Characterisation of an area of coexistent visceral and cutaneous leishmaniasis transmission in the State of Piauí, Brazil.

BACKGROUND: In Brazil, transmission of visceral and cutaneous leishmaniasis has expanded geographically over the last decades, with both clinical forms occurring simultaneously in the same area. OBJECTIVES: This study characterised the clinical, spatial, and temporal distribution, and performed entomological surveillance and natural infection analysis of a leishmaniasis-endemic area. METHODS: In order to characterise the risk of leishmaniasis transmission in Altos, Piauí, we described the clinical and socio-demographic variables and the spatial and temporal distribution of cases of American visceral leishmaniasis (AVL) and American cutaneous leishmaniasis (ACL) cases and identified potential phlebotomine vectors. FINDINGS: The urban area concentrated almost 54% of ACL and 86.8% of AVL cases. The temporal and spatial distribution of AVL and ACL cases in Altos show a reduction in the number of risk areas, but the presence of permanent disease transmission foci is observed especially in the urban area. 3,808 phlebotomine specimens were captured, with Lutzomyia longipalpis as the most frequent species (98.45%). Of the 35 females assessed for natural infection, one specimen of Lu. longipalpis tested positive for the presence of Leishmania infantum and Leishmania braziliensis DNA. MAIN CONCLUSION: Our results indicate the presence of risk areas for ACL and AVL in the municipality of Altos and highlight the importance of entomological surveillance to further understand a possible role of Lu. longipalpis in ACL transmission.

Characterisation of an area of coexistent visceral and cutaneous leishmaniasis transmission in the State of Piauí, Brazil

BACKGROUND In Brazil, transmission of visceral and cutaneous leishmaniasis has expanded geographically over the last decades, with both clinical forms occurring simultaneously in the same area. OBJECTIVES This study characterised the clinical, spatial, and temporal distribution, and performed entomological surveillance and natural infection analysis of a leishmaniasis-endemic area. METHODS In order to characterise the risk of leishmaniasis transmission in Altos, Piauí, we described the clinical and socio-demographic variables and the spatial and temporal distribution of cases of American visceral leishmaniasis (AVL) and American cutaneous leishmaniasis (ACL) cases and identified potential phlebotomine vectors. FINDINGS The urban area concentrated almost 54% of ACL and 86.8% of AVL cases. The temporal and spatial distribution of AVL and ACL cases in Altos show a reduction in the number of risk areas, but the presence of permanent disease transmission foci is observed especially in the urban area. 3,808 phlebotomine specimens were captured, with Lutzomyia longipalpis as the most frequent species (98.45%). Of the 35 females assessed for natural infection, one specimen of Lu. longipalpis tested positive for the presence of Leishmania infantum and Leishmania braziliensis DNA. MAIN CONCLUSION Our results indicate the presence of risk areas for ACL and AVL in the municipality of Altos and highlight the importance of entomological surveillance to further understand a possible role of Lu. longipalpis in ACL transmission.

Clinical, hematological, biochemical, and histopathological evaluations in domestic cats (Felis catus) infected by Leishmania infantum.

A high frequency of feline leishmaniasis has been reported in several countries. However, much information about disease progression in cats still needs to be clarified. This study aimed to verify the occurrence of clinicopathological changes in cats infected with Leishmania infantum. A total of 60 cats were divided into three groups of 20 animals each: control, suspects, and infected. All 60 cats underwent blood count and biochemical analyses. Serum samples from 20 animals with leishmaniasis were also used to diagnose feline immunodeficiency virus and feline leukemia virus. A total of five of the infected animals underwent necropsy for a histopathological study. The main clinical findings in cats with leishmaniasis were lymphadenomegaly (65%), alopecia (55%), ulcerative skin lesions and weight loss (40%), skin nodules (25%), a significant reduction in red blood cells (p=0.0005) and hematocrit (p=0.0007), hyperplasia in spleen 4/5(80%), presence of Leishmania in the spleen 2/5(40%), hepatitis 3/5(60%), liver degeneration 4/5(80%) and inflammatory nephropathy 3/5(60%). It was concluded that cats with leishmaniasis presented significant clinical, hematological, and histopathological alterations compatible with L. infantum infection. The observation of lymphadenomegaly, weight loss, skin lesions and low concentration of red blood cells, contributes significantly to the diagnosis and analysis of progression of feline leishmaniasis.

