Insecticidal properties of essential oils from <Emphasis Type="Italic">Lippia turbinata</Emphasis> and <Emphasis Type="Italic">Lippia polystachya</Emphasis> (Verbenaceae) against <Emphasis Type="Italic">Culex quinquefasciatus</Emphasis> (Diptera: Culicidae)

Mosquitoes are important vectors of diseases to humans and domestic animals. Chemical control of vectors remains a main resource for the prevention and control of vector-borne diseases. Due to the development of insecticide resistance and risks to human health and the environment of synthetic compounds, the search for alternative pesticides is encouraged. This work assessed the insecticidal activity of essential oils (EOs) from Lippia turbinata and L. polystachya from Argentina on Culex quinquefasciatus mosquitoes. EOs were extracted by hydro-distillation and analyzed with gas–liquid chromatography and mass spectrometry. The insecticidal activity against mosquito larvae, pupae, and adults were evaluated according to World Health Organization protocols. Concentrations ranking from 10 to 160 ppm were assessed at 1, 2, 3, and 24 h posttreatment. The composition of the EO of L. polystachya and L. turbinata were qualitatively similar, with α-thujone and carvone as main constituent; differences were mostly due to the proportion of each component. β-caryophyllene was also an important constituent of the EO of L. turbinata. Both EO were larvicidal at concentrations of 80 ppm or higher, but only L. turbinata was adulticidal. No pupal mortality was detected. The potential of these EOs for vector control is discussed.     

Winter activity of <Emphasis Type="Italic">Phlebotomus</Emphasis> (<Emphasis Type="Italic">Transphlebotomus</Emphasis>) <Emphasis Type="Italic">mascittii</Emphasis>, Grassi 1908 (Diptera: Psychodidae) on the island of Corsica

In the first week of February 2008 on the island of Corsica, 55 Phlebotomus (Transphlebotomus) mascittii, 39 males and 16 females, were caught in a railway tunnel. This is the first proof of the winter activity of the adult P. mascittii in Europe.     

<Emphasis Type="Italic">Trypanosoma</Emphasis> (<Emphasis Type="Italic">Herpetosoma</Emphasis>) <Emphasis Type="Italic">kuseli</Emphasis> sp. n. (Protozoa: Kinetoplastida) in Siberian flying squirrels (<Emphasis Type="Italic">Pteromys volans</Emphasis>)

All trypanosome species classified in the subgenus Herpetosoma in sciurid hosts have been recorded from ground and tree squirrels to date, but not from any flying squirrels. We describe in this paper a novel trypanosome species, Trypanosoma (Herpetosoma) kuseli sp. n., from Siberian flying squirrels (Pteromys volans) imported from China, and compare it with T. (H.) otospermophili in Richardson's ground squirrels (Spermophilus richardsonii) and Columbian ground squirrels (Spermophilus columbianus) from the USA. Due to a short free flagellum, the new species appeared stumpy compared with T. otospermophili (length of free flagellum 7.0 +/- 0.8 microm, total length 32.1 +/- 0.8 microm, n = 13 and length of free flagellum 15.5 +/- 1.6 microm, total length 35.9 +/- 1.0 microm, n = 13, respectively). Another conspicuous morphological feature of the new species was an anteriorly positioned kinetoplast, found approximately at the midpoint between the nucleus and the posterior end. These characters have not been recorded from any squirrel Herpetosoma trypanosome species. Comparison of the nucleotide sequences of the small and large subunit rRNA genes indicated that T. kuseli sp. n. was more homologous to T. otospermophili than murid Herpetosoma species, such as T. grosi, T. lewisi, T. musculi, T. microti and T. evotomys.     

Cysteine proteinases from promastigotes of <Emphasis Type="Italic">Leishmania</Emphasis> (<Emphasis Type="Italic">Viannia</Emphasis>) <Emphasis Type="Italic">braziliensis</Emphasis>

