Aspirin treatment exacerbates oral infections by Trypanosoma cruzi.

Oral transmission of the protozoan parasite Trypanosoma cruzi, the etiological agent of Chagas disease, has been documented in Latin American countries. The reported cases of infection were due to the ingestion of contaminated fresh fruit, juices, or sugar cane juice. There have been few studies on the physiopathology of the disease in oral transmission cases. Gastritis is a common ailment that can be caused by poor dietary habits, intake of alcohol or other gastric irritants, bacterial infection, or by the widespread use of non-steroidal anti-inflammatory drugs (NSAIDs). This study investigated in a mouse model whether gastric mucosal injury, induced by aspirin, would affect the course of disease in animals infected with T. cruzi by the oral route. The CL14 and G strains of T. cruzi, both of low infectivity, were used. To this end, groups of BALB/c mice were treated during 5 days with aspirin (100 mg kg(-1)) before oral infection with T. cruzi metacyclic forms (4 × 10(5) or 5 × 10(7) parasites/mouse). Histological analysis and determination of nitric oxide and TNF-α were performed in gastric samples obtained 5 days after infection. Parasitemia was monitored from the thirteenth day after infection. The results indicate that aspirin treatment of mice injured their gastric mucosa and facilitated invasion by both CL14 and G strains of T. cruzi. Strain CL14 caused more severe infection compared to the G strain, as larger numbers of amastigote nests were found in the stomach and parasitemia levels were higher. Our study is novel in that it shows that gastric mucosal damage caused by aspirin, a commonly used NSAID, facilitates T. cruzi infection by the oral route.

[Oral transmission of Trypanosoma cruzi : a new epidemiological scenario for Chagas' disease in Colombia and other South American countries]. / Transmisión oral de Trypanosoma cruzi: una nueva situación epidemiológica de la enfermedad de Chagas en Colombia y otros países suramericanos.

Many cases of infection caused by the oral transmission of Trypanosoma cruzi have been reported during the last decade. These have been due to the contamination of food by faeces from sylvatic triatomines or by leakage from reservoirs in areas where domiciliated vectors have been controlled or where there has been no prior background of domiciliation. The United Nations Food and Agriculture Organization (FAO) and the World Health Organization (WHO) have used epidemiological, clinical and socioeconomic criteria for ranking parasites transmitted by the contamination of food in different areas of the world; T. cruzi was placed tenth in importance amongst a group of 24 parasites in such ranking. Environmental changes such as deforestation and global warming have affected ecotopes and the behaviour of T. cruzi vectors and reservoirs so that these have become displaced to new areas, thereby leading to such new transmission scenario caused by the contamination of food, which requires evaluation in Colombia. The current review deals with the oral transmission of Chagas' disease, emphasising studies aimed at identifying the pertinent risk factors, the triatomine species involved, the physiopathology of oral infection, the parasite's genotypes implicated in this type of transmission in Colombia and other Latin American regions, as well as the need for ongoing epidemiological surveillance and control policies.

Transmisión oral de Trypanosoma cruzi: una nueva situación epidemiológica de la enfermedad de Chagas en Colombia y otros países suramericanos / Oral transmission of Trypanosoma cruzi : a new epidemiological scenario for Chagas´ disease in Colombia and other South American countries

Durante la última década se han reportado numerosos casos de infección por Trypanosoma cruzi por vía oral, debidos a la contaminación de alimentos con heces de triatominos silvestres o con secreciones de reservorios en áreas donde los vectores domiciliados han sido controlados o no hay antecedentes de domiciliación. Con base en criterios epidemiológicos, clínicos y socioeconómicos, la Organización de las Naciones Unidas para la Agricultura y la Alimentación (FAO) y la Organización Mundial de la Salud (OMS) establecieron una clasificación de los parásitos transmitidos por contaminación de alimentos en diferentes regiones del mundo, en la cual T. cruzi ocupó el décimo lugar de importancia en un grupo de 24 parásitos. Los cambios ambientales, como la deforestación y el calentamiento global, han afectado los ecotopos y el comportamiento de los vectores y de los reservorios de T. cruzi , de manera que estos se han desplazado a nuevas zonas, generando una nueva forma de transmisión por contaminación de alimentos que requiere su evaluación en el país. La presente revisión aborda la transmisión oral de la enfermedad de Chagas con énfasis en los estudios orientados a identificar los factores de riesgo, las especies de triatominos involucrados, la fisiopatología de la infección oral y los genotipos del parásito que están implicados en esta forma de transmisión en Colombia y en otras regiones de América Latina, así como la necesidad de adoptar políticas para su control y vigilancia epidemiológica.

