The Extraordinary Case of a Woman with a 30-Year-Long Diffuse Leishmaniasis Cured with One Single Ampoule of Intranasal Pentavalent Antimoniate.

Infection with Leishmania amazonensis and L. mexicana may lead to diffuse cutaneous leishmaniasis. The cure is exceptional, especially for the strange case of this lady. Case report: The patient acquired the disease in childhood and remained with lesions for over 30 years, albeit several treatments. She worsened after a pregnancy, developing disseminated lesions. Miltefosine with amphotericin B and pentamidine resulted in remission. Lesions reappeared after one year, accompanied by intra-nasal infiltration of the disease. The nasal spraying of a single ampoule of pentavalent antimoniate resulted in the sustained disappearance of the nasal symptoms and all the cutaneous lesions. After over eight years, she remains disease-free, albeit under renal replacement therapy. The high nasal mucosal antimonial concentration may explain the long-lasting cure via new MHC class I epitope-specific CD8+ cell clones against L. amazonensis present in the nasal mucosa.

Scorpion envenoming in Morona Santiago, Amazonian Ecuador: Molecular phylogenetics confirms involvement of the Tityus obscurus group.

Scorpion envenoming by species in the genus Tityus is hereby reported from rural locations in the Amazonian province of Morona Santiago, southeastern Ecuador. Twenty envenoming cases (18 patients under 15 years of age) including one death (a 4-year-old male) were recorded at the Macas General Hospital, Morona Santiago, between January 2015 and December 2016 from the counties of Taisha (n=17), Huamboyo (n=1), Palora (n=1), and Logroño (n=1). An additional fatality from 2014 (a 3-year-old female from Nayantza, Taisha county) is also reported. Leukocytosis and low serum potassium levels were detected in most patients. We observed a significant negative correlation between leukocytosis and hypokalemia. Scorpions involved in three accidents from Macuma, Taisha County, were identified as genetically related to Tityus obscurus from the Brazilian Amazonian region based on comparison of mitochondrial DNA sequences encoding cytochrome oxidase subunit I. These cases, along with previously reported envenoming from northern Manabí, reinforce the notion that scorpionism is a health hazard for children in Ecuador and emphasizes the need to supply effective antivenoms against local species, which are not currently available. The genetic affinity of the Ecuadorian specimens with T. obscurus may underlay toxinological, clinical, and venom antigenic relationships among Amazonian scorpions that deserves further exploration for designing therapeutic strategies to treat scorpionism in the region.

Evaluation of agreement between tests for the diagnosis of leprosy / Avaliação de concordância entre testes para diagnóstico da hanseníase

ABSTRACT Introduction: Leprosy is a chronic infectious disease caused by the intracellular parasite Mycobacterium leprae. The diagnosis is essentially clinical, based on symptoms, skin exam, peripheral nerves and epidemiological history. Laboratory tests are carried out to complement the result of clinical diagnosis, or even serving as a confirmatory method. Objective: To investigate the positivity and agreement between skin smear, enzyme-linked immunosorbent assay (ELISA) with synthetic antigen ND-O-BSA, ML Flow test and polymerase chain reaction (PCR) for detection of Mycobacterium leprae in new cases of leprosy. Methods: We conducted a case series study assessing a convenience sample of 39 new cases of leprosy and a control group of 18 household contacts in Belém (PA) and in Igarapé-Açu (PA) from March 2014 to September 2015. Results: The agreement between ELISA, ML Flow and PCR tests combinations showed slight to absent reproducibility (Kappa ≤ 0.24). The results showed greater sensitivity in PCR assay, with higher positivity in multibacillary cases. The ELISA test showed low positivity, even in multibacillary cases, resulting in no reaction to paucibacillary cases and household contacts. Conclusion: The high sensitivity of PCR decreases the agreement with other tests.

