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Marina A Costa,1 Layane M Teodoro,1 Gustavo H Bahia-de-Oliveira,2 Ana Paula N Nunes,3 Ricardo A Barata1 1Laboratório de Parasitologia, Departamento de Ciências Biológicas, Universidade Federal dos Vales dos Jequitinhonha e Mucuri, Diamantina, MG, Brazil; 2Laboratório de Doenças Parasitárias, Departamento de Farmácia, Universidade Federal dos Vales dos Jequitinhonha e Mucuri, Diamantina, MG, Brazil; 3Laboratório de Bioestatística e Epidemiologia, Departamento de Ciências Básicas, Universidade Federal dos Vales dos Jequitinhonha e Mucuri, Diamantina, MG, Brazil Background: Banknotes are objects of great turnover and diffusion among the population, and can be efficient mechanisms in the dissemination of several intestinal parasites. This study investigated the presence of biological forms of intestinal parasites present in circulating cash banknotes in the city of Diamantina, Minas Gerais, Brazil, aiming to propose interventions aimed at improving local public health. Materials and methods: Between February and April 2017, samples of banknotes collected in five commercial establishments of the city were analyzed, namely: 1) gas station, 2) supermarket, 3) butchery, 4) pharmacy and 5) free fair. Ten circulating banknotes of each value (R$2.00, R$5.00, R$10.00, R$20.00, R$50.00 and R$100.00) were exchanged for new banknotes, washed with Tween 80 and, after spontaneous sedimentation, analyzed under an optical microscope with a magnification of 10× and 40×. Results: Cysts of Entamoeba coli (74.60%), cysts of Endolimax nana (16.94%), Giardia lamblia cysts (4.44%), unidentified nematode larvae (2.02%), Hymenolepis sp. (0.81%), Taenia sp. (0.81%) and Ancylostomidae larvae (0.40%) were found. Conclusions: The results revealed the need for improvements in basic sanitation, health and education activities that emphasized the importance of proper hand hygiene. Keywords: intestinal parasites, banknotes, Diamantina

Sabrina S Andrade,1 Layane M Teodoro,1 Daniel JS Viana,1 Egleise M Canuto-Sales,2 Gustavo H Bahia-de-Oliveira,2 Suedali Villas Bôas,3 Ricardo A Barata1 1Department of Biological Sciences, 2Department of Pharmacy, 3Department of Basic Sciences, Universidade Federal dos Vales do Jequitinhonha e Mucuri, Diamantina, Minas Gerais, Brazil Background: Intestinal parasites’ eggs, larvae, or cysts can be carried in public transport buses, and contribute to the increased incidence of diseases. This study aimed to detect biological forms of intestinal parasites in samples from public buses in the town of Diamantina, Minas Gerais, in order to know the local situation and propose interventions to improve public health. Materials and methods: In November 2014, six samples were obtained in buses of the two stations by using Graham method, in duplicate, by affixing a 6×5 cm clear tape, six times on each collection site of the bus, in an area of ~30 cm2. Then, each tape was positioned longitudinally on a slide microscope, and the identification of the biological forms of the parasites was performed with the aid of a 40× objective optical microscope. Results: A total of 216 slides were analyzed, of which 86 (39.8%) were positive for at least one intestinal parasite. Cysts of Entamoeba coli were the most frequently found in this study (52.1%), followed by Endolimax nana cysts (30.7%), Iodamoeba butschlii (6.5%), helminth larvae (4.7%), Giardia lamblia cysts (3.6%), Hymenolepis nana eggs (1.2%), Enterobius vermicularis eggs (0.6%), and Entamoeba histolytica cysts (0.6%). Top right handrails and right stanchions had the highest occurrence of biological forms, with 18.3% and 14.8%, respectively. Conclusion: The results indicated the need for better cleaning of the buses and better personal hygiene by users, since pathogenic and non-pathogenic intestinal parasites were found, suggesting fecal contamination of these sites, representing a risk to public health. Keywords: intestinal parasites, enteric parasite infection, public transport, Diamantina

