Genotypic and Epidemiologic Profiles of Giardia duodenalis in Four Brazilian Biogeographic Regions

Human infections with gut protozoan parasites are neglected and not targeted by specific control initiatives, leading to a knowledge gap concerning their regional diversity and epidemiology. The present study aims to explore Giardia duodenalis genetic diversity and assess the epidemiologic scenario of subclinical infections in different Brazilian biogeographic regions. Cross-sectional surveys (n = 1334 subjects) were conducted in four municipalities in order to obtain fecal samples and socioenvironmental data. Microscopy of non-diarrheal feces and nucleotide sequencing of a ß-giardin gene fragment were performed. From a total of 51 samples that could be sequenced, 27 (52.9%) ß-giardin sequences were characterized as assemblage A and 24 (47.1%) as assemblage B. In the Amazon, assemblage B was the most frequently detected, predominantly BIII, and with two novel sub-assemblages. Assemblage A predominated in the extra-Amazon region, with five novel sub-assemblages. Prevalence reached 17.8% (64/360) in the Amazon, 8.8% (48/544) in the Atlantic Forest, 7.4% (22/299) in Cerrado and 2.3% (3/131) in the Semiarid. People living in poverty and extreme poverty presented significantly higher positivity rates. In conclusion, subclinical giardiasis is endemic in Brazilian communities in different biogeographic regions, presenting high genetic diversity and a heterogeneous genotypic distribution.

Molecular detection of Metastrongylus salmi eggs from pigs in low-resource communities in the state of Piauí, northeastern Brazil.

Metastrongylosis is an infection of the respiratory tract of pigs caused by parasites of the genus Metastrongylus, whose eggs are similar to other Strongylida through light microscopy; species-specific identification can be performed with molecular tools. We explored the species composition and the genetic diversity of Metastrongylus infecting pigs in close contact with humans in impoverished rural communities in the state of Piauí, in northeastern Brazil. Fecal samples (n = 78) were collected for parasitologic tests. Egg morphometry and molecular characterization, using the cytochrome c oxidase subunit 1 (cox1) gene, were performed. For strongyliform eggs, 62 of 78 (80%) pigs were positive and 6 of 99 (6%) eggs had dimensions compatible with Metastrongylus. Of the 37 samples submitted to PCR, 10 were identified as M. salmi. We found 3 M. salmi haplotypes, including 2 new and 1 described previously in Europe. Overall, M. salmi demonstrated lower intraspecific genetic diversity: diversity index (H) ± SD = 0.318 ± 0.164, n = 12, compared with published M. pudendotectus sequences (1.000 ± 0.272, n = 3). To our knowledge, M. salmi DNA sequences have not been published previously from pigs in South America.

A community-based, cross-sectional study to assess interactions between income, nutritional status and enteric parasitism in two Brazilian cities: are we moving positively towards 2030?

BACKGROUND: This study assessed the interactions between income, nutritional status and intestinal parasitism in children in Brazil. METHODS: A cross-sectional study (n = 421 children aged 1 to 14 years living in the states of Piauí (rural communities in the city of Teresina) and Rio de Janeiro (rural and periurban communities in the city of Cachoeiras de Macacu) was performed in order to obtain income and anthropometric data, as well as fecal samples for parasitological analyses through the Ritchie technique. RESULTS: Children infected with Ascaris lumbricoides had significantly lower means of height-for-age z scores (- 1.36 ± 0.75 vs. - 0.11 ± 1.02; p < 0.001), weight-for-age z scores (- 1.23 ± 0.74 vs. 0.09 ± 1.15; p = 0.001), and weight-for-height z scores (- 0.68 ± 0.44 vs. 0.23 ± 1.25; p = 0.006) when compared with uninfected children. Infection with hookworm was also associated with lower means of height-for-age z scores (- 1.08 ± 1.17 vs. - 0.12 ± 1.02; p = 0.015) and weight-for-age z scores (- 1.03 ± 1.13 vs. 0.08 ± 1.15; p = 0.012). Children infected with Entamoeba coli presented significantly lower means of height-for-age z scores (- 0.54 ± 1.02 vs. - 0.09 ± 1.02; p = 0.005) and weight-for-age z scores (- 0.44 ± 1.15 vs. 0.12 ± 1.15; p = 0.002). The multivariate multiple linear regression analysis showed that height-for-age z scores are independently influenced by monthly per capita family income (ß = 0.145; p = 0.003), female gender (ß = 0.117; p = 0.015), and infections with A. lumbricoides (ß = - 0.141; p = 0.006) and Entamoeba coli (ß = - 0.100; p = 0.043). Weight-for-age z scores are influenced by monthly per capita family income (ß = 0.175; p < 0.001), female gender (ß = 0.123; p = 0.010), and infections with A. lumbricoides (ß = - 0.127; p = 0.012), and Entamoeba coli (ß = - 0.101; p = 0.039). Monthly per capita family income (ß = 0.102; p = 0.039) and female gender (ß = 0.134; p = 0.007) positively influences mid upper arm circumpherence. CONCLUSIONS: Intestinal parasitism and low family income negatively influence the physical development of children in low-income communities in different Brazilian regions.

