@portal.ufgd.edu.br
Professor
Universidade Federal da Grande Dourados
Infectious Diseases, Parasitology, Microbiology
Scopus Publications
Eduardo José de Arruda, Glenda Biasotto, Marisa Masumi Beppu, Fernando Jorge Monteiro, Pedro L. Granja, Maria Rangel, Andreia Leite, Isaías Cabrini, Tiago Santos, Daniel A. Gonçalves,et al.
Elsevier BV
Carmem Cícera Maria da Silva, Carolina Rangel de Lima Santos, Eliomar Pivante Céleri, David Salles, Julia Miranda Fardin, Kamily Fagundes Pussi, Daniel Claudio de Oliveira Gomes, Vinicius de Oliveira Ribeiro, Leila Cristina Konrad-Moraes, Herintha Coeto Neitzke-Abreu,et al.
Ubiquity Press, Ltd.
Background: The World Health Organization declared the end of the COVID-19 pandemic in May 2023, three years after the adoption of global emergency measures. Monitoring of SARS-CoV-2 in sewage underscores its importance due to its effectiveness and cost-effectiveness, highlighting the need to prioritize research on water resources and sanitation. Objectives: The aim of this study was to conduct an epidemiological assessment of SARS-CoV-2 in the sewage system of a higher education institution located in Vitória Espírito Santo State, Maruípe campus. Methods: Over a period of 66 days, from February 6 to April 12, 2023, 15 samples were collected. Each sample consisted of 1 L, collected in 1 hour, with 250 mL collected every 15 minutes. The samples were characterized by assessing their appearance, and pH was measured using a Horiba U-50 multiparameter probe. The extracted RNA was subjected to RT-qPCR using the Allplex™ 2019-nCovAssay Seegene kit. Results: The samples exhibited a cloudy appearance with impurities, and the pH ranged from 6.35 to 8.17. Among the evaluated samples, SARS-CoV-2 RNA was detected in two, and, by comparing this with the epidemiological bulletin issued by the State Health Department, an increase in cases in the state was observed during the collection period of these samples. Conclusions: Sewage monitoring proved to be an important tool in this post-pandemic period, serving as an alert and prevention mechanism for the population in relation to new outbreaks. Furthermore, it represents a low-cost mapping strategy and extensive testing of a population, aligning with the studies presented at the beginning of the pandemic. We recommend specific adjustments considering distinct populations.
Carolina Rangel de Lima Santos, Kamily Fagundes Pussi, Iara Beatriz Andrade de Sousa, Manoel Sebastião da Costa Lima Junior, and Herintha Coeto Netitzke-Abreu
Universidade de São Paulo. Agência de Bibliotecas e Coleções Digitais
In February 2023, according to the World Health Organization, there had been more than 700 million cases of COVID-19 worldwide. Maternal immunization has become a hotspot in researchon the transplacental transfer of antibodies to the fetus. Vaccinations administered during pregnancy are intended to protect the mother from serious illness. Case presentation: In this report, we describe a case wherein the transfer of anti-SARS-CoV-2 antibodies to the fetus, baby, and infant was observed through immunization and breastfeeding. Maternal antibodies were detected in the umbilical cord at birth, in breast milk for up to 10 mo after birth, and in the child’s blood for up to 1 y of age, which suggests the importance of vaccination during pregnancy as well as maternal and neonatal protection. Conclusion: When an vaccine was administered to the pregnant patient, effective immunological results were obtained in terms of both placental transfer and breastfeeding, for up to 1 y of age.
