Interaction of avirulent Leishmania species with rat peritoneal macrophages

An "in vitro" system has been developed for study of host cell-parasite interaction in visceral and cutaneous leishmaniasis. Avirulent promastigotes of L. brasiliensis and L. donovani, from strains originally isolated from human cases and mantained by serial culture in Davis' Medium were allowed to infect cultured macrophages from rat peritoneal exudate. Challenge of the macrophages by parasites took place in 199 medium, at 33ºC for L. brasiliensis and at 37ºC for L. donovani. Although the rat is resistant to infections by Leishmania spp., the promastigotes not only invaded the host cells, but transformed into amastigotes and later mutiplied, from 10 min after challenge to 24 hours later.

Functional analysis of the murine T lymphocyte immune response to a protozoan parasite, Leishmania tropica

The results presented in this review summarize a seirs of experiments designed to characterize the murine T cell imune response to the protozoan parasite Leishmania tropica. Enriched T cell populations and T cell clones specific for L. tropica antigens were derived from lymph nodes of primed mice and maintained in continous culture in vitro. These T lymphocytes were shown (A) to express the Lyt 1+ 3- cell surface phenotype, (B) to proliferate specifically in vitro in response to parasite antigens, together with a source of irradiated syngeneic macrophages, (C) to transfer antigen-specific delayed-type hypersensitivity (DTH) responses to normal syngeneic mice, (D) to induce specific activation of parasitized macrophages in vitro resulting in the destruction of intracellular parasites, (E) to provide specific helper activity for antibody responses in vitro in a hapten-carrier system. Protection studies using these defiened T cell populations should allow the characterization of parasite antigen(s) implicated in the induction of cellular immune responses beneficial for the host.

Interactions between Leishmania mexicana mexicana promastigotes and amastigotes and murine peritoneal macrophages in vitro

Unstimulated adherent mouse peritoneal cells were cultured in vitro and infected with equal numbers of a single strain of Leishmania m. mexicana amastigotes (AM), virulent promastigotes (VP), avirulent promastigotes (AVP) and fixed promastigotes (FP). Duplicate May-Grünwald-Giemsa stained coverslips were examined at time intervals up to 13 days. By 3 hr post infection, the number of macrophages containing parasites varied between 60.5% (VP) and 84% (AM) for macrophages exposed to living parasites, compared to 6.5% for macrophages exposed for FP. However, variable numbers of parasites showed degenerative changes by 3 hr, and the number of macrophages containing morphologically intact parasites varied significantly between cells infected with AM (84%) and those infected with VP (42%) or AVP(40%). The mean number on intacte parasites/macrophage also differed significantly between AM-infected cells and living or fixed promastigotes-infected cells. Quantitation of intact and degenerated parasites indicated parasite multiplication, as well as destruction, in VP-infected cells and parasite survival and multiplication in AM-infecte monolayers; in contrast no evidence of parasite multiplication was seen in AVP-infected cells. Changes in the mono layer itself (cell loss and macrophage vacuolization) were also evaluated. These results suggest that crucial events determining the outcome of infection occur in the host-parasite relationship during the fist 24 hours of infection. These events are apparently influenced not only by parasite or host strain but by environmentally induced variation within a given strain.

Presence of Psychodopygus wellcomei (Diptera: Psychodidae), a proven vector of Leishmania braziliensis braziliensis, in Ceara State

Psychodopygus wellcomei, a proven vector of (muco-)cutaneous leishmaniasis, has been found for the first time outside of the Amazon Basin, in Ceará State. Parasitological and entomological evidence suggests that the Leishmania braziliensis braziliensis/Ps. wellcomei zoonosis is widespread on the Brazilian Shield.

Reproductive isolation between different forms of Lutzomyia longipalpis (Lutz & Neiva), (Diptera: Psychodidae), the vector of Leishmania donovani chagasi Cunha & Chagas and its significance to kala-azar distribution in South America

The males of the sandfly Lutzomyia longipalpis occur in two forms, one which bears a single pair of pale spots on tergite 4 and another in which an additional pair of spots characterizes tergite 3. In crosses between laboratory reared stocks of the two forms originating from allopatric and sympatric sites in Brazil nearly all males of one form fail to inseminate females of the other. In addition, insemination failure between some allopatric populaytions of Lu. longipalpis with similar tergal spot patterns is recorded, indicating the existence of additional forms in an apparent species complex. The possibility that Lu. longipalpis sensu latu represents more than a single taxon is discussed and the relevance of these findings to future epidemiological studies on kala-azar is considered.

Leishmania infections in Lutzomyia longipalpis (Diptera: Psychodidae) on the Island of Sao Luis, Maranhao State, Brazil

Dissection of Lutzomyia longipalpis, captured in the São Luis focus of visceral Leishmaniasis revealed a 1.8% promastigote infection rate.