The identification of parasite fauna can contribute to the assessment of the health status of sea turtles, and thus make it possible to develop effective management and treatment strategies aimed at conserving these animals. The present study aimed to investigate the parasite fauna in sea turtles rescued in six sections of the coast of Santa Catarina and Paraná (Brazil) from January 2020 to December 2022 through the recording of exam reports available in the Information System of Aquatic Biota Monitoring. In total, records of 334 sea turtles were analyzed during this period, of which 219 (65.6 %) were infected with at least one parasite. Chelonia mydas was the most frequent species with 86.5 % of rescues and presented the highest species richness. At least 43 species belonging to 36 different genera have been recorded parasitizing sea turtles. The most common helminths were Cricocephalus albus, Metacetabulum invaginatum, Pronocephalus obliquus, with 12.3 %, 8.7 % and 8.4 %, respectively, while protozoa of the genus Entamoeba were the most prevalent in the turtles analyzed (8.1 %). An interesting finding was the unprecedented finding of Hymenolepis sp. eggs in faeces of some turtle species. The results showed that the parasite fauna was quite diverse, with several species known to be capable of harming the physical health and well-being of sea turtles. In view of this, it is necessary to adopt strategies for monitoring the health of the different rescued species with a view to conserving sea turtles in Brazilian territory., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Porteirinha is endemic for visceral leishmaniasis (VL), with intense disease transmission of the disease. We evaluated the impact of canine euthanasia as a single control measure on the incidence of VL in humans and canines., Methods: A prospective observational cohort study was carried out over four years (1998-2002) in 8 of the 12 neighborhoods of the city. The dynamics of canine visceral leishmaniasis (CVL) transmission were evaluated for 2 years, before beginning the screening-culling intervention. The comparative morbidity index (CMI) was used to stratify areas with the greatest risk of CVL, and the spatial distribution of human and canine VL cases was compared using univariate and bivariate K-functions., Results: Human cases conglomerated in three neighborhoods. Spatial clusters were detected for CVL in 1998, 2000, and 2001, but not in 1999, when greater spatial dispersion occurred. The screening and culling intervention reduced the number of human VL cases and decreased the incidence of CVL, mainly in neighborhoods with a high CMI., Conclusions: The systematic euthanasia of seropositive dogs was shown to be an effective control action of the Program for Control of Visceral Leishmaniasis (PCLV) in Brazil. The fundamental role of domestic dogs in the epidemiological chain of VL was reaffirmed.

Background : The present study was carried out in the rural and urban area of Diamantina/Minas Gerais (MG), an endemic municipality for visceral leishmaniasis (VL) in Brazil. Methods : Patient notification records, canine prevalence, and phlebotomine fauna were evaluated. Results : In the period from 2016 to 2018, eight human cases were confirmed, with three deaths, predominantly in males. In the same period, a total of 1,388 dogs resided in the rural and urban area of the municipality were submitted to the DPP® and ELISA, with a percentage of confirmed canine cases of 29.9% and 29.4%, respectively. The entomological study conducted in the municipality revealed the presence of 10 species of sand flies, with a predominance of Lutzomyia longipalpis (55.75%), mainly in the rural area. Conclusions : Unlike what is happening in urban centers, the results of this study suggest that the VL in Diamantina is in the process of urbanization, given the high percentage of confirmed canine cases and the high density of Lu. longipalpis in the rural area of the municipality. These risk factors warn about the need for continuous surveillance and the need to control actions of VL in this area., (Copyright ©2021, Yale Journal of Biology and Medicine.)

