Isolation and isoenzyme characterization of Leishmania (Viannia) braziliensis from a case of human cutaneous leishmaniasis in northeast centre of the state of São Paulo

The diagnosis of human cutaneous leishmaniasis in small towns is sometimes made without the species identification of the Leishmania, even in areas without previous epidemiological surveys. Here we report the isolation of a Leishmania strain from a patient of Rincão, state of São Paulo, that was identified by isoenzyme characterization as L. (Viannia) braziliensis. Sand fly collections were made in the area where the patient live in order to investigate the likely vector species.

Description of Pintomyia (Pifanomyia) brazilorum sp. nov. a new fossil species from the Dominican Republic (Diptera: Psychodidae: Phlebotominae)

A sand fly fossil was found in amber, a vegetal resin, which allows all the external phlebotomine structures to be seen. The piece that contains the new species is 14 mm long ´ 8 mm wide ´ 3 mm high. All the structures from the head, thorax, and abdomen were examined under the microscope and measured with a calibrated micrometric eye-piece. The morphological aspects of the new species suggest its inclusion in the Pintomyia genus, Pifanomyia subgenus though it is not possible to include it in any of the series known for this subgenus. The presence of two atrophied spines on the gonostyles and gonocoxites without tufts of setae permit the exclusion of the new species from the other species of the subgenus Pifanomyia. The new species is named Pintomyia (Pifanomyia) brazilorum sp. nov.

Phlebotomines (Diptera: Psychodidae) in forested areas of the Serra da Bodoquena, state of Mato Grosso do Sul, Brazil

Investigation was undertaken on the behaviour of the phlebotomine fauna in caves, forests, and anthropic environments of the Serra da Bodoquena, between January 1998 and January 2000. This paper reports on the phlebotomines captured in forested areas with automatic light traps (ALT), Shannon traps (ST), aspiration (AN), at natural resting sites and by human attractiveness (HA) during 24 h. The diversity and abundance of the species were investigated with ALT installed at 16 points (ground level) and 6 in the canopy. Natural infection by flagellates was investigated in females captured with ST, AN, and HA. The sand fly fauna was represented by 23 species. Twenty-two of these were captured with ALT, 15 of them on the western side, and 20 on the eastern. Lutzomyia longipalpis and Nyssomyia whitmani were the most abundant on the former and this species together with Lutzomyia almerioi on the latter side. On the eastern side the ecotopes located close to caves rendered a significantly greater number (P < 0.01) of specimens than did more distant sites. On this side Lu. almerioi contributed with 56% of the total number of specimens. Lu. almerioi females were predominantly attracted by humans (96.4%) and by ST (93.2%) and three of the 2173 dissected (0.138%) presented natural infection by flagellates. The attraction of Lu. almerioi to humans occurred during all seasons, predominantly in the summer, and in nocturnal and diurnal periods. Thus it is bothersome to inhabitants of and visitors to the Bodoquena ridge and a potential vector of flagellates.

Abundance of Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae) and urban transmission of visceral leishmaniasis in Campo Grande, state of Mato Grosso do Sul, Brazil

The outspread and urbanization of visceral leishmaniasis (VL) in Campo Grande, state of Mato Grosso do Sul, lead us to undertake the present study over diversity and abundance of sand flies in the urban area to compare with previous search carried out during 1999/2000, before the identification of the disease in the human population.The captures were carried out with automatic light traps, weekly, from February 2004 to February 2005 on three sites including a forested area (Zé Pereira), two peridomicilies (shelters of domestic animals and cultivation areas), and intradomicilie. In the present study 110 collections were obtained during 13 months for 1320 h of collections, resulting in 5004 specimens, 3649 males and 1355 females belonging to the 20 following species: Brumptomyia avellari, Brumptomyia sp., Bichromomyia flaviscutellata, Evandromyia lenti, E. termitophila, E. cortelezzii, E. borrouli, Lutzomyia sp., L. longipalpis, Micropygomyia quinquefer, N. antunesi, N. whitmani, Pintomyia christenseni, Pi. damascenoi, Psathyromyia aragaoi, Ps. campograndensis, Ps. hermanlenti, Ps. shannoni, Pychodopygus claustrei, and Sciopemyia sordellii. L. longipalpis was the most abundant species in the anthropic environment with 92.22% of the captures. This shows an increase of sixty times in the density of L. longipalpis compared to the last sand fly evaluation in 1999/2000. The high density of L. longipalpis in Campo Grande is the main factor of risk in transmission of the disease to human in the urban area. The capture of N. antunesi, typical specie from Amazonian region, in Mato Grosso do Sul is reported for the first time.