Helton Krisman de Araújo, Silvana de Oliveira Castro, Maria Joelma Alves da Silva Valejo, Manoel Sebastião da Costa Lima Junior, and Herintha Coeto Neitzke-Abreu
American Society of Parasitologists
Conventional PCR provides Leishmania species characterization with even a small amount of biological material. Species-specific primers have been a widely used alternative; however, nonspecific amplifications are a reality, interfering with PCR efficiency. In endemic areas with multiple etiological agents for leishmaniasis, there is a requirement for higher specificity of primers. This study evaluates 3 pairs of primers described for the identification and characterization of Leishmania infantum. Primers RV1/RV2, LEISH1/LEISH2, and FLC2/RLC2 were used with the DNA of L. infantum, Leishmania amazonensis, and Leishmania braziliensis. An initial temperature curve was performed (52-62 C) to determine the optimal annealing temperature, followed by a dilution curve of Leishmania DNA (500 pg/μl, 50 pg/μl, 5 pg/μl, 500 fg/μl, 50 fg/μl, 5 fg/μl, and 0.5 fg/μl) to be used for analytical sensitivity. RV1/RV2 PCR amplified L. infantum and L. amazonensis at all analyzed temperatures; LEISH1/LEISH2 PCR amplified all 3 species of Leishmania, although at some temperatures L. infantum was specifically amplified, and, finally, FLC2/RLC2 PCR amplified only L. infantum at all temperatures analyzed. In terms of sensitivity, RV1/RV2 PCR detected 1 fg of L. infantum DNA and 100 pg of L. amazonensis DNA; LEISH1/LEISH2 PCR detected 1 fg of L. infantum DNA, 100 fg of L. amazonensis DNA, and 10 fg of L. braziliensis DNA; and FLC2/RLC2 PCR detected 10 fg of L. infantum DNA. Thus, PCR with FLC2/RLC2 primers is best suited for the molecular characterization of L. infantum, especially in areas where there is an incidence of more than 1 Leishmania species, such as South America.
Milena Menezes Corrêa Pederiva, Sidney Mariano dos Santos, Liz Graciela Sanabria Rivarola, Victor Jorge Guerreiro, Katiana Simões Lopes, Manoel Sebastião da Costa Lima Junior, and Herintha Coeto Neitzke-Abreu
Elsevier BV
Herintha Coeto Neitzke-Abreu, Georgia Medeiros de Castro Andrade, Paulo Silva de Almeida, Gilmar Cipriano Ribeiro, Thaís Alves Ribeiro, D'Angela Maciel Barrios, Kamily Fagundes Pussi, José Dilermando Andrade Filho, Felipe Dutra-Rêgo, and Fredy Galvis Ovallos
FapUNIFESP (SciELO)
ABSTRACT Background: Here, Leishmania presence in sand flies from Três Lagoas, Mato Grosso do Sul, Brazil, after visceral leishmaniasis (VL) was investigated. Methods: In April 2022, two light traps were deployed within and around the residence for two days post-VL case report. Results: A total of 120 Lutzomyia longipalpis were collected. Suprapyloric flagellates were found in a female sand fly with eggs and residual blood during midgut dissection. Sequencing of ITS1 and cytb fragments confirmed Leishmania infantum DNA and identified Homo sapiens as the blood source, respectively. Conclusions: This study emphasizes the importance of monitoring sand flies in VL endemic areas.
Herintha Coeto Neitzke-Abreu, Gabriel Barbosa Costa, Milena Nunes da Silva, Edith Palacio, Alexandre da Silva Cardoso, Paulo Silva de Almeida, and Manoel Sebastião da Costa Lima-Junior
Springer Science and Business Media LLC
Abstract Background In the State of Mato Grosso do Sul, Brazil, sand flies and cases of visceral (VL) and cutaneous (CL) leishmaniases have been reported in almost all municipalities. The aim of this study was to analyze the geographic distribution of VL and CL in relation the sand fly species found in the municipalities of Mato Grosso do Sul. Methods We analyzed VL and CL cases from 2001 to 2018 using data from the Notifiable Diseases Information System (SINAN). Data collected since 2003 on the presence of sand fly vectors (proven or suspected) were provided by the State Health Secretariat. Results A total of 3566 and 3030 cases of VL and CL, respectively, were reported from 2001 to 2018. The municipalities with the most reported cases of VL were Campo Grande (2495), Três Lagoas (442), Corumbá (140) and Aquidauana (136); and those for CL were Campo Grande (635) and Bodoquena (197). The following sand fly species with vector potential were found in 59 municipalities (74.7%): Lutzomyia longipalpis, Lutzomyia cruzi, Nyssomyia whitmani, Migonemyia migonei, Nyssomyia neivai, Pintomyia pessoai, Bichromomyia flaviscutellata and Pintomyia fischeri. Sand flies were present in six municipalities where no cases of VL were reported and in two municipalities where no cases of CL were reported. Conclusions Our results indicate that the geographical distribution of VL and CL in Mato Grosso do Sul expanded during the study period, and highlight the presence of sand fly vectors in municipalities where these diseases are currently considered to be non-endemic. Graphical Abstract
Sarfaraz Ahmad Ejazi, Samiran Saha, Anirban Bhattacharyya, Sonali Das, Nathália Lopes Fontoura Mateus, Manoel Sebastião da Costa Lima, Herintha Coeto Neitzke-Abreu, Ivete Lopes de Mendonca, Carlos Henrique Nery Costa, Otoni Alves de Oliveira Melo,et al.