This work shows a promising, environmentally friendly and greener alternative for the production and application of electrochemically produced Graphene Oxide (GO) for the adsorptive removal of Methylene Blue (MB) dye in an aqueous medium. During the adsorption tests, GO produced via electrochemical route reached the equilibrium in only 10 min of contact, exhibiting a percentage removal of MB over 97%. It could also be observed that the experimental data better fitted to the pseudo-second order kinetic model. By analysing the isotherms, it was verified the maximum adsorptive capacity was 500 mg g -1 (303.15 K) and that in overall, adsorptive capacity decreases with the increase in temperature. Experimental equilibrium data were better fitted to the Freundlich isotherms in all temperatures studied (303.15, 318.15 and 333.15 K). The thermodynamic analysis confirmed the exothermic nature of the process, and that MB adsorption onto GO occurs spontaneously. ΔH◦ and ΔG◦ values suggested that physisorption occurred, which is mainly due to π-π interactions and electrostatic interactions between MB and oxygen functional groups on the GO surface. Cost-effectiveness analysis showed there is a lower cost involved in the production of electrochemical GO, as compared to the Hummers method; and in the reusability study, even after 5 cycles GO removed ≥ 90% MB. Thus, the electrochemically produced GO seems to be an efficient, cost-effective and environmentally friendly alternative for colour removal from water, as it uses less hazardous and expensive reagents when compared to those applied in the traditional GO synthesis, without losing, however, the efficiency in colour removal from water.

Intestinal parasites are a major public health problem in developing countries, most prevalent in areas where sanitation is poor and the population's hygiene is inadequate. They affect people of all ages, although school-age children are the most susceptible. In this study, we investigated the presence of intestinal parasites in the shared environments of a public school in the town of Diamantina, Minas Gerais State. From December 2012 to February 2013, samples were collected for three months (once a month) by using the Graham method (1941), in duplicate, by affixing a 6 x 5 cm clear tape, six times in each collection site, in a space of about 30 cm2. Then, each tape was positioned longitudinally on a microscope slide and the identification of the biological forms of the parasites was performed with the aid of a 40X objective from an optical microscope. Eleven sites were selected for sampling. Cysts of Entamoeba coli were the most frequently found in this study (50%), followed by Hymenolepis diminuta eggs (27.6%), Iodamoeba butschllii cysts (5.6%), Ascaris lumbricoides eggs (5.6%), Taenia species eggs (5.6%) and hookworm eggs (5.6%). The highest positivity rates were found in the samples drawn from the cafeteria's eating table. The results have indicated the need to improve the cleaning in the school environments, as well as the development of educational practices that may help in the preservation of public health.

The aim of this study was to identify blood meals of female sandflies captured in the municipality of Governador Valadares, an endemic area of visceral and cutaneous leishmaniasis, in the State of Minas Gerais, Brazil. From May 2011 to January 2012, captures were performed using HP light traps in four districts. There were 2,614 specimens (2,090 males and 524 females) captured; 97 engorged females were identified belonging to the species Lutzomyia longipalpis(82.1%) and Lutzomyia cortelezzii(17.9%). Considering simple and mixed feeding, the enzyme-linked immunosorbent assay revealed a predominance of chicken blood (43.6%) in Lutzomyia longipalpis, showing the important role that chickens exert around the residential areas of Governador Valadares. This finding increases the chances of sandflies contact with other vertebrates and consequently the risk of leishmaniasis transmission.

Background: Sand fly collections were performed to study ecological aspects of the Phlebotominae fauna of the Xakriabá Indigenous Reserve, an area with endemic cutaneous leishmaniasis, located in the state of Minas Gerais, Brazil., Methods: The collections were performed in peridomicile areas and along trails previously selected for the study of wild and synanthropic Leishmania hosts. Differences in the distribution patterns of the sand fly species as well as in species richness and abundance between the different ecotopes were investigated during both rainy and dry seasons over the course of the study period., Results: A total of 8,046 sand flies belonging to 11 genera and 28 species were collected. Lutzomyia longipalpis and Nyssomyia intermedia were the most abundant species in peridomicile areas, whereas Martinsmyia minasensis and Lutzomyia cavernicola were the most abundant species among the different trail ecotopes., Conclusion: The different composition of the sand fly fauna observed in the peridomicile areas and in the trails during the study, reinforces the importance of sampled different areas in a phlebotomine fauna survey. The presence of Lutzomyia longipalpis and Ny. Intermedia most abundant in peridomicile can be important to Leishmania infantum and Leishmania braziliensis transmission in the Imbaúbas native village.