Many cases of infection caused by the oral transmission of Trypanosoma cruzi have been reported during the last decade. These have been due to the contamination of food by faeces from sylvatic triatomines or by leakage from reservoirs in areas where domiciliated vectors have been controlled or where there has been no prior background of domiciliation. The United Nations Food and Agriculture Organization (FAO) and the World Health Organization (WHO) have used epidemiological, clinical and socioeconomic criteria for ranking parasites transmitted by the contamination of food in different areas of the world; T. cruzi was placed tenth in importance amongst a group of 24 parasites in such ranking. Environmental changes such as deforestation and global warming have affected ecotopes and the behaviour of T. cruzi vectors and reservoirs so that these have become displaced to new areas, thereby leading to such new transmission scenario caused by the contamination of food, which requires evaluation in Colombia. The current review deals with the oral transmission of Chagas´ disease, emphasising studies aimed at identifying the pertinent risk factors, the triatomine species involved, the physiopathology of oral infection, the parasite´s genotypes implicated in this type of transmission in Colombia and other Latin American regions, as well as the need for ongoing epidemiological surveillance and control policies.

Trypanosoma cruzi genotyping supports a common source of infection in a school-related oral outbreak of acute Chagas disease in Venezuela.

Trypanosoma cruzi I, a discrete typing unit (DTU) found in human infections in Venezuela and other countries of the northern region of South America and in Central America, has been recently classified into five intra-DTU genotypes (Ia, Ib, Ic, Id, Ie) based on sequence polymorphisms found in the spliced leader intergenic region. In this paper we report the genotype identification of T. cruzi human isolates from one outbreak of acute orally acquired Chagas disease that occurred in a non-endemic region of Venezuela and from T. cruzi triatomine and rat isolates captured at a guava juice preparation site which was identified as the presumptive source of infection. The genotyping of all these isolates as TcId supports the view of a common source of infection in this oral Chagas disease outbreak through the ingestion of guava juice. Implications for clinical manifestations and dynamics of transmission cycles are discussed.

As manifestações de ECG do maior surto da doença de Chagas devido a infecção oral na América Latina / ECG manifestations of the biggest outbreak of Chagas disease due to oral Infection in Latin-America

FUNDAMENTO: A doença de Chagas afeta mais de 15 milhões de pessoas em todo o mundo. Embora a transmissão vetorial tenha diminuído, a transmissão oral tornou-se relevante. Recentemente, nosso grupo publicou as características clínicas e epidemiológicas do maior surto relatado até hoje da doença de Chagas transmitida oralmente. OBJETIVO: Descrever alterações eletrocardiográficas que ocorrem na população de estudo durante o surto causado pela ingestão de suco de goiaba contaminado. MÉTODOS: Avaliamos 103 casos positivos, dos quais 76 (74%) tinham <18 anos de idade (média das idades: 9,1 ± 3,1 anos) e 27 (26%) tinham >18 anos (média das idades: 46 ± 11,8 anos). Todos os pacientes foram submetidos a avaliações clínicas e ECG. Caso os pacientes apresentassem palpitações ou alterações evidentes do ritmo na linha basal, o monitoramento de ECG ambulatorial seria realizado. RESULTADOS: Um total de 68 casos(66%;53 crianças e15 adultos) apresentaram anormalidades no ECG. Além disso, 69,7% (53/76) daqueles com idade < 18anos e 56% (15/27) daqueles com idade >18 anos apresentaram alguma alteração no ECG(p = ns). Anormalidades de ST-T foram observadas em 37,86% casos (39/103) e arritmias foram evidente sem 28,16% casos(29/103). Alterações de ST ocorreram em 72% daqueles com idade < 18 anos em comparação aos de > 18 anos (p < 0,0001). CONCLUSÃO: Este estudo relata o maior número de casos no mesmo surto de doença de Chagas causada por contaminação oral, com ECGs relatados. As alterações no ECG que sugerem miocardite aguda e arritmias foram as anormalidades encontradas com maior frequência.