RESUMO Introdução: A hanseníase é uma doença infecciosa crônica causada pelo parasita intracelular Mycobacterium leprae. O diagnóstico é essencialmente clínico, com base em sintomas, exame da pele, nervos periféricos e história epidemiológica. Testes laboratoriais são realizados para complementar o resultado de diagnóstico clínico, ou mesmo servindo como método de confirmação. Objetivo: Investigar a positividade e a concordância da baciloscopia, do ensaio de imunoadsorvente ligado à enzima (ELISA) com o antígeno sintético ND-O-BSA, do ML Flow e da reação em cadeia da polimerase (PCR) para a detecção de Mycobacterium leprae em casos novos de hanseníase. Métodos: Foi realizada uma série de casos, avaliando uma amostra de conveniência de 39 novos casos de hanseníase e um grupo-controle de 18 contatos domiciliares em Belém (PA) e Igarapé-Açu (PA) a partir de março 2014 a setembro de 2015. Resultados: A concordância entre as combinações ELISA, ML Flow e PCR mostrou reprodutibilidade leve a ausente (Kappa ≤ 0,24). Os resultados mostraram maior sensibilidade no ensaio de PCR, com maior positividade em casos multibacilares. O teste ELISA mostrou baixa positividade, mesmo em casos multibacilares, resultando em nenhuma reação nos casos paucibacilares e contatos domiciliares. Conclusão: A alta sensibilidade da PCR diminui a concordância com outros testes.

Bone marrow parasite burden among patients with New World kala-azar is associated with disease severity.

Kala-azar or visceral leishmaniasis, found mostly throughout the Indian Subcontinent, East Africa, and Brazil, kills 20,000-40,000 persons annually. The agents, Leishmania donovani and Leishmania infantum, are obligatory intracellular protozoa of mononuclear phagocytes found principally in the spleen and bone marrow. Protracted fever, anemia, wasting, hepatosplenomegaly, hemorrhages, and bacterial co-infections are typical features. One hundred and twenty-two (122) in-hospital patients were studied to verify if higher bone marrow parasite load estimated by quantitative polymerase chain reaction is associated with severe disease. The estimated median parasite load was 5.0 parasites/10(6) human nucleated cells. It is much higher in deceased than among survivors (median 75.0 versus 4.2). Patients who lost more weight had a higher parasite burden, as well as patients with epistaxis, abdominal pain, edema, and jaundice. This study suggests that higher parasite load is influenced by wasting, which may lead to more severe disease.

Adherence to Plasmodium vivax malaria treatment in the Brazilian Amazon Region.

BACKGROUND: Patients' adherence to malaria treatment is an important factor in determining the therapeutic response to anti-malarial drugs. It contributes to the patient's complete recovery and prevents the emergence of parasite resistance to anti-malarial drugs. In Brazil, the low compliance with malaria treatment probably explains the large number of Plasmodium vivax malaria relapses observed in the past years. The goal of this study was to estimate the proportion of patients adhering to the P. vivax malaria treatment with chloroquine + primaquine in the dosages recommended by the Brazilian Ministry of Health. METHODS: Patients who were being treated for P. vivax malaria with chloroquine plus primaquine were eligible for the study. On the seventh day of taking primaquine, they were visited at their home and were interviewed. The patients were classified as probably adherent, if they reported having taken all the medication as prescribed, in the correct period of time and dosage, and had no medication tablets remaining; probably non-adherent, if they reported not having taken the medication, in the correct period of time and dosage, and did not show any remaining tablets; and certainly non-adherent, if they showed any remaining medication tablets. RESULTS: 242 of the 280 patients reported having correctly followed the prescribed instructions and represented a treatment adherence frequency (CI95%) of 86.4% (81.7%-90.1%). Of the 38 patients who did not follow the recommendations, 27 (9.6%) were still taking the medication on the day of the interview and, therefore, still had primaquine tablets left in the blister pack. These patients were then classified as certainly non-adherent to treatment. Although 11 patients did not show any tablets left, they reported incorrect use of the prescribed therapy regimen and were considered as probably non-adherent to treatment. CONCLUSIONS: Compliance with the P. vivax malaria treatment is a characteristic of 242/280 patients in the surveyed region. However, the group of non-adherent patients can have an impact on the magnitude of transmission and relapses of P. vivax infections currently observed in the studied area. Simple practices can be introduced in the healthcare services in order to improve compliance with the treatment prescribed.

