Domestic feline cutaneous leishmaniasis in the municipality of Ribas do Rio Pardo, Mato Grosso do Sul state, Brazil: a case report

Cutaneous leishmaniases are anthropozoonoses that involve many species of Leishmania and a wide variety of wild mammalian hosts, thus presenting high importance to public health. This study reports the second case of feline leishmaniasis in Mato Grosso do Sul state, in which Leishmania (Leishmania) amazonensis was found in a domestic cat from Ribas do Rio Pardo. Clinical signs were similar to those observed in other diseases commonly diagnosed in cats, such as cryptococcosis and sporotrichosis. Cutaneous leishmaniasis should, therefore, be added to differential diagnoses by feline veterinary practitioners, and also adequate investigations should be carried out to verify the relevance of domestic cats as L. amazonensis reservoirs.

The aetiological agents of American cutaneous leishmaniasis in the municipality of Monte Negro, Rondônia state, western Amazonia, Brazil.

Although American cutaneous leishmaniasis (ACL) is one of the most important endemic diseases in the Brazilian state of Rondônia, there is very little information on the species of parasite involved. The objective of the present study was to identify the Leishmania species causing ACL in the Monte Negro municipality of the state. Over a 6-year period (1997-2002), the skin lesions of 233 patients were examined while the patients were attending an outpatients' clinic at the University of São Paulo's Advanced Research Unit in Monte Negro. ACL was diagnosed in 137 (58.8%) of the patients and leishmanial parasites were successfully isolated from 14 of the ACL cases. Using a panel of 24 monoclonal antibodies, 12 of the 14 isolates were identified, as L. (Viannia) braziliensis (seven), L. (V.) lainsoni (one), a L. (V.) lainsoni-like species (two), a L. (V.) guyanensis-like species (one), or a L. (Viannia) species that was different from all named species (one). These are the first records of human infection with L. (V.) braziliensis and L. (V.) lainsoni in Rondônia.

Leishmania (Viannia) utingensis n. sp., a parasite from the sandfly Lutzomyia (Viannamyia) tuberculata in Amazonian Brazil.

A leishmanial parasite isolated in 1977 from a specimen of the sandfly Lutzomyia tuberculata from Pará State, Amazonian Brazil, has been characterized following its comparison with other species of Leishmania from the same region, using isoenzyme profiles, monoclonal antibodies and characterization of the miniexon gene repeat, using the polymerase chain reaction technique (PCR). It is described here under the name of Leishmania (Viannia) utingensis n. sp.

Genetic variation in populations of Leishmania species in Brazil.

The population structure of strains of Leishmania (Viannia) braziliensis sensu lato from Pará State and Paraná State in Brazil, of L. (V.) shawi and L. (Leishmania) amazonensis from Pará State, and the relationships of type strains of the subgenera L. (Viannia) and L. (Leishmania) were examined by the random-amplified polymorphic deoxyribonucleic acid (RAPD) technique. Four different primers (M13-40, QG1, L15996 and delta gt11R) were used. The bands were analysed using the neighbor-joining (NJ) and unweighted pair-group method with arithmetic averages (UPGMA) algorithms of the MEGA package. The topology of the NJ and UPGMA trees was very similar but they were not always identical. Both trees differentiated the standard strains of the different species. Strains from the same location were grouped together only in the UPGMA phenogram of the M13-40 primer. L. (V.) braziliensis isolates from Paraná State were genetically closer to those from Paragominas, Pará State than to those from the Amazonian regions of Carajás in Pará State and Peru. The relationship was not dependent on geographical distance. It is postulated that the groups arose from different origins, in which the Amazonian stocks were related to Psychodopygus sand flies while the Paraná strains originated from a gene pool transmitted by Lutzomyia sand flies such as Lutzomyia (Nyssomyia) whitmani. Transmission by Ps. complexus in Paragominas is considered to be a secondary adaptation from the Lutzomyia leishmanial gene pool. Although the vectors of L. (V.) braziliensis are poorly known in the Amazon region, there is strong evidence that the major vectors are all Psychodopygus spp. There was a high degree of genetic variability amongst the L. (V.) shawi strains and there was no clear grouping according to the strains' origins. The genetic variability amongst L. (L.) amazonensis strains from the same locations was much lower but they formed 2 groups which coincided with their origin. Our results support the clonal population structure of Leishmania isolates and suggest that their distribution is related to the origin of the gene pool as well as to present vector and reservoir movements.