Antileishmania and immunomodulatory potential of cashew nut shell liquid and cardanol.

Conventional treatments for leishmaniasis have caused serious adverse effects, poor tolerance, development of resistant strains. Natural products have been investigated as potential therapeutic alternatives. The cashew nut shell liquid (CNSL) is a natural source of phenolic compounds with several biological activities, where cardanol (CN) is considered one of the most important and promising compounds. This study aimed to evaluate antileishmanial, cytotoxic and immunomodulatory activities of CNSL and CN. Both showed antileishmanial potential, with IC50 for CNSL and CN against Leishmania infantum: 148.12 and 56.74 µg/mL; against Leishmania braziliensis: 85.71 and 64.28 µg/mL; against Leishmania major: 153.56 and 122.31 µg/mL, respectively. The mean cytotoxic concentrations (CC50) of CNSL and CN were 37.51 and 31.44 µg/mL, respectively. CNSL and CN significantly reduced the percentage of infected macrophages, with a selectivity index (SI) >20 for CN. CNSL and cardanol caused an increase in phagocytic capacity and lysosomal volume. Survival rates of Zophobas morio larvae at doses of 3; 30 and 300 mg/kg were: 85%, 75% and 60% in contact with CNSL and 85%, 60% and 40% in contact with CN, respectively. There was a significant difference between the survival curves of larvae when treated with CN, demonstrating a significant acute toxicity for this substance. Additional investigations are needed to evaluate these substances in the in vivo experimental infection model.

Development of Immunological Assays Based on Leishmania donovani Antigen for Diagnosis of Canine Visceral Leishmaniasis and Their Multicenter Evaluation in Brazil and Italy.

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.

Methods of macrophages activation and their modulation for the prospection of new antileishmania drugs: a review

Leishmaniasis are a group of parasitic zoonoses provoked by protozoa from Leishmania genus and belonging to the group of neglected tropical diseases. The search and development for new drugs is necessary not only to investigate the activity against only the parasite, but also to investigate the possible synergistic effect of new drugs with the immune response of the host. In the present review, macrophages are pointed out as potential targets of the investigation of new antileishmanial drugs, and some methodologies in order to assess their activation as response to Leishmania-infected cells are presented. Macrophages are an important role in the cellular immune response, since they are cells from mononuclear phagocytic system, the first line of defense of the host, against parasites from Leishmania genus. Phagocytic capacity, lysosomal activity, increase of nitric oxide and intracellular calcium levels are parameters regarding assessment of macrophages activation which allow them to be more hostile in order to solve the infection and lead the patient to cure. In this context, we bring 19 substances already investigated and that activate macrophages, what makes them promising in the antileishmanial treatment. Therefore, assessment of macrophages activation, are important tools for discovery of immunomodulatory compounds which have potential to act in synergism with host immune response. Such compounds might be promising as monotherapy in the treatment of leishmaniasis, as well as being used as adjuvants in vaccines and/or in combination with conventional drugs.

Transmission of Leishmania infantum from cats to dogs.

Leishmania infantum infection in cats has been reported in several countries, including Brazil. However, the transmission of visceral leishmaniasis (VL) from cats to another host has not been proven yet. Therefore, the aim of this study was to verify the possibility of L. infantum transmission from cats to dogs. In order to verify the possibility of VL transmission from the cat to the dog, xenodiagnosis was carried out in a VL-positive cat, using 55 female Lutzomyia longipalpis. Five days later, 40 insects were dissected to verify Leishmania infection. The remaining 15 females were fed in a healthy dog. The potential infection of the dog was verified through clinical, serological, parasitological examinations, and PCR, at three, six, and twelve months post-infection. All 55 L. longipalpis females became visibly engorged. Leishmania promastigotes were detected in 27.5% of the dissected insects. Leishmania infection in the dog was confirmed upon first evaluation. DNA sequencing of the parasite isolated from the cat confirmed L. infantum infection and showed 99% similarity with the L. infantum DNA sequences from the dogs. Through this study, it was possible to confirm the L. infantum experimental transmission from a domestic cat to a domestic dog through its biological vector L. longipalpis.

