Non-invasive sampling in Itatiaia National Park, Brazil: wild mammal parasite detection.

BACKGROUND: Non-invasive sampling through faecal collection is one of the most cost-effective alternatives for monitoring of free-living wild mammals, as it provides information on animal taxonomy as well as the dynamics of the gastrointestinal parasites that potentially infect these animals. In this context, this study aimed to perform an epidemiological survey of gastrointestinal parasites using non-invasive faecal samples from carnivores and artiodactyls identified by stool macroscopy, guard hair morphology and DNA sequencing in Itatiaia National Park. Between 2017 and 2018, faeces from carnivores and artiodactyls were collected along trails in the park. The host species were identified through macroscopic and trichological examinations and molecular biology. To investigate the parasites, the Faust, Lutz and modified Ritchie and Sheather techniques and enzyme immunoassays to detect Cryptosporidium sp. antigens were used. RESULTS: A total of 244 stool samples were collected. The species identified were Chrysocyon brachyurus, Leopardus guttulus, Canis familiaris, Cerdocyon thous, Puma yagouaroundi, Leopardus pardalis, Puma concolor and Sus scrofa. There were 81.1% samples that were positive for parasites distributed mainly in the high part of the park. Helminths, especially eggs of the family Ascarididae, were more frequently detected in carnivore faeces (70.9%). Protozoa, especially Cryptosporidium sp., represented the highest frequency of infection in artiodactyl faeces (87.1%). This zoonotic protozoon was detected in eight mammalian species, including in a wild boar. High values of structural richness and Shannon and Simpson diversity indices were observed for the parasites, especially in the faeces of C. brachyurus. Significant differences in parasite diversity were observed between wild and domestic animals, such as C. brachyurus and C. familiaris, respectively, and between taxonomically distant species, such as C. brachyurus and S. scrofa. The highest values for parasite similarity were found among the species that frequented similar areas of the park, such as C. brachyurus and L. guttulus. CONCLUSIONS: The animals and parasite infections were identified through the combination of three techniques. High frequency parasite structures were diagnosed. Zoonotic protozoa were found and mainly occurred in samples from introduced species.

Trypanoxyuris (Trypanoxyuris) minutus (Schneider, 1866) among Alouatta guariba clamitans (Cabrera, 1940) in the state of Rio de Janeiro, Brazil.

This study reports on infection by Trypanoxyuris (Trypanoxyuris) minutus among Alouatta guariba clamitans through biometry on adults and eggs. A total of 58 adult helminths were recovered from the small segment of the animal's large intestine. This study is the first report of this parasite among A. guariba clamitans in Rio de Janeiro.

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

BACKGROUND: 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. METHODS: Fecal samples were collected from primates, of which 960 came from the Research Center (Cecal/Fiocruz) and 115 from the Primate Center (CPRJ/Inea). The study involved species of the New World (NW) primates and of the Old World (OW). RESULTS: The estimated prevalence was 56.7%, of which 91.3% presented protozoans and 7.4% presented helminths. Statistical difference between the nurturing centers occurred in the overall value of parasitosis and in the isolated frequency of Balantidium coli and Entamoeba sp., especially in the samples of OW primates living in Cecal. CONCLUSIONS: These results demonstrated the need for implements of sanitation programs in the sites for captive primates nurturing.

Cryptosporidium species in non-human animal species in Latin America: Systematic review and meta-analysis.