An outbreak of cutaneous leishmaniasis among soldiers in Belém, Pará State, Brazil, caused by Leishmania (Viannia) lindenbergi n. sp. A new leishmanial parasite of man in the Amazon region.

Eight cases of cutaneous leishmaniasis are recorded among soldiers of the Brazilian Forest Infantry stationed in Belém, State of Pará, north of Brazil. The infections, all acquired during manoeuvres in nearby degraded primary forest, are attributed to a new member of the subgenus Viannia, Leishmania (V.) lindenbergi n. sp. A further infection by this parasite was encountered in a woman, who lived very close to the same piece of forest. The new parasite has been characterised and differentiated from other known species of the subgenus Viannia following the combined use of enzyme electrophoresis and monoclonal antibodies techniques. The eco-epidemiology of L. (V.) lindenbergi is discussed: by far the most abundant anthropophilic sandfly in the type locality was identified as Lutzomyia (Nyssomyia) antunesi (Coutinho), and this remains high on the list of possible vectors.

American visceral leishmaniasis: wild animal hosts.

In Colombia it has been suggested that the rodent Proechimys canicollis may be a reservoir-host of Leishmania (L.) chagasi, the cause of American visceral leishmaniasis, based on polymerase chain reaction/hybridization tests. We have detected no infection in laboratory-bred specimens of another species, P. guyannensis, after their inoculation with promastigotes or amastigotes of this parasite from Amazonian Brazil.

Characterization of Leishmania sp. strains isolated from autochthonous cases of human cutaneous leishmaniasis in Santa Catarina State, southern Brazil.

Four Leishmania sp. samples were isolated from autochthonous human cases of American cutaneous leishmaniasis (ACL) in Santa Catarina State, southern Brazil. These strains were characterized using indirect immunofluorescence with a panel of Leishmania-specific monoclonal antibodies (MAbs), and by PCR amplification and hybridization assay of the mini-exon gene with group specific probes. The results obtained with the MAbs were in agreement with the genetic marker. Two isolates (MHOM/BR/89/JSC89-H1 and MHOM/BR/89/JSC89-H2) were identified as L. (Leishmania) amazonensis and two (MHOM/BR/96/LSC96-H3 and MHOM/BR/97/LSC97-H4) as L. (Viannia) braziliensis. The southernmost autochthonous cases of ACL in Brazil are due to two different Leishmania sp. species, confirming the spreading of ACL on the American continent.

A mitochondrial DNA phylogeny indicates close relationships between populations of Lutzomyia whitmani (Diptera: Psychodidae, Phlebotominae) from the rain-forest regions of Amazônia and northeast Brazil.

Phylogenetic analysis of all 31 described mitochondrial (cytochrome b) haplotypes of Lutzomyia whitmani demonstrated that new material from the State of Rondônia, in southwest Amazônia, forms a clade within a lineage found only in the rain-forest regions of Brazil. This rain-forest lineage also contains two other clades of haplotypes, one from eastern Amazônia and one from the Atlantic forest zone of northeast Brazil (including the type locality of the species in Ilhéus, State of Bahia). These findings do not favour recognizing two allopatric cryptic species of L. whitmani, one associated with the silvatic transmission of Leishmania shawi in southeast Amazônia and the other with the peridomestic transmission of Le. braziliensis in northeast Brazil. Instead, they suggest that there is (or has been in the recent past) a continuum of inter-breeding populations of L. whitmani in the rain-forest regions of Brazil.

