging process in the nitrergic neurons of the sSC does not lead to significant changes in the synthesis of NO from the constitutive NOS isoforms.