BACKGROUND: Chagas disease affects more than 15 million people worldwide. Although vector-borne transmission has decreased, oral transmission has become important. Recently, our group published the clinical and epidemiological characteristics of the largest outbreak of orally transmitted Chagas disease reported till date. OBJECTIVE: To describe electrocardiographic changes occurring in the study population during the outbreak caused by ingestion of contaminated guava juice. METHODS: We evaluated 103 positive cases, of which 76 (74%) were aged <18 years (average age: 9.1 ± 3.1 years) and 27 (26%) were aged > 18 years (average age: 46 ± 11.8 years). All patients underwent clinical evaluations and ECG. If the patients had palpitations or evident alterations of rhythm at baseline, ambulatory ECG monitoring was performed. RESULTS: A total of 68 cases (66%; 53 children and 15 adults) had ECG abnormalities. Further, 69.7% (53/76) of those aged < 18 years and 56% (15/27) of those aged >18 years showed some ECG alteration (p = ns). ST-T abnormalities were observed in 37.86% cases (39/103) and arrhythmias were evident in 28.16% cases (29/103). ST alterations occurred in 72% of those aged <18 years compared with 19% of those aged >18 years (p < 0.0001). CONCLUSIONS: This study reports the largest number of cases in the same outbreak of acute Chagas disease caused by oral contamination, with recorded ECGs. ECG changes suggestive of acute myocarditis and arrhythmias were the most frequent abnormalities found.

ECG manifestations of the biggest outbreak of Chagas disease due to oral infection in Latin-America.

BACKGROUND: Chagas disease affects more than 15 million people worldwide. Although vector-borne transmission has decreased, oral transmission has become important. Recently, our group published the clinical and epidemiological characteristics of the largest outbreak of orally transmitted Chagas disease reported till date. OBJECTIVE: To describe electrocardiographic changes occurring in the study population during the outbreak caused by ingestion of contaminated guava juice. METHODS: We evaluated 103 positive cases, of which 76 (74%) were aged ≤ 18 years (average age: 9.1 ± 3.1 years) and 27 (26%) were aged > 18 years (average age: 46 ± 11.8 years). All patients underwent clinical evaluations and ECG. If the patients had palpitations or evident alterations of rhythm at baseline, ambulatory ECG monitoring was performed. RESULTS: A total of 68 cases (66%; 53 children and 15 adults) had ECG abnormalities. Further, 69.7% (53/76) of those aged ≤ 18 years and 56% (15/27) of those aged >18 years showed some ECG alteration (p = ns). ST-T abnormalities were observed in 37.86% cases (39/103) and arrhythmias were evident in 28.16% cases (29/103). ST alterations occurred in 72% of those aged ≤ 18 years compared with 19% of those aged >18 years (p < 0.0001). CONCLUSIONS: This study reports the largest number of cases in the same outbreak of acute Chagas disease caused by oral contamination, with recorded ECGs. ECG changes suggestive of acute myocarditis and arrhythmias were the most frequent abnormalities found.

[Survival of Trypanosoma cruzi in experimentally contaminated drinks]. / Supervivencia de Trypanosoma cruzi en bebidas experimentalmente contaminadas.

