High intestinal parasite infection detected in children from Región Autónoma Atlántico Norte (R.A.A.N.) of Nicaragua.

There is a lack of epidemiological information concerning intestinal parasitic infections, and especially in soil-transmitted helminths, occurring in some departments of Nicaragua. Up to now, this is the first study involving two nearby areas (Puerto Cabezas and Siuna municipalities) of the Región Autónoma Atlántico Norte (R.A.A.N.). One stool sample was analyzed by Kato-Katz, formaldehyde-ethyl acetate concentration method and modified Ziehl-Neelsen technique, and a simple questionnaire concerning demographic, sanitary and behavioral data was distributed among 735 children and evaluated. Overall prevalence of infection reached 97.0%, being the highest prevalences detected in all Nicaragua. The higher protozoan prevalence appears in Siuna (94.5%), a rural interior municipality, with a typical tropical monsoon climate, while the higher helminths rates were reached in Puerto Cabezas (92.8%), the urbanized coastal capital, with a typical tropical rainforest climate. No statistical differences were found with regard to sex. However, the 6-11-year age-group children presented the highest prevalences. Most T. trichiura infections (59.4%) were of light intensity, while 51.7% of Ascaris lumbricoides were of moderate intensity. Multivariable logistic regression analysis indicated that those who drink rainwater and walk barefoot were 2.9 and 2.5 times more likely to have helminth infections, respectively. Results from one geographical setting might not be applied to other nearby with different climatic conditions. The use of anthelmintic drugs only will not be sufficient to bring prevalence to low levels. It is necessary to design geographically more specific intervention, with communication and interaction between different disciplines (e.g. parasitology, biochemistry, molecular biology, epidemiology, public health, etc.) being imperative to reduce STH infection.

Anemia and undernutrition in intestinally parasitized schoolchildren from Gakenke district, Northern Province of Rwanda.

BACKGROUND: Rwanda is a sub-Saharan country, where intestinal parasite infections, anemia and undernutrition coexist. The purpose of this research is to study the relationship between intestinal parasite infections and undernutrition/anemia to clarify the priorities of intervention in the rural area of Gakenke district in the Northern Province of Rwanda. MATERIALS AND METHODS: A total of 674 students from Nemba I School, participated in a cross-sectional study, in which their parasitological and nutritional status were analysed. Statistical analysis was performed by χ2 test, univariate analysis and Odds ratios (OR). RESULTS: A total of 95.3% of children presented intestinal parasitism, most of whom (94.5%) infected by protozoa and 36.1% infected by soil-transmitted helminths (STH), with Trichuris trichiura (27.3%) being the most prevalent. Multiple infections were found to be high (83.8%), with protozoa and STH co-infections in 30.6%. STH infections were mainly of low/moderate intensity. Neither infection nor STH infection of any intensity profile, was significantly related to anemia. In addition, STH infection, regardless of the intensity profile, was not associated with stunting, underweight or thinness. There was no difference between genders nor among ages in odds of anemia and nutritional status in STH-infected schoolchildren. CONCLUSION: Multiparasitism remains high among Rwandan schoolchildren and is likely to cause nutritional problems. This work emphasizes the importance of keeping up health programs to reduce the prevalence of infection.

Evidence for Transmission of Taenia solium Taeniasis/Cysticercosis in a Rural Area of Northern Rwanda.

Cysticercosis is a parasitic infection caused by the metacestode larval stage (cysticercus) of Taenia solium. In humans, cysticercosis may infect the central nervous system and cause neurocysticercosis, which is responsible for over 50,000 deaths per year worldwide and is the major cause of preventable epilepsy cases, especially in low-income countries. Cysticercosis infection is endemic in many less developed countries where poor hygiene conditions and free-range pig management favor their transmission. A cross-sectional study was conducted in 680 children from a rural primary school in Gakenke district (Northern province of Rwanda). Stool samples were collected from participants and analyzed using the Kato-Katz method (KK), formol-ether concentration (FEC), and/or copro-antigen enzyme-linked immunosorbent assay (CoAg-ELISA) to detect taeniasis. Blood samples were collected and analyzed using enzyme-linked immunoelectrotransfer blot (EITB) and antigen enzyme-linked immunosorbent assay (Ag-ELISA) to detect human cysticercosis. The overall proportion of taeniasis positivity was 0.3% (2/680), and both cases were also confirmed by CoAg-ELISA. A total of 13.3% (76/572) of the children studied were positive to cysticercosis (T. solium-specific serum antibodies detected by EITB), of whom 38.0% (27/71) had viable cysticercus (T. solium antigens by Ag-ELISA). This study provides evidence of the highest cysticercosis prevalence reported in Rwanda in children to date. Systematic investigations into porcine and human cysticercosis as well as health education and hygiene measures for T. solium control are needed in Gakenke district.

