Immuno-enzymatic evaluation of the recombinant TSSA-II protein of Trypanosoma cruzi in dogs and human sera: a tool for epidemiological studies.

The rTSSA-II (recombinant Trypomastigote Small Surface II) antigen was evaluated by ELISA to detect anti-Trypanosoma cruzi antibodies in sera from naturally infected dogs and humans. For this evaluation ELISA-rTSSA-II was standardized and groups were classified according to the results obtained through xenodiagnosis, ELISA and PCR. Sensitivity (Se), Specificity (Sp), Kappa index (KI) and area under curve (AUC) were determined. The Se was determined by using 14 sera from dogs infected with T. cruzi VI (TcVI) whereas Sp was determined by using 95 non-chagasic sera by xenodiagnosis, ELISA-Homogenate and PCR. The performance of ELISA-rTSSA-II in dog sera was high (AUC=0·93 and KI=0·91). The Se was 92·85% (1 false negative) and Sp was 100%. Two sera from dogs infected with TcI and 1 with TcIII were negative. For patients infected with T. cruzi, reactivity was 87·8% (36/41), there was only 1 indeterminate, and Sp was 100%. Fifty-four sera from non-chagasic and 68 sera from patients with cutaneous leishmaniasis did not react with rTSS-II. ELISA-rTSSA-II showed a high performance when studying sera from naturally infected dogs and it also presented 100% Sp. This assay could be an important tool to carry out sero-epidemiological surveys on the prevalence of T. cruzi circulating lineages in the region.

The identification of sandfly species, from an area of Argentina with endemic leishmaniasis, by the PCR-based analysis of the gene coding for 18S ribosomal RNA.

The area around Río Blanco, in the Orán department in the north of the Argentinian province of Salta, is endemic for American tegumentary leishmaniasis. In an attempt to facilitate the identification of the Lutzomyia species in this area, sequences of the gene coding for the 18S ribosomal RNA (rRNA) of sandflies caught in a Shannon trap were explored, by a combination of PCR and analysis of restriction-fragment-length polymorphism (RFLP). The products from the PCR, which employed two primers developed specifically for this study (Lu.18S 1S and Lu.18S AR), were cloned into a commercial vector (pGEM-T Easy) so that their nucleotide sequences could be investigated. In the RFLP analysis, the products of single and double digestion with the AfaI and HapII restriction enzymes were separated by electrophoresis in 3% or 4% agarose. Taken together with the results of a morphological investigation of the flies, the resultant DNA fragment patterns were sufficient to identify most of the sandflies caught as Lu. neivai. Although two other species, Lu. cortelezzii and Lu. sallesi, were collected, they were relatively rare and only identified morphologically. A single digestion of the 18S-rRNA gene sequences with AfaI or HapII appeared sufficient and useful for the identification of Lu. neivai from the north of Salta province, and for several other Lutzomyia species.

Characterization of human infection by Leishmania spp. in the Northwest of Argentina: immune response, double infection with Trypanosoma cruzi and species of Leishmania involved.

The aims of this study were to characterize human American tegumentary leishmaniasis, which includes cutaneous, mucocutaneous and mucosal leishmaniasis, in Northwest Argentina, to determine the prevalence of double infection with Trypanosoma cruzi and to identify the species of Leishmania in this area. Most of the 330 leishmaniasis patients presented cutaneous ulcers (96.1%), 2.4% mucocutaneous and 1.5% the mucosal form ('espundia'). The aetiological agents, determined by isoenzyme electrophoresis, were identified as Leishmania (Viannia) braziliensis in 16 out of 20 isolates and in the remaining 4 as Leishmania (Leishimania) amazonensis, the first ever-documented in Argentina. Sera analysed by ELISA and IFA using complex antigen from both T. cruzi and L. braziliensis showed a very high percentage of positives (66.3-78.2%). When antigens for specific diagnosis of Chagas' disease were used, 40.9% of the leishmaniasis patients were also found to be infected by T. cruzi. These results indicate that the strong immune response against T. cruzi gave no protection to Leishmania, in spite of the serological cross-reaction between these parasites. In addition, we showed that more than 40% of the patients would be misdiagnosed as chagasic if complex antigens, as epimastigotes or soluble fraction from epimastigotes, were used in IFA or ELISA. This is of paramount importance not only because patients' treatment would be associated to misdiagnosis but the fact that in many countries in Central and South America, a positive test for Chagas' disease means a rejection for those seeking employment.