Leishmania (Viannia) braziliensis is the major causative agent of American tegumentary leishmaniasis, a disease that has a wide geographical distribution and is a severe public health problem. The cysteine proteinase B (CPB) from Leishmania spp. represents an important virulence factor. In this study, we characterized and localized cysteine proteinases in L. (V.) braziliensis promastigotes. By a combination of triton X-114 extraction, concanavalin A-affinity, and ion exchange chromatographies, we obtained an enriched fraction of hydrophobic proteins rich in mannose residues. This fraction contained two proteinases of 63 and 43 kDa, which were recognized by a CPB antiserum, and were partially sensitive to E-64 in enzymatic assays with the peptide Glu-Phe-Leu. In confocal microscopy, the CPB homologues localized in the peripheral region of the parasite. This data together with direct agglutination and flow cytometry assays suggest a surface localization of the CPB homologues. The incubation of intact promastigotes with phospholipase C reduced the number of CPB-positive cells, while anti-cross-reacting determinant and anti-CPB antisera recognized two polypeptides (63 and 43 kDa) derived from phospholipase C treatment, suggesting that some CPB isoforms may be glycosylphosphatidylinositol-anchored. Collectively, our results suggest the presence of CPB homologues in L. braziliensis surface and highlight the need for further studies on L. braziliensis cysteine proteinases, which require enrichment methods for enzymatic detection.     

Ultrastructure of eggshell of<Emphasis Type="Italic"> Chrysomya nigripes</Emphasis> Aubertin (Diptera: Calliphoridae)

We examined the eggshell structure of the blowfly, Chrysomya nigripes Aubertin, a species of forensic importance, using scanning electron microscopy. A relatively wide plastron region was located dorsally. It extended almost the entire length of the egg and bifurcated slightly, close to the micropyle, which is a deep cavity surrounded by an elevated, fine, wavy rim. The chorionic sculpture had an hexagonal pattern, with a smooth and elevated boundary. The ruptured eggshell showed the outermost exochorion, membranous outer endochorion and pillar layer, which also had an hexagonal pattern on a perforated mesh pillar layer. This study provides a greater database on the egg of this forensically significant blowfly.     

Redescription of <Emphasis Type="Italic">Lemuricola</Emphasis> (<Emphasis Type="Italic">Madoxyuris</Emphasis>) <Emphasis Type="Italic">bauchoti</Emphasis> (Nematoda,Oxyuridae) from <Emphasis Type="Italic">Lemur catta</Emphasis> in Madagascar

Lemuricola (Madoxyuris) bauchoti Chabaud, Brygoo et Petter, 1965 is redescribed from material collected from the ring-tailed lemur, Lemur catta, from the Beza Mahafaly Special Reserve in Madagascar using the scanning electron microscope. This is a new host record and the first oxyurid reported from the ring-tailed lemur. Previously, records of each species of the subgenus Madoxyuris have been restricted to a single host species, but the close relationship between these nematodes and their Strepsirrhini hosts will only be proven when additional records fill in the gaps in their distribution.     

First report on susceptibility of wild <Emphasis Type="Italic">Aedes aegypti</Emphasis> (Diptera: Culicidae) using <Emphasis Type="Italic">Carapa guianensis</Emphasis> (Meliaceae) and <Emphasis Type="Italic">Copaifera</Emphasis> sp. (Leguminosae)

Oils of Carapa guianensis and Copaifera spp. are well known in the Amazonian region as natural insect repellent, and studies have reported their efficiency as larvicide against some laboratory mosquito species. However, in wild populations of mosquitoes, these oils have not yet been evaluated. Thus, the objective of this study was to investigate their efficiency as larvicide in wild populations of Aedes aegypti with a history of exposure to organophosphate. The susceptibility of larvae was determined under three different temperatures, 15°C, 20°C, and 30°C. For each test, 1,000 larvae were used (late third instar and early fourth instar—four replicates of 25 larvae per concentration). Statistical tests were used to identify significant differences. The results demonstrated that as the laboratory A. aegypti, the wild populations of A. aegypti were also susceptible to C. guianensis and Copaifera sp. oils. The lethal concentrations for Copaifera sp. ranged from LC₅₀ 47 to LC₉₀ 91 (milligrams per liter), and for C. guianensis, they were LC₅₀ 136 to LC₉₀ 551 (milligrams per liter). In relation to different temperature, the effectiveness of the oils on larvae mortality was directly related to the increase of temperature, and better results were observed for temperature at 25°C. The results presented here indicate the potential larvicidal activity of C. guianensis and species of Copaifera, in populations of A. aegypti from the wild. Therefore, the results presented here are very important since such populations are primarily responsible for transmitting the dengue virus in the environment.     