Frontiers Media SA
Canine visceral leishmaniasis (CVL) due to Leishmania infantum infection is a zoonotic disease prevalent in the areas of South America and the Mediterranean. Infected dogs as reservoirs can contribute to disease transmission and can be a scourge to public health. Therefore, early diagnosis of infected dogs may play a pivotal role in circumscribing disease progression. Invasive tissue aspiration and insufficient serological methods impair a single assay for prompt CVL diagnosis. In the present study, we aimed to evaluate the potential of Leishmania donovani isolated membrane protein, LAg, for the diagnosis of CVL through immunological assays. Initially, enzyme-linked immunosorbent assay was done with Brazilian dog sera to evaluate the performance of LAg in diagnosing CVL and found sensitivity and specificity of 92.50% and 95%, respectively. The study further confirmed the diagnostic efficacy of LAg in a dipstick format. The dipstick test of canine sera from three centers in Brazil and one center in Italy collectively showed sensitivity values in the range of 53.33% to 100% in recognizing symptomatic dogs and specificity values between 75% and 100% to rule out healthy dogs. Moreover, a rapid immunochromatographic test was developed and optimized using LAg. This test was able to identify 94.73% of CVL of Brazilian origin with specificity of 97.29%. The current results highlight the reactive potential of the L. donovani antigen, LAg, for L. infantum CVL diagnosis and support our previous findings, which suggest the utility of LAg for the diagnosis of both L. donovani and L. infantum human VL in a variety of endemic regions. LAg as a diagnostic candidate may be employed to identify comprehensive CVL cases in epidemiological areas.
Manoel Sebastião da Costa Lima-Junior, Paulo Silva de Almeida, José Oliveira da Silva, Rosana Alexandre da Silva, Odival Faccenda, Dioelen Virgínia Borges Souza de Aquino Coelho, Gabriel Barbosa Costa, Agruslávia Rezende de Souza, Marcos Gino Fernandes, and Herintha Coeto Neitzke-Abreu
Oxford University Press (OUP)
AbstractStudies of the geographic distribution of sand flies and the factors associated with their occurrence are necessary to understand the risk of leishmaniasis transmission. The objective of this study was to characterize the sand fly fauna, particularly the spatial distribution of Lutzomyia longipalpis (Lutz & Neiva), and correlate these with climate factors in the Dourados municipality, Brazil. The collection of sand flies was carried out with CDC Light Traps over two periods: at six sites for three consecutive nights each month from August 2012 to July 2013; and at four other sites for two consecutive nights each month from April 2017 to February 2018. We collected 591 sand flies in the first period and 121 in the second period for a total of 712 sand flies; 697 of the total collected were Lu. longipalpis. The minimum and maximum sand fly infestation rate (sites with vector presence) was 11.1% and 83.33% in the first period, and 0% and 50.0% in the second period. No sand flies with Leishmania were identified via PCR. Lu. longipalpis presented an aggregate disposition with excellent adjustment. Rainfall and relative humidity were the abiotic factors that influenced the vector infestation level. The aggregate distribution for this species was predicted by the environmental factors that favor the proliferation of Lu. longipalpis. The results of this study should assist in devising measures to control sand flies in Dourados, Brazil.
Karen Araújo Magalhães, Kamily Fagundes Pussi, Hélton Krisman de Araújo, Silvia Barbosa do Carmo, Elisabete Friozi, Lidiane Schultz Branquinho, Manoel Sebastião da Costa Lima Junior, and Herintha Coeto Neitzke-Abreu
FapUNIFESP (SciELO)
Abstract The dog is the main domestic reservoir of Leishmania and font of infection for the vector, constituting an important host for the transmission of the parasite to humans. Non-invasive collection of swab samples for leishmaniasis diagnosis has been a promising alternative. This study analyzed the positivity of polymerase chain reaction (PCR) for the diagnosis of canine leishmaniasis in conjunctiva samples. DNA extraction was performed using SDS 20% and PCR was performed using 13A/13B primers that amplify 120-bp of Leishmania kDNA. Of the 77 dogs analyzed, 50 (64.93%) had ocular changes: 25 (32.47%) dogs had periocular lesion, 41 (53.25%) dogs had purulent eye discharge, and 17 (22.08%) dogs had both signals. PCR was positive in 35 dogs (45.45%), and there was no significant difference between dogs with and without ocular signals (p=0.4074). PCR positivity was significant higher in dogs without periocular injury (p=0.0018). Conjunctive PCR, a less invasive, fast, and painless collection technique, is indicated to complement the diagnosis, especially in dogs without periocular injury, independent of the presence of purulent eye discharge.
