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

Abstract

The diagnosis of human cutaneous leishmaniasis in small towns is sometimes made without the species identification of the Leishmania, even in areas without previous epidemiological surveys. Here Leishmaniasis is a zoonotic infection affecting 12 million people worldwide in 88 countries. The annual incidence is 2 million, with 1.5 million presenting the cutaneous form and 0.5 million the visceral form. In Brazil, American cutaneous leishmaniasis (ACL) is a widespread disease (Marzochi & Marzochi 1994). Since the beginning of the last century to nowadays, changes on the environment of the state of São Paulo have altered the epidemiological pattern of ACL from an epidemic to an endemic one (Tolezano 1994 (Grimaldi et al. 1989, Tolezano 1994. During the medical routine of health centres in small towns the diagnosis of ACL suspects is carried out by standard tests such as Montenegro's skin test and histopatological analysis, without the species identification. However, the species and the strain identification is important for eco-epidemiological evaluations and to monitor the entrance of new Leishmania species in a local focus (Lainson et al. 1994). Seven ACL cases were notified from 1998 to 2004 in the municipality of Rincão (map reference) which it is located in the northeast centre of the state of São Paulo, at the boarder of the western Paulista Plateau. The area is crossed by MogiGuaçu river and its landscape is composed by residual forest and sugar cane plantations. To identify the Leishmania species circulating on that region, attempts were made to isolate a strain from one patient. Both punch biopsy and aspirates were made from the lesion. A fragment was taken and triturated in saline plus antibiotics (1000 U/ ml penicillin and 1 mg/ml streptomycin). The aspirates were transfered into a tube containing saline and antibiotics and centrifuged after 3 h (1000 g for 1 min). Fragments and the pellet of the aspirated were inoculated separately into a biphasic culture medium (NNN-agar rabbit blood with LIT medium). These cultures were kept at 28 o C. Promastigotes forms were observed after eight days but only in the tubes inoculated with aspirates. The Leishmania isolate was analyzed by multi-locus enzyme electrophoresis approach as previously described (Cupolillo et al. 1994). The strain was identified as L. (V.) braziliensis zymodeme IOC/Z27. This same zymodeme has been found in other endemic ACL areas of Southeast Brazil (Cupolillo et al. 2003).Entomological surveys were performed about 50 m from the edge of Mogi-Guaçu river, area where the patient live (21 o 35'13''S, 48 o 04'15''W). Shannon trap and CDC light traps were used in two nights from 18 to 22h. The only sand fly species captured was Lutzomyia neivai (Pinto, 1926) (149 females and 79 males). This species was previously designated as a junior synonym of L. intermedia (Lutz & Neiva, 1912). Marcondes (1996) revalidated the name and Andrade Filho et al. (2003) redescribed both species, adopting the genus Nyssomyia instead of Lutzomyia according to Gala...

Octenol as Attractant to Nyssomyia neivai (Diptera: Psychodidae: Phlebotominae) in the Field

“…The area is used for Þshing and is considered endemic for human cutaneous leishmaniasis. Leishmania braziliensis (Vianna) was isolated from a local resident (Pinto et al 2005).…”

Antimicrobial activity of RP-1 peptide conjugate with ferrocene group

“…The number of intracellular amastigotes was counted under a microscope. At least 200 macrophages were counted in each of the three slides and the determination of the ratio between infected and uninfected macrophages, enabling the determination of the IC 50 -AMA in relation to drug-free control [3]. At the end, the inhibitory concentration for 50% of the amastigote forms (IC 50 -AMA) in BioEstat software was determined by nonlinear regression, the data was expressed as mean ± SD of two independent replicates, and the obtained results converted to the infection index values, which were obtained as the ratio between the percentage of infected macrophages and the average number of amastigotes per macrophage.…”

“…The world population has been increasing exponentially and might reach around 8.6 billion people by 2030 [1]. In developing countries, a fraction of the expanding population lives in areas with poor sanitation and infrastructure, which contributes to elevate the risk of contracting infectious diseases such as vector-borne diseases and food/ water-borne illness [2][3][4][5], among others. Leishmaniasis, a vector-borne disease caused by Leishmania spp., is endemic in 98 countries and its cutaneous form presents an incidence of 0.7 to 1.2 million new cases per year [6].…”

Bioactive compounds against neglected diseases isolated from macroalgae: a review

“…Furthermore, growth in human population is causing unplanned and poorly managed urbanization, mainly in developing countries, putting millions of people to live in areas with poor infrastructure. It is estimated that the percentage of people living in urban areas will increase from 54% in 2015 to 66% by 2050 (WHO 2014), contributing to the worsening infrastructure problems, climate changes, and emergence of vector-borne diseases (i.e., leishmaniasis/malaria), soil-transmitted helminthic infections (i.e., ascaridiasis/strongyloidiasis), and food-and water-borne illness (i.e., amoebiasis/giardiasis), among other NDs caused by bacteria and fungi (yaws/mycetoma) (Foley et al 2005;Pinto et al 2005Pinto et al , 2011Oryan and Akbari, 2016) (Fig. 1).…”