Clinical status and parasitic infection in a Wichí Aboriginal community in Salta, Argentina.

In a study, carried out in 2000, of the clinical and parasitological status of a Wichi Aboriginal community living in the suburbs of Tartagal, northern Salta, Argentina, 154 individuals were screened for parasitic infections. Ninety-five faecal samples were also obtained from the same population. Ninety-three percent of the subjects were positive for 1 or more of the parasites investigated by direct test and 70.5% of them had parasitic superinfection. The most frequent helminths were Strongyloides stercoralis (50.5%) and hookworm (47.4%). We found low reinfection rates and a long reinfection period after treatment and provision of safe water and sanitation. Serum reactivity of these patients was analysed by enzyme-linked immunosorbent assay and indirect immunofluorescent assay and 22.1% of them had anti-Toxocara antibodies, 16.2% were positive for a complex antigen of Leishmania braziliensis, 29.9% were positive for a complex Trypanosoma cruzi antigen, and 17.5% were positive for a specific Trypanosoma cruzi antigen, Ag 163B6/cruzipain.

[Mucocutaneous leishmaniasis in naturally infected dogs in Salta, Argentina]. / Leishmaniosis mucocutánea en perros naturalmente infectados en Salta, Argentina.

The objective of the present study is to describe two cases of dogs with mucocutaneous lesions caused by Leishmania spp. Both dogs presented destruction of the nasal septum, hyperemia with soft palate edema and barking alteration due to laryngeal compromise. Biopsies were taken from the lesion border and Leishmania spp. amastigotes were seen in the imprints. The dogs presented positive serology when complex soluble antigen from Leishmania mexicana was used. One of the dogs was also suspected to be infected by Trypanosoma cruzi as suggested by its positive reaction with a purified specific antigen, Ag163B6-cruzipain. Most of the studies concerning leishmaniosis in dogs have described the cutaneous form of this disease in close association with human cases of Leishmania infection instead of the mucocutaneous form described herein. The presence of dogs with mucocutaneous leishmaniosis alerts on an increase of the prevalence of this form in humans, which can cause deforming lesions, alterations of the speech and even an inadequate nutrition due to difficulties in deglutition.

Leishmaniosis mucocutánea en perros naturalmente infectados en Salta, Argentina / Mucocutaneous leishmaniasis in naturally infected dogs in Salta, Argentina

The objective of the present study is to describe two cases of dogs with mucocutaneous lesions caused by Leishmania spp. Both dogs presented destruction of the nasal septum, hyperemia with soft palate edema and barking alteration due to laryngeal compromise. Biopsies were taken from the lesion border and Leishmania spp. amastigotes were seen in the imprints. The dogs presented positive serology when complex soluble antigen from Leishmania mexicana was used. One of the dogs was also suspected to be infected by Trypanosoma cruzi as suggested by its positive reaction with a purified specific antigen, Ag163B6-cruzipain. Most of the studies concerning leishmaniosis in dogs have described the cutaneous form of this disease in close association with human cases of Leishmania infection instead of the mucocutaneous form described herein. The presence of dogs with mucocutaneous leishmaniosis alerts on an increase of the prevalence of this form in humans, which can cause deforming lesions, alterations of the speech and even an inadequate nutrition due to difficulties in deglutition.

Leishmaniosis mucocutánea en perros naturalmente infectados en Salta, Argentina / Mucocutaneous leishmaniasis in naturally infected dogs in Salta, Argentina

The objective of the present study is to describe two cases of dogs with mucocutaneous lesions caused by Leishmania spp. Both dogs presented destruction of the nasal septum, hyperemia with soft palate edema and barking alteration due to laryngeal compromise. Biopsies were taken from the lesion border and Leishmania spp. amastigotes were seen in the imprints. The dogs presented positive serology when complex soluble antigen from Leishmania mexicana was used. One of the dogs was also suspected to be infected by Trypanosoma cruzi as suggested by its positive reaction with a purified specific antigen, Ag163B6-cruzipain. Most of the studies concerning leishmaniosis in dogs have described the cutaneous form of this disease in close association with human cases of Leishmania infection instead of the mucocutaneous form described herein. The presence of dogs with mucocutaneous leishmaniosis alerts on an increase of the prevalence of this form in humans, which can cause deforming lesions, alterations of the speech and even an inadequate nutrition due to difficulties in deglutition.(AU)

[Zoonotic parasitosis transmitted by dogs in the Chaco Salteño, Argentina]. / Parasitosis zoonóticas transmitidas por perros en el Chaco Salteño.