Cryptosporidiosis is an infection caused by a protozoon that inhabits the gastrointestinal tract. More than forty valid species have been described in the genus Cryptosporidium, infecting a broad range of hosts around the world, some with zoonotic transmission and others with predominant anthroponotic transmission. Prevalence studies conducted in Latin American countries have been specific, without consolidating information on species prevalences. Thus, the aim of this study was to perform a systematic review and meta-analysis addressing the prevalence of Cryptosporidium species in animals in Latin America. The estimated pooled prevalence rate for cryptosporidiosis in animals, by means of meta-analysis with a random-effects model, based on species identification, was 18.0% (95% CI 11.0%-27.0%) with high heterogeneity. The estimated overall prevalence was 20.3% (36/177) in pets, 19.9% (1309/6573) in livestock animals and 23.9% (954/3995) in exotic/captive animals. Evidence of circulation of 16 Cryptosporidium species was found in five Latin American countries: Brazil, Colombia, Chile, Argentina and Mexico. Through meta-analysis with a random-effects model, the pooled prevalence rate for Cryptosporidium parvum was 0.7% (95% CI 0.2%-2.4%). Cryptosporidium felis (8.5%) was the most prevalent species in pets, C. parvum (10.3%) in livestock animals and Cryptosporidium galli (17.6%) in exotic/captive animals. C. parvum was the species with the greatest geographical dispersion, which can be explained by its eurixenic and zoonotic potential. Few studies on cryptosporidiosis in animals in Latin America were found, which shows that there is a need for investment in and expansion of studies on this parasite. The pooled prevalence of C. parvum in Latin America and its wide circulation are similar to what has been observed in other developing regions, which reaffirms the importance of this species as the cause of a neglected, emerging and zoonotic parasitosis.

Microscopic Detection, Hematological Evaluation and Molecular Characterization of Piroplasms from Naturally Infected Dogs in Rio de Janeiro, Brazil.

PURPOSE: This study aimed to analyze the frequency of piroplasmids in the blood of dogs in Rio de Janeiro, compare the performance of microscopic techniques, assess the risk factors associated with infections and also molecularly and morphologically characterize the piroplasmids identified. METHODS: In all, 407 blood samples were collected from dogs between 2018 and 2019. These were subjected to microscopic parasitological techniques for thin and thick smears, stained with Giemsa and using a rapid staining kit. The slides were read under an optical microscope and the protozoa were characterized morphometrically. In addition, the blood samples were subjected to molecular characterization for diagnosing piroplasmid species using primers that amplified the gene 18S rRNA. RESULTS: Piroplasmids were detected in 38 (9.3%) samples. Of these, 33 samples presented nucleotide sequences compatible with Babesia vogeli. Most of the positive samples were young, male, defined breeds dogs that had been attended in clinics in São Gonçalo city. Thrombocytopenia and leukopenia were the hematological alterations more observed in positive samples, but positive samples without alterations were also detected. The sex was the only variable that showed statistical differences. Males dogs being more often infected than females (p < 0.05). The microscope slides mostly showed piriform and oval merozoites measuring greater than 2.5 µm in length, which were compatible with B. vogeli. However, smaller forms were also identified, thus demonstrating the polymorphic nature of this parasite. CONCLUSION: Babesia vogeli was detected in blood samples from dogs in the metropolitan cities of Rio de Janeiro by molecular techniques in different parasite morphotypes.

Detection of Dirofilaria immitis using microscopic, serological and molecular techniques among dogs in Cabo Frio, RJ, Brazil.

Heartworm disease is a health problem for dogs and cats, especially in tropical and subtropical coastal regions of the world. Some studies have compared the efficacy of the diagnostic techniques used to detect this parasitosis. Therefore, the aim of this study was to compare parasitological optical microscopy (POM), serological and molecular techniques for diagnosing canine heartworm infection. Samples were collected between July 2015 and April 2016 from 103 dogs in Cabo Frio, RJ, Brazil. The wet fresh blood, thick smears, thin smears and modified Knott's test were used to detect microfilariae. ELISA (Snap™ 4Dx ® IDEXX) was used to detect antigens and the polymerase chain reaction (PCR) was used to detect DNA and enable sequencing for species differentiation and confirmation. 19.4% of samples were positive according to microscopy. Through PCR, 15.5% of the total were positive. Using ELISA, the positivity rate was 29.1%. Occult heartworm infection was detected in 11.6% of the samples. ELISA sensitivity was shown to be higher than PCR or microscopy (P = 0.001). Sequencing of samples confirmed the presence of Dirofilaria immitis and Acanthocheilonema reconditum . ELISA was more effective for serological diagnosis canine heartworm and should be used in clinical and epidemiological studies.

Molecular evaluation of piroplasms and hematological changes in canine blood stored in a clinical laboratory in Niterói, Rio de Janeiro.