Reduced prevalence of soil-transmitted helminths and high frequency of protozoan infections in the surrounding urban area of Curitiba, Paraná, Brazil.

Human populations living in the surrounding urban areas of large Brazilian cities have increased vulnerability to intestinal parasites. However, the epidemiological scenario of soil-transmitted helminths (STH) in Curitiba, Paraná's main city, remains largely unknown. To bridge this gap of knowledge, this study aims to determine the prevalence of intestinal parasites and to investigate potential transmission pathways of the most prevalent species detected. We conducted a cross-sectional epidemiological study between July and September 2014 among schoolchildren in urban and peri-urban (deprived) areas of the municipality of Campo do Tenente, Curitiba. A total of 549 stool samples were used for coproparasitological diagnosis. Microscopy-positive samples of the most common species found were re-assessed by PCR and sequencing methods at the small subunit rRNA gene. Prevalence of infection by any given enteroparasite was 24.8%, but soil-transmitted helminths were only detected in 3.5% of the examined samples. Frequency of protozoan infections reached 90% and 97.8% in single and multiple infections, respectively. Blastocystis sp. (38.9%) was the most frequently species found in the surveyed schoolchildren population. A total of 41 Blastocystis-positive samples were unambiguously typed as ST1 (36.4%), ST2 (21.2%), ST3 (39.4%), and ST1 + ST3 mixed infection (3.0%). These results indicate that Blastocystis transmission is primarily anthroponotic in origin. This data highlights the importance of maintaining the anthelminthic control programs currently in place and of improving sanitary disposal of human excreta in poor-resource settings.

Soil-Transmitted Helminth Infections and Anemia in Schoolchildren from Corn Island Archipelago (RAAS, Nicaragua).

The prevalence and intensity of soil-transmitted helminth (STH) infections in schoolchildren from Corn Islands (Nicaragua) were examined to detect mono- or poly-STH infected children, measuring different intensity levels, and to elucidate measurably increased odds of being anemic. A total of 341 stool samples provided by 2- to 15-year-old children were examined using a concentration technique and a Kato-Katz slide. Intensity of infection was expressed as eggs per gram (epg) of feces to classify light, moderate, or heavy intensity infection. A finger-prick blood sample was obtained from each student in the field. Soil-transmitted helminth prevalence was 54.3%, with Trichuris trichiura as the most prevalent species (48.9%). The combination T. trichiura/Ascaris lumbricoides (12.6%) was the most common. When T. trichiura or A. lumbricoides appeared as a single infection, light or moderate intensity infections were seen, whereas when multiple species were identified, heavy infections were present. Anemia was detected in those with any kind of STH infection (42.7%), with statistically significant differences (P = 0.004) when compared with uninfected individuals (28.2%). Polyparasite infection with one parasite species at moderate intensity and the other parasite species at light intensity or absent was found to be a significant factor for the odds of being anemic (odds ratio = 2.07). The present study reveals a high level of STH transmission requiring a deworming control program in Corn Islands and pointing to the need of improving the education and sanitary conditions of the population to avoid environmental contamination and reinfection.

Enteroparasites in Preschool Children on the Pacific Region of Nicaragua.