T cell responses to crude and defined leishmanial antigens in patients from the lower Amazon region of Brazil infected with different species of Leishmania of the subgenera Leishmania and Viannia.

Amazonian localized cutaneous leishmaniasis (LCL) is caused by parasites of the subgenera Leishmania and Viannia. Respectively, these parasites may cause diffuse cutaneous leishmaniasis (DCL) and mucocutaneous leishmaniasis (MCL). This, together with differing skin test responses, suggests some species-specificity in cell mediated immunity. In this study, T cell responses (proliferative and interferon-gamma) to crude and defined antigens were examined in paired samples pre and post chemotherapy. Untreated L. (L.) amazonensis LCL patients showed lower responses to crude leishmanial antigens than the L. (V.) spp. group. L. (V.) braziliensis antigen was a more potent stimulator of T cell responses than L. (L.) amazonensis antigen in all patient groups. Few positive responses were seen to the L. (L.) amazonensis glycoprotein GP46. A substantial proportion of LCL patients did respond to the L. (L.) pifanoi amastigote antigens A2, and the surface membrane glycoprotein P8. DCL patients were poor responders to all leishmanial antigens, except GP46. In contrast, MCL patients were good responders to all antigens except GP46 and A2. A significant rise in the response to P8 and A2 antigen was seen post treatment across all LCL and MCL patients, indicating that these antigens might provide suitable vaccine candidates.

The dermal leishmaniases of Brazil, with special reference to the eco-epidemiology of the disease in Amazonia

Six species of Leishmania are at present known to cause cutaneous and/or mucocutaneous leishamniasis in Brazil, and they are all to be found in the Amazon region of this country. The eco-epidemiology of each is discussed, with the observation that the Amazonian leishmaniases are all zoonoses, with their source in silvatic mammals and phlebotomine sandfly vectors. With man's destruction of the natural forest in southern Brazil, some sandfly species have survived by adapting to a peridomestic or domiciliary habitat in rural areas. Some domestic animals, such as dogs and equines are seemingly now involved in the epidemiology of the disease. No such process has yet been reported in the Amazon region, but may well take place with the continuing devastation of its forest.

The dermal leishmaniases of Brazil, with special reference to the eco-epidemiology of the disease in Amazonia.

Six species of Leishmania are at present known to cause cutaneous and/or mucocutaneous leishmaniasis in Brazil, and they are all to be found in the Amazon region of this country. The eco-epidemiology of each is discussed, with the observation that the Amazonian leishmaniases are all zoonoses, with their source in silvatic mammals and phlebotomine sandfly vectors. With man's destruction of the natural forest in southern Brazil, some sandfly species have survived by adapting to a peridomestic or domiciliary habitat in rural areas. Some domestic animals, such as dogs and equines are seemingly now involved in the epidemiology of the disease. No such process has yet been reported in the Amazon region, but may well take place with the continuing devastation of its forest.

Cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis in Amazonian Brazil, and the significance of a negative Montenegro skin-test in human infections.

The clinical and epidemiological features of 62 cases of cutaneous leishmaniasis due to Leishmania (Leishmania) amazonensis, from Pará State, Amazonian Brazil, are discussed. The parasite, isolated in hamster skin and/or blood-agar culture medium, was in each case identified by both biological characteristics and a monoclonal antibody specific for promastigotes of L. (L.) amazonensis. Of the 62 patients, 46 (74.2%) presented with a single cutaneous lesion, and on no occasion was evidence found indicating metastatic spread to either the naso-pharyngeal mucosae or the viscera. Recent claims that this parasite may be responsible for both mucocutaneous leishmaniasis and typical visceral leishmaniasis are discussed. Meglumine antimoniate (Glucantime) proved highly efficient in the treatment of all patients. Of the 62 patients examined by the Montenegro skin test, only 32 (51.6%) gave a positive reaction. The significance of this finding is considered and the hypothesis made that the parasite itself may induce an immunoinhibition. Field studies amply confirmed the role of Lutzomyia flaviscutellata as the major sandfly vector of L. (L.) amazonensis in Amazonia.