Gallic and Ellagic Acids Are Promising Adjuvants to Conventional Amphotericin B for the Treatment of Cutaneous Leishmaniasis.

In this study, we demonstrated the potential associative effect of combining conventional amphotericin B (Amph B) with gallic acid (GA) and with ellagic acid (EA) in topical formulations for the treatment of cutaneous leishmaniasis in BALB/c mice. Preliminary stability tests of the formulations and in vitro drug release studies with Amph B, GA, Amph B plus GA, EA, and Amph B plus EA were carried out, as well as assessment of the in vivo treatment of BALB/c mice infected with Leishmania major After 40 days of infection, the animals were divided into 6 groups and treated twice a day for 21 days with a gel containing Amph B, GA, Amph B plus GA, EA, or Amph B plus EA, and the negative-control group was treated with the vehicle. In the animals that received treatment, there was reduction of the lesion size and reduction of the parasitic load. Histopathological analysis of the treatments with GA, EA, and combinations with Amph B showed circumscribed lesions with the presence of fibroblasts, granulation tissue, and collagen deposition, as well as the presence of activated macrophages. The formulations containing GA and EA activated macrophages in all evaluated parameters, resulting in the activation of cells of the innate immune response, which can generate healing and protection. GA and EA produced an associative effect with Amph B, which makes them promising for use with conventional Amph B in the treatment of cutaneous leishmaniasis.

Evaluation of the serum biochemistry and histopathology of kidney and bladder of dogs with Leishmania sp. in their urine / Avaliação da bioquímica sérica e histopatologia de rim e bexiga de cães com Leishmania sp. na urina

The visceral establishment of Leishmania infantum in dogs may result in kidney and bladder tissue injury, with L. infantum ending up in urine. This study therefore aimed at investigating the presence of Leishmania sp. in urinary sediments, and correlating the results with those from renal and bladder serum biochemistry and histopathology. Thirty dogs with negative Nested-Polymerase Chain Reaction (PCR) for E. canis were used in the experiment, and were divided into three groups: control group (10 dogs), neither leishmaniasis nor clinical changes; group I (15 dogs), leishmaniasis but no Leishmania sp. in urine; and group II (5 dogs), leishmaniasis, as well as Leishmania sp. in urine. All animals were submitted to clinical, serological, and parasitological diagnosis for leishmaniasis, biochemical exams, and kidney and bladder histopathology. The parasite was also detected in the bladder imprint of one group II dog. Group II dogs presented with very low albumin concentrations, low albumin/globulin ratios, and kidney and bladder lesions. In the kidneys, hydropic degeneration, thickened Bowman's capsule, and thickening of the tubular capsule were detected in all dogs with positive urinary sediment. However, no significant difference in these renal changes was observed between groups. The intensity and distribution of bladder inflammatory infiltrates were significantly (p-value < 0.05, Kruskal-Wallis' and Dunn's tests) higher in group II dogs, compared with those of the other groups. The presence of Leishmania sp.in the urine of infected dogs appeared to be related to low serum albumin concentrations and more severe bladder lesions