Cercarial shedding from <Emphasis Type="Italic">Galba</Emphasis><Emphasis Type="Italic">truncatula</Emphasis> infected with <Emphasis Type="Italic">Fasciola</Emphasis><Emphasis Type="Italic">gigantica</Emphasis> of distinct geographic origins

Galba truncatula snails were experimentally infected with either of two different isolates of Fasciola gigantica, originating from Egypt or China, to determine the influence of these isolates on the characteristics of snail infections. The survival rates of G. truncatula on day 30 post-exposure were 90.0% and 60.2% in the Egyptian and Chinese groups, respectively. The frequency of cercaria-shedding snails within the Egyptian group was 79.8%, whereas in the Chinese group it was 22.4%. The parasite origin had a significant effect on the durations of the prepatent and patent periods. The mean number of cercariae shed from the Egyptian group was significantly greater than that shed from the Chinese group (a mean of 275.5 per cercaria-shedding snail compared with 29.0). These results could be explained by the fact that G. truncatula might be a natural intermediate host for F. gigantica in Egypt, and the greater adaptability of the Egyptian miracidia of F. gigantica to unusual snail hosts. These results demonstrate the influence of the geographic origin of the parasite on the success of trematodes infecting snails.     

Antiviral biflavonoids from <Emphasis Type="Italic">Radix Wikstroemiae</Emphasis> (<Emphasis Type="Italic">Liaogewanggen</Emphasis>)

Background  

Radix Wikstroemiae is a common Chinese herbal medicine. The ethyl acetate fraction of the ethanolic extract of W. indica possesses potent in vitro antiviral activity against respiratory syncytial virus (RSV). This study aims to identify the antiviral components of the active fraction.     

Isolation and identification of bioactive compounds in <Emphasis Type="Italic">Andrographis paniculata</Emphasis> (<Emphasis Type="Italic">Chuanxinlian</Emphasis>)

Andrographis paniculata (Burm. f.) Nees (Acanthaceae) is a medicinal plant used in many countries. Its major constituents are diterpenoids, flavonoids and polyphenols. Among the single compounds extracted from A. paniculata, andrographolide is the major one in terms of bioactive properties and abundance. Among the andrographolide analogues, 14-deoxy-11,12-didehydroandrographolide is immunostimulatory, anti-infective and anti-atherosclerotic; neoandrographolide is anti-inflammatory, anti-infective and anti-hepatotoxic; 14-deoxyandrographolide is immunomodulatory and anti-atherosclerotic. Among the less abundant compounds from A. paniculata, andrograpanin is both anti-inflammatory and anti-infective; 14-deoxy-14,15-dehydroandrographolide is anti-inflammatory; isoandrographolide, 3,19-isopropylideneandrographolide and 14-acetylandrographolide are tumor suppressive; arabinogalactan proteins are anti-hepatotoxic. The four flavonoids from A. paniculata, namely 7-O-methylwogonin, apigenin, onysilin and 3,4-dicaffeoylquinic acid are anti-atherosclerotic.     

Crossing experiment of <Emphasis Type="Italic">Anopheles maculatus</Emphasis> form K and <Emphasis Type="Italic">Anopheles willmori</Emphasis> (James) (Diptera: Culicidae)

We recently reported crossing experiments between Anopheles maculatus form K and five members of the Maculatus group to support the specific status of form K. In the present study, we performed further crosses between form K and a sixth species of the Maculatus group, Anopheles willmori (James). Low viability was observed in hybrid males and females. All hybrid males were sterile with atrophied testes, or partially sterile with abnormal spermatozoa. The hybrid females showed normal ovaries. The ovarian nurse cell polytene chromosomes of the F₁ hybrid females displayed complete asynapsis. Backcrossing showed low viability. All males were sterile with atrophied testes or partially sterile with abnormal spermatozoa, and the females showed varying degrees of atrophied ovaries. The results provide clear evidence that form K is not conspecific to A. willmori supporting previous studies that form K represents another species of the Maculatus group.     

Infections by the Yeast<Emphasis Type="Italic"> Kodomaea</Emphasis> (<Emphasis Type="Italic">Pichia</Emphasis>)<Emphasis Type="Italic"> ohmeri</Emphasis>: Two Cases and Literature Review

A 14-year-old boy who was neutropenic following chemotherapy for leukemia developed fungemia caused by the yeast Kodomaea ohmeri (Pichia ohmeri). The infection was cured by catheter removal and the use of fluconazole. A 74-year-old man who had undergone surgeries for a subcutaneous tumor developed polymicrobic cellulitis involving Kodomaea ohmeri. Despite surgical debridement and antibiotic therapy, the patient died of complications. Including these 2 cases, there have been 10 Kodomaea ohmeri infections reported thus far, all occurring in patients with pre-existing conditions. There have been seven cases of fungemia and one case each of peritonitis, funguria, and cellulitis. The treatment employed varied depending on the site/source of infection. Seven patients recovered and three died. The microbiological data available suggest that Kodomaea ohmeri can be identified definitively by biochemical tests and is susceptible to amphotericin B and either susceptible to or dose dependently susceptible to itraconazole and fluconazole.     