Lutzomyia (Nyssomyia) whitmani s.l . (Antunes & Coutinho, 1939)(Diptera: Psychodidae: Phlebotominae): geographical distribution and the epidemiology of American cutaneous leishmaniasis in Brazil Mini-review

The aim of the present study, in view of the widespread geographical distribution of Lutzomyia (Nyssomyia) whitmani s.l. in Brazil, in close association with the regions of transmission of Leishmania (Viannia) braziliensis and L. (V.) shawi, is to investigate the distribution of this sand fly species and American cutaneous leishmaniasis (ACL) in relationship to vegetation and landscape ecology throughout Brazil. Thematic maps were elaborated by the MapInfo programme, giving information on the spatial distribution of L. whitmani s.l., in accordance with types of vegetation and foci of ACL. With regards to the known areas of transmission of ACL in Brazil, it is notable that L. (N.) whitmani s.l. occurs in most of them, where it has been implicated as a possible vector of L. (V.) braziliensis. The presence of L. whitmani s.l. has been registered in 26 states, the one exception being Santa Catarina; in some states such as Roraima, Acre, Tocantins, and Mato Grosso do Sul this sand fly species has been recorded in a large number of municipalities. L. whitmani s.l. has been found in association with a variety of vegetation types, including the Amazonian forest, savanna ("campos cerrados"), and northeastern savanna ("caatingas nordestinas" or "savana estépica").

Cloning and characterization of a V-ATPase subunit C from the American visceral leishmaniasis vector Lutzomyia longipalpis modulated during development and blood ingestion

Visceral leishmaniasis (VL) is a serious tropical disease that affects approximately 500 thousand people worldwide every year. In the Americas, VL is caused by the parasite Leishmania (Leishmania) infantum chagasi mainly transmitted by the bite of the sand fly vector Lutzomyia longipalpis. Despite recent advances in the study of interaction between Leishmania and sand flies, very little is known about sand fly protein expression profiles. Understanding how the expression of proteins may be affected by blood feeding and/or presence of parasite in the vector's midgut might allow us to devise new strategies for controlling the spread of leishmaniasis. In this work, we report the characterization of a vacuolar ATPase subunit C from L. longipalpis by screening of a midgut cDNA library with a 220 bp fragment identified by means of differential display reverse transcriptase-polymerase chain reaction analysis. The expression of the gene varies along insect development and is upregulated in males and bloodfed L. longipalpis, compared to unfed flies.

Effects of timber harvest on phlebotomine sand flies (Diptera: Psychodidae) in a production forest: abundance of species on tree trunks and prevalence of trypanosomatids

The Amazon forest is being exploited for timber production. The harvest removes trees, used by sand flies as resting sites, and decreases the canopy, used as refuges by some hosts. The present study evaluated the impact of the timber harvest, the abundance of sand flies, and their trypanosomatid infection rates before and after selective logging. The study was accomplished in terra-firme production forest in an area of timber harvest, state of Amazonas, Brazil. Sand fly catches were carried out in three areas: one before and after the timber harvest, and two control areas, a nature preservation area and a previously exploited area. The flies were caught by aspiration on tree trunks. Samples of sand flies were dissected for parasitological examination. In the site that suffered a harvest, a larger number of individuals was caught before the selective extraction of timber, showing significant difference in relation to the number of individuals and their flagellate infection rates after the logging. The other two areas did not show differences among their sand fly populations. This fact is suggestive of a fauna sensitive to the environmental alterations associated with selective logging.