O estabelecimento visceral de Leishmania infantum em cães pode resultar em lesões nos tecidos dos rins e da bexiga, favorecendo a chegando do parasito até a urina. Portanto, este estudo teve como objetivo investigar a presença de Leishmania sp. em sedimentos urinários e correlacionar os resultados com os achados de quantificações bioquímicas séricas e histopatologia de rim e bexiga. Trinta cães com Nested-Reação em Cadeia da Polimerase (PCR) negativa para E. canis foram utilizados no experimento e foram divididos em três grupos: grupo controle (10 cães), negativos para leishmaniose e sem alterações clínicas; grupo I (15 cães), com leishmaniose, mas sem Leishmania sp. na urina; e grupo II (5 cães), com leishmaniose e com Leishmaniasp. na urina. Todos os animais foram submetidos a diagnóstico clínico, sorológico e parasitológico para leishmaniose, exames bioquímicos e histopatologia de rim e bexiga. O parasito foi detectado no imprimt de bexiga de um cão do grupo II. Os cães do grupo II apresentaram concentrações muito baixas de albumina, baixa relação albumina/globulina e lesões nos rins e na bexiga. Nos rins, foram detectadas degeneração hidrópica, espessamento da cápsula de Bowman e espessamento da cápsula tubular, em todos os cães com sedimento urinário positivo. No entanto, nenhuma diferença significativa nessas alterações renais foi observada entre os grupos. A intensidade e a distribuição dos infiltrados inflamatórios da bexiga foram significativamente (p-valor < 0,05, testes de Kruskal-Wallis e Dunn) maiores nos cães do grupo II, em comparação com a dos outros grupos. A presença de Leishmania sp. na urina de cães infectados parece estar relacionada a baixa concentração sérica de albumina e a lesões mais graves na bexiga.

Infection of Lutzomyia longipalpis in cats infected with Leishmania infantum.

The clinical manifestations most frequently observed in cats with leishmaniasis caused by Leishmania infantum are cutaneous alterations, which suggest a high parasitic load in the skin and the possibility of infecting a vector. This study evaluated the infectiousness of to phlebotomine sand flies cats infected with L. infantum. A total of 12 cats with infection by L. infantum from the city of Teresina, Piauí, Brazil, were included in the study. Cats were diagnosed by direct visualization of the parasite. Laboratory-bred insects, free from infection by Leishmania spp. were offered a blood meal for 60 min on cats infected with L. infantum. On the fifth and sixth day after the blood meal, flies were dissected to assess promastigote forms of the parasite in the digestive system. Eight cats (67 %) were able to infect the vectors. The frequency of infected insects per cat ranged 0.0-94.4%. The mean frequency of insects feeding on cats was 95.2 %. Large numbers of the parasite were observed per insect, but were not quantified. The result confirm that cats are able to infect L. longipalpis, indicating that cats are part of the epidemiological chain of VL, acting as reservoir of the disease.

Transmission of Leishmania infantum from cats to dogs / Transmissão de Leishmania infantum de gatos para cães

Abstract Leishmania infantum infection in cats has been reported in several countries, including Brazil. However, the transmission of visceral leishmaniasis (VL) from cats to another host has not been proven yet. Therefore, the aim of this study was to verify the possibility of L. infantum transmission from cats to dogs. In order to verify the possibility of VL transmission from the cat to the dog, xenodiagnosis was carried out in a VL-positive cat, using 55 female Lutzomyia longipalpis. Five days later, 40 insects were dissected to verify Leishmania infection. The remaining 15 females were fed in a healthy dog. The potential infection of the dog was verified through clinical, serological, parasitological examinations, and PCR, at three, six, and twelve months post-infection. All 55 L. longipalpis females became visibly engorged. Leishmania promastigotes were detected in 27.5% of the dissected insects. Leishmania infection in the dog was confirmed upon first evaluation. DNA sequencing of the parasite isolated from the cat confirmed L. infantum infection and showed 99% similarity with the L. infantum DNA sequences from the dogs. Through this study, it was possible to confirm the L. infantum experimental transmission from a domestic cat to a domestic dog through its biological vector L. longipalpis.

Resumo A infecção por Leishmania infantum em gatos tem sido relatada em vários países, incluindo o Brasil. No entanto, a transmissão da leishmaniose visceral (LV) de gatos para outro hospedeiro ainda não foi comprovada. Portanto, o objetivo deste estudo foi verificar a possibilidade de transmissão de L. infantum de gatos para cães. Para verificar a possibilidade de transmissão da LV do gato para o cão, foi realizado xenodiagnóstico em um gato com LV, utilizando-se 55 fêmeas de Lutzomyia longipalpis. Cinco dias depois, 40 insetos foram dissecados para verificar a infecção por Leishmania. As 15 fêmeas restantes foram alimentadas em um cão saudável. A possível infecção no cão foi verificada por meio de exames clínicos, sorológicos, parasitológicos e PCR, três, seis e doze meses após a infecção. Todas as 55 fêmeas de L. longipalpis ficaram visivelmente ingurgitadas. Promastigotas de Leishmania foram detectadas em 27,5% dos insetos dissecados. A infecção por Leishmania no cão foi confirmada na primeira avaliação. O sequenciamento do DNA do parasito isolado do gato confirmou a infecção por L. infantum e apresentou 99% de similaridade com sequências de DNA de L. infantum de cães. Através deste estudo, foi possível confirmar a transmissão experimental de L. infantum de um gato doméstico para um cão doméstico através do seu vetor biológico L. longipalpis.