INTRODUCTION: Trypanosoma cruzi is the causative agent of Chagas disease, transmitted primarily by triatomine insects. However, in 2005, oral transmission was documented in countries where the disease is endemic for Chagas disease. This trend may also occur in Colombia, a situation that motivated epidemiological alerts and the necessity for exploring the risk level of oral, human-to-human infection by T. cruzi. OBJECTIVE: Survival times were established for the T. cruzi strain DS using juices involved in the outbreak of Lebrija County (Cesar, Colombia) in 2008. MATERIALS AND METHODS: Survival of the T. cruzi strain was evaluated as defined by vitality (forward movement) and viability (growth in isolation medium Novy, McNeal and Nicolle/liver infusion tryptose). This strain was molecularly characterized as TCLA, isolated from a patient associated with an outbreak in Aguachica County (Santander, very near Lebrija). Its survival was tested in tangerine juice, guava, soursop (guanábana), water and sugar water. RESULTS: The T. cruzi strain DS remained vital in mandarin at room temperature for 72 hr, at refrigerated temperatures for 36 hr;, the soursop (guanábana) for 48 hr at room temperature and 384 hr under refrigeration; and guava at both temperatures 24 hr. This strain was viable 2 and 24 hours post-infection in each of the other juices at the two temperature conditions. CONCLUSIONS: The DS T. cruzi strain survived in all drinks for more than 24 hours post-infection, with a survival time of 384 hr in the juice of soursop (guanábana) under refrigeration.

Strongyloides stercoralis in common vegetables and herbs in Kota Bharu, Kelantan, Malaysia.

Transmission of soil-transmitted helminthes infection is by faecal oral route, and is influenced by food preference. Kelantanese love to consume ulam which are raw vegetables and herbs. Some of the herbs grow on grounds with high humidity and are abundant near drainage areas, these are also places with higher likelihood of harbouring viable parasite ova. The aim of this study was to determine the prevalence of soiltransmitted helminthes in vegetables, herbs and fruits found in our local setting. The results by microscopy showed that there was no helminthes ovum or protozoan parasite in the samples. However, Strongyloides stercoralis rhabdatiform larvae were identified in water samples used to wash pegaga, kesum and water spinach, and the number of larvae observed were 152, 9 and 16 respectively. Analysis by real-time PCR confirmed the microscopic observation of this helminth. This study highlighted that vegetables and herbs are likely sources of Strongyloides stercoralis infection in Kota Bharu, Kelantan. Thus vegetable sellers as well as the food handlers are the two important groups who are at high risk of acquiring the infection.

Cryptosporidium recovered from Musca domestica, Musca sorbens and mango juice accessed by synanthropic flies in Bahirdar, Ethiopia.

The study was conducted to determine the role of house flies, Musca domestica and Musca sorbens to carry Cryptosporidium species in natural environment and filth flies potential for contamination of food item they visited using acid-fast stain technique. Cryptosporidium was identified from flies collected in dairy cow barns, butchery, market and defecating grounds. Musca domestica captured from dairy cow barns and M. sorbens from defecating ground were found carrying more oocyst of Cryptosporidium parvum. Oocyst load per fly for M. domestica and M. sorbens was 5.84 and 3.42, respectively. Flies' population dynamics in each month had little relation to the monthly oocyst frequency, r = 0.06 and 0.02 for M. domestica and M. sorbens, respectively. Cryptosporidium species oocysts were isolated from frozen mango juice, which filth flies visited in dairy farm barn. Load of oocysts in the mango juice was dependent on time contact of flies with mango juice and more oocysts were recovered (P < 0.05) in mango juice samples accessed by filth flies for longer period. Role of filth flies to carry and deposit Cryptosporidium species oocyst for development of food-borne cryptosporidiosis is signified.

Detection of some intestinal protozoa in commercial fresh juices.