Toxocara canis and Ancylostoma spp. are geohelmints that parasites dogs and can eventually affect humans, mainly children, causing visceral and cutaneous larva migrans respectively, constituting a serious public health problem. This study was carried out in two towns located in the xerophilous forest Chaco salteño where humans live closely with many animals, including dogs. Hematological values and anti-Toxocara canis antibodies, determined by ELISA in serum, were evaluated in 98 children from this area. Thirty-six children presented with eosinophilia of 10% or higher in peripheral blood. Twenty out of 98 (20.4%) children had antibodies against antigen from L2 larvae of Toxocara canis. A high percentage (55.6%) of the children with eosinophilia presented anti-Toxocara canis antibodies. Nine children had multiple serpiginous lesions typical of cutaneous larva migrans. Feces from dogs were collected in the area where children lived, in order to search for parasite contamination. Three different techniques of stool examination were employed and eggs were counted. Out of the 106 feces examined, parasites were found in 82 samples (77.4%). Ancylostoma spp eggs were found in 74 (69.8%) samples and eggs from Toxocara canis were found in 19 (17.2%). The average number of T. canis and Ancylostoma spp eggs/gr of feces, were 200 and 3,871 respectively. Giardia spp (14.5%), Trichuris vulpis (7.6%), Genus Endamoeba (2.8%) and Taenia spp (1.9%) were also identified in the stools. Sanitary control and health education in order to control these parasitoses are emphasized.

Parasitosis zoonóticas transmitidas por perros en el Chaco Salteño. / [Zoonotic parasitosis transmitted by dogs in the Chaco Salteño, Argentina]

Toxocara canis and Ancylostoma spp. are geohelmints that parasites dogs and can eventually affect humans, mainly children, causing visceral and cutaneous larva migrans respectively, constituting a serious public health problem. This study was carried out in two towns located in the xerophilous forest Chaco salteño where humans live closely with many animals, including dogs. Hematological values and anti-Toxocara canis antibodies, determined by ELISA in serum, were evaluated in 98 children from this area. Thirty-six children presented with eosinophilia of 10

or higher in peripheral blood. Twenty out of 98 (20.4

) children had antibodies against antigen from L2 larvae of Toxocara canis. A high percentage (55.6

) of the children with eosinophilia presented anti-Toxocara canis antibodies. Nine children had multiple serpiginous lesions typical of cutaneous larva migrans. Feces from dogs were collected in the area where children lived, in order to search for parasite contamination. Three different techniques of stool examination were employed and eggs were counted. Out of the 106 feces examined, parasites were found in 82 samples (77.4

). Ancylostoma spp eggs were found in 74 (69.8

) samples and eggs from Toxocara canis were found in 19 (17.2

). The average number of T. canis and Ancylostoma spp eggs/gr of feces, were 200 and 3,871 respectively. Giardia spp (14.5

), Trichuris vulpis (7.6

), Genus Endamoeba (2.8

) and Taenia spp (1.9

) were also identified in the stools. Sanitary control and health education in order to control these parasitoses are emphasized.

Leishmaniosis mucocutánea en perros naturalmente infectados en Salta, Argentina. / [Mucocutaneous leishmaniasis in naturally infected dogs in Salta, Argentina]

The objective of the present study is to describe two cases of dogs with mucocutaneous lesions caused by Leishmania spp. Both dogs presented destruction of the nasal septum, hyperemia with soft palate edema and barking alteration due to laryngeal compromise. Biopsies were taken from the lesion border and Leishmania spp. amastigotes were seen in the imprints. The dogs presented positive serology when complex soluble antigen from Leishmania mexicana was used. One of the dogs was also suspected to be infected by Trypanosoma cruzi as suggested by its positive reaction with a purified specific antigen, Ag163B6-cruzipain. Most of the studies concerning leishmaniosis in dogs have described the cutaneous form of this disease in close association with human cases of Leishmania infection instead of the mucocutaneous form described herein. The presence of dogs with mucocutaneous leishmaniosis alerts on an increase of the prevalence of this form in humans, which can cause deforming lesions, alterations of the speech and even an inadequate nutrition due to difficulties in deglutition.