Atractis thapari (Nematoda, Atractidae) parasitizing Chelonoidis carbonarius and C. denticulatus (Testudinidae) in the state of Piauí, Brazil / Atractis thapari (Nematoda, Atractidae) parasitando Chelonoidis carbonarius e C. denticulatus (Testudinidae) no estado do Piauí, Brasil

Abstract Atractis thapari Petter, 1966, an atractid nematode, was collected parasitizing the large intestine of tortoises of the species Chelonoidis carbonarius (Spix, 1824) (Cc) and C. denticulatus (Linnaeus, 1766) (Cd) (Testudinidae) in the Zoobotanical Park of the municipality of Teresina, state of Piauí, Brazil. Taxonomic identification was based on morphological and morphometrical features, and using bright-field and scanning electron microscopy. The present study adds new observations on the morphology of A. thapari, mainly relating to mouth papillae, the excretory pore, deirids, and male and female posterior ends. The parasitic indices of prevalence (P), mean intensity (MI), mean abundance (MA) and range of infection (RI) for A. thapari in these two tortoise species were: P = 100%, MI = 154,667, MA = 154,667, RI = 5,500-588,500 (Cc); P = 100%, MI = 93,639, MA = 93,639, RI = 1,000-224,500 (Cd). This report confirms the occurrence of A. thapari in Neotropical region, South America, Brazil, and extends its occurrences to a new host, the tortoise C. carbonarius. Adjustment of host management with the aim of improving hygiene and health conditions is suggested.

Resumo Atractis thapari Petter, 1966, um nematoide atractídeo, foi coletado parasitando o intestino grosso de jabutis, Chelonoidis carbonarius (Spix, 1824) (Cc) e C. denticulatus (Linnaeus, 1766) (Cd) no Parque Zoobotânico, município de Teresina, estado do Piauí, Brasil. A identificação taxonômica foi baseada nos caracteres morfológicos e morfométricos usando microscopias de campo claro e eletrônica de varredura. O presente estudo adiciona novas observações na morfologia, principalmente relacionadas as papilas bucais, poro excretor, deirídeos, e extremidade posterior de machos e fêmeas. Os índices parasitários de prevalência (P), intensidade média (IM), abundância média (AM) e amplitude de variação de infecção (AI) de A. thapari em ambos jabutis foram P = 100%, IM = 154.667, AM = 154.667, AI = 5.500-588.500 (Cc); P = 100%, I M= 93.639, AM = 93.639, AI = 1.000-224.500 (Cd). Este registro confirma a ocorrência de A. thapari na região Neotropical, América do Sul, Brasil, e expande a um novo hospedeiro, o jabuti C. carbonarius. É sugerido um ajuste no manejo dos hospedeiros objetivando melhora das suas condições higiênico sanitárias.

Atractis thapari (Nematoda, Atractidae) parasitizing Chelonoidis carbonarius and C. denticulatus (Testudinidae) in the state of Piauí, Brazil.