Piroplasm species were analyzed by molecular tools in total 31 blood samples from positive dogs, previously checked by stained slides, stored until DNA extraction between 2016 to 2018 in the laboratory Clinical Analyzes in Niterói, Rio de Janeiro. The piroplasms were identified by PCR, targeting the 18S rRNA gene and sequencing. From the total number of samples only 24 (77.4%) were positive and show adequate nucleotide sequences for interpretation with identity between 93%-100% with Babesia vogeli in compared to the sequences isolated of infected dogs from other states in Brazil deposited on GenBank. Most of dogs infected with B. vogeli had anemia (62.5%) and thrombocytopenia (95.8%). The findings of this study are compatible with previous reports in the literature and highlight B. vogeli as the most incriminated species in canine piroplasmosis in Brazil, and thrombocytopenia the hematological alteration most frequently identified in this infection. It is important to note that this is the first study involving the molecular characterization of piroplasms in the metropolitan region of Rio de Janeiro, based on PCR followed by sequencing.

Oncicola venezuelensis (Marteau, 1977) (Acanthocephala: Oligacanthorhynchidae) in Puma concolor in Rio de Janeiro, Brazil.

Specimens of Oncicola venezuelensis (Marteau, 1977) were recovered from fragments of intestinal tissue of a female Puma concolar (Linn, 1771) found dead in Petrópolis, Rio de Janeiro in 2017. A total of 140 helminths were recovered. Five males and 5 females of the helminths were analyzed morphologically as well as 50 parasite eggs recovered in intestinal contents. Morphologically, these helminths were compatible with the genus Oncicola, because of the size and shape of the proboscis, the size and disposition of the lemnisci and the morphometry of the eggs, in which the external membrane of the shell was delicate and clear. From histopathology, the helminths were deeply embeded in the mucosa reaching up to the muscle layer. One specimen was also identified molecularly with universal primers that amplified the eukaryote region ITS1-5.8S-ITS2. The helminth showed 99% identity with the gene sequence of O. venezuelensis deposited in GenBank. It is important to emphasize, this parasite has been very little reported in the literature, which reinforces the importance of this report.

Gastrointestinal Parasites in Captive Animals at the Rio de Janeiro Zoo.

BACKGROUND: Gastrointestinal parasites may determine diarrhea, dysentery or even death in captive mammals. These animals tend to be more susceptible to parasitic infections due to confinement and stress. Purpose To increase the information about these etiological agents in captive animals in Brazil, the gastrointestinal parasites of the captive mammals of the Rio de Janeiro Zoo were investigated. METHODS: From 2016 to 2018, 180 fecal samples were collected from animals housed in the Rio de Janeiro Zoo: 63 from animals of the order Primates, 26 of Carnivora, 78 of Artiodactyla, 9 of Perissodactyla and 4 of the order Rheiformes. The feces were processed by direct examination and by the techniques of Faust et al., Sheather, Ritchie, Lutz, and smears were stained with safranin. Immunoenzymatic assays were also performed to investigate antigens of Giardia duodenalis, Cryptosporidium spp., Entamoeba histolytica/Entamoeba dispar. RESULTS: Parasite positivity was identified in 68.3% of the fecal samples, with a parasite positivity rate of 68.2% among primates, 65.3% among carnivores, 69.2% among artiodactyls, 33.3% among perissodactyls, and 100% among rheiformes. The most frequently detected parasite was Entamoeba histolytica/E. dispar antigens, which showed a statistically significant positivity rate (33.3%; p = 0.000), particularly in the feces of carnivores (30.7%) and artiodactyls (53.8%). A statistically significant positivity rate of Balantioides coli (11.1%; p = 0.001) was also detected in feces from nonhuman primates, tapirs, collared peccaries and rheas. The positivity of Cryptosporidium sp. antigens in feces of the orders Carnivora, Artiodactyla and Primates was also statistically significant (7.2%, p = 0.010). Oocysts compatible with Cryptosporidium spp. were detected in 6.3% from primates. The helminths most frequently detected were thin-shelled eggs of nematodes (17.7%, p = 0.000), nematode larvae (15.5%, p = 0.000) and Trichuris trichiura eggs (6.1%, p = 0.018). CONCLUSION: The positivity rate for gastrointestinal parasites demonstrates the need for a sanitation management program to be implemented in the zoo, including routine diagnostic parasitology tests followed by specific treatment for each parasitosis.

