Chagas disease in the Chilean altiplane. Epidemiological, parasitological, and clinical aspects]

Chagas' disease is endemo-enzootic in periurban and rural sections of the first seven (18 degrees 30'-34 degrees 36' South lat.), out of the thirteen political-administrative regions of Chile. Permanent inhabited localities situated at different altitudes over the sea level (OSL) are classified in: coastal settlement (0-1,499 m OSL), intermediate settlement (1,500-3,000 m OSL) and altiplano settlement (> 3,000 m OSL). A total of 1,558 dwellings and their occupants from regions I and II were surveyed in 1982-1984 for different epidemiological Trypanosoma cruzi infection parameters. Hence, 261 dwellings, 189 domestic mammals and 517 people (ages 5- > or = 60 years) from the Altiplano settlement were studied. The results obtained were: 4(1.5%) and 2(0.8%) of dwellings infested with Triatoma infestans, according to occupants information and by observation of direct evidences respectively; 8(4.2%) animals (1 dog, 4 goats and 3 sheep) with a positive hemagglutination test (IHAT) for Chagas' disease; 21 people (4.1%) with a positive IHAT; 4(19.0%) and 3(14.3%), out of the 21 IHAT positive individuals with all types of electrocardiogram (EKG) abnormalities and EKG abnormalities suggestive of a chagasic etiology respectively, whereas 65(13.1%) and 15(3.0%), out of the 496 IHAT negative ones presented the same kinds of EKG abnormalities.(ABSTRACT TRUNCATED AT 250 WORDS)     

Overview of the epidemiology of Chagas' disease in Chile]

Chile is a long and narrow country located in the south western coast of South America. Chagas' disease exists in the seven first (18 degrees 30'-34 degrees 36' South lat.) of the total of thirteen administrative regions of the country. In the 1982-1990 period a series of studies considering different epidemiological aspects of this parasitic zoonosis has been carried out with the following results: 5,601 rural of periurban dwellings were surveyed for the presence of Triatoma infestans (the most important and almost exclusive vector of Trypanosoma cruzi in Chile). 37.4% of the dwellings were infested according to the inhabitants and 29.4% were found infested according to the presence of tracks or insects captured. In 659 (17.2%) out of 3,822 T. infestans captured and examined T. cruzi was found in their abdominal contents. The most common sources of T. infestans feeding were mammals (89.0%), including man, and birds (9.5%). An indirect hemagglutination test (IHAT) for Chagas' disease, a very sensitive and specific reaction, was performed to 5,050 domestic mammals, resulting positive 7.9% of cats, 7.0% of dogs, 7.0% of goats, 4.9% of sheep and 4.1% of rabbits. 2,579 (16.9%) out of 15,418 persons were positive for the IAHT for Chagas' disease. The rates of infection were rather similar in males (17.5%) and females (16.2%) with an increase in infection rates in accordance with increase of age of individuals. The overall frequency of ECG abnormalities in positive IHAT persons was 18.7% against 8.8% in those with negative IHAT, whereas ECG abnormalities considered as suggestive of a chagasic etiology were 6.8% and 2.2% respectively. The esophageal motility in 311 persons with a positive IHAT and in 150 with a negative IHAT was found altered in 42.8% and 18.7% respectively. In the corresponding urban sectors of the 7 regions mentioned 2.7% of blood donors, 2.3% of delivering mothers, 2.6% of newborns and 0.6% of school children had positive IHAT. 646 chagasic women and 709 non-chagasic women in their reproductive span of life, and the products of the pregnancies that they had in a 6-year period were followed-up. No significant differences were found neither in the number nor in the evolution of pregnancies in both groups of mothers. Xenodiagnosis of children from chagasic mothers resulted positive in 6.3-8.9%, showing the transmission of T. cruzi by the placental route. Recently, 3 cases of congenital Chagas' disease of second generation have been demonstrated.(ABSTRACT TRUNCATED AT 400 WORDS)     

Household chagasic endemia detected as a consequence of a case of congenital infection]