The aim of this study was to determine for the first time the prevalence of enteroparasites in preschool children originating from the seven departments of the Pacific region in Nicaragua. One stool sample of each of 1,217 children, from 6 months to 5 years of age, was collected and personal data were recorded on delivery of the container. Samples fixed in 10% formalin were processed by a formol-acetate concentration and a modified Ziehl-Neelsen technique. The overall prevalence of enteroparasite infections was 68.2% with a total of at least 20 species. Blastocystis hominis (45.5%), Giardia intestinalis (31.7%), Trichuris trichiura (8.2%), and Ascaris lumbricoides (5.2%) were the most prevalent protozoa and helminth species in the total study as well as in all departments. Protozoan prevalence presented a statistically significant difference by gender (male: 69.6%; female: 46.7%; P < 0.001), and males presented a higher T. trichiura infection rate than females (male: 9.9%; female 6.4%; P < 0.035). Protozoan prevalence increased with age with a statistically significant difference (P < 0.001). Helminths were always more prevalent in urban areas (P < 0.0002). Protozoan infections result statistically higher than helminth infections so that water-based transmission could be suspected. Based on the differences with respect to species and parasite prevalence between the seven departments in the Pacific region of Nicaragua, the exploration of local factors associated with the transmission of enteroparasites should also be considered to reduce infection.

Intestinal symptoms and Blastocystis load in schoolchildren of Paranaguá Bay, Paraná, Brazil.

The symptomatology of Blastocystis cannot be attributed to any particular subtype, although can be related to a high Blastocystis infection load. One stool sample from each of 217 schoolchildren of Vale de Sol Paranaguá Bay (Paraná, Brazil) was collected. Three milliliters of each fixed stool sample were processed applying the formalin-ether concentration technique. After obtaining the overall prevalence of intestinal parasites, quantification was carried out in Blastocystis positive samples. A total of 75/217 (34.6%) children suffered from intestinal symptoms (abdominal pain and/or persistent diarrhea), of whom 41.3% (31/75) presented moderate/heavy Blastocystis load with a statistically significant risk to present intestinal symptoms (OR 0.039 [0.006-0.15]; p<0.001) Moreover, those symptomatic schoolchildren monoparasitized only by Blastocystis (10/75, 13.3%), and those polyparasitized by Blastocystis with other non-pathogenic species (15/75, 20%) with moderate/heavy loads, also entail a statistically significant risk of intestinal symptomatology, both in monoparasitism (12%, OR 0.10 [0.004-0.63]; p=0.021) and in polyparasitism with a non-pathogenic parasite (18.6%, OR 0.059 [0.002-0.35]; p=0.001). For the first time in Brazil, using data from schoolchildren of Paranaguá Bay, we demonstrated that moderate/ heavy loads of Blastocystis could be related to intestinal symptoms.

Association between enteric protozoan parasites and gastrointestinal illness among HIV- and tuberculosis-infected individuals in the Chowke district, southern Mozambique.

Human immune deficiency virus (HIV) and tuberculosis (TB) infections remain major public health issues globally, particularly in sub-Saharan Africa. Impairment of both cell-mediated and humoral immunity by HIV and/or TB infections may limit the host's defences against other pathogens, including the diarrheagenic protozoan Cryptosporidium spp., Giardia intestinalis, and Entamoeba histolytica. During September-December 2015 a cross-sectional study was conducted to assess the prevalence and molecular diversity of these enteric parasites among HIV- and/or TB-infected patients at a medical reference centre in Chowke district, southern Mozambique. A total of 99 stool specimens were initially screened by direct microscopy and further confirmed and characterised by molecular methods. DNA sequence analyses of the genes encoding the small subunit ribosomal RNA and the 60-kDa glycoprotein were used for the typing and sub-typing of Cryptosporidium isolates, respectively. G. intestinalis-positive isolates by real-time PCR were subsequently typed at the glutamate dehydrogenase locus. Differential diagnosis of E. histolytica/dispar was achieved by real-time PCR. G. intestinalis (8.1%) was the enteric protozoan more frequently detected, followed by Cryptosporidium spp. (7.1%), and Entamoeba histolytica/dispar (6.1%). Two HIV-infected (but not TB-infected) patients harbour G. intestinalis and Cryptosporidium spp. co-infections. Two (29%) G. intestinalis isolates were successfully characterised, revealing the presence of known AII and novel BIV genotypes. Four (57%) Cryptosporidium isolates were unmistakeable assigned to C. hominis, identifying two (IbA10G2 and IdA22) sub-types. Cryptosporidium infections were not associated to diarrhoea in HIV-positive patients, probably because improved immune function in the affected individuals due to antiretroviral therapy. G. intestinalis was considered a non-opportunistic pathogen, whereas the presence of E. histolytica could not be confirmed by molecular methods. Based on their common presence in the studied clinical population, we recommend the effective diagnosis and treatment of these enteropathogens for improving the management of HIV and TB patients.

