Otilio Machado Pereira Bastos

Paleoparasitology: Perspectives with New Techniques

Paleoparasitology is the study of parasites found in archaeological material. The development of this field of research began with histological identification of helminth eggs in mummy tissues, analysis of coprolites, and recently through molecular biology. An approach to the history of paleoparasitology is reviewed in this paper, with special reference to the studies of ancient DNA identified in archaeological material.

Contaminação por enteroparasitas em hortaliças comercializadas nas cidades de Niterói e Rio de Janeiro, Brasil

The aim of this study was to evaluate the parasitological contamination of vegetables to be consumed raw and commercialized in Niteroi and Rio de Janeiro cities. We studied 128 samples of vegetables ¾ lettuce (Lactuca sativa) and watercress (Nasturtium officinale) ¾ from supermarkets, greengrocer shops and self-service restaurants. Only 6.2% of the samples were positive for parasitic structures with morphological aspects similar to those of animal parasites. We detected acharids, acharid eggs, insects, nematode larvae and ciliated protozoa in most of the samples (96.1%), including those from restaurants. This high percentage suggests a risk of human infection since parasite structures capable of infecting man may exist in association with these agents.The aim of this study was to evaluate the parasitological contamination of vegetables to be consumed raw and commercialized in Niteroi and Rio de Janeiro cities. We studied 128 samples of vegetables — lettuce (Lactuca sativa) and watercress (Nasturtium officinale) — from supermarkets, greengrocer shops and self-service restaurants. Only 6.2% of the samples were positive for parasitic structures with morphological aspects similar to those of animal parasites. We detected acharids, acharid eggs, insects, nematode larvae and ciliated protozoa in most of the samples (96.1%), including those from restaurants. This high percentage suggests a risk of human infection since parasite structures capable of infecting man may exist in association with these agents. Key-words: Intestinal parasite. Vegetables. Vanessa C.L. Mesquita, Cathia M.B. Serra, Otílio M.P. Bastos e Cláudia M.A. Uchôa Intestinal parasites contamination from vegetables comercialized in Niterói and Rio de Janeiro cities, Brazil As enteroparas i toses por possuí rem ecossistema infectivo com mecanismos de infecção passivo oral e/ou ativo cutâneo, apresentam-se com distribuição cosmopolita, porém com possibil idade de variação na prevalência7. Geralmente, tanto em áreas rurais quanto urbanas dos países de terceiro mundo devido as baixas condições sanitárias, as parasi toses intest inais são amplamente difundidas, sendo as hortaliças citadas como um dos veículos de suas estruturas infectantes5 9. A principal forma de contaminação dessas hortaliçasThe aim of this study was to evaluate the parasitological contamination of vegetables to be consumed raw and commercialized in Niterói and Rio de Janeiro cities. We studied 128 samples of vegetables--lettuce (Lactuca sativa) and watercress (Nasturtium officinale)--from supermarkets, greengrocer shops and self-service restaurants. Only 6.2% of the samples were positive for parasitic structures with morphological aspects similar to those of animal parasites. We detected acarids, acarid eggs, insects, nematode larvae and ciliated protozoa in most of the samples (96.1%), including those from restaurants. This high percentage suggests a risk of human infection since parasite structures capable of infecting man may exist in association with these agents.

Prevalence and risk factors for Toxoplasma gondii infection among pregnant and postpartum women attended at public healthcare facilities in the City of Niterói, State of Rio de Janeiro, Brazil.

INTRODUCTION To determine the prevalence of immunoglobulin G (IgG) and immunoglobulin M (IgM) anti-Toxoplasma gondii antibodies among pregnant and postpartum women attended within the public healthcare system in Niterói, State of Rio de Janeiro,and to detect possible exposure factors associated with T. gondii infection in this population. METHODS IgM and IgG anti- T. gondii antibodies were investigated in 276 pregnant and 124 postpartum women by using the indirect immunofluorescence (IFAT) and immunoenzymatic assay (ELISA) techniques. The participants were selected by convenience sampling. All these 400 patients filled out a free and informed consent statement, answered an epidemiological questionnaire and were informed about the disease. RESULTS Among the 400 samples analyzed, 234 (58.5%) were reactive to IgG anti-T. gondii antibodies, according to the IFAT and/or ELISA assay. One pregnant woman was found to be reactive to IgM anti- T. gondii antibodies, with an intermediate IgG avidity test. Risk factor analysis showed that seropositivity was significantly associated (p<0.05) with age, contact with cats and presence of rodents at home. Through a logistic regression model, these associations were confirmed for age and contact with cats, while education at least of the high school level was found to be a protective factor. CONCLUSIONS The prevalence rate of IgG anti-T. gondii antibodies in the city of Niterói was high and the risk factors for infection detected after multivariate analysis were: age over 30 years, contact with cats and education levels lower than university graduate level.

Detection of Toxoplasma gondii DNA by Polymerase Chain Reaction in Experimentally Desiccated Tissues

Despite toxoplasmosis being a common infection among human and other warm-blooded animals worldwide, there are no findings about Toxoplasma gondii evolutionary forms in ancient populations. The molecular techniques used for amplification of genetic material have allowed recovery of ancient DNA (aDNA) from parasites contained in mummified tissues. The application of polymerase chain reaction (PCR) to paleoparasitological toxoplasmosis research becomes a promising option, since it might allow diagnosis, acquisition of paleoepidemiological data, access to toxoplasmosis information related origin, evolution, and distribution among the ancient populations. Furthermore, it makes possible the analysis of parasite aDNA aiming at phylogenetic studies. To standardize and evaluate PCR applicability to toxoplasmosis paleodiagnostic, an experimental mummification protocol was tested using desiccated tissues from mice infected with the ME49 strain cysts, the chronic infection group (CIG), or infected with tachyzoites (RH strain), the acute infection group (AIG). Tissues were subjected to DNA extraction followed by PCR amplification of T. gondii B1 gene. PCR recovered T. gondii DNA in thigh muscle, encephalon, heart, and lung samples. AIG presented PCR positivity in encephalon, lungs, hearts, and livers. Based on this results, we propose this molecular approach for toxoplasmosis research in past populations.