During a study on prevalence of parasitic and viral serological markers in pregnant adolescents, a 17-year-old primipara from Polpaico, village near Santiago, gave birth to a normal male newborn in a Santiago hospital. As both of them presented positive an indirect hemagglutination test (IHAT) for Chagas' disease and the corresponding xenodiagnosis (XD), the infection in the infant was considered to be acquired through the placental route. According to recent epidemiological surveys Polpaico is an endemo-enzootic chagasic rural settlement, where 14.7% of dwellings were infested with Triatoma infestans, while triatominae, persons and domestic mammals were found infected with Trypanosoma cruzi. One month later the adolescent mother, her son and other 11 consanguineous members of the family were visited in their homes in order to submit each of them to a physical examination and to IHAT for Chagas' disease, and XD to those whose IHAT resulted positive. Thus, in 7 (53.8%) the IHAT was positive and in 4 (57.1%) out of these 7 presented positive the XD, results that as a whole yielded a household chagasic endemics.     

Epidemiology of toxoplasmosis in Chile. VI. Prevalence of human infection, investigated by means of an indirect hemagglutination test, in regions VII, VIII and IX

In presecuting the investigations on the epidemiology of toxoplasmosis in Chile, a new series of serological surveys has been performed during 1982-1989 in 10 urban and 25 periurban-rural localities from the regions VII, VIII and IX of the country (34 degrees 41'-39 degrees 38' South lat.). In 9,758, randomly selected apparently healthy persons, and indirect hemagglutination test (IHAT) for toxoplasmosis was carried out. The age of these individuals (4,203 males and 5,555 females) varied between 4 and 84 years. The examined persons represent a 0.33% of the total population of the three studied regions. IHAT titers of 1: > or = 16 were regarded as positive. The global prevalence for positive IHAT was 45.5% (50.5% in men and 41.7% in women). A higher proportion of positive tests was observed in urban areas (47.0%) than in periurban-rural sections (33.3%). An increasing prevalence with age was also observed. Only 5 (0.05%) persons had IHAT titers higher than 1:1000.     

Comparison of the yield of indirect hemagglutination reaction and complement fixation reaction in the diagnosis of neurocysticercosis

Two variations of an indirect hemagglutination test (IHAT) and a complement fixation test (CFT) for the diagnosis of human cysticercosis were compared and evaluated. For the IHAT, a cysticerci crude total saline extract (SE) and a cysticerci lyophylized and delipidized veronal bicarbonate saline buffer (VBS) extract were used, comparing their diagnosis yieldings with that of a CFT in 57 confirmed cysticercosis patients: 45 serum samples and 32 cerebrospinal fluid (CSF). Sera and CSF from 29 patients with other neurological diseases and 25 sera from healthy volunteers were also compared. Both types of methods presented an overall average concordance of 91.5% and 97.0% with CSF and sera respectively. With respect to the sensitivity observed with CFT was 85.2% and 93.3% for CSF and sera, whereas that of IHAT was 96.9% in CSF and 97.8% in sera, when SE antigen was used; with the VBS antigen for IHAT 96.9% and 95.6% were detected in CSF and sera respectively. In order to determine the specificity of the IHAT, besides the study in healthy volunteers, in patients with other neurological diseases and in 156 serum samples from individuals with other parasitoses, such as hydatidosis (43), trichinosis (56), fascioliasis (31) and Chagas' disease (26) were also tested. A high reactivity with the hydatidosis group was found. The specificity, using a titre > or = 1:16 as a diagnostic value and without considering hydatidic sera was 99.4% for RHAI (SE), 100.0% for RHAI (VBS). The use of IHAT and CFT in diagnosis of human cysticercosis is discussed.     