Species diversity of Leishmania (Viannia) parasites circulating in an endemic area for cutaneous leishmaniasis located in the Atlantic rainforest region of northeastern Brazil.

OBJECTIVES: To identify the aetiological agents of cutaneous leishmaniasis and to investigate the genetic polymorphism of Leishmania (Viannia) parasites circulating in an area with endemic cutaneous leishmaniasis (CL) in the Atlantic rainforest region of northeastern Brazil. METHODS: Leishmania spp. isolates came from three sources: (i) patients diagnosed clinically and parasitologically with CL based on primary lesions, secondary lesions, clinical recidiva, mucocutaneous leishmaniasis and scars; (ii) sentinel hamsters, sylvatic or synanthropic small rodents; and (iii) the sand fly species Lutzomyia whitmani. Isolates were characterised using monoclonal antibodies, multilocus enzyme electrophoresis (MLEE) and polymerase chain reaction-restriction fragment length polymorphism of the internal transcribed spacer region rDNA locus. RESULTS: Seventy-seven isolates were obtained and characterised. All isolates were identified as Leishmania (Viannia) braziliensis serodeme 1 based on reactivity to monoclonal antibodies. MLEE identified 10 zymodemes circulating in the study region. Most isolates were classified as zymodemes closely related to L. (V.) braziliensis, but five isolates were classified as Leishmania (Viannia) shawi. All but three of the identified zymodemes have so far been observed only in the study region. Enzootic transmission and multiclonal infection were observed. CONCLUSIONS: Our results confirm that transmission cycle complexity and the co-existence of two or more species in the same area can affect the level of genetic polymorphism in a natural Leishmania population. Although it is not possible to make inferences as to the modes of genetic exchange, one can speculate that some of the zymodemes specific to the region are hybrids of L. (V.) braziliensis and L. (V.) shawi.

Recent observations on the sand fly (Diptera: Psychodidae) fauna of the State of Rondônia, Western Amazônia, Brazil: the importance of Psychdopygus davisi as a vector of zoonotic cutaneous leishmaniasis.

Sand flies were collected in the central region of the state of Rondônia (W 64 degrees 30' to 63 degrees 00' and S 10 degrees 00'to 11 degrees 00') using Shannon and CDC light traps from October 1997 to August 2000. A total of 85,850 specimens representing 78 named species were captured. Of these 14 were new records for Rondônia. The proportion of males/females was 1/1.131. Trypanosomatids, that are presently being identified, were detected in 11 species. Leishmania (Viannia) naiffi was recorded from Psychodopygus davisi and P. hirsutus. In the present study the dominant species was P. davisi (39.6%) followed by Lutzomyia whitmani (13.1%), P. carrerai (11.6%), and P. hirsutus (10.2%). The importance of P. davisi as a vector of zoonotic cutaneous leishmaniasis is discussed.

Recent observations on the sand fly (Diptera: Psychodidae) fauna of the State of Rondônia, Western Amazônia, Brazil: the importance of Psychdopygus davisi as a vector of zoonotic cutaneous leishmaniasis

Sand flies were collected in the central region of the state of Rondônia (W 64 30' to 63 00' and S 10 00'to 11 00') using Shannon and CDC light traps from October 1997 to August 2000. A total of 85,850 specimens representing 78 named species were captured. Of these 14 were new records for Rondônia. The proportion of males/females was 1/1.131. Trypanosomatids, that are presently being identified, were detected in 11 species. Leishmania (Viannia) naiffi was recorded from Psychodopygus davisi and P. hirsutus. In the present study the dominant species was P. davisi (39.6 percent) followed by Lutzomyia whitmani (13.1 percent), P. carrerai (11.6 percent), and P. hirsutus (10.2 percent). The importance of P. davisi as a vector of zoonotic cutaneous leishmaniasis is discussed

Wild and synanthropic hosts of Leishmania (Viannia) braziliensis in the endemic cutaneous leishmaniasis locality of Amaraji, Pernambuco State, Brazil.