Atractis thapari Petter, 1966, an atractid nematode, was collected parasitizing the large intestine of tortoises of the species Chelonoidis carbonarius (Spix, 1824) (Cc) and C. denticulatus (Linnaeus, 1766) (Cd) (Testudinidae) in the Zoobotanical Park of the municipality of Teresina, state of Piauí, Brazil. Taxonomic identification was based on morphological and morphometrical features, and using bright-field and scanning electron microscopy. The present study adds new observations on the morphology of A. thapari, mainly relating to mouth papillae, the excretory pore, deirids, and male and female posterior ends. The parasitic indices of prevalence (P), mean intensity (MI), mean abundance (MA) and range of infection (RI) for A. thapari in these two tortoise species were: P = 100%, MI = 154,667, MA = 154,667, RI = 5,500-588,500 (Cc); P = 100%, MI = 93,639, MA = 93,639, RI = 1,000-224,500 (Cd). This report confirms the occurrence of A. thapari in Neotropical region, South America, Brazil, and extends its occurrences to a new host, the tortoise C. carbonarius. Adjustment of host management with the aim of improving hygiene and health conditions is suggested.

Serological tests fail to discriminate dogs with visceral leishmaniasis that transmit Leishmania infantum to the vector Lutzomyia longipalpis.

INTRODUCTION: The control of reservoirs for Leishmania infantum -induced zoonotic visceral leishmaniasis requires the identification of dogs posing a population risk. Here, we assessed the performance of several assays to identify Lutzomyia longipalpis infectious dogs. METHODS: We evaluated 99 dogs that were positive for visceral leishmaniasis based on parasite identification. Serological analyses were performed using an enzyme-linked immunosorbent assay, immunofluorescence antibody tests in 1:40 and 1:80 dilutions, rapid dual path platform tests, immunochromatographic assay with a recombinant rK39 antigen, fast agglutination screening tests, and direct agglutination tests. We also performed PCR to analyze peripheral blood and xenodiagnosis. RESULTS: Forty-six dogs infected at least one L. longipalpis specimen. Although the serological test sensitivities were above 85% for detecting L. longipalpis infectious dogs, none showed a satisfactory performance, as both specificity (0.06 to 13%) and the area under the receiver operating characteristic curve (45 to 53%) were low. The PCR results were also weak, with a sensitivity of 30%, specificity of 72%, and an area under the receiver operating characteristic curve of 51%. The infected L. longipalpis proportion was higher among asymptomatic dogs than symptomatic dogs. Among the symptomatic dogs, those with ulceration-free skin diseases were more infectious, with an odds ratio of 9.3 (confidence interval of 1.10 - 428.5). The larger the number of insects fed, the greater the detected infectiousness. CONCLUSIONS: Our study supports the imperative to develop novel technologies for identifying the infectious dogs that transmit L. infantum for the benefit of public health.

Chapiniella variabilis (Nematoda) parasitizing Chelonoidis carbonarius and C. denticulatus (Testudinidae) in the state of Piauí / Chapiniella variabilis (Nematoda) parasitando Chelonoidis carbonarius e C. denticulatus (Testudinidae) do estado do Piauí

Abstract Chapiniella variabilis (Chapin, 1924), a strongylid nematode, was collected parasitizing the large intestine of the tortoises Chelonoidis carbonarius (Spix, 1824) (Cc) and C. denticulatus (Linnaeus, 1766) (Cd) in the Zoobotanical Park of the municipality of Teresina, state of Piauí, Brazil. The taxonomic identification was based on morphological and morphometric features, using bright-field and scanning electron microscopy. The present study adds new observations on the morphology, mainly relating to the mouth papillae, external and internal leaf-crown elements, excretory pore, deirids and male and female posterior end. The parasitic indices of prevalence (P), mean intensity (MI), mean abundance (MA) and range of infection (RI) of C. variabilis in these two tortoise species were: P = 100%, MI = 833.3, MA = 833.3, RI = 500-1,500 (Cc); P = 100%, MI = 472.2, MA = 472.2, RI = 333-500 (Cd). This record expands occurrences of C. variabilis to a new host, C. carbonarius, and to another state in Brazil, in the Neotropical region of South America. Adjustment to host management with the aim of improving hygiene and health conditions is suggested.