Benznidazole decreases the risk of chronic Chagas disease progression and cardiovascular events: A long-term follow up study.

BACKGROUND: Chagas disease (CD) remains an important endemic disease in Latin America. However, CD became globalized in recent decades. The majority of the chronically infected individuals did not receive etiologic treatment for several reasons, among them the most conspicuous is the lack of access to diagnosis. The impact of trypanocidal treatment on CD chronic phase, without cardiac involvement (indeterminate form ICF), is yet to be determined. We aimed to evaluate the effect of trypanocidal treatment with benznidazole (BZN) on the rate of progression to Chagas heart disease in patients with ICF. METHODS: This is a retrospective cohort observational study including patients with ICF treated with BZN and compared to a group of non-treated patients matched for age, sex, region of origin, and the year of cohort entry. We reviewed the medical charts of all patients followed from May 1987 to June 2020 at the outpatient center of the Evandro Chagas National Institute of Infectious Diseases (INI) of the Oswaldo Cruz Foundation (Fiocruz), Rio de Janeiro, Brazil. Patients' follow-up included at least one annual medical visit and one annual electrocardiogram (ECG). Echocardiographic exams were performed at baseline and during the follow-up. Disease progression from ICF to cardiac form was defined by changes in baseline ECG. Cumulative incidence and the incidence rate were described in the incidence analysis. Cox proportional hazards models were used to estimate hazard ratios and 95% confidence intervals for the association between BZN and CD progression, cardiovascular events or death. FINDINGS: One hundred and fourteen treated patients met the study inclusion criteria. A comparison group of 114 non-treated patients matched for age, sex, region of origin, and the year of cohort entry was also included, totalizing 228 patients. Most patients included in the study were male (70.2%), and their mean age was 31.3 (+7.4) years. Over a median follow-up of 15.1 years (ranging from 1.0 to 32.4), the cumulative CD progression incidence in treated patients was 7.9% vs. 21.1% in the non-treated group (p = 0.04) and the CD progression rate was 0.49 per 1.000 patients/year in treated patients vs. 1.10 per 1.000 patients/year for non-treated patients (p = 0.02). BZN treatment was associated with a decreased risk of CD progression in both unadjusted (HR 0.46; 95%CI 0.21 to 0.98) and adjusted (HR 0.43; 95%CI 0.19 to 0.96) models and with a decreased risk of occurrence of the composite of cardiovascular events only in the adjusted (HR 0.15; 95%CI 0.03 to 0.80) model. No association was observed between BZN treatment and mortality. INTERPRETATION: In a long-term follow-up, BZN treatment was associated with a decreased incidence of CD progression from ICF to the cardiac form and also with a decreased risk of cardiovascular events. Therefore, our results indicate that BZN treatment for CD patients with ICF should be implemented into clinical practice.

Infections with Giardia duodenalis and Entamoeba histolytica/Entamoeba dispar as Hidden and Prevalent Conditions in Periurban Communities in the State of Rio de Janeiro, Brazil.