Fresh fruit juices are popular, but not always safe. For assessing the likelihood of infection with newly emerging intestinal protozoa, commercial fresh orange, lemon, sugar cane, strawberry, and mango juices were screened by wet mounts, Weber's modified trichrome and modified Ziehl-Neelsen stains. Protozoa viability was done by fluorescein-diacetate/propidium-iodide staining, and infectivity was performed in Swiss albino mice. Results showed that 35.43% were contaminated with one or more of Cryptosporidia, Microsporidia, and Cyclospora, as well as Giardia spp. Strawberry was the most contaminated juice (54.28%), while orange was the slightest (22.86%). Cryptosporidia was the highest contaminant (61.29%), and Cyclospora was the least (14.52%). Microsporidia spp. was the most robust contaminant which retained its viability and infectivity in juices in which it was detected. Moderately acidic strawberry and mango juices and alkaline sugar cane juice pose a possible threat, due to harboring the highest viable and infectious protozoa. Regarding highly acidic juices, viability and infectivity decreased in lemon, yet was not still risk free. Orange juice was comparatively safe, as viability dramatically declined, while infectivity was completely abolished. Hence consumers, especially high risk group, are placed at hazard of contracting intestinal protozoa infections, especially through moderately acidic and alkaline juices.

Large urban outbreak of orally acquired acute Chagas disease at a school in Caracas, Venezuela.

BACKGROUND: Trypanosoma cruzi oral transmission is possible through food contamination by vector's feces. Little is known about the epidemiology and clinical features of microepidemics of orally acquired acute Chagas disease (CD). METHODS: A case-control, cohort-nested, epidemiological study was conducted during an outbreak of acute CD that affected a school community. Structured interviews were designed to identify symptoms and sources of infection. Electrocardiograms were obtained for all patients. Specific serum antibodies were assessed by immunoenzimatic and indirect hemagglutination tests. In some cases, parasitemia was tested directly or by culture, animal inoculation, and/or a polymerase chain reaction technique. RESULTS: Infection was confirmed in 103 of 1000 exposed individuals. Of those infected, 75% were symptomatic, 20.3% required hospitalization, 59% showed ECG abnormalities, parasitemia was documented in 44, and 1 child died. Clinical features differed from those seen in vectorial transmission. The infection rate was significantly higher among younger children. An epidemiological investigation incriminated contaminated fresh guava juice as the sole source of infection. CONCLUSIONS: This outbreak was unique, because it affected a large, urban, predominantly young, middle-class, otherwise healthy population and resulted in an unprecedented public health emergency. Rapid diagnosis and treatment avoided higher lethality. Food-borne transmission of T. cruzi may occur more often than is currently recognized.

Chagas' disease as a foodborne illness.

Various researchers have studied the importance of the oral transmission of Chagas' disease since the mid-20th century. Only in recent years, due to an outbreak that occurred in the Brazilian State of Santa Catarina in 2005 and to various outbreaks occurring during the last 3 years in the Brazilian Amazon basin, mainly associated with the consumption of Amazonian palm berry or açaí (Euterpe oleracea Mart.) juice, has this transmission route aroused the attention of researchers. Nevertheless, reports published in the 1960s already indicated the possibility of Chagas' disease transmission via food in Brazil, mainly in the Amazonian region. Recently, in December 2007, an outbreak of Chagas' disease occurred in Caracas, Venezuela, related to ingestion of contaminated fruit juices. The objective of this article is to point out the importance of foodborne transmission in the etiology of Chagas' disease, on the basis of published research and Brazilian epidemiology data.

[FTA/PCR detection for Cryptosporidium added in beverage].

An established FVA/PCR protocol was applied to detect Cryptosporidium oocysts which were added in beverage, and proved to be as sensitive as 1.0 oocyst/ml detected within 6 hours. The study indicates that this technique is sensitive, time-saving and easy to perform.

Evaluation of two DNA template preparation methods for post-immunomagnetic separation detection of Cryptosporidium parvum in foods and beverages by PCR.

Cryptosporidium parvum oocysts were recovered by immunomagnetic separation from six artificially contaminated foods. Two DNA isolation methods were subsequently evaluated by PCR. The FTA Concentrator-PS filter provided rapid and reproducible detection, although variability increased at lower inoculum levels (88% and 15% detection in high- and low-inoculum-level samples, respectively). Total DNA extraction generated consistent results at all oocyst levels but resulted in longer analysis time (100% and 59% detection in high- and low-inoculum-level samples, respectively). Also reflected in this study was that the matrix played an important role in the ability to recover oocysts, as sample turbidity, pH, and PCR inhibitors all influenced detection.