Epidemiology of toxoplasmosis in Chile. V. Prevalence of the infection in humans and domestic and wild animals, studied by indirect hemagglutination reaction, in the Juan Fernández Archipelago. V Region

A serological study utilizing an indirect hemagglutination test (IHAT) for toxoplasmosis was carried out in 222 humans and in 58 domestic animals (31 dogs, Canis familiaris; 27 cats, Felis catus), and in 62 wild mammals distributed into 50 rabbits, Oryctolagus cuniculus and 12 goats, Capra hircus. This survey was performed in the Juan Fernández Archipelago, formed by three islands: Robinson Crusoe, Santa Clara and Alejandro Selkirk (80 degrees 47'-78 degrees 47' west long., and 33 degrees 36'-33 degrees 47' south lat.). Blood samples were collected in filter paper and IHAT with titres greater than or equal to 1:16 were considered positive. This survey showed a prevalence of 42.3% in humans with no difference between men (43.0%) and women (41.5%). A high prevalence was found within groups of young individuals (0 to 19 years old), men and women. Regarding the domestic animal population, 44.8% resulted positive, distributed as follows: dogs 9.7% and cats 85.2%. Twenty one percent of wild animals were positive, distributed as follows: rabbits 8.0% and goats 75.0%. The global prevalence of toxoplasmosis in animals (domestic and wild) was 32.5%. All titres in humans and animals were less than or equal to 1:512. Toxoplasmosis is well extended among the human and animal population of the Juan Fernández Archipelago.     

Results of serological reactions in patients with presumptive diagnosis of toxoplasmosis]

Even though Toxoplasma gondii is an ubiquitous parasite that can effect most of human structures and organs, not all clinical manifestations suggestive of being produced by it are caused by this protozoon. For these reasons sera samples of patients suspected of having toxoplasmosis are sent to the laboratory for detecting specific antibodies which would facilitate the differential diagnosis. Thus, 716 sera from suspected patients, mainly from the Metropolitan Region of Chile, were sent to the Parasitology Laboratory of Chile University in order to carry out in them, specific serological tests for toxoplasmosis: indirect hemagglutination test (IHAT), Sabin Feldman reaction (SFT) and complement fixation test (CFT). Were considered positive: IHAT and/or SFT with titers > or = 1:16 and CFT with titer > or = 1:5. The pathologies for demanding these serological tests were obstetrical problems 210 (29.3%), congenital problems 193 (27.0%), ophthalmopathies 81 (11.3%), adenopathies 77 (10.8%), AIDS 67 (9.4%), myocardiopathies 46 (6.4%) and miscellaneous 42 (5.9%). The positivity found in these sera was higher in ophthalmopathies (61.7%), followed by obstetrical problems, miscellaneous problems, myocardiopathies and AIDS (50.7-52.4%), less frequent was the positivity in adenopathies (35.1%) and congenital problems (23.1%). In general, the 43.7% of positivity for toxoplasmosis found in these patients is higher than the 37.0% found in the general population. High titers of IHAT and SFT plus positive CFT was found in 13-fold higher proportion than in the general population.     

Immunoenzyme assay using micro Dot on nitrocellulose (Dot-ELISA) in the diagnosis of Chagas' disease. I. Comparative study of 2 antigenic preparations of Trypanosoma cruzi

Using the Dot-ELISA technique, two antigenic preparations of Trypanosoma cruzi epimastigote forms have been compared for the diagnosis of Chagas' disease: (1) The cytoplasmic fraction (cytoplasmic antigen) and (2) whole formalin fixed epimastigotes (integral antigen). There was been used sera from 95 chagasic patients with chronic cardiomyopathy, positive conventional serology and either positive or negative xenodiagnosis; 74 subjects with negative conventional serology, and either clinically normal or presenting cardiomyopathy; 74 patients with different diseases including syphilis, toxoplasmosis, leishmaniasis or autoantibodies such as rheumatoid factor and antinuclear antibodies. By defining the diagnostic titers (cut off): 1:512 for cytoplasmic antigen and 1:128 for the integral antigen, a sensitivity of 100% has been obtained with both antigenic preparations, being the specificity of 96% for the former and 100% for the latter when leishmaniasis sera were not included. A comparative study with conventional serology was carried out using 147 sera from a Laboratory of Chagas' diagnosis; Dot-ELISA with cytoplasmic antigen showed co-positivity index of 1.0, co-negativity 0.989 and efficiency of 0.993, and Dot-ELISA with integral antigen 1.0, 0.979 and 0.986 respectively. According to this evaluation, Dot-ELISA using whole formalin fixed epimastigotes might be a practical alternative for the serological diagnosis of Chagas' disease.     