Comparison of Four Parasitological Techniques for Laboratory Diagnosis of Eggs from Spirometra spp. in Wild Mammal Fecal Samples.

PURPOSE: This study aimed to compare the performance of four different microscopic coproparasitological techniques in relation to egg recovering and the frequency of alterations in the eggs observed through each technique. METHODS: A total of 213 fecal samples from free-living carnivorous mammals were collected between 2017 and 2018 in Itatiaia National Park, RJ, Brazil. Faust and modified Sheather floatation techniques as well as Lutz and modified Ritchie sedimentation techniques were applied. RESULTS: The total positivity rate for Spirometra spp. eggs was 24.4%, and these were detected mainly through Lutz and modified Ritchie sedimentation techniques, with substantial agreement (kappa = 0.73; p = 0.00 < 0.05; McNemar p value = 1.0; Fisher's exact test p = 0.616). Faust and modified Sheather flotation techniques did not present good egg recovery, with frequencies of 6.6% and 7.5%, respectively. Eggs with morphological alterations were mostly observed through Faust (17.3%) and modified Sheather (13.5%). Both flotation techniques presented statistically significant frequencies of deformed eggs, in comparison with the sedimentation techniques (p = 0.00). Low frequencies of deformed eggs were observed when the samples were analyzed through modified Ritchie and Lutz sedimentation techniques. CONCLUSIONS: From these results, sedimentation techniques such as modified Ritchie and Lutz methods were more efficient for diagnosing the eggs of this helminth in fecal material from free-living carnivores and should always be used when analyzing fecal samples from hosts of different species.

Balantioides coli: morphological and ultrastructural characteristics of pig and non-human primate isolates.

Balantioides coli is a ciliated protozoon that inhabits the intestine of pigs, non-human primates and humans. Light microscopy studies have described over 50 species of the genus Balantioides but their validity is in doubt. Due to the limited information about this genus, this study is aimed to identify morphological characteristics of Balantioides coli isolated using fluorescence microscopy and both scanning (SEM) and transmission electron microscopy (TEM). Trophozoites isolated from the feces of pig and macaque were washed and subjected to centrifugation. These cells were fixed with paraformaldehyde for immunofluorescence. Other aliquots of these trophozoites were fixed with glutaraldehyde, post fixed with osmium tetroxide and processed for SEM and TEM. Immunofluorescence studies revealed microtubules with a longitudinal distribution to the main axis of the parasite and in the constitution of cilia. SEM demonstrated a high concentration of cilia covering the oral apparatus and a poor presence of such structures in cytopyge. TEM revealed in the plasma membrane, several associated structures were observed to delineate the cellular cortex and mucocysts. The cytoskeleton of the oral region was observed in detail and had an organization pattern consisting of microtubules, which formed files and nematodesmal networks. Organelles such as hydrogenosomes like and peroxisomes were observed close to the cortex. Macronuclei were observed, but structures that were consistent with micronuclei were not identified. Ultrastructural morphological analysis of isolates confirms its similarity to Balantioides coli. In this study were identified structures that had not yet been described, such as hydrogenosomes like and cytoskeletal structures.

Comparative study of three xenic media culture for cultivation of Balantidium coli strains.

The aim of the present study was to evaluate the growth rate of Balantidium coli in three xenic media cultures. Between 2013 and 2015, 10 B. coli isolates obtained from feces of Cynomolgus macaques, and 30 isolates from feces of pigs were studied. An inoculum of 500 trophozoites was transferred to tubes containing LES, TYSGM-9 and Pavlova media. These cultures were evaluated at incubation times of 24, 48, 72 and 96 hours. In most of strains analyzed wasn't showed significant difference in the growth rate comparing TYSGM-9 and Pavlova media (Wilcoxon p>0.016). In Pavlova medium, the trophozoites showed a maximum growth at 72 hours with significant difference when compared with the times of 24 h and 96 h (Wilcoxon <0.008). In LES, viable trophozoites were observed until 24 hours, with a significant difference (Friedman p<0.05, Wilcoxon p<0.016) in the number of parasite cells compared with Pavlova and TYSGM-9 media cultures. Thus, LES medium seemed to be less adequate than the other media for maintenance of B. coli. Despite the satisfactory results in TYSGM-9, Pavlova medium was considered ideal for the maintenance of this protozoan strain, guaranteeing the viability of the parasite with subculture every three days, presenting lower costs.