Evidence of Leishmania infection was found in small mammals captured between 1996 and 2000 in the Amaraji region, Pernambuco State, Brazil. The kDNA polymerase chain reaction (PCR), using primers specific for subgenus L. (Viannia), was positive for 43/153 water rats (Nectomys squamipes), 13/81 black rats (Rattus rattus), 15/103 grass mice (Bolomys lasiurus), 1/14 marsh mice (Holochilus scieurus), 2/50 field mice (Akodon arviculoides), 2/12 woolly opossums (Marmosa sp.), and 5/37 common opossums (Didelphis albiventris). This same kDNA PCR was positive for 12/61 dog and 8/58 horse skin samples. In paired PCR tests of 203 small mammals, 18.7% were positive with the kDNA primers and 18.2% with rDNA primers. Amastigotes were seen in 26/460 and L. (V.) braziliensis was isolated from 5 grass mice and 1 black rat. We concluded that small mammals, particularly rodents, are infected with parasites of the subgenus L. (Viannia). The isolation of L. (V.) braziliensis zymodeme IOC/Z74 from 6 rodents and the fact that all the other described L. (Viannia) species that commonly infect humans have never been found in rodents or marsupials leads us to suggest that the positive PCRs indicate infections of L. (V.) braziliensis. The isolation of zymodeme IOC/Z74 from humans reinforces our hypothesis that small, ground-loving mammals, such as rodents are the primary reservoirs of L. (V.) braziliensis.

Nota sobre leishmaniose canina no noroeste do Estado do Paraná, sul do Brasil / Leishmaniasis in dogs in southern Brazil

Em área endêmica de leishmaniose tegumentar americana no Município de Jussara, Estado do Paraná, Brasil, detectaram-se três cäes domésticos infectados por Leishmania (Viannia) brasiliensis

Leishmaniose cutânea experimental: II. Aspectos evolutivos da infecçäo no primata Cebus apella (Cebidae) pela Leishmania (V.) braziliensis e L. (L.) amazonensis / Experimental skin leishmaniasis: II. Course of the infection in the Cebus apella primate (Cebidae) caused by Leishmania (V.) braziliensis and L. (L.) amazonensis

Objetivando avaliar o potencial do primata C. apella como modelo experimental da leishmaniose cutânea produzida pela L. (V.) brasiliensis e L. (L.) amazonensis, inocularam-se, via intradérmica, 3 X10**6 de promastigotas dessas leishmanias, em 8 sítios da cauda de 10 espécimens desse desse primata, 5 deles com a L. (V.) braziliensis e outros 5 com a L. (L.) amazonensis. Posteriormente, às inoculaçöes, o exame semanal dos animais e biópsias mensais, revelaram os seguintes resultados relativos a cada parasita: a) L. (V.) brasiliensis: o periodo de incubaçäo foi d e15-20 dias; aos 30 dias evidenciaram-se lesöes pápulo-eritematosas, que evoluiram para nódulo ao fim de 60 dias; no 3§ mês, notou-se espontânea destas lesöes e, no 4§ mês, deu-se o inicio da reparaçäo das lesöes ulceradas, culminando com a cura em um dos animais após 5 meses, em dois após 6 meses, noutro após 7 meses e, no ultimo, após 10 meses. Quanto ao parasitismo nas lesöes, foi demonstrado nos 5 animais, até 90 dias; depois disto, somente em 2 até 120 dias e, por fim, até 180 dias apenas naquele que curou depois de 10 meses. b) L. (L.) amazonensis: o período de incubaçäo foi de 20 dias; aos 30 dias notou-se lesöes pápulo-eritematosas, que também evoluiram para nódulos ao fim de 60 dias, porém, a partir do 3§ mês, estas lesöes regrediram rapidamente ao fim de 90 dias, quando näo mais detectou-se o parasita na pele dos animais. Em relaçäo aos testes de Montenegro, somente 2 dos 5 animais infectados com a L. (V.) braziliensis reagiram ao teste, 6 e 90 dias após as inoculaçöes. Os resultados observados permitiram confirmar a infectividade do C. apella a estas leishmanias e, também, reforçar a indicaçäo desse primata como modelo experimental da leishmaniose cutânea por estes parasitas