Cryptosporidiosis associated with ozonated apple cider.

We linked an outbreak of cryptosporidiosis to ozonated apple cider by using molecular and epidemiologic methods. Because ozonation was insufficient in preventing this outbreak, its use in rendering apple cider safe for drinking is questioned.

Microbiological evaluation of sugarcane juice sold at street stands and juice handling conditions in São Carlos, São Paulo, Brazil.

Fresh sugarcane juice is sold by street vendors without any heat treatment in São Carlos, São Paulo, Brazil. Twenty-four samples of point-of-sale juice were tested by standard methods to determine heterotrophic bacteria, total and thermo-tolerant coliform counts, Salmonella, and parasites in the juice. 25% of samples showed poor sanitary conditions, with thermo-tolerant coliform levels higher than allowed by Brazilian standards. Salmonella spp. and parasites were absent in all samples. Thermo-tolerant coliforms were detected on the hands of 37% of juice handlers, and heterotrophic bacterial counts reached 2.0 x 10(3) cfu/per hand. Escherichia coli was detected in one hand sample, and no Salmonella spp. was detected. Screening questionnaires were used to interview the vendors, and 62% of interviewees were either unfamiliar with or failed to adopt adequate hygiene for food handling.

Potential sources of microbial contamination in unpasteurized apple cider.

A study was conducted to identify possible sources of microbial contamination and to assess the effect of good cleaning and sanitation practices on the microbial quality and safety of unpasteurized apple cider. Raw unwashed apples, washed apples, cleaning water, fresh cider, and finished cider samples were collected from five Ontario producers over 4 months and microbiologically tested. Total coliforms were found in 31, 71 and 38% of the unwashed apple, water, and washed apple samples, respectively. Escherichia coli was found in 40% of the water samples from one producer alone. The washing step was identified as a potential source of contamination, possibly due to water in the dump tanks seldom being refreshed, and because scrubbers, spray nozzles, and conveyors were not properly cleaned and sanitized. Higher total coliform counts (P < 0.0001) and prevalence (P < 0.0001) in fresh cider compared with those in unwashed apples and washed apples indicated considerable microbial buildup along the process, possibly explained by the lack of appropriate equipment sanitation procedures. Results showed that producers who had better sanitary practices in place had lower (P < 0.001) total coliform prevalence than the rest of the producers. Overall results show that good sanitation procedures are associated with improved microbial quality of fresh cider in terms of total coliforms and that operators who pasteurize and/or UV treat their product should still be required to have a sound good manufacturing practices program in place to prevent recontamination. Cryptosporidium parvum, an important pathogen for this industry, was found in different sample types, including washed apples, water, and fresh and finished cider.

Risk factors for Fasciola hepatica infection in children: a case-control study.

We conducted a case-control study to clarify the risk factors for human fascioliasis in the Northern Peruvian Altiplano. Cases (n=61) were individuals who were diagnosed as having chronic fascioliasis by faecal and/or serologic (Fas2-ELISA) studies. Controls (n=61) had negative serologic and coprological results for Fasciola hepatica. We identified an association between fascioliasis and four variables (40 analysed): the habit of drinking alfalfa juice (OR=4.5; 95% CI 1.8-11.1; P<0.001); familiarity with aquatic plants (OR=4.3; 95% CI 1.8-10.6; P<0.001); dog ownership (OR=5; 95% CI 1.7-15.1; P=0.002); and raising more than five sheep (OR=0.3; 95% CI 0.1-0.8; P=0.01). According to clinical presentation and laboratory studies, dizzy spells (P=0.01), history of jaundice (P=0.01), peripheral eosinophilia (P=0.005) and Ascaris lumbricoides in stools (P=0.001) were associated with fascioliasis. The principal exposure factor for F. hepatica infection was drinking alfalfa juice. In conclusion, we suggest that human fascioliasis in Peru should be suspected in patients from livestock-rearing areas, who present with recurrent episodes of jaundice and who have a history of consumption of alfalfa juice or aquatic plants, or who have eosinophilia.