Paulo Silva de Almeida, Rosilene Francisca Moreira, José Oliveira da Silva, Talita Moreira Silva, Pedro Catarino da Costa Filho, Claucio Junior De Souza Carrijo, Herintha Neitzke-Abreu, Marcos Takashi Obara, Andreia Fernandes Brilhante, Marcia Bicudo de Paula,et al.
Pensoft Publishers
Aedes (Ochlerotatus) lepidus (Cerqueira & Paraense, 1945) is reported from a cave in Templo dos Pilares Municipal Park, Mato Grosso do Sul state, Brazil. A single exploratory collection of culicids was carried out using CDC light traps on 16 November 2015. Four larvae and 11 adults (1♂, 10♀) were collected. The larvae were foundin a water-filled hole in a stone. This is the first record of the species in Mato Grosso do Sul, which increases to 63 the number of culicids species recorded in the state.
Aurora Diotallevi, Gloria Buffi, Marcello Ceccarelli, Herintha Coeto Neitzke-Abreu, Laisa Vieira Gnutzmann, Manoel Sebastião da Costa Lima Junior, Alice Di Domenico, Mauro De Santi, Mauro Magnani, and Luca Galluzzi
Elsevier BV
Lucas Portela Silva, Silvia Montenegro, Roberto Werkauser, Kamila Gaudêncio da Silva Sales, Fábia Carla Silva Soares, Vlaudia Maria Assis Costa, Ana Cristina Bezerra, Maria Betania do Amaral Pinto, Suzany Maria Ferreira, Herintha Coeto Neitzke-Abreu,et al.
FapUNIFESP (SciELO)
ABSTRACT This study has estimated the risk of Leishmania transmission via blood transfusion in one of the largest blood banks in Northeastern Brazil, where visceral leishmaniasis is endemic. Five hundred blood samples from donors were tested for circulating Leishmania spp. DNA by real-time PCR. Positive samples were tested by a species-specific conventional PCR targeting Leishmania infantum . Overall, 6.2% (95% CI: 4.1–8.3%) of the samples carried Leishmania DNA and in one sample the species was confirmed as L. infantum . No statistically significant differences were found in relation to gender, sex, education level, incomeas well as the place of residence between positive and negative blood donors. Our results confirm the presence of asymptomatic Leishmania carriers among blood donors in a large blood bank in Northeastern Brazil. Considering the studied population, we estimate that for every 1,000 blood donors screened, 41 to 83 will be positive for Leishmania DNA. This finding reinforces the urgent need for elaborating specific Blood bank guidelines to allow the early detection of asymptomatic Leishmania carriers among blood donors before their blood products are transfused to uninfected individuals.
Herintha Coeto Neitzke-Abreu, Kárin Rosi Reinhold-Castro, Vanderson Carvalho Fenelon, Rubens Massafera, Jessé Truppel, Flávio Haragushiku Otomura, and Ueslei Teodoro
FapUNIFESP (SciELO)
The occurrence of cutaneous leishmaniasis (CL) in the municipalities of Jaboti, Japira, Pinhalão, and Tomazina in the North Pioneer of the State of Paraná, where this disease is endemic, prompted the investigation of sandfly fauna, their population changes throughout the year, and their behavior in and around human accommodation. In these municipalities, the collection of sandflies was conducted using Falcão traps from 7 pm to 6 am once a month. 32,994 sandflies, 18,442 males and 14,552 females, were collected and represented by the species Brumptomyia brumpti, Brumptomyia cunhai, Evandromyia correalimai, Evandromyia cortelezzii, Expapillata firmatoi, Micropygomyia ferreirana, Migonemyia migonei, Nyssomyia neivai, Nyssomyia whitmani, Pintomyia fischeri, Pintomyia monticola, Pintomyia pessoai, and Psathyromyia bigeniculata. Ny. neivai (SISA = 0.9808) predominated in Japira (46.9%), Pinhalão (53.1%), and Tomazina (38.9%), while Ny. whitmani (SISA = 0.9423) predominated in Jaboti (53.8%). Most sandflies were captured in domestic animal shelters, with a peak in September. The high number of sandflies collected in domestic animal shelters, species richness, and presence of species involved in the epidemiology of CL reveal the need for permanent entomological surveillance in the municipalities mentioned. The use of phlebotomine control measures in these municipalities should be considered in conjunction with the anthropogenic actions that have resulted in changes to sandfly fauna and behavior. A R T I C L E I N F O Article history: Received 30 July 2020 Accepted 04 October 2020 Available online 06 November 2020 Associate Editor: Diana Grisales Ochoa