Isolation and molecular identification of Leishmania ( Viannia ) peruviana from naturally infected Lutzomyia peruensis (Diptera: Psychodidae) in the Peruvian Andes

Leishmania (Viannia) peruviana was isolated from 1/75 Lutzomyia peruensis captured during May 2006 in an endemic cutaneous leishmaniasis region of the Peruvian Andes (Chaute, Huarochiri, Lima, Peru). Sand fly gut with promastigotes was inoculated into a hamster and the remaining body was fixed in ethanol. L. (Viannia) sp. was determined by polymerase chain reaction (PCR), and Leishmania species through molecular genotyping by PCR-restriction fragment length polymorphism analyses targeting the genes cpb and hsp70, resulting L. (V.) peruviana. The infected sand fly appeared 15 days after the rains finished, time expected and useful real time data for interventions when transmission is occurring.

Incidence of myiasis in Panama during the eradication of Cochliomyia hominivorax (Coquerel 1858, Diptera: Calliphoridae) (2002-2005)

We present the results of a study on myiasis in Panama during the first years of a Cochliomyia hominivorax eradication program (1998-2005), with the aim of investigating the behavior of the flies that produce myiasis in animals and human beings. The hosts that registered positive for myiasis were cattle (46.4%), dogs (15.3%), humans (14.7%), birds (12%), pigs (6%), horses (4%), and sheep (1%). Six fly species caused myiasis: Dermatobia hominis (58%), Phaenicia spp. (20%), Cochliomyia macellaria (19%), Chrysomya rufifacies (0.4%), and maggots of unidentified species belonging to the Sarcophagidae (3%) and Muscidae (0.3%). With the Dubois index, was no evidence that the absence of C. hominivorax allowed an increase in the cases of facultative myiasis.

Phlebotomine sand flies (Diptera: Psychodidae) in the municipality of Várzea Grande: an area of transmission of visceral leishmaniasis in the state of Mato Grosso, Brazil

Visceral leishmaniasis (VL) has been naturally transmitted in periurban areas due to the emergence and reemergence of its vectors in such areas. Aimed to further knowledge on ecological aspects affecting the occurrence of phlebotomine sand flies in VL transmission areas in the municipality of Várzea Grande, state of Mato Grosso (MT), Brazil, sand fly captures were carried out. Monthly collections of sand flies were undertaken with CDC light-traps, which were left in both intradomiciliary and peridomiciliary areas of ten residences during four consecutive days between January 2004 and June 2006. Twenty-two species of genus Lutzomyia and one of Brumptomyia were captured. The most abundant species was Lutzomyia longipalpis (65.23%), followed by L. evandroi (16.26%), L. lenti (7.69%), L. whitmani (4.92%), L. sallesi (2.34%) and L. termitophila (1.32%). The highest density of the main VL vector, L. longipalpis, was found in peridomiciliary areas, mostly males. No significant correlation was found between environment (temperature, air relative humidity and rain fall) and phlebotomine density; although a slight increase in sand fly density has been observed in the period following rainfalls, particularly L. longipalpis. No correlation was observed between distribution and density of L. longipalpis, prevalence of human VL cases and the presence of serologically positive dogs. The presence of infected dogs, increased vector density, susceptibility rate and interruption of epidemiological surveillance may raise the risk of VL transmission to man in Várzea Grande.

Diptera Brachycera found inside the esophagus of a mummified adult male from the early XIX century, Lisbon, Portugal

Fly puparia and adult fragments of diptera muscid were found inside the esophagus of a mummified body from the early XIX century, buried inside the crypt of the Sacrament Church (Lisbon, Portugal). The identification of the material revealed a monospecific colonization by Ophyra capensis (Wiedemann) (Diptera: Muscidae), a species known to invade corpses in the ammoniacal fermentation wave. This species can be found in corpses kept indoors, not available to the early waves of blowflies (Diptera: Calliphoridae). In the present case, the number of pupae and their developmental stage suggest that the female invaded the mummified corpse through the partially opened mouth and the oviposition took place directly inside the esophagus. This is the first case of O. capensis infesting internal organs of an intact corpse. The use of chemical products for the embalming process probably explains why external colonization did not occur.