In the present study we surveyed the fauna of phlebotomine sand flies and small mammals in peridomestic areas from a Brazilian municipality where the American cutaneous leishmaniasis (ACL) is endemic. A total of 608 female phlebotomine sand flies were captured during nine months in 2009 and 2010. Seven different species were represented with 60% of them being Lutzomyia intermedia and Lu. whitmani, both incriminated vectors of ACL. Lu. longipalpis, a proven vector of visceral leishmaniasis (VL) was also captured at high proportion (12.8%). Genomic DNA analysis of 136 species-specific pools of female sand flies followed by molecular genotyping showed the presence of Leishmania infantum DNA in two pools of Lu. longipalpis. The same Leishmania species was found in one blood sample from Rattus norvegicus among 119 blood and tissue samples analysed. This is the first report of Le. infantum in R. norvegicus in the Americas and suggests a possible role for this rodent species in the zoonotic cycle of VL. Our study coincided with the reemergence of VL in Governador Valadares.

This study was developed in the urban area of Governador Valadares, a reemerging focus of intense transmission of visceral leishmaniasis (VL) in Brazil, presenting 86 human cases of VL from 2008 to 2011. The disease prevailed in males (73.2%) with most patients between 0 and 9 years (44.1%) and a lethality rate of 16.2%. A canine survey was carried out on 16,529 domestic dogs in 35 districts in the area and it showed that 30.2% of them (4,992 dogs) were positive for VL by serum assays. Prevalence ratios for canine VL varied between 13.6% and 53.4%. The clinical exam of 343 seropositive dogs showed that 49.9% of them were considered symptomatic, with larger prevalence of canine VL being in short-furred animals (90%). The entomological survey was performed in eight districts, where 2,539 phlebotomines were captured, preferentially in the peridomicile (84.5%). Lutzomyia longipalpis was the predominant species (90%) suggesting its participation in the VL transmission in the area. The correlation between canine prevalence and L. longipalpis density was evaluated.

A new species of Brazilian phlebotomine sandfly found in Brazil, municipality of Diamantina, state of Minas Gerais, is described based on males and females collected in a quartzite cave. The body of spermathecae is continuous to the individual duct, lanky and tapering at the end, with conical shaped, not striated and presenting the head with dense setae. The male presents gonostyle with four spines and a small subterminal seta and gonocoxite with one group of persistent setae. The paramere is simple with a group of small setae on the dorsal apex. The morphological features of this new species permit its inclusion in the migonei group.

This study investigated the sandfly fauna of two quartzite caves in the Espinhaço Mountain Range, located in the municipality of Diamantina, state Minas Gerais. From August 2010-July 2011, entomological sampling was performed in the caves of Salitre and Monte Cristo with two HP light traps exposed in the photic and aphotic zones of each cave. The sandfly fauna consisted of 17 species, among which Lutzomyia cipoensis was predominant (54.76%). The male/female ratio in the total captures was 1:2.5. The aphotic zone showed the highest frequency of specimens captured (65%). A greater density of sandflies occurred during the summer (January-February), coinciding with the period of higher temperatures, humidity and rainfall. The presence of Lutzomyia longipalpis, Lutzomyia whitmani and Lutzomyia pessoai, proven or suspected vectors of leishmaniasis, is of concern because the area is visited by many tourists.

The environmental changes resulting from the construction of hydroelectric dams may affect the fauna of insect vectors and consequently the epidemiology of the diseases they transmit. This work examined the mosquito and sand fly fauna in the area of the Aimorés hydroelectric power plant, analyzing the seasonal distribution and the degree of species synanthropy in different ecotopes. Between November, 2008 and September, 2009, entomological captures were performed with the help of HP light traps in the rural, urban, and forest areas of Aimorés, Ituêta, Resplendor, and Baixo Guandu counties. The fauna proved to be quite diversified. Twenty-two species of mosquitoes and 11 species of sand flies were found. Culex quinquefasciatus was predominant among mosquitoes (76.7%), while Lutzomyia intermedia prevailed among sand flies (34.5%). Some of the captured species have medical interest. Supported by the high degree of synanthropy, those species reinforce the need for epidemiological surveillance., (© 2012 The Society for Vector Ecology.)