Herintha Coeto Neitzke-Abreu, Victor Bruno Duarte Vieira, Ana Paula Stefanello da Silveira, Leticia Surian Batalini, Laísa Vieira Gnutzmann, Silvana de Oliveira Castro, Silvia Barbosa do Carmo, Elisabete Friozi, and Manoel Sebastião da Costa Lima-Junior
ResearchersLinks Ltd
| The objective of this study was to evaluate a polymerase chain reaction (PCR) for the diagnosis of Canine leishmaniasis (CanL) using less invasive sample collection. PCR-based detection of Leishmania infantum using ear peripheral blood collected from dogs with symptomatic CanL and coinfection with Ehrlichia sp. in the city of Campo Grande (Brazil) was investigated. A total of 268 CanL seropositive dogs were sampled. Of the 268 dogs tested, 70.90% were PCR positive for CanL, 72.43% of the dogs with ear lesion/s (185) were PCR positive, whereas only 67.47% of dogs without ear lesion/s (83) were PCR positive. Additionally, 35.82% of dogs tested were positive based on parasitological diagnosis for Ehrlichia sp. It was concluded that ear blood PCR may be an alternative for the diagnosis of CanL. Because it is more rapid collect and less invasive sample, this makes ear-blood PCR a potentially useful supplement to traditional diagnostic methods.
Aurora Diotallevi, Gloria Buffi, Marcello Ceccarelli, Herintha Coeto Neitzke-Abreu, Laisa Vieira Gnutzmann, Manoel Sebastião da Costa Lima, Alice Di Domenico, Mauro De Santi, Mauro Magnani, and Luca Galluzzi
Elsevier BV
Paulo Silva de Almeida, Marcia Bicudo de Paula, Andreia Fernandes Brilhante, Antônio Ralph Medeiros-Sousa, Herintha Coeto Neitzke-Abreu, Claucio Junior de Souza Carrijo, Pedro Catarino da Costa Filho, and Eunice Aparecida Bianchi Galati
Elsevier BV
Paulo Silva de Almeida, Talita Moreira Silva, Rosilene Francisca Moreira, Vanessa Fukuda Mariano, Miguel Prudencio Oliveira Neto, Dioelen Virginia Borges Souza de Aquino, Herintha Coeto Neitzke-Abreu, Gustavo Mayr de Lima Carvalho, and Jose Dilermando Andrade Filho
Universidade Federal de Goias
The purpose of this study was to search for Lutzomyia longipalpis and Lutzomyia cruzi in the urban area of the Água Clara Municipality, Mato Grosso do Sul State, Brazil. Two CDC traps were placed in the intradomicile and the peridomicile areas in four residences once a month, for three consecutive nights during 24 months. Were collected 11 species of sand flies totaling 3,418 specimens, 2,762 males and 656 females. Lu. longipalpis was the dominant species and there was no statistical difference between the peridomicile and intradomicile areas. Lu. cruzi was not found. The high capture rate for Lu. longipalpis in the urban area of Água Clara is cause for concern regarding necessary preventive measures to avoid future cases of visceral leishmaniasis.
 KEY WORDS: Leishmaniasis; Lutzomyia longipalpis; Phlebotominae; calazar.
Ana Paula Stefanello da Silveira, Victor Bruno Duarte Vieira, Leticia Surian Batalini, Silvia Barbosa do Carmo, Elisabete Friozi, Eduardo José de Arruda, Manoel Sebastião da Costa Lima Junior, and Herintha Coeto Neitzke-Abreu
FapUNIFESP (SciELO)
INTRODUCTION
Peripheral blood of 400 dogs infected with Leishmania and Ehrlichia were analyzed using polymerase chain reaction (PCR), and clinical signs were characterized.
METHODS
PCR and parasitological tests were conducted.
RESULTS
PCR was positive for Leishmania in 84.75%, and parasitological tests showed that 63.25% and 31.75% were positive for Leishmania and Ehrlichia, respectively. All animals showed more than three clinical signs. PCR results were negative for Leishmania in 15.25% of the samples.
