Extraction of Trypanosoma cruzi DNA from food: a contribution to the elucidation of acute Chagas disease outbreaks

Abstract: INTRODUCTION: Before 2004, the occurrence of acute Chagas disease (ACD) by oral transmission associated with food was scarcely known or investigated. Originally sporadic and circumstantial, ACD occurrences have now become frequent in the Amazon region, with recently related outbreaks spreading to several Brazilian states. These cases are associated with the consumption of açai juice by waste reservoir animals or insect vectors infected with Trypanosoma cruzi in endemic areas. Although guidelines for processing the fruit to minimize contamination through microorganisms and parasites exist, açai-based products must be assessed for quality, for which the demand for appropriate methodologies must be met. METHODS: Dilutions ranging from 5 to 1,000 T. cruzi CL Brener cells were mixed with 2mL of acai juice. Four Extraction of T. cruzi DNA methods were used on the fruit, and the cetyltrimethyl ammonium bromide (CTAB) method was selected according to JRC, 2005. RESULTS: DNA extraction by the CTAB method yielded satisfactory results with regard to purity and concentration for use in PCR. Overall, the methods employed proved that not only extraction efficiency but also high sensitivity in amplification was important. CONCLUSIONS: The method for T. cruzi detection in food is a powerful tool in the epidemiological investigation of outbreaks as it turns epidemiological evidence into supporting data that serve to confirm T. cruzi infection in the foods. It also facilitates food quality control and assessment of good manufacturing practices involving acai-based products.

FOOD CONCENTRATION AND TEMPERATURE EFFECTS ON LIFE CYCLE CHARACTERISTICS OF TROPICAL CLADOCERA (Daphnia gessneri Herbst, Diaphanosoma sarsi Richard, Moina reticulata (Daday)): I. DEVELOPMENT TIME

The effects of food concentration and temperature on embryonic and postem-bryonic duration of three tropical species, Daphnia gessneri(1.5mm), Diaphanosoma sarsi(1.2mm) and Moina reticulata(0.8mm), were investigated as part of life cycle studies which included growth, body size and reproduction. These are the very first experimental studies undertaken on these species. The long-term growth experiments were performed under controlled laboratory conditions at all combinations of temperature (22"C, 27"C and 32"C) and constant food concentration (0.03, 0.05, 0.10, 0.25, 0.50 and 1.00 mgC/L) of the unicellular green alga Scenedesmus acutus.Animals were examined twice daily throughout their life cycle from the neonate to third adult instar. In all three species, temperature exerted the most powerful influence on embryonic duration but there was also a smaller food effect. In D. gessneri,postembry-onic durations remained more or less the same at food levels 0.25 mgC/L but were influenced by temperature. At food concentrations of 0.1 mgC/L or lower, postembryonic durations became increasingly prolonged, particularly at high temperatures. This threshold concentration is affected by temperature: in D. gessneri,it was 0.1 mgC/L at 22oC and 27oC but higher at 32oC (between 0.25 and 0.50 mgC/L). At the same temperature of 27oC, the food threshold level varied between species: it was higher (0.25 mgC/L) for D. sarsiand lower (0.05 mgC/L) for M. reticulatacompared with D. gessneri(0.1 mgC/L). In both embryonic and postembryonic durations there is a body size effect as the absolute durations were longest in the largest species and shortest in the smallest species In all three species, prolongation of postembryonic duration at combinations of high temperature and lowered food levels was accompanied by increased number of juvenile instars.

The food spectrum of the cardinal - tetra (Paracheirodon axelrodi, Characidae) in its natural habitat

The cardinal tetra (Paracheirodon axelrodi) is the most intensively commercialized ornamental fish from the Rio Negro Basin (Amazonas State, Brasil). Analysis of the stomach and gut contents of fish caught in their natural habitats show conclusively that the cardinal is essentially a predator, feeding on the mesofauna that adheres to submerged litter, roots and waterplants. Microcrustacea and chironomid larvae (Diptera) were the most frequently ingested prey, while algae intake was relatively infrequent. It is argued that the relatively small size of the cardinals captured in their natural habitat is due to the annual migrations imposed by the inundation cycles, rather than to resource limitation, because it is known from earlier investigations of similar habitats, that these plant substrates are densely colonized by the aquatic mesofauna. Cardinals raised in captivity are larger and have higher rates of growth.