Montes Claros in Minas Gerais State, Brazil, was considered an intense transmission area for visceral leishmaniasis. This study evaluated sand fly fauna after insecticide application. Captures were performed in 10 districts from September 2005 to August 2006 with CDC light traps inside and outside each residence. Cypermethrin was sprayed in two cycles during November/2005 and May/2006. The 636 specimens collected, belonging to 10 species, were predominantly Lutzomyia longipalpis (79%), and most frequently males (70%). The highest percentage of specimens were captured in areas surrounding domiciles (85.8%). The main species were observed to be sensitive to treatment with the insecticide. The results showed a reduction in the number of sand flies collected after use of cypermethrin in homes and annexes, and with residual effect lasting from two to four months.

Introduction: A study on the phlebotomine sandfly fauna was carried out in an endemic area for American tegumentary leishmaniasis in the municipality of Governador Valadares, in the State of Minas Gerais, Brazil., Methods: Captures were undertaken using HP light traps in four districts, on three nights per month, for one year (from January to December 2008). Correlations between climatic factors (temperature, relative air humidity and rainfall) and the numbers of sandflies collected was observed., Results: 5,413 phlebotomine specimens were caught and were identified as belonging to 12 species. Of these specimens, 2,851 (52%) were females and 2,562 (48%) were males., Conclusions: Lutzomyia intermedia predominated (29.9% of the species caught), thus suggesting that they were responsible for transmission of American tegumentary leishmaniasis, together with L. whitmani, which was also found in the area (4.3%). The presence of L. longipalpis (11.9%), the main vector for visceral leishmaniasis in Brazil, is an important finding, which makes rigorous entomological surveillance of the area necessary.

The vectorial capacity of Rhipicephalus sanguineus in the transmission of canine visceral leishmaniasis has been evaluated through a laboratory-controlled experiment. One healthy Leishmania-free dog and two dogs naturally infected with Leishmania were infested with R. sanguineus in various stages of development. Engorged larvae, unfed nymphs, engorged nymphs, unfed adults, engorged female adults and fed male adults were collected from the experimental animals and examined for Leishmania infection by optical microscopy, polymerase chain reaction (PCR) and parasite culture. Leishmania forms were not detected in any of the 433 smears prepared from engorged colonies nor in any of the 118 smears prepared from unfed colonies. However, one flagellate structure was identified in one of the smears. All pools of R. sanguineus that had fed on the infected dogs tested PCR-positive for Leishmania DNA, with the single exception of the pool of engorged larvae. In contrast, all pools of ticks that had fed on the Leishmania-free dog were PCR-negative. Leishmania growth was not observed in any of the tick colonies following incubation on culture medium. Considering that no Leishmania forms were identified in any of the meticulously analysed smears derived from engorged colonies of R. sanguineus, it appears somewhat unlikely that the maintenance and multiplication of Leishmania occurs within the tick.

In the present paper, we evaluate the relationship between climate variables and population density of Lutzomyia longipalpis in Montes Claros, an area of active transmission of American visceral leishmaniasis (AVL) in Brazil. Entomological captures were performed in 10 selected districts of the city, between September 2002-August 2003. A total of 773 specimens of L. longipalpiswere captured in the period and the population density could be associated with local climate variables (cumulative rainfall, average temperature and relative humidity) through a mathematical linear model with a determination coefficient (Rsqr) of 0.752. Although based on an oversimplified statistical analysis, as far as the vector is concerned, this approach showed to be potentially useful as a starting point to guide control measures for AVL in Montes Claros.