The indirect hemagglutination reaction in the diagnosis of trichinosis

An indirect hemagglutination test (IHAT) for trichinosis using a Melcher's antigen was evaluated and compared with a precipitin test (PT) and a bentonite flocculation test (BFT). One hundred and forty eight serum samples from patients from the whole country confirmed or suspected to have trichinosis by clinical or epidemiological evidences were studied: 117 (79.1%) samples resulted positive by IHAT, 138 (93.2%) by PT and 65 (43.9%) by BFT. Sixty three serum samples from patients with strong clinical suspect of trichinosis presented the PT and the BFT positive and were compared with the IHAT for sensitivity study. IHAT was positive in 60 (95.2%) serum samples. In order to determine the specificity of IHAT 25 serum samples from healthy volunteers and 124 serum samples from individuals with other parasitoses, such as cysticercosis (48), hydatidosis (45) and fascioliasis (31) were studied. The specificity, using a titre > or = 1:16 as a possible diagnostic value was 96%. The use of IHAT with RP and BFT in the diagnosis of human trichinosis is discussed.     

Seroepidemiological survey of human toxoplasmosis in Osorno, Region X, Chile, 1998]

A serological survey was carried out in Osorno X Region, Chile (40 degrees 21'-40 degrees 46' South lat, and 73 degrees 26' -72 degrees-46' West long.). An indirect hemagglutination test (IHAT) for toxoplasmosis was performed to 305 persons--160 blood donors and 145 with sexually transmitted disease (STD). The age of the surveyed persons (167 males and 138 females) varied between 10 and 72 years. IHAT titers > 16 were considered as positive. The general prevalence was 20.3%--21.2% in blood donors and 19.3% in persons with some STD--with no differences between males and females. However, significant differences between males and females with STD were found (35.6% and 8.1% respectively). No differences between urban and rural inhabitants were found with a slight higher prevalence in the urban ones. No antibodies against Toxoplasma gondii were detected in six AIDS patients. The importance of performing toxoplasmosis immunodiagnosis and individual prophylaxis to avoid the infection in high risk group are recommended.     

Yielding of xenodiagnosis, according to the number of boxes used in 1,181 persons with chronic chagasic infection diagnosed with indirect hemagglutination reaction]

A study for evaluation of the yielding of xenodiagnosis (XD) in 1,181 persons with a previous positive indirect hemagglutination test (IHAT) for Trypanosoma cruzi infection was carried out. The infection of these people was detected during epidemiological surveys performed in rural-periurban and urban sections of the endemo-enzootic area of Chagas' disease in Chile, which involves the first seven, out of the thirteen political-administrative regions of the country. The sex distribution was 75.0% females and 25.0% males, varying the ages between 2 and 80 years. According to individual and geographical possibilities each person was submitted to 1-8 XD, which consisted in cylindrical wooden boxes containing seven third instar nymphs, laboratory reared, of Triatoma infestans. The boxes, covered with a piece of tulle fixed with a rubber band were applied on the skin of the posterior side of the arm of the subject to be examined, held with a linen bracelet during 25-30 min. After the insects were fed the boxes were maintained in the laboratory at 27 degrees C and 85% relative environmental humidity. Posteriorly, all the nymphs of each box were examined at 30, 60 and 90 days after the application. A drop of abdominal content of each of them, homogenized with a drop of saline, was examined at the microscope looking for T. cruzi. XD resulted positive in 503 (42.6%) people. The positiveness of XD showed a trend of increasing according to the number of boxes used, from 11.4% with one to 51.6% with six. Under a practical point of view, the simultaneous application of four XD boxes seems to be advisable.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of ROP18 alleles in human toxoplasmosis