Decapod crustaceans used as food by the Yanomami Indians of the Balawa-ú village, State of Amazonas, Brazil

The Yanomami are a group of South American Indians that live in the rainforest along the borderlands of Brazil and Venezuela. They depend on hunting, gardening and wild food for survival; crustaceans are a highly prized food item in their diet. Taxonomical and ethnozoological aspects of the Yanomami Indians of the Balawa-ú village, state of Amazonas, Brazil, related to the crustaceans are described. Information and specimens were obtained from August to December, 2003. Interviews were conducted with residents of the village and focused on questions about species exploited, indigenous names, modes of capture and use of the species. One shrimp species of the family Palaemonidae (Macrobrachium brasiliense) and two crab species of Trichodactylidae (Sylviocarcinus pictus, Valdivia serrata) as well as two of Pseudothelphusidae (Fredius fittkaui, F. platyacanthus) were recorded. The indigenous names applied to these species are: shuhu, for shrimp, oko and peimatherimi for each of the two pseudothelphusid crabs, and hesiki tôtôrema for both trichodactylid crabs.

Food consumption of adolescents. A simplified questionnaire for evaluating cardiovascular risk

OBJECTIVE: To develop a simplified questionnaire for self-evaluation by adolescents of foods associated with the risk of coronary diseases. METHODS: Frequency questionnaires about 80 foods were answered by representative samples of 256 adolescents aged 12 to 19 from Rio de Janeiro as part of the Nutrition and Health Research project. The dependent variable was the serum cholesterol predicting equation as influenced by diet, and the independent variables were the foods. The variables were normalized and, using Pearson's correlation coefficient, those with r>0.10 were selected for the regression model. The model was analyzed for sex, age, random sample, and total calories. Those food products that explained 85% of the cholesterol variation equation were present in the caloric model, and contained trans fatty acids were selected for the questionnaire. RESULTS: Sixty-five food products had a statistically significant correlation (P<0.001) with the dependent variable. The simplified questionnaire included 9 food products present in all tested models: steak or broiled meat, hamburger, full-fat cheese, French fries or potato chips, whole milk, pies or cakes, cookies, sausages, butter or margarine. The limit of the added food points for self-evaluation was 100, and over 120 points was considered excessive. CONCLUSION: The scores given to the food products and the criteria for the evaluation of the consumption limits enabled the adolescents to get to know and to balance their intake.

Effects of intraspecific competition and food deprivation on the immature phase of Ascia monuste orseis (Lepidoptera, Pieridae)

The effect of intraspecific competition for food on larvae and of food deprivation for 24 h on 2nd and 4th instars of Ascia monuste orseis (Godart, 1819) was investigated. Intraspecific competition for food during the immature phase leads to long pupation time, high larval mortality, reduced adult weight, and reduced number of eggs per female. In food deprivation experiments, the major differences in A. monuste orseis performance were long pupation time in the group that was deprived during the 2nd instar; and a negative effect on reproduction in the group that was deprived during the 4th instar, with reduced adult weight. Both food deprived periods tested are critical, and deprivation during the 2nd instar seems to have an effect as drastic as during the 4th instar because it directly affects larvae survival. Immatures can resist food deprivation for 24 h during the 2nd and 4th instars (low mortality), have a compensatory behaviour (high ingestion and biomass gain) during the 5th instar, and do not demonstrate cannibalistic behaviour during food deprivation.