In Brazil, visceral leishmaniasis (VL) is caused by Leishmania chagasi parasites that are transmitted to man through the bites of infected females of Lutzomyia longipalpis sand flies. In order to evaluate transmission risk and to clarify the epidemiology of this tropical disease, studies focused on the vector and favorable environmental conditions are of fundamental importance. In this work, we surveyed the phlebotomine sand fly fauna in Janaúba, a Brazilian municipality that is endemic for VL. During a two-year period, entomological captures were performed monthly in 15 districts with high, moderate and low profiles of VL transmission. A total of 14,591 phlebotomine sand flies were captured (92% L. longipalpis), with a predominance of males. Most specimens were captured in the peri-domicile setting, although the number of specimens captured in the intra-domicile setting emphasises the anthropophilic behaviour of this insect. The population density of L. longipalpis was modulated by climate variations, particularly with clear increases immediately after the rainy season. However, the pattern of distribution did not coincide with the occurrence of human or canine cases of VL. This suggests that the eco-epidemiology of VL is particular to each area of transmission and must be taken into account during the design of public health control actions.

Visceral leishmaniasis (VL) is a growing zoonosis with an increasing number of new cases and a rapid geographical spreading of the disease. In the present study, a canine survey was carried out in the city of Montes Claros (320,000 inhabitants), an endemic area of American visceral leishmaniasis in the state of Minas Gerais, Brazil. A total number of 4795 dogs were examined by serology, which showed a rate of seropositivity of 5%. Isoenzymatic analysis confirmed Leishmania infantum chagasi as the local aetiological agent of CVL. Canine tissues were assayed for the presence of Leishmania parasite DNA using different techniques. The infectivity of asymptomatic, oligosymptomatic and symptomatic seropositive dogs was tested by xenodiagnosis using laboratory reared Lutzomyia longipalpis. Rates of infection of 5.4%, 5.1% and 28.4% were found for the phlebotomine sand flies that fed in asymptomatic, oligosymptomatic and symptomatic dogs, respectively. Our results indicate that, under experimental conditions, symptomatic dogs are about four times more infective to VL vectors than oligosymptomatic or asymptomatic animals. The lower infectivity rates of dogs displaying any of the last two clinical forms of leishmaniasis, however, must be taken into account in the epidemiology of CVL.

A study of Lutzomyia longipalpis (Lutz and Neiva, 1912) (Diptera: Psychodidae), the primary vector of American visceral leishmaniasis (AVL), and the canine form of the disease, was carried out in Porteirinha. The city is situated in the northern part of the Brazilian State of Minas Gerais and is an endemic area of AVL. Systematic phlebotomine captures were performed in seven districts with previously reported cases of canine visceral leishmaniasis, during 2 years (January 2000--December 2001). A total of 2328 specimens of L. longipalpis were captured. The association between the local climate variables and the population density of L. longipalpis was evaluated and rainfall was determined to be a major factor, with increased populations during the rainy season (October--March). At the same time period, blood samples from every dog domiciled in the same seven districts, in total 14,077 animals, were analyzed for infection by viscerotropic Leishmania using indirect immunofluorescence assay (IFA). Accumulated incidence rates of canine VL per district varied from 3.40 to 14.34 for the 2-year period. A positive correlation between the population density of L. longipalpis and the canine cases of visceral leishmaniasis in Porteirinha was observed.

A study on the phlebotomine sand fly fauna in Belo Horizonte city, state of Minas Gerais, Brazil, was carried out. From April 2001 to March 2003, monthly systematic collections were performed in three houses from each of the nine regions of the city, using CDC light traps for four consecutive days. The traps were set into the houses and in peridomestic areas totaling 54 traps. A number of 3871 sand fly specimens of the genera Lutzomyia and Brumptomyia were collected. Sixty eight percent of the specimens were L. longipalpis and 16% L. whitmani, insect vectors of visceral and American cutaneous leishmaniasis, respectively. Environmental factors such as temperature, humidity, and frequency of precipitation suggest that the number of insects increases after rainy periods. During the same period mentioned above, seasonal captures were carried out in parks and green areas of Belo Horizonte, using Shannon trap. A total of 579 phlebotomine sand flies were collected from which 398 (68.7%) were females with the predominance of L. whitmani and L. monticola. Those specimens were used for natural infection examination, by polymerase chain reaction. No Leishmania DNA was present in any of the specimens tested.