Hookworm-like eggs in children's faecal samples from a rural area of Rwanda.

BACKGROUND: Hookworm eggs identification and quantification is usually carried out by Kato-Katz method. However various structures present in the smear may be confused with eggs of such parasites. OBJECTIVE: To document the presence of structures in Kato-Katz slides that could initially be misinterpreted as hookworm eggs. METHOD: 497 faecal samples were analysed by Kato-Katz technique, diphasic concentration technique, agar-plate coprocultive and larvae obtained were analysed by PCR and characterized by sequencing. RESULT: Hookworm-like eggs were found in 159 (32%) of the samples by Kato-Katz, finally identified as Caenorhabditis elegans by PCR technique. CONCLUSION: The diagnosis of human hookworm eggs, only by the use of Kato-Katz technique can lead to false positives because of similarities with eggs of other free-living worms, from wet soils like those of Rwanda that could contaminate stool samples.

Strongyloidiasis with emphasis on human infections and its different clinical forms.

Strongyloidiasis (caused by Strongyloides stercoralis, and to a lesser extent by Strongyloides fuelleborni) is one of the most neglected tropical diseases with endemic areas and affecting more than 100 million people worldwide. Chronic infections in endemic areas can be maintained for decades through the autoinfective cycle with the L3 filariform larvae. In these endemic areas, misdiagnosis, inadequate treatment and the facilitation of the hyperinfection syndrome by immunosuppression are frequent and contribute to a high mortality rate. Despite the serious health impact of strongyloidiasis, it is a neglected disease and very little is known about this parasite and the disease when compared to other helminth infections. Control of the disease is difficult because of the many gaps in our knowledge of strongyloidiasis. We examine the recent literature on different aspects of strongyloidiasis with emphasis in those aspects that need further research.

Analysis of the tegument of Zygocotyle lunata (Trematoda: Paramphistomidae) adults by scanning electron microscopy.

Paramphistomiasis is a parasitic disease caused by various species of the Paramphistomidae. These species mainly affect domestic and wild ruminants; the economic impact of these diseases is often underestimated. Traditionally, the identification of paramphistomes has been difficult and has been based on morphological aspects such as the body shape, the position of the esophagus and cecae, or the tegumental papillae. Despite the many investigations regarding the tegumental papillae for other paramphistomes, very few efforts have been made using Zygocotyle lunata, partly because many authors differ with respect to the importance of papillae in the classification of paramphistomids. Herein, we characterize by scanning electron microscopy new tegumental papillae not previously described on the tegument of 3-wk-old Z. lunata adults obtained from mice. Three morphologically different papillae (rosette, ciliated, and conical papillae) were observed and are described. Based on these results, we assert that the newly recognized morphological features should be used for species differentiation in the future in addition to the new molecular techniques.

Hyperendemic human fascioliasis in Andean valleys: an altitudinal transect analysis in children of Cajamarca province, Peru.

A coprological survey including 476 2-18 year old school children from six rural localities between 2627 and 3061 m altitude was performed in Cajamarca province, Peru. Prevalences of fascioliasis ranging from 6.7 to 47.7% (mean 24.4%) proved to be the highest so far recorded in that human hyperendemic area. Higher prevalences in females and in the 2-5 year old group were not significant. Intensities ranged from 24 to 864 eggs per gram (arithmetic mean: 113; geometric mean: 68), the majority shedding less than 100, and without significant differences according to gender or age group. Fasciola hepatica was the most common helminth within a spectrum of 11-12 protozoan and 9-11 helminth species, 97.3% of the children showing infection with at least one parasite. The highest levels corresponded to coinfection with seven different species in females and subjects older than 5 years. Fascioliasis prevalence correlation with altitude appeared significant. An epidemiological characterisation of the valley transmission pattern of fascioliasis in Cajamarca is made by comparison with other better known hyperendemic areas. Results suggest that human fascioliasis may be widespread throughout different parts of Cajamarca province, even far away from the city, and that long-term fascioliasis chronicity and superimposed repetitive infections may be probably frequent.