Parasitological and immunological diagnoses from feces of captive-bred snakes at Vital Brazil Institute

Fecal samples from 56 snakes at the Vital Brazil Institute, in the city of Niterói, Rio de Janeiro, were tested using the sedimentation and flotation techniques to investigate the evolutionary forms of parasites such as helminths and protozoa, and using enzyme immunoassay techniques to detect antigens of Cryptosporidium sp. and Giardia sp. Among the animals tested, 80.3% were positive for parasites. Out of these, there were 16 Bothrops jararaca, 16 B. jararacussu and 13 Crotalus durissus. The prevalence of parasitic nematodes was 41.1%, and nematodes were found in all three snake species. Among these, the most frequent finding was eggs of Kalicephalus sp., which were diagnosed in 25% of the snakes. The positivity for protozoa detected using parasite concentration techniques was 75%, including oocysts of Caryospora sp. in 75%, cysts with morphology similar to Giardia sp. 3.6%, amoeboid cysts in 41.1% and unsporulated coccidia oocysts in 8.9%. Immunoassays for Cryptosporidium sp. antigens produced positive findings in 60.7%. Pseudoparasites were detected in 64.3%. These results show that there is a need to improve the sanitary handling of captive-bred snakes, and also for the animal house that supplies rodents to feed them. The results also highlight that diagnostic tests should be performed periodically on stool specimens from captive-bred snakes.

Isolation and maintenance of Balantidium coli (Malmsteim, 1857) cultured from fecal samples of pigs and non-human primates.

Balantidium coli is a protozoa that can determine dysentery in humans, pigs and non-human primates having zoonotic potential. The lack of standardization in isolation and maintenance hinders the development of research on its biology and epidemiology. This study is aimed to standardize the isolation and maintenance of this parasite from animal feces, in culture medium, Pavlova modified. From 2012 to 2014, 1905 fecal samples were collected from captive animals of Rio de Janeiro. Were selected for isolation samples with a minimum of 10 trophozoites and/or 30 cysts of B. coli, totaling 88 pigs, 26 Cynomolgus and 90 rhesus macaques. In the presence of cysts, the sample was homogenized in saline solution, 500 μL was removed and inoculated into culture medium. The material that contained trophozoites the inoculum was made from 240 μL of fecal solution. All inoculate tubes with the subcultures were kept at 36°C, and sterile rice starch was always added to the medium. The parasites isolate from pigs, 34%, and from Cynomolgus 38.4% were maintained in vitro for a period of more than 24 months. These procedures proved to be adequate for isolation and maintenance of B. coli from different animals, they were found to be inexpensive and easy to perform.

Trichuris trichiura in a post-Colonial Brazilian mummy.

Trichuris trichiura is a soil-transmitted helminth which is prevalent in warm, moist, tropical and subtropical regions of the world with poor sanitation. Heavy whipworm can result either in Trichuris dysenteric syndrome - especially in children - or in a chronic colitis. In heavy infections, worms can spread proximally and may cause ileitis. Here we provide first microscopic evidence for a T. trichiura adult worm embedded in the rectum of a post-Colonial Brazilian adult mummy. During Colonial and post-Colonial times, many European chroniclers described a parasitic disease named Maculo whose symptomatology coincides with heavy helminthiasis. Based on our findings and on comparison of ancient textual evidence with modern description of heavy whipworm, we feel confident in considering that the two syndromes are expressions of the same pathological condition.

Balantidium coli and other gastrointestinal parasites in captives non-human primates of the Rio de Janeiro, Brazil

Parasites are agents of diarrhea in captive non‐human primates (NHP). To broaden information about those etiological agents in captive animals in Brazil, gastrointestinal parasites in captive NHP were researched in nurturing Centers of Rio de Janeiro State.

Parasites in stool samples in the environment of Ilha da Marambaia, Rio de Janeiro, Brazil: an approach in public health

This research aimed to describe the frequency of parasites in stool samples in the environment of Ilha da Marambaia, Rio de Janeiro, Brazil. One hundred and five stool samples were collected and processed by the coproparasitological techniques ethyl acetate sedimentation and centrifuge-flotation using saturated sugar solution. Parasites were detected in 81.9% of the samples, hookworm being the most prevalent, followed by Trichuris vulpis. Ascaris sp. eggs were also found. A high level of evolutive forms of parasites with public health risk was found in stool samples of the environment studied. We propose that health education programs, allied to an improvement of human and animal health care, must be employed to reduce the environmental contamination.

TOXOPLASMOSIS-RELATED KNOWLEDGE AMONG PREGNANT AND POSTPARTUM WOMEN ATTENDED IN PUBLIC HEALTH UNITS IN NITERÓI, RIO DE JANEIRO, BRAZIL

The present study conducted a toxoplasmosis-related knowledge level survey with 400 pregnant and puerperal women attended in public health units in the municipality of Niterói, Rio de Janeiro. Only 111 (27.8%) women claimed to know about the disease. Most of them (n = 289; 72.2%) had never heard about toxoplasmosis nor knew how to prevent the infection by Toxoplasma gondii. A significant difference (p = 0.013) regarding the presence of anti-T. gondii IgG was observed between women who claimed to know about the disease and those who had never heard about it. These results highlight the importance of a systematic serological screening process for toxoplasmosis, as well as the importance of primary prevention by accurate information during prenatal care, an important Public Health action to be implemented.