A study of the phlebotomine sand fly fauna was carried out in an endemic area of American visceral leishmaniasis (AVL) in the municipality of Porteirinha, in the Brazilian state of Minas Gerais. Captures were performed with CDC light traps in 7 districts, 5 days per month, during 2 consecutive years (January 2000 to December 2001). A total of 3240 sand flies were captured and identified. Sixteen species were found, among which 15 belonged to the genus Lutzomyia and one to the genus Brumptomyia. Lutzomyia longipalpis, a proven vector of AVL, was the predominant species (71.85%) throughout the time period. The interference of climatic factors (temperature, humidity, and rainfall) over the populational dynamics of the sand flies was determined. Statistical analysis of the data showed a significant correlation among the number of phlebotomine sand flies collected, rainfall, and humidity, whereas the effect of temperature was negligible, in that particular region. The amount of collected phlebotomine, the number of human cases, and the prevalence of canine AVL in the districts of Porteirinha are discussed.

An alkaline serineprotease, capable of hydrolyzing Nalpha-benzoyl- dl arginine p-nitroanilide, was secreted by Fusarium oxysporum var. lini grown in the presence of gelatin as the sole nitrogen and carbon source. The protease was purified 65-fold to electrophoretic homogenity from the culture supernatant in a three-step procedure comprising QSepharose chromatography, affinity chromatography, and FPLC on a MonoQ column. SDS-PAGE analysis of the purified protein indicated an estimated molecular mass of 41 kDa. The protease had optimum activity at a reaction temperature of 45 degrees C and showed a rapid decrease of activity at 48 degrees C. The optimum pH was around 8.0. Characterization of the protease showed that Ca2+ and Mg2+ cations increased the activity, which was not inhibited by EDTA or 1,10-phenanthroline. The enzyme activity on Nalpha-benzoyl-DL arginine p-nitroanilide was inhibited by 4-(2-aminoethyl)-benzenesulfonyl fluoride hydrochloride, p-aminobenzamidine dihydrochloride, aprotinin, 3-4 dichloroisocoumarin, and N-tosyl-L-lysine chloromethyl ketone. The enzyme is also inhibited by substrate concentrations higher than 2.5 x 10(-4)M. The protease had a Michaelis-Menten constant of 0.16 mM and a V(max) of 0.60 mumol released product.min(-1).mg(-1) enzyme when assayed in a non-inhibiting substrate concentration. The activity on Nalpha-benzoyl- dl arginine p-nitroanilide was competitively inhibited by p-aminobenzamidine dihydrochoride. A K(i) value of 0.04 mM was obtained.

A morphometric survey examined adult specimens of Lutzomyia whitmani (Antunes & Coutinho) captured at 5 municipalities in southeastern and northeastern Brazil to compare the populations. The localities were Ilhéus (Bahia), Martinho Campos (Minas Gerais), Corte de Pedra (Bahia), Baturité (Ceará), and Amaraji (Pernambuco): all are known foci of American cutaneous leishmaniasis. Fifteen males and 15 females from each population were analyzed morphometrically for 42 and 37 characters, respectively. Statistical data alone were insufficient to discriminate among the 5 populations. Further analysis generated phenograms that indicated there were 2 spatial clusters: the 1st was composed of specimens from Ilhéus (Bahia) and Baturité (Ceará) and the 2nd of specimens from Martinho Campos (Minas Gerais), Corte de Pedra (Bahia), and Amaraji (Pernambuco). Although insufficient to define the taxonomic status of the populations studied, the results delineated the existence of biogeographical structuring within L. whitmani. Complementary studies on the susceptibility to Leishmania braziliensis infection in the 5 populations are in progress to clarify the relationship between the 2 biogeographical clusters and American cutaneous leishmaniasis transmission in those Brazilian regions.