<Emphasis Type="Italic">Phlebotomus langeroni</Emphasis> Nitzulescu (Diptera,Psychodidae) a new vector for <Emphasis Type="Italic">Leishmania infantum</Emphasis> in Europe

Burrows of the wild rabbit, Oryctolagus cuniculus, a lagomorph that has been recently suggested as a Leishmania infantum reservoir, constitute an unspoilt biotope in phlebotomine studies in Europe. We hypothesize that Phlebotomus langeroni, a proven vector of L. infantum in North Africa, is associated with rabbits and may have been overlooked in Europe. Sandfly captures were carried out with CDC light traps in an L. infantum endemic area of southern Spain with a high density of lagomorphs and a large numbers of burrows. The stable, permanent, and highly abundant presence of P. langeroni was assessed. After morphological identification, this sandfly species was characterized by comparing it with P. perniciosus and other P. langeroni populations from North Africa through molecular techniques. P. langeroni had not been found in southern Spain to date, despite being a highly investigated area, except for this particular biotope. Its activity period turned out to begin in mid-July, ending in late October, accounting for a maximum activity during this month. This study shows that P. langeroni is associated with the existence of rabbit burrows and has been overlooked in Europe. L. infantum DNA was found in almost half of the female specimens (47.6%) captured inside a biotope where wild rabbits are infected as well.     

Subcellular localization of an extracellular serine protease in<Emphasis Type="Italic"> Leishmania</Emphasis> (<Emphasis Type="Italic">Leishmania</Emphasis>)<Emphasis Type="Italic"> amazonensis</Emphasis>

Extracellular proteolytic activity was detected in a Leishmania (L.) amazonensis culture supernatant and a 56-kDa protein was purified using (NH₄)₂SO₄ precipitation followed by affinity chromatography on aprotinin–agarose. A rabbit serum obtained against the 56-kDa extracellular serine protease was used in order to analyze its location in L. (L.) amazonensis parasites. Immunocytochemistry studies revealed that the enzyme is mainly found in the flagellar pocket and cytoplasmic vesicles of promastigote forms, whereas in amastigotes, it is located in electron-dense structures resembling megasomes. These results indicate that the 56-kDa serine protease is released into the extracellular environment through the flagellar pocket; and its intracellular location suggests either a correlated enzymatic activity or intracellular trafficking.     

<Emphasis Type="Italic">Amaranthus oleracea</Emphasis> and <Emphasis Type="Italic">Euphorbia hirta</Emphasis>: natural potential larvicidal agents against the urban Indian malaria vector, <Emphasis Type="Italic">Anopheles stephensi</Emphasis> Liston (Diptera: Culicidae)

Malaria control in developing countries is based largely on vector eradication by the use of mosquito larvicides which is an ideal method for controlling mosquito and the related epidemics. On account of ecohazardous nature, nontarget specificity of chemical insecticides and evidences of developing resistance against them in the exposed species, currently, importance of secondary plant metabolites has been acknowledged. Insecticides of plant origin are environmentally safe, degradable, and target specific. In view of this fact, the present work highlights the larvicidal property of extracts of Amaranthus oleracea and Euphorbia hirta against the third instar larvae of Anopheles stephensi, the urban malaria vector. LC₅₀ values for the carbon tetrachloride fraction of A. oleracea against larvae are 17,768.00 and 13,780.00 ppm after 24 and 48 h of exposure accordingly. For the methanol extract of the same, LC₅₀ values are 15,541.00 and 10,174.00 ppm after 24 and 48 h of exposure. In the case of petroleum ether extract, LC₅₀ values after 24 and 48 h of exposure are 848.75 and 311.50 ppm. LC₅₀ values for carbon tetrachloride extracts of E. hirta against the larvae are 11,063.00 and 10,922.00 ppm after 24 and 48 h of exposure, respectively. For methanol extract of the same extract, the LC₅₀ values are 19,280.00 and 18,476.00 ppm after 24 and 48 h of exposure. In the case of petroleum ether extract, LC₅₀ values after a 24- and 48-h exposure period are 9,693.90 and 7,752.80 ppm. The results obtained for petroleum extracts of A. oleracea are encouraging and there are probabilities that the active principle contained in this extract may be more effective than its crude form and may serve as ecofriendly mosquito larvicide.     