Abundance, breeding and food of the Little Blue Heron Egretta caerulea (Aves, Ardeidae) in the Patos Lagoon estuary, a recently colonized area in southern Brazil

We document the expansion of the breeding distribution of the Little Blue Heron Egretta caerulea (Linnaeus, 1758) to 850 km beyond its previous southern limit in South America. In addition we present data on abundance, breeding biology and food of the species in the Patos Lagoon estuary, the area which the species recently colonized. The maximum abundance recorded in the breeding colony and in a nocturnal roosting site was 53 and 49 individuals respectively. Nesting occurred from September to March. Birds nested in a mixed breeding colony together with about 3,000 breeding pairs of seven other species of Pelecaniformes, in a swampy forest near the margin of the estuary. Five nests were between 1.5 and 4.3 m from the ground, on the shrub Daphnopsis racemosa (Thymelaeaceae), on the trees Sebastiana brasiliensis (Euphorbiaceae) and Mimosa bimucronata (Leguminosae), or on the bamboo Bambusa sp. (Poaceae). Four nests produced two fledglings each, while one nest was abandoned. Of 13 grouped samples of food regurgitated by five nestlings, Pink Shrimp Farfantepenaeus paulensis (Perez-Farfante, 1967) constituted 70% in mass, while total length of ingested fishes and shrimps varied mostly between 20 and 50 mm. Estuarine prey items represented 99% of the total food mass. The recent southward expansion of the breeding range of the Little Blue Heron in South America may be a response to climate warming of the Patos Lagoon estuary. Degradation of estuaries in the southwestern Atlantic may also be forcing the birds to breed in areas outside previous geographical range.

Toxicological and toxicogenetic effects of plants used in popular medicine and in cattle food

Toxicological and toxicogenetic effects of aqueous (tea) and hexanica fruit extract of Indigofera suffruticosa Mill, and hydroalcoholic root extract od Solanum agrarium Stendt. Were evaluated in Balb C male mice intraperitoneally exposed. A hepatotoxic effect was observed just for animals treated with aqueous fruit extract of I. suffruticosa. In relation to the toxicogenetic effect, just the group trreated with 12.5% of toxic dose of aqueous fruit extract of I. suffruticosa showed a statistically significant increase in the frequency of cells with chromosome aberrations (cytogenetic effect), although a slight increase was also observed for the highest dose (25% of LF50_ of hydroalcoholic root extract of S. agrarium. The results obtanied show that before S. agrarium is used as medicine and before the wide use of I. suffruticosa in cattle food, careful evaluation must be done.

Theoretical estimates of consumable food and probability of acquiring food in larvae of Chrysomya putoria (Diptera: Calliphoridae)

An indirect estimate of consumable food and probability of acquiring food in a blowfly species, Chrysomya putoria, is presented. This alternative procedure combines three distinct models to estimate consumable food in the context of the exploitative competition experienced by immature individuals in blowfly populations. The relevant parameters are derived from data for pupal weight and survival and estimates of density-independent larval mortality in twenty different larval densities. As part of this procedure, the probability of acquiring food per unit of time and the time taken to exhaust the food supply are also calculated. The procedure employed here may be valuable for estimations in insects whose immature stages develop inside the food substrate, where it is difficult to partial out confounding effects such as separation of faeces. This procedure also has the advantage of taking into account the population dynamics of immatures living under crowded conditions, which are particularly characteristic of blowflies and other insects as well.

Dynamics of evolution and resistance to starvation of Triatoma vitticeps (Stal 1859) (Reduviidae: Triatominae), submitted to two different regimens of food deprivation

The resistance to starvation of Triatoma vitticeps has been analyzed comparatively, according to different regimens of food deprivation under laboratory conditions. One cohort, composed of 100 specimens of each evolutionary nymphal stage, was submitted to continuous fasting until death; the second group, arranged in the same way, was fed once on chicken. Through this work, it was possible not only to compare the results obtained from the first group with other authors' results but, also, to analyze the dynamics of evolution, molting, longevity and the increase in insect longevity, in the second group. The average values recorded for survival time revealed statistical differences between the two groups. Among the important results detected, there is one that deserves to be emphasized: the incredible increase in longevity among insects that received only one feed - an average increase in survival time that reached 2.95 to 3.30 times in nymphs of 3rd and 4th stages, respectively. One 5th stage nymph survived for up to 350 days and the females may triplicate their survival rate, what represents an important epidemiological factor. The knowledge about this type of biological characteristic of T. vitticeps may contribute to prevent the domiciliation of this species, what seems to be incipient in some municipal districts, in Brazil.