This study aims to assess the prevalence, distribution, and etiological profile of intestinal parasitism in children living in periurban areas in Cachoeiras de Macacu, Rio de Janeiro, Brazil. A community-based cross-sectional survey (n = 479) was carried out. Prevalence of infection with G. duodenalis and E. histolytica/E. dispar was 8.6% (n = 41) and 13.4% (n = 64), respectively. Infection with G. duodenalis was significantly more frequent among children living in poor families (24/187 (12.8%) vs. 16/272 (5.9%); prevalence ratio (PR) = 2.18; 95% confidence interval (CI) = 1.19-3.99; p=0.011). This difference was also significant for infection with any pathogenic parasite (43/187 (23%) vs. 40/272 (14/7%); PR = 1.56; 95% CI = 1.06-2.30; p=0.026). In addition, people residing in houses with more than four inhabitants showed significantly higher positivity for infections with G. duodenalis and with E. histolytica/E. dispar (22/138 (15.9%) vs. 16/311 (5.1%); PR = 3.09; 95% CI = 1.68-5.71; p < 0.001 for G. duodenalis and 32/138 (23.2%) vs. 30/311 (9.6%); PR = 2.40; 95% CI = 1.52-3.79; p < 0.001 for E. histolytica/E. dispar). Laboratory diagnosis of protozoan enteric infections and effective drugs for their treatment are unmet goals in the primary health care system. Therefore, giardiasis and amebiasis are neglected conditions.

Genetic diversity of Ascaris spp. infecting humans and pigs in distinct Brazilian regions, as revealed by mitochondrial DNA.

In this study, we assessed the genetic diversity of Ascaris lumbricoides / Ascaris suum circulating in humans and pigs, exploring potential zoonotic cycles in endemic areas in Brazil. We carried out cross-sectional surveys in four municipalities: Santa Isabel do Rio Negro (SIRN-AM) (n = 328); Nossa Senhora de Nazaré (NSN-PI) and Teresina (TER-PI) (n = 605 and n = 297, respectively); and Cachoeiras de Macacu (CAM-RJ) (n = 543). We also studied 61 fecal samples/adult worms obtained from pigs (n = 53 in NSN-PI and n = 8 in TER-PI). A ~450 bp fragment of the Ascaris cytochrome c oxidase subunit 1 (cox1) and ~400 bp of the NADH dehydrogenase subunit 1 (nad1) were amplified and sequenced. Maximum-likelihood (ML) tree and Median-joining (MJ) haplotype network analyses were performed. We also performed scanning electron micrographs of adult specimens. Positivity rates were 93/328 (28.4%) in SIRN-AM, 6/297 (2.0%) in TER-PI, 0/605 (0%) in NSN-PI, and 6/543 (1.1%) in CAM-RJ. In NSN-PI it reached 11/53 (20.7%) in pigs. The MJ network based on cox1 locus (383 bp) revealed three main clusters, one centered around haplotypes H01/H28/H32 and the other around H07/H11. The cox1 haplotypes had a heterogeneous distribution, showing no pattern by geographic region, and high haplotype diversity. The ML trees based on cox1 and nad1 loci showed a similar topology with each other, and with the haplotype networks. Three distinct clusters were observed. Sequences of cox1 and nad1 from humans and animals were distributed throughout the tree and it was not possible to differentiate specimens of human and swine origin. Ascaris populations obtained from humans and swine in different Brazilian regions are not discriminable through the genetic markers used, which indicates the potential for zoonotic transmission and the need for better control of these infections in swine herds, mainly when created in a peridomestic environment.

Spatial and Molecular Epidemiology of Giardia intestinalis Deep in the Amazon, Brazil.

BACKGROUND: Current control policies for intestinal parasitosis focuses on soil-transmitted helminths, being ineffective against Giardia intestinalis, a highly prevalent protozoon that impacts children's nutritional status in developing countries. The objective of this study was to explore spatial and molecular epidemiology of Giardia intestinalis in children of Amerindian descent in the Brazilian Amazon. METHODOLOGY/PRINCIPAL FINDINGS: A cross sectional survey was performed in the Brazilian Amazon with 433 children aged 1 to 14 years. Fecal samples were processed through parasitological techniques and molecular characterization. Prevalence of G. intestinalis infection was 16.9% (73/433), reaching 22.2% (35/158) among children aged 2-5 years, and a wide distribution throughout the city with some hot spots. Positivity-rate was similar among children living in distinct socioeconomic strata (48/280 [17.1%] and 19/116 [16.4%] below and above the poverty line, respectively). Sequencing of the ß-giardin gene revealed 52.2% (n = 12) of assemblage A and 47.8% (n = 11) of assemblage B with high haplotype diversity for the latter. The isolates clustered into two well-supported G. intestinalis clades. A total of 38 haplotypes were obtained, with the following subassemblages distribution: 5.3% (n = 2) AII, 26.3% (n = 10) AIII, 7.9% (n = 3) BIII, and 60.5% (n = 23) new B genotypes not previously described. CONCLUSIONS/SIGNIFICANCE: Giardia intestinalis infection presents a high prevalence rate among Amerindian descended children living in Santa Isabel do Rio Negro/Amazon. The wide distribution observed in a small city suggests the presence of multiple sources of infection, which could be related to environmental contamination with feces, possibly of human and animal origin, highlighting the need of improving sanitation, safe water supply and access to diagnosis and adequate treatment of infections.