Efecte de la freqüència d'incendis sobre l'estructura i composició de les comunitats vegetals a la península del Cap de Creus

L’efecte de la freqüència d’incendis sobre la dinàmica de les comunitats vegetals mediterrànies ha estat estudiada a la península del cap de Creus. S’han realitzat observacions en 24 parcel·les per percebre canvis en l’estructura i composició de les comunitats vegetals en funció de la freqüència de foc (de 1 a 5 incendis en una mateixa parcel·la) i de les estratègies regeneratives postincendi que presenten les espècies (rebrotació i germinació). Una elevada freqüència d’incendis va associada a una disminució de l’abundància de les espècies llenyoses rebrotadores obligades i de les espècies llenyoses germinadores obligades. D’aquestes últimes, les espècies del gènere Cistus acaben desapareixent a freqüències altes de foc. Les espècies rebrotadores facultatives es mantenen indiferents a la freqüència d’incendis, fet que els permet ser les dominants als estrats arbustius de les comunitats a freqüències elevades de foc. S’observa una tendència a l’increment d’espècies herbàcies, incloses les gramínies, a major números de focs. Brachypodium retusum és dominant en totes les àrees, demostrant una gran capacitat de colonització i regeneració postincendi. Aquests resultats suggereixen una transformació de les comunitats vegetals dominades per espècies arbustives llenyoses a estructures més simplificades dominades per herbàcies, establint-se un mecanisme de retroalimentació entre aquestes i el foc.

Estudi i revalorització dels paisatges culturals de l'alta muntanya pirinenca: la vall del Madriu (Andorra), patrimoni de la humanitat de la UNESCO

Estudi arqueològic integrat de la vall del Madriu-Perafita-Claror (Andorra) iniciat al 2004 sota la coordinació de l’ICAC. L’estudi s’insereix en la declaració de la vall com a Patrimoni de la Humanitat (UNESCO). Las recerques s’enfoquen des de l’Arqueologia del Paisatge, de manera diacrònica i pluridisciplinar, fent especial atenció a les relacions de les societats amb el medi. Les restes arqueològiques treballades s’associen a activitats ramaderes, metal•lúrgiques i de carboneig. La recerca paleoecològica se centra en sediments lacustres, torbosos i arqueològics. L’any 2006, s’han analitzat a alta resolució temporal dues seqüencies palinològiques: l’estany Blau (2471 m) i la torbera de Bosc dels Estanyons (2180 m), obtenint les principals fases d’explotació i antropització d’aquest espai altimontà, així com l’evolució del paisatge en els darrers 11000 anys . La campanya de camp s’ha centrat en l’obtenció de mostres de pluja pol•línica referencial a la vall i en el sondatge de l’estany Forcat (2539 m). Un total de tres mostres han estat enviades a datar per C14. Els treballs arqueològics s’han centrat en l’excavació de sondejos de diagnòstic als jaciments de Basses de Setut (2325 m) i Pleta de les Bacives ( 2530 m) i en la realització de prospeccions a les capçaleres de les valls de Madriu, Perafita i Claror. S’han documentat estructures arqueològiques i s’han realitzat estudis antracològics dels sediments excavats en aquestes. S’ha realitzat un transecte altitudinal de carboneres al llarg de la vall per tal de enregistrar una major diversitat cronològica dels espais de carboneig. S’han enviat datacions C14 per tal de determinar la cronologia de les estructures. Les datacions del 2005 mostren una intensa ocupació ramadera i de carboneig a partir del segle XIV, en època baixmedieval i moderna, així com un establiment metal·lúrgic d’època romana.

El "Status" taxonomico de leishmania garnhami, indicado por su patron de desarrollo en el Vector

Bajo condiciones experimentales, se estabelece el de desarrollo primario de Leishmania garnhami em su vector Lutzomyia townsendi, siguiendo disecciones diarias entre el Iro y 12mo dia post-imnfección. Se demuestra que L. garnhami puede lograr el estabelecimiento inicial de la infección en cualquiera de las regiones del intestino y en los túbulos de Malpighi del insecto vector. Un comportamiento similar fue observado en ejemplares de Lu. longipalpis. Se discute la susceptibilidad de Lu. townsendi a las infecciones por L. mexicana, L. braziliensis y L. garnhami. Se concluye que L. garnhami, a juzgar por su comportamiento en el vector, debe considerarse distinta a las especies del complejo L.mexicana y que su patrón de desarrollo difiere del de las especies de la Sección Suprapylaria, ya que invaden el triângulo pilórico, y del de la Peripylaria por su invasión a los túbulos de Malpighi.