Resumo Chapiniella variabilis (Chapin, 1924), um nematoide estrongilídeo, foi coletado parasitando o intestino grosso de jabutis, Chelonoidis carbonarius (Spix, 1824) e C. denticulatus (Linnaeus, 1766) do Parque Zoobotânico, município de Teresina, estado do Piauí, Brasil. A identificação taxonômica foi baseada nos caracteres morfológicos e morfométricos usando microscopias de campo claro e eletrônica de varredura. O presente estudo adiciona novas observações na morfologia, principalmente relacionadas as papilas bucais, elementos externos e internos da coroa-foliar, poro excretor, deirídeos, e extremidade posterior de machos e fêmeas. Os índices parasitários de prevalência (P), intensidade média (IM), abundância média (AM) e amplitude de variação de infecção (AI) de C. variabilis em ambos jabutis foram P = 100%, IM = 833,3, AM = 833,3, AI = 500-1.500 (Cc); P = 100%, IM = 472,2, AM = 472,2, AI = 333-500 (Cd). Este registro aumenta a ocorrência de C. variabilis a um novo hospedeiro, C. carbonarius, e a um outro estado do Brasil, na região Neotropical da América do Sul. É sugerido um ajuste no manejo dos hospedeiros objetivando melhora das suas condições higiênico sanitárias.

Serological tests fail to discriminate dogs with visceral leishmaniasis that transmit Leishmania infantum to the vector Lutzomyia longipalpis

Abstract INTRODUCTION The control of reservoirs for Leishmania infantum -induced zoonotic visceral leishmaniasis requires the identification of dogs posing a population risk. Here, we assessed the performance of several assays to identify Lutzomyia longipalpis infectious dogs. METHODS We evaluated 99 dogs that were positive for visceral leishmaniasis based on parasite identification. Serological analyses were performed using an enzyme-linked immunosorbent assay, immunofluorescence antibody tests in 1:40 and 1:80 dilutions, rapid dual path platform tests, immunochromatographic assay with a recombinant rK39 antigen, fast agglutination screening tests, and direct agglutination tests. We also performed PCR to analyze peripheral blood and xenodiagnosis. RESULTS Forty-six dogs infected at least one L. longipalpis specimen. Although the serological test sensitivities were above 85% for detecting L. longipalpis infectious dogs, none showed a satisfactory performance, as both specificity (0.06 to 13%) and the area under the receiver operating characteristic curve (45 to 53%) were low. The PCR results were also weak, with a sensitivity of 30%, specificity of 72%, and an area under the receiver operating characteristic curve of 51%. The infected L. longipalpis proportion was higher among asymptomatic dogs than symptomatic dogs. Among the symptomatic dogs, those with ulceration-free skin diseases were more infectious, with an odds ratio of 9.3 (confidence interval of 1.10 - 428.5). The larger the number of insects fed, the greater the detected infectiousness. CONCLUSIONS Our study supports the imperative to develop novel technologies for identifying the infectious dogs that transmit L. infantum for the benefit of public health.

Occurrence of Lutzomyia longipalpis Lutz & Neiva 1912 and Cerdocyon thous Linnaeus 1977, in a visceral leishmaniasis endemic area in Brazil.

Cerdocyon thous presents a wide geographic distribution in Brazil and its role as a possible Leishmania infantum reservoir in a visceral leishmaniasis (VL) transmission cycle regardless of dogs (Canis familiaris) has been discussed. From this perspective, this work describes the occurrence and use of the habitat by Cerdocyon thous in a Lutzomyia longipalpis occurrence area Teresina (Piaui - Brazil), VL endemic region. Three specimens of C. thous were monitored with the use of radio telemetry and trails and footprints, seeking to find possible natural dens in order to collect the sanflies from the site. Luminous CDC and Damasceno traps were simultaneously installed at the visited sites, where two specimens of L. longipalpis and one L. termitophila were captured. The identification of the dens and trails, allows us to infer that the dens are not used only by the C. thous. Finding the VL vector in natural C. thous natural dens, reinforces the hypothesis of transmission of Le. infantum in the outskirts of the large urban centers, in a cycle that independs from dogs.

The performance of serological tests for Leishmania infantum infection screening in dogs depends on the prevalence of the disease.