<Emphasis Type="Italic">Leishmania</Emphasis> (<Emphasis Type="Italic">Viannia</Emphasis>) <Emphasis Type="Italic">shawi</Emphasis> purified antigens confer protection against murine cutaneous leishmaniasis

Objective  

Leishmania (Viannia) shawi was characterized only recently, and few studies concerning the immunogenic and protective properties of its antigens have been performed. The present study aimed to evaluate the protective potential of the five antigenic fractions isolated from L. (V.) shawi promastigotes in experimental cutaneous leishmaniasis.     

Occurrence of <Emphasis Type="Italic">Cryptosporidium hominis</Emphasis> in pigeons (<Emphasis Type="Italic">Columba livia</Emphasis>)

This study demonstrated the presence of Cryptosporidium hominis in pigeons for the first time. Previously, C. hominis had been cited only in another bird species, Branta canadiensis. The present findings suggest that pigeons may act as mechanical vectors for this protozoan.     

Larvicidal efficacy of <Emphasis Type="Italic">Jatropha curcas</Emphasis> and bacterial insecticide, <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis>, against lymphatic filarial vector<Emphasis Type="Italic">, Culex quinquefasciatus</Emphasis> Say (Diptera: Culicidae)

The present study explored the effects of Jatropha curcas leaf extract and Bacillus thuringiensis israelensis larvicidal activity against the lymphatic filarial vector, Culex quinquefasciatus. Wights were selected for investigating the larvicidal potential against the first to fourth instar larvae of the laboratory-reared mosquito species, C. quinquefasciatus Say, in which the major lymphatic filariasis was used. The medicinal plants were collected from the area around Bharathiar University, Coimbatore. The dried plant materials were powdered by an electric blender. From the powder, 100 g of the plant materials was extracted with 300 ml of organic solvents of methanol for 8 h, using a Soxhlet apparatus, and filtered. The crude plant extracts were evaporated to dryness in a rotary vacuum evaporator. The plant extract showed larvicidal effects after 24 h of exposure; however, the highest larval mortality was found in the leaf extract of methanol J. curcas against the first to fourth instar larvae of values LC₅₀ = 1.200%, 1.290%, 1.358%, and 1.448% and LC₉₀ = 2.094%, 2.323%, 2.444%, and 2.544% and B. thuringiensis israelensis against the first to fourth instar larvae of values LC₅₀ = 9.332%, 9.832%, 10.212%, 10.622% and LC₉₀ = 15.225%, 15.508%, 15.887%, and 15.986% larvae of C. quinquefasciatus, respectively. No mortality was observed in the control. These results suggest methanol extracts of J. curcas and B. thuringiensis israelensis have potential to be used as an ideal eco-friendly approach for the control of the major lymphatic filarial vector, C. quinquefasciatus.     

In vitro efficacy of plant extracts and synthesized substances on <Emphasis Type="Italic">Rhipicephalus</Emphasis> (<Emphasis Type="Italic">Boophilus</Emphasis>) <Emphasis Type="Italic">Microplus</Emphasis> (Acari: Ixodidae)

Herbal drugs have been widely evaluated as an alternative method of parasite control, aiming to slow development of resistance and obtain low-cost biodegradable parasiticides. This study evaluated the in vitro efficacy on Rhipicephalus (Boophilus) microplus of extracts from Carapa guianensis seed oil, Cymbopogon martinii and Cymbopogon schoenanthus leaf essential oil, and Piper tuberculatum leaf crude extract and similar synthesized substances. In the immersion test, engorged females were evaluated in five dilutions ranging from 10% to 0.030625% concentration. In the larval test on impregnated filter paper, the concentration ranged from 10% to 0.02%. The treatments and controls were done in three replicates. Chemical analysis of the oils was performed by gas chromatography. The main compounds were oleic acid (46.8%) for C. guianensis and geraniol for C. martinii (81.4%), and C. schoenanthus (62.5%). The isolated and synthesized substances showed no significant effect on larvae and adult. C. martinii and P. tuberculatum showed the best efficacy on the engorged females. The LC₅₀ and LC₉₀ were 2.93% and 6.66% and 3.76% and 25.03%, respectively. In the larval test, the LC₅₀ and LC₉₀ obtained for C. martinii, P. tuberculatum, and C. schoenanthus were 0.47% and 0.63%, 0.41% and 0.79%, 0.57% and 0.96%, respectively. The fact that geraniol is present in greater quantities in C. martinii explains its higher activity in relation to C. shoenanthus. It is necessary to validate the in vivo use of safe and effective phytoparasiticidal substances. Efforts should be focused on developing formulations that enhance the efficacy in vivo and lengthen the residual period.