The retained capacity of Lutzomya longipalpis (Lutz & Neiva) to transmit Leishmania chagasi (Cunha & Chagas) after eight years (64 generations in a closed laboratory colony

A closed Lutzomyia longipalpis colony, from Ceará has been used to transmit Leishmania chagasi isolated from a fox in Pará state. The last time this colony was successfully used in similar transmission experiments was eight years (64 generations) ago indicating that this colony of Lu. longipalpis has fully maintained its vectorial capacity in spite of such a long period of maintainance in the laboratory.

Activación de celulas supresoras de la respuesta de hipersensibilidad tardía en ratones BALB/c infectados o vacunados con Leishmania mexicana pifanoi

Los ratones BALB/c inmunizados SC con promastigotes no viables de Leishmania mexicana pifanoi, desarrollaron respuestas de hipersensibilidad tardia (RHT) contra el parásito. En contraposición, los ratones BALB/c infectados crónicamente con L.m. pifanoi y aquellos inmunizados IV con una dosis supraóptima de parásitos muertos, mostraron inhibición de la RHT. La supresión de la RHT en los animales infectados, estuvo precedida por un estado transitorio de inmunidad celular, manifestado durante la fase inicial del desarrollo de las lesiones (4-12 semanas). La inhibición de la RHT observada en los animales infectados o inmunizados con dosis supraóptimas, fue causada por un mecanismo activo, transferible a receptores singénicos a través de las células esplénicas de los donantes suprimidos. La población de células supresoras fue no adherente a "nylon", sensible al tratamiento con un suero anti-timocitos de ratón y C', y de acción específica sobre la RHT contra antígenos leishmánicos. Se propone que la ocurrencia de células T supresoras de la RHT en los animales infectados crónicamente o inmunizados con dosis supraóptimas, es causada por el exceso de carga antigénica. Esta última, en el caso de los animales infectados, secundaria a una falla primaria en el control inmunológico de la población parasitaria.

Interleukin- 2 production during murine infection by Leishmania mexicana amazonensis

Highly susceptible BALB/c mice, resistant C57B1/6 and their F1 progeny (BDF1) were infected subcutaneously in the foot pad with Leishmania mexicana amazonenesis. At various times after infection, spleen or draining popliteal lymph node cells were assayed for their capacity to generate Interleukin-2 (I1-2) by Concanavalin A (ConA) stimulation. In both BALB/c and C57B1/6 strains there was a transient increase in their capacity to produce I1-2, from the 3rd to the 10th week post-infection. Return to pre-infection levels ocurred between 13th to 16th week post-infection in all three strains. BALB/c mice always produced higher titers of 11-2 than C57B1/6, but such differences were statistically significant only at 3 and 10 weeks post-infection. BDF1 mice had titers similar to those observed in BALB/c mice. I1-2 production by ConA-stimulated lymph node cells was lower as compared to the spleen, but with a similar pattern among the three mice strains. Our data show that susceptibility to infection by l. mexicana amazonenesis is not associated with deficient ConA-stimulated I1-2 production.

Infecção experimental de Lutzomyia whitmani em cães infectados com Leishmania braziliensis braziliensis

Lutzomyia (N.) whitmani foi infectada em lesões leishmanióticas de três de nove cães parasitados por Leishmania braziliensis braziliensis . Os índices de infecção desses flebotomíneos foram 8,3% (1/12), 7,1% (1/14) e 1,8% (3,160), respectivamente. Por outro lado, 180 Lu. whitmani que se alimentaram em áreas não-ulceradas em um dos cães foram negativos, após dissecação. É discutida a potencialidade de Lu. whitmani como vector de L.B. braziliensis na região endêmica de Três Braços, Bahia, Brasil.

The susceptibility of wild caught sand flies to infection by a subspecies of Leishmania mexicana isolated from proechimys iheringi denigratus (Rodentia, echimyidae)

Several species of wild caught sand flies were collected in the same site where a subspecies of leishmania mexicana was isolated from the rodent Proechimys iheringi denigratus. The absence of natural infection in these sand flies permitted us to test, with relative assurance, the susceptibility of wild caught females to infection by this parasite. the success obtained in these experimental infections suggest that one or more of the sand fly species encountered in high numbers in the same site where the infected rodents were captured may be the vector(s) of this subspecies of l. mexicana.

Active cutaneous leishmaniasis in Brazil, induced by Leishmania donovani chagasi

L.d. chagasi was isolated from active cutaneous leishmaniasis in both human and canine infections in an endemic area in Rio de Janeiro, Brazil. Both isolates were identified by molecular and immunological characterization of the parasite using three different methods: electrophoretic mobility of isoenzymes; restriction endonuclease fragment analysis of kDNA and serodeme analysis using monoclonal antibodies. This seems to be the first well documented case in the New World of a "viscerotropic" Leishmania inducing a case of cutaneous leishmaniasis. This observation emphasizes that the diagnosis of the etiologic agent of human or canine visceral leishmaniasis based solely upon clinical and epidemiological critwria may lead to erroneous conclusions.