CONCLUSIONS
Conventional PCR of peripheral blood can be used for diagnosing canine visceral leishmaniasis in combination with other techniques, especially in uncertain cases that need species identification.
Bárbara A. dos Santos, Kárin R. Reinhold-Castro, Carolina C. Conter, Herintha C. Neitzke-Abreu, Edilson C. Cristóvão, Thaís G.V. Silveira, and Ueslei Teodoro
Mary Ann Liebert Inc
Leishmaniases are classified as tegumentary leishmaniasis (TL) and visceral leishmaniasis (VL). Brazil is among the countries with the highest number of TL and VL cases. This study was undertaken to standardize the multiplex polymerase chain reaction (PCR) for the detection of the genus Leishmania in sandflies of endemic regions, on islands in the Upper Paraná River, northwestern Paraná. The sandflies were collected on 10 islands, from November 2012 to November 2014, with Falcão light traps, identified and conserved in tubes containing isopropanol, for subsequent DNA extraction. Two pairs of primers were used for multiplex PCR: A1/A2 and 5Llcac/3Llcac. Nyssomyia neivai was the predominant species of the collected specimens. A total of 3870 samples of female sandflies were analyzed and submitted to multiplex PCR, for the validation of the technique. All pools showed the 220 bp fragment for sandfly DNA detection, but no ∼120 bp fragment of Leishmania DNA was found. Although no natural infection of Ny. neivai by Leishmania was found in this study, the interaction of sandflies with Leishmania and its natural reservoirs continues in these Paraná River islands, despite the low diversity of the sandfly fauna. Some of these islands have permanent residents and are frequented by tourists.
Jaqueline de Carvalho Gasparotto, Magda Clara Vieira da Costa-Ribeiro, Vanete Thomaz-Soccol, Sandra Mara Rodrigues da Silva Liebel, Herintha Coeto Neitzke-Abreu, Kárin Rosi Reinhold-Castro, Edilson Colhera Cristovão, and Ueslei Teodoro
FapUNIFESP (SciELO)
ABSTRACT The genetic study of sandfly populations needs to be further explored given the importance of these insects for public health. Were sequenced the NDH4 mitochondrial gene from populations of Nyssomyia neivai from Doutor Camargo, Lobato, Japira, and Porto Rico, municipalities in the State of Paraná, Brazil, to understand the genetic structure and gene flow. Eighty specimens of Ny. Neivai were sequenced, 20 from each municipality, and 269 base pairs were obtained. A total of 27 haplotypes and 28 polymorphic sites were found, along with a haplotypic diversity of 0.80696 and a nucleotide diversity of 0.00567. Haplotype H5, with 33 specimens, was the most common among the four populations. Only haplotypes H5 and H7 were present in all four populations. The population from Doutor Camargo showed the highest genetic diversity, and only this population shared haplotypes with those from the other municipalities. The highest number of haplotypes was sheared with Lobato which also had the highest number of unique haplotypes. This probably occurred because of constant anthropic changes that happened in the environment during the first half of the twentieth century, mainly after 1998. There was no significant correlation between genetic and geographical distances regarding these populations. However, the highest genetic and geographical distances, and the lowest gene flow were observed between Japira and Porto Rico. Geographical distance is a possible barrier between these municipalities through the blocking of haplotype sharing.
Kárin Rosi Reinhold-Castro, Jaqueline de Carvalho Gasparotto, Herintha Coeto Neitzke-Abreu, and Ueslei Teodoro
Society for Vector Ecology
ABSTRACT:
We report the results of an investigation of natural larval sand fly habitats in the Recanto Marista, Doutor Camargo municipality, Paraná state, Brazil, from May, 2010 to August, 2012. We used Alencar emergence traps (AT), experimental traps (ET), and soil samples incubated in a biochemical oxygen demand incubator. Eight sand flies were collected with ATs. One specimen was collected with an ET and 21 were collected in soil samples. The collected species were Brumptomyia brumpti, Micropygomyia ferreirana, Migonemyia bursiformis, Migonemyia migonei, Nyssomyia neivai, Nyssomyia whitmani, and Pintomyia pessoai. The laval habitats of sand flies were located in the Recanto Marista, especially between tree roots, but the number of adults that emerged in the traps and soil samples was small despite the high density of sand flies that has been recorded in the Recanto Marista.