Chapter 3. Recent advances in the biology of echinostomes.

This chapter examines the significant literature on the biology of echinostomes. The members of the family Echinostomatidae are medically and veterinary-important parasitic flatworms that invade humans, domestic animals and wildlife and also parasitize in their larval stages numerous invertebrate and cold-blooded vertebrate hosts. All echinostomes possess a complicated lifecycle expressed by: (i) alternation of seven generations known as the adult, egg, miracidium, sporocyst, redia, cercaria and metacercaria, and (ii) inclusion of three host categories known as the definitive host and first and second intermediate hosts. Moreover, echinostomes have served as experimental models in parasitology at all levels of organization. We discuss recent advances in several areas of the biological sciences that feature studies on echinostomes. Initially, we consider aspects of the lifecycle, development and systematics of selected members of the Echinostomatidae. We then highlight host-parasite interactions between echinostomes and their intermediate and definitive hosts with emphasis on the application of novel techniques to these topics.

Euparyphium albuferensis and Echinostoma friedi (Trematoda: Echinostomatidae): experimental cercarial transmission success in sympatric snail communities.

Euparyphium albuferensis and Echinostoma friedi cercarial infectivity to four species of sympatric snails was examined under single- or multiple-choice laboratory conditions to show the level of parasite-snail host compatibility. Radix peregra, Lymnaeafuscus, Physella acuta and Gyraulus chinensis act as second intermediate hosts of both parasite species although different cercarial transmission success (CTS) was observed. In single-host experiments, R. peregra and P. acuta showed a high degree of compatibility with E. albuferensis, while only P. acuta in the case of E. friedi. In two-choice snail communities, a snail with high CTS increased the values of another with low compatibility, in both parasite species. In multiple-choice snail communities, high CTS of some hosts decreased, while low CTS of other hosts increased. The degree of parasite-host compatibility of each snail species could be determined by the presence of other snails in the community.

Effect of Echinostoma friedi (Trematoda: Echinostomatidae) experimental infection on longevity, growth and fecundity of juvenile Radix peregra (Gastropoda: Lymnaeidae) and Biomphalaria glabrata (Gastropoda: Planorbidae) snails.

The effect of Echinostoma friedi experimental infection on longevity, growth and fecundity of two susceptible first intermediate host snails, Radix peregra and Biomphalaria glabrata, was studied to contrast the level of compatibility. 120 R. peregra and 150 B. glabrata snails were used exposed to one, three or five miracidia and divided in three categories: INF (snails exposed and infected); ENI (exposed but not infected) and C (control or not miracidial-exposed snails). R. peregra INF snails' death process starts sooner, but in a prolonged extension, while B. glabrata INF snails have a much shorter life span. The infection and the miracidial exposure are able to reduce R. peregra normal development (stunting). B. glabrata INF snails' growth exceeds that of C snails (gigantism). E. friedi produces a total parasitic castration of R. peregra and B. glabrata INF snails. R. peregra would be considered as the required snail host, while B. glabrata only as an adequate snail host.

Echinostoma caproni: identification of enolase in excretory/secretory products, molecular cloning, and functional expression.

In order to investigate molecules that could be involved in host-trematode relationships, we have analysed the excretory/secretory products (ESP) of Echinostoma caproni following a proteomic approach. Actin, Gluthathione S-transferase (GST) and enolase have been identified in the ESP. Enolase, observed to be one of the most abundant proteins, was further characterized. The molecular cloning and in vitro expression in Escherichia coli of E. caproni enolase allowed us to determine that the protein contains 431 amino acids and a theoretical MW of 46272 Da. E. caproni enolase shows high homology to other trematode enolases. The recombinant protein binds specifically to human plasminogen in vitro, as observed for the native protein, confirming its properties as a host-interacting molecule.