The role of the virulent gene ROP18 polymorphisms is not known in human toxoplasmosis. A total of 320 clinical samples were analyzed. In samples positive for ROP18 gene, we determined by an allele specific PCR, if patients got the upstream insertion positive ROP18 sequence Toxoplasma strain (mouse avirulent strain) or the upstream insertion negative ROP18 sequence Toxoplasma strain (mouse virulent strain). We designed an ELISA assay for antibodies against ROP18 derived peptides from the three major clonal lineages of Toxoplasma. 20 clinical samples were of quality for ROP18 allele analysis. In patients with ocular toxoplasmosis, a higher inflammatory reaction on eye was associated to a PCR negative result for the upstream region of ROP18. 23.3%, 33% and 16.6% of serums from individuals with ocular toxoplasmosis were positive for type I, type II and type III ROP18 derived peptides, respectively but this assay was affected by cross reaction. The absence of Toxoplasma ROP18 promoter insertion sequence in ocular toxoplasmosis was correlated with severe ocular inflammatory response. Determination of antibodies against ROP18 protein was not useful for serotyping in human toxoplasmosis.     

Chagas' disease and AIDS

Chagas' disease caused by Trypanosoma cruzi is an opportunistic infection in the setting of HIV/AIDS. Some individuals with HIV and chronic T. cruzi infection may experience a reactivation, which is most commonly manifested by meningoencephalitis. A reactivation myocarditis is the second most common manifestation. These presentations may be difficult to distinguish from toxoplasmosis in individuals with HIV/AIDS. The overlap of HIV and Trypanosoma cruzi infection occurs not only in endemic areas but also in non-endemic areas of North America and Europe where the diagnosis may be even more difficult. The pathological features, diagnosis and the role of cytokines in the pathogenesis of the disease are discussed.     

Seroepidemiological study of human hydatidosis and trichinosis, with the indirect hemagglutination reaction, in San Juan de la Costa County. Osorno, X Region, Chile. 1990-1991]

San Juan de la Costa County (40 degrees 45' South lat., 73 degrees 19' West long.) is located in the Osorno province, South of Chile. Its population is 8,486 inhabitants. The basic economic activities are agriculture, cattle raising, timber production and manufacture of wood and coal. According to official reports, the incidence of human hydatidosis and trichinosis in this locality in 1989 were 24 and 59 per 100,000 respectively. In order to contribute to a better knowledge of the epidemiology of human hydatidosis and trichinosis in San Juan de la Costa County, an indirect hemagglutination test (IHAT) for these parasitoses was performed to 511 randomized people. Nine (1.8%) individuals resulted positive for hydatidosis and twenty four (4.7%) were positive for trichinosis. Some considerations on the corresponding prophylactic measures are proposed.     

HIV antibodies in beggar blood donors in Rio de Janeiro, Brazil

The prevalence of HIV antibodies, as well as evidence of hepatitis B, syphilis, and Chagas' disease, was tested in 87 male and 13 female clients of a church-funded medical clinic in Rio de Janeiro who often donated blood to commercial blood banks. 5 individuals were seropositive for HIV, 2 homosexuals, 1 bisexual, and 2 heterosexuals. 21 had evidence of hepatitis B, including 2 with HBsag antibodies. 13 tested positive for syphilis, and 5 were positive for T. cruzi (Chagas' disease). The high incidence of positive tests for hepatitis B and Chagas' disease was possibly due to donation by plasmapheresis, which has been suspected to cause outbreaks of non-A, non-B hepatitis and malaria in this area. The practice of selling contaminated blood to unsuspecting recipients should be prevented no matter how high the cost.     

The prevalence of Toxoplasma gondii antibodies in wild and captive cetaceans from Japan

The seroprevalence of Toxoplasma gondii was investigated in wild and captive cetaceans from Japan. Antibodies against T. gondii were examined by both latex agglutination test (LAT) and indirect hemagglutination test (IHAT) for 77 serum or plasma samples obtained from 59 individuals of 6 species, including 2 hybrids. Antibody titers greater than 1:64 in LAT and greater than 1:640 in IHAT, indicative of the presence of T. gondii, were found in 11.9% of 59 individuals. In 7 samples that showed a positive reaction by IHAT, T. gondii titers were examined for each immunoglobulin (Ig) fraction separated by sucrose gradient centrifugation. The antibody peaks in each fraction were divided into 3 types, thought to be a reaction of IgM (type 1), IgG (type 2), and IgM with IgG (type 3). Type 1 was found in serum from a bottle-nosed dolphin (Tursiops truncatus) and a killer whale (Orcinus orca) sampled soon after capture off the Japanese coast in 1988; it was concluded that infection in the wild had occurred less than 15 yr before the study was performed. The prevalence of putative IgM and IgG antibodies from a captive-bred T. truncatus suggested that T. gondii infection also occurred in the aquarium.     