Investigation of Chagas disease in four periurban areas in northeastern Brazil: epidemiologic survey in man, vectors, non-human hosts and reservoirs.

Chagas disease was investigated in four periurban areas of Ceará state, northeastern Brazil, through serological, parasitological and molecular methods in humans, reservoirs and vectors. A cross-sectional survey revealed a seroprevalence rate of 1.2% (13/1076 residents, six also proving positive through PCR). Trypanosoma cruzi infection was not detected in children under 10 years old. Triatoma pseudomaculata prevailed in the peridomiciles: 63 specimens, 69% (34/49) infected with trypanosomatids. Rhodnius nasutus was captured in Copernicia prunifera palm trees (n=280; 25.0% infected with trypanosomatids) and inside dwellings (n=8, all uninfected). Trypanosoma cruzi seropositive reservoirs, represented by Didelphis albiventris (n=27), Rattus rattus (n=24), Thrichomys laurentius (n=2), Mus musculus (n=1) and Monodelphis domestica (n=1), were identified. Among domestic dogs (n=96) seroprevalence reached 21.9%. Miniexon multiplex PCR assays characterized TcI in triatomines. Both TcI and TcII were detected in wild mammal hosts. We conclude that Trypanosoma cruzi circulates within a domestic zoonotic cycle, requiring continuous surveillance. Insecticide application to domiciles does not appear to prevent continuous reintroduction of wild triatomine specimens, presenting a challenge to authorities involved in Chagas disease control.

Bartonella spp. infection in HIV positive individuals, their pets and ectoparasites in Rio de Janeiro, Brazil: serological and molecular study.

BACKGROUND: Bartonella is the agent of cat-scratch disease, but is also responsible for more severe conditions such as retinitis, meningoencephalitis, endocarditis and bacillary angiomatosis. Its seroprevalence is unknown in Brazil. METHODS: Patients in an AIDS clinic, asymptomatic at the time of the study, were enrolled prospectively. They answered a structured questionnaire and had blood taken for serological and molecular assays. Cat breeder's pets were tested serologically and collected ectoparasites were tested by molecular biology techniques. Blood donors, paired by age and sex, were tested for Bartonella IgG antibodies. RESULTS: 125 HIV positive patients with a median age of 34 were studied; 61 were male and 75% were on HAART. Mean most recent CD4 count was 351-500 cells/mm(3). A high rate of contact with ticks, fleas and lice was observed. Bartonella IgG seroreactivity rate was 38.4% in HIV positive individuals and breeding cats was closely associated with infection (OR 3.6, CI 1.1-11.9, p<0.05). No difference was found between the sexes. Titers were 1:32 in 39 patients, 1:64 in seven, 1:128 in one and 1:256 in one. In the control group, IgG seroreactivity to Bartonella spp. was 34%, and female sex was correlated to seropositivity. Fourteen of 61 (23%) males vs 29/64 (45.3%) females were seroreactive to Bartonella (OR 2.8, CI 1.2-6.5, p<0.01). Titers were 1:32 in 29 patients, 1:64 in ten and 1:128 in four. CONCLUSIONS: Bartonella spp. seroprevalence is high in HIV positive and in blood donors in Rio de Janeiro. This may be of public health relevance.

Pseudoparasitism by Calodium hepaticum (syn. Capillaria hepatica; Hepaticola hepatica) in the Negro River, Brazilian Amazon.

We report the finding of eggs of Calodium spp. (syn. Capillaria spp.; Hepaticola spp.) in a fecal sample from an old woman living in a riverine community in the Negro River Basin and describe the associated epidemiological investigation. The case probably does not represent true parasitism; the eggs, which were compatible with the species Calodium hepaticum, were most likely ingested upon consumption of infected tapir (Tapirus terrestris) liver, subsequently passing through the gut and being eliminated. The evolution of these eggs to infective stages in the environment, given the poor sanitation background, could provide the risk of occurrence of hepatic disease in humans.