Urban distribution of Phlebotominae in a cutaneous leishmaniasis focus, Argentina

Urbanization and vector domestication are currently proposed as factors that contributed to the recent increase of American cutaneous leishmaniasis (ACL). Is likely also urban transmission? Oran is the main city in the Argentinean hyper-endemic area of ACL, and human cases in urban residences are usually reported. In order to assess the spatial distribution of risk, phlebotomine traps were located in different environments of Oran. A total of 7,787 sand flies were captured: Lutzomyia neivai (98.1%), Lutzomyia migonei (1.2%), Lutzomyia cortelezzii (0.7%), and one Lutzomyia shannoni. During the season of transmission (April-May) a single sand fly was obtained in one out of five urban sites, while a trap in a peri-urban pigsty captured up to 2,985 Lu. neivai/night. Captures performed in the other season of vector activity (September-October) revealed that small-scale changes in the pigsty environment resulted in noticeable changes in the abundance of Lu. neivai. In addition, in a new neighbourhood, on the fringe of the city, 1,073 Lu. neivai/site were captured in the forested edge but one in the yard of the houses. Therefore, in this urban ACL focus the human-vector effective contact risk is still associated with peri-urban vegetation and ecotone modifications despite the urban residence of the cases.

Are light traps baited with kairomones effective in the capture of Lutzomyia longipalpis and Lutzomyia intermedia? An evaluation of synthetic human odor as an attractant for phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae)

Phlebotomine sand flies are often captured with human bait and/or light traps, either with or without an animal bait. More recently, synthetic attractants have been used as bait in traps to improve the capture of phlebotomine sand flies as well as other insects of medical and veterinary importance. The aim of the present study was to evaluate the effects of the kairomone 1-octen-3-ol (octenol) and the synthetic human odor BG-Mesh LureTM (BGML - lactic acid, caproic acid and ammonia) baited in modified CDC light traps on the capture of phlebotomine sand flies. The experiments followed the 5x5 Latin square design. Among the species caught, Lutzomyia intermedia apparently presented a dose-dependent response to octenol. The response obtained with the BGML, alone or in combination with octenol (5 mg/h), indicated some degree of attractiveness of these baits to different phlebotomine sand fly species. Octenol seems to be more attractive to L. intermedia than to Lutzomyia longipalpis, while the BGML presented a higher success in capturing L. longipalpis. When the components of the BGML were used separately, there was no increase in catching the female of L. intermedia. Apparently, there was no synergistic effect between the octenol and the BGML. In conclusion, the octenol and the BGML were demonstrated to be possible baits to attract some phlebotomine sand fly species.

Description of Micropygomyia brandaoi sp. n. (Diptera: Psychodidae: Phlebotominae), a fossil phlebotomine from the Dominican Republic

The description of Micropygomyia brandaoi, a new species of fossil phlebotomine sand fly, is based on one male specimen obtained from Dominican amber of the Miocene period (20 million years). In this new species, the fifth palpal segment is long, the coxite lacks a setal tuft and the style shows four well-developed spines. This set of characters allowed us to place the new species in the genus Micropygomyia Barretto.

Serine protease activities in Oxysarcodexia thornax (Walker) (Diptera: Sarcophagidae) first instar larva

We report for the first time the expression of multiple protease activities in the first instar larva (L1) of the flesh fly Oxysarcodexia thornax (Walker). Zymographic analysis of homogenates from freshly obtained L1 revealed a complex proteolytic profile ranging from 21.5 to 136 kDa. Although some activities were detected at pH 3.5 and 5.5, the optimum pH for most of the proteolytic activities was between pH 7.5 and 9.5. Seven of 10 proteases were completely inactivated by phenyl-methyl sulfonyl-fluoride, suggesting that main proteases expressed by L1 belong to serine proteases class. Complete inactivation of all enzymatic activities was obtained using N-p-Tosyl-L-phenylalanine chloromethyl ketone (100 µM), a specific inhibitor of chymotrypsin-like serine proteases.