Leishmaniose cutanea experimental: I-sobre a susceptibilidade do primata cebus apella (Cebidae) a infecçäo pela Leishmania (Viannia) Lainsoni Silveira, Shaw, Braga e Ishikawa, 1987 / Experimental cutaneous leishmaniasis: I-on the susceptibility of the primate cebus apella (Cebidae) to the infection caused by Leishmania (Viannia) Lainsoni Silveira, Shaw, Braga and Ishikawa, 1987

Foi investigada a susceptibilidade do primata Cebus apella (Cebidae) à infecçäo experimentada pela Leishmania (Viannia) lainsoni, com o objetivo de estudar a patogenia desse parasita, ainda pouco conhecido para o homem. Desse forma, cinco espécimes jovens daquele primata, 2 machos e 3 fêmeas, foram inoculados, itraderme, em oito sítios diferentes da regiäo dorsal da cauda com 3 x 10***6 de promastigotas do parasita (MHOM/BR/81/M6426, Benevides, Pará), obtidas de cultura da fase estacionária. Em seguida às inoculaçöes, a infecçäo experimental no animais foi comprovada, näo só pela presença de amastigotas do parasita na pele dos animais inoculados, mas, também, pela concomitância desse achado associado ao desenvolvimento de lesäo cutânea nos pontos da pele onde o parasita foi inoculado. Diante desses resultados, ficou demonstrada a susceptibilidade do primata Cebus apella à infecçäo experimental pela leishmania lainsoni, cujo período de infecçäo durou quese quatro meses, suficiente para testar drogas antileishmanióticas e estudar a patogênese da doença causada por este parasita

Sobre a sensibilidade da cultura de leucócitos circulantes na detecçäo de Leishmania no sangue periférico de pacientes com leishmaniose tegumentar / Sensitivity of the culture of circulating leucocytes in the detection of Leishmania in the peripheral blood of patients with tegumentary leishmaniasis

Foi investigada a presença de Leishmani, através da cultura de leucócitos circulantes, no sangue periférico de 60 pacientes portadores de leishmaniose tegumentar americana, nas suas diferentes formas clínicas, assim como nas principais fases evolutivas da doença. Biópsias de lesöes cutâneas e/ou de mucosa desses pacientes foram obtidas com a finalidade de isolar e caracterizar os parasitas, através da técnica de anticorpos monoclonais. Dos 60 pacientes examinados, foram isoladas 40 amostras de Leishmania das lesöes biopsiadas, sendo 5 de Leishmania (V.) brasiliensis, 3 de L. (V.) guyanensis, 1 de L. (V.) lainsoni, 13 de L. (L.) amazonensis e 18 näo puderam ser caracterizados a nível específico, porém, reagiram com anticorpos monoclonais do grupo brazilensis. Quanto à pesquisa através das culturas de leucócitos circulantes, esta revelou resultados completamente negativos. Com base nesses achados, os autores concluíram ser pouco consciente atribuir valor à cultura de leucócitos para o diagnóstico da leishmaniose tegumentar