Risk presented by Copernicia prunifera palm trees in the Rhodnius nasutus distribution in a Chagas disease-endemic area of the Brazilian northeast.

With the aid of live-bait traps, we studied the risk that Copernicia prunifera palm trees, present in both periurban and rural localities of an endemic Brazilian northeast Chagas disease region, represent to domestic infestation by Rhodnius nasutus. In this area, this important vector has been encountered harboring and transmitting Trypanosoma cruzi, the etiologic agent of the American trypanosomiasis, to mammals, possibly including humans. Results indicate that this bug colonizes C. prunifera palm trees of both regions, mainly in dry seasons, and is infected with high levels of T. cruzi. Although more triatomines were captured in rural areas, proportionally the number of infected bugs from peri-urban regions was much higher. Herein we address the epidemiologic implications and challenge for the Brazilian health authorities to control the disease in this region, where the native palm trees have been largely destroyed causing a severe disturbance in the environmental equilibrium.

Characterization of rickettsia rickettsii in a case of fatal Brazilian spotted fever in the city of Rio de Janeiro, Brazil.

A lethal case of Brazilian spotted fever (BSF) is presented. Clinical features were initially of gastrointestinal involvement and evolved with progression to septic shock, meningoencephalitis and death on the 6th day of illness. Indirect immunofluorescence assay (IFA) for spotted fever group rickettsia (SFGR) was non-reactive. Diagnosis was confirmed by the polymerase chain reaction (PCR) and the nucleotide sequencing of a fragment of the ompA gene showed 100% homology to Rickettsia rickettsii. BSF has not been reported in the city of Rio de Janeiro in the last three decades, and the present description should alert the clinicians to its presence in urban Rio de Janeiro, and to the differential diagnosis with dengue fever, gastroenteritis, leptospirosis and bacterial septic shock, among others.

Characterization of rickettsia rickettsii in a case of Fatal Brazilian spotted fever in the city of Rio de Janeiro, Brazil

A lethal case of Brazilian spotted fever (BSF) is presented. Clinical features were initially of gastrointestinal involvement and evolved with progression to septic shock, meningoencephalitis and death on the 6th day of illness. Indirect immunofluorescence assay (IFA) for spotted fever group rickettsia (SFGR) was non-reactive. Diagnosis was confirmed by the polymerase chain reaction (PCR) and the nucleotide sequencing of a fragment of the ompA gene showed 100 percent homology to Rickettsia rickettsii. BSF has not been reported in the city of Rio de Janeiro in the last three decades, and the present description should alert the clinicians to its presence in urban Rio de Janeiro, and to the differential diagnosis with dengue fever, gastroenteritis, leptospirosis and bacterial septic shock, among others.

Estudo das parasitoses intestinais e da infecção chagásica no Município de Novo Airão, Estado do Amazonas, Brasil / Cross-sectional study of intestinal parasites and Chagas' disease in the Municipality of Novo Airão, State of Amazonas, Brazil

Foi realizado um estudo seccional descritivo em uma amostra sistemática por conglomerado de uma em cada dez famílias residentes na sede do Município de Novo Airão, na meso-região norte do Estado do Amazonas, a 250 quilômetros de Manaus por via fluvial. De cada um dos residentes nos 89 domicílios da amostra foram solicitadas fezes para exame pelos métodos de sedimentação de Lutz e pelo Baermann-Moraes-Coutinho e colhido sangue por punção venosa para testes de ELISA e imunofluorescência para pesquisa de anticorpos anti-Trypanosoma cruzi. O exame de 316 amostras de fezes mostrou 87,6 por cento com um ou mais parasitos: 35,1 por cento com Ascaris lumbricoides, 29,1 por cento com Entamoeba histolytica, 17,4 por cento com Giardia lamblia e outros parasitos com menor prevalência. Estes resultados foram correlacionados com a falta de saneamento e de suprimento de água tratada. De 346 soros examinados 16 (4,6 por cento) foram reativos para anticorpos anti-Trypanosoma cruzi, mas somente em três deles houve correlação entre este resultado e o contacto das pessoas com triatomíneos silvestres, conhecidos localmente como "piolho da piaçava"