Cutaneous leishmaniasis of man due to Leishmania (Viannia) shawi Lainson, de Souza, Póvoa, Ishikawa & Silveira, in Pará State, Brazil.

Leishmania (Viannia) shawi Lainson, Braga, de Souza, Póvoa, Ishikawa & Silveira, 1989, was originally recorded from monkeys (Cebus apella and Chiropotes satanas), sloths (Choloepus didactylus and Bradypus tridactylus) and coatis (Nasua nasua) and the sandfly, Lutzomyia whitmani. With a panel of Leishmania specific monoclonal antibodies, it was found that 30.5% of the Leishmania strains from patients, who had contracted cutaneous leishmaniasis in Pará State, were similar, but not identical to L. (V.) guyanensis. The enzyme profiles of 18 of these strains were determined, and it was found that 12 isolates belonged to the same zymodeme as the type strain of L. (V.) shawi. The other 6 belonged to a second L. (V.) shawi zymodeme that only differed from the other by having a slightly faster PEP band. These results are the first records of infections of L. (V.) shawi in man and suggest that this parasite is probably common in areas of Pará State to the south of the Amazon River.

Cutaneous leishmaniasis of man due to Leishmania (Viannia) naiffi Lainson and Shaw, 1989.

Until recently mammalian hosts of Leishmania (Viannia) naiffi Lainson & Shaw, 1989 have been limited to armadillos, Dasypus novemcinctus, in Amazonian Brazil. Past evidence for human cutaneous leishmaniasis due to this parasite is reviewed, and a recent infection in man reported, with identification of the parasite by isoenzyme profiles and a specific monoclonal antibody. The role of some phlebotomine sandfly species as vectors is discussed.

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

As a means of assessing the usefulness of the monkey Cebus apella as an experimental model for the study of cutaneous leishmaniasis, 5 of these animals were inoculated intradermally at 8 sites along the tail with 3 X 10(6) promastigotes of L. (V.) braziliensis, while a further 5 monkeys received similar inoculations with 3 X 10(6) promastigotes of L. (L.) amazonensis. Following the inoculations, weekly examinations and monthly biopsies showed evolution of resulting skin lesions to be as follows: a) L. (V.) braziliensis: lesions were first visible 15-20 days post-inoculation (p.i), and at 30 days they were clearly of an erythematous-papular nature, which assumed a nodular form at 60 days; after 3 months a spontaneous ulceration of these lesions was noted and, at 4 months, the initiation of healing. In one animal total healing was apparent 5 months p.i; in two others at 6 months, in another monkey after 7 months, and in the last animal at 10 months p.i. Amastigotes were demonstrated in smears from the lesions of all monkeys up to 90 days p.i; up to 120 days in two animals, and at 180 days p.i. in the monkey which showed resolution of the lesions after 10 months. b) L. (L.) amazonensis lesions were first apparent after 20 days p.i; at 30 days they were of an erythematous-papular nature, developing into nodules at 60 days. From the third month of infection onwards, however, the lesions diminished rapidly and, at 90 days p.i. amastigotes were no longer detectable in the skin.(ABSTRACT TRUNCATED AT 250 WORDS)

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

The possible presence of Leishmania in the peripheral blood of 60 patients with American cutaneous leishmaniasis was investigated by the culture of circulating leucocytes. Patients were selected with a variety of clinical forms of the disease and in different evolutionary stages of infection. Biopsies of skin and/or mucosal lesions were made in order to isolate the parasites, which were identified using monoclonal antibodies. 40 isolations were obtained, including 5 of Leishmania (Viannia) braziliensis, 3 L. (V.) guyanesis, 1 L. (V.) lainsoni, 13 L. (Leishmania) amazonensis and 18 which could only be identified as parasites of the braziliensis complex. Cultures of circulanting leucocytes were consistently negative, and the authors conclude that this method is of little use in diagnosis of cutaneous or mucocutaneous leishmaniasis.