Dogs are considered the main reservoir of Leishmania infantum. This protozoan causes visceral leishmaniasis (VL), an uncontrolled urban zoonosis in Brazil. Serological tests and polymerase chain reaction (PCR) on peripheral blood were performed to identify infected dogs in scenarios of higher and lower prevalence of the disease (Teresina and Vitória). One-hundred infected and 57 non-infected animals from Teresina and 100 non-infected animals from Vitória were studied. Animal selection was not dependent on previous serology. The sensitivity (Teresina) and specificity (Teresina and Vitória) were as follows: indirect antibody fluorescence (IFAT) cut-off of 1:40 (IFAT 1:40): 96%, 18%, and 76%; IFAT 1:80: 90%, 33%, and 93%; direct agglutination test (DAT): 96%, 33%, and 98%; fast agglutination screening test (FAST): 93%, 68%, and 100%; immunochromatographic assay with a recombinant rK39 antigen (rK39): 88%, 74%, and 98%; enzyme linked immunosorbent assay (ELISA): 91%, 79%, and 98%; rapid dual-path platform test (TR DPP®): 98%, 60%, and 98%; and blood PCR: 29%, 93%, and 97%, respectively. In the high transmission area, none of the tests adequately discriminated L. infantum-infected from non-infected dogs. However, in the high transmission city, the area under the receiver operating characteristic (ROC) curve of FAST, DAT, ICrK39, ELISA and TR DPP® was high.

Gallic and ellagic acids: two natural immunomodulator compounds solve infection of macrophages by Leishmania major.

Leishmaniasis is a complex of parasitic protozoan diseases caused by more than 20 different species of parasites from Leishmania genus. Conventional treatments are high costly, and promote a sort of side effects. Besides, protozoan resistance to treatments has been reported. Natural products have been investigated as a source of new therapeutic alternatives, not only acting directly against the parasite but also being able to synergistically act on the host immune system in order to control parasitemia. Gallic acid (GA) and ellagic acid (EA) are plant-derived phenolic compounds which are able to induce antiinflammatory, gastroprotective, and anticarcinogenic activities. Therefore, the antileishmania, cytotoxic, and immunomodulatory activities of GA and EA were evaluated in this study. Both GA and EA were able to inhibit the growth of Leishmania major promastigotes (effective concentration (EC50) values 16.4 and 9.8 µg/mL, respectively). The cytotoxicity against BALB/c murine macrophages for GA and EA was also assessed (CC50 values 126.6 and 23.8 µg/mL, respectively). Interestingly, GA and EA also significantly reduced the infection and infectivity of macrophages infected by L. major (EC50 values 5.0 and 0.9 µg/mL, respectively), with selectivity index higher than 20. Furthermore, both GA and EA induced high immunomodulatory activity evidenced by the increase of phagocytic capability, lysosomal volume, nitrite release, and intracellular calcium [Ca2+i] in macrophages. Further investigations are reinforced in order to evaluate the therapeutic effects of GA and EA in in vivo experimental infection model of leishmaniasis.

Chapiniella variabilis (Nematoda) parasitizing Chelonoidis carbonarius and C. denticulatus (Testudinidae) in the state of Piauí.

Chapiniella variabilis (Chapin, 1924), a strongylid nematode, was collected parasitizing the large intestine of the tortoises Chelonoidis carbonarius (Spix, 1824) (Cc) and C. denticulatus (Linnaeus, 1766) (Cd) in the Zoobotanical Park of the municipality of Teresina, state of Piauí, Brazil. The taxonomic identification was based on morphological and morphometric features, using bright-field and scanning electron microscopy. The present study adds new observations on the morphology, mainly relating to the mouth papillae, external and internal leaf-crown elements, excretory pore, deirids and male and female posterior end. The parasitic indices of prevalence (P), mean intensity (MI), mean abundance (MA) and range of infection (RI) of C. variabilis in these two tortoise species were: P = 100%, MI = 833.3, MA = 833.3, RI = 500-1,500 (Cc); P = 100%, MI = 472.2, MA = 472.2, RI = 333-500 (Cd). This record expands occurrences of C. variabilis to a new host, C. carbonarius, and to another state in Brazil, in the Neotropical region of South America. Adjustment to host management with the aim of improving hygiene and health conditions is suggested.