Immunological and structural characterization of an epitope from the Trypanosoma cruzi KMP-11 protein

The K1 peptide is an HLA-A*0201-restricted cytotoxic epitope derived from the Trypanosoma cruzi KMP-11 protein, this being the etiological agent of Chagas' disease. This work describes the K1 peptide's secondary structure and its recognition by sera from chagasic patients. Circular dichroism and NMR spectroscopy analysis revealed that the K1 peptide adopts an alpha-helical conformation. Fifty-six percent of individuals had anti-K1 and 86% anti-KMP-11 antibodies by ELISA in the chronic Chagas' group and 28 and 68% in the indeterminate Chagas' group, respectively. By contrast, no reactivity was observed in sera from healthy individuals and tuberculosis patients. Antibody response subclass specificity to the K1 peptide was IgG1 and IgG3. Taken together these results support the idea that the K1 peptide acts as a B-cell-inducer epitope during Chagas' disease.     

Ocular toxoplasmosis: evaluation of lacrimal-specific secretory IgA levels in both patients with active and inactive phases of the disease

Ocular toxoplasmosis can result in recurrent uveitis. Studies have shown that a correlation between active ocular toxoplasmosis and the presence of anti-Toxoplasma gondii secretory IgA (SIgA) in tears. This study compares anti-T. gondii SIgA levels in patients' tears during the acute and inactive phases of toxoplasmic uveitis. Twenty-nine positive tear specific SIgA for T. gondii patients with acute toxoplasmic uveitis were selected and were followed-up for at least two years, when the anti-T. gondii SIgA tears levels were determined. Specific SIgA for T. gondii was negative in 22 patients (75.86%) and positive in seven patients (24.13%) of whom six (85.7%) were followed over three years. Average SIgA levels during the acute phase are 1.54 and decrease significantly to 0.72 (p = 0.0001) during the inactive phase of disease. Because anti-T. gondii SIgA in the tear is negative in 75.86% of patients after the acute phase of infection, T. gondii SIgA levels may be used as a complementary diagnostic marker for active ocular toxoplasmosis.     

Usefulness of the ELISA-IgG test in sera and filter paper blood eluates in the Chagas disease immunodiagnosis]

An ELISA-IgG test using a soluble extract of Trypanosoma cruzi antigen has been developed. One hundred and seventy eight sera (74 healthy people, 44 chagasic patients with positive xenodiagnosis and 60 with clinical symptoms), and 64 paper filter blood eluates (35 healthy people, 10 chagasic patients and 19 with clinical symptoms) were tested by ELISA-IgG and an indirect hemagglutination test. Both types of methods presented an overall average concordance of 98.3% and 100.0% with sera and filter paper blood eluates respectively. A cut off (average of optical density plus 3 standard deviations) of 0.365 and 0.379 for sera and eluates was found. The use of ELISA-IgG in Chagas' disease clinical diagnosis and seroepidemiological studies is discussed.     

High correlation between Chagas' disease serology and PCR-based detection of Trypanosoma cruzi kinetoplast DNA in Bolivian children living in an endemic area

Abstract The detection of Trypanosoma cruzi kinetoplast DNA by polymerase chain reaction (PCR) amplification is a potentially powerful tool for the parasitological diagnosis of Chagas' disease. We have applied this technique in a field situation in Bolivia, where 45 children from a primary school were subjected to serological testing, buffy coat analysis and PCR diagnosis. 26 of the 28 serology-positive individuals were also positive by PCR. In addition, two serology-negative children gave a positive result by PCR, including one who was positive in the buffy coat test. These results suggest that PCR detection of T. cruzi DNA in blood can be a very useful complement to serology in Chagas' disease diagnosis in Bolivia.