Gastrointestinal parasites among felids inhabiting the Serra dos Órgãos National Park, Rio de Janeiro, Brazil.

This study aimed to investigate the species of felids that inhabit the Serra dos Órgãos National Park (Parnaso) and gastrointestinal parasites at various stages of their life cycles in the feces of these animals. Between 2013 and 2015, felid feces were collected from trails in Parnaso. The sampling points were georeferenced. A total of 82 fecal samples were processed, of which 79 were collected on the ground, two from captured felids and one from a necropsied animal. All samples underwent coproparasitological techniques. Samples collected from the environment underwent additional trichological analysis. Fur patterns corresponding to Leopardus guttulus, Leopardus pardalis, Leopardus wiedii and Puma yagouaroundi were observed in 32 of the samples collected on the soil. High frequency of potentially parasitic evolving forms (88.6%) was observed in felid feces, particularly eggs of the family Diphyllobothriidae (68.6%). Besides, were also detected, eggs of superfamily Ascaridoidea (42.9%), nematode larvae (28.6%), eggs of order Strongylida (28.6%), Capillaria sp. (8.6%), Trichuris sp. (8.6%), eggs of order Spirurida (2.9%), unsporulated coccidian oocysts (8.6%) and Eimeria sp. (2.9%). Felid feces presented higher frequency of polyparasitism (60%) than monoparasitism (28.6%).

Gastrointestinal parasites in stray and shelter cats in the municipality of Rio de Janeiro, Brazil.

The increasingly urban nature of the population has led many people to choose independent pets, such as cats. This situation has also made it possible for these animals to be abandoned, thus increasing the numbers of cats on the streets and in shelters. These animals can act as a source of infection for other hosts. Between 2014 and 2015, the frequency of gastrointestinal parasites in captive and stray cats in the municipality of Rio de Janeiro was analyzed. Ninety-one fecal samples were collected from captive cats and 172 from stray cats. Centrifugal sedimentation and flotation techniques were used. The frequency of parasites among the stray cats was 77.3%, and this was significantly higher than the frequency observed in captive cats (49.5%). Helminths were detected more frequently, and hookworms were the parasites most detected. Toxocara cati, Cystoisospora sp. and Dipylidium caninum were also detected. No statistical difference in the frequency of parasites was observed between the sexes among the captive cats. However, among the stray cats, males (85.5%) presented higher positivity than females (71.8%). The high frequency of hookworms, which are the agent for "cutaneous larva migrans" in humans, shows the need to control parasitic infections among the cats studied.

First molecular characterization of Balantioides coli (Malmsten, 1857) isolates maintained in vitro culture and from feces of captive animals, Rio de Janeiro, Brazil.

Ciliate protozoa of the genus Balantioides can parasitize a variety of animals. The morphology of the evolutionary forms of the parasite and the host species affected have long been the only characteristics used to taxonomically identify the species of these protozoa, but these variables are not very precise. To confirm species identity, molecular biology tools are currently used. In this context, this study aimed to analyze protozoan isolates maintained in culture medium and from fecal samples from captive animals in Rio de Janeiro, Brazil, by means of molecular tools. Forty isolates maintained in Pavlova modified medium (30 were isolated from feces of pigs and 10 from feces of cynomolgus macaques) were analyzed. In addition, 34 fecal samples (8 from pigs, 8 from cynomolgus macaques and 18 from rhesus macaques) containing Balantioides coli-like cysts were analyzed. All samples were subjected to DNA extraction and the polymerase chain reaction (PCR) to amplify the fragment ITS1 - 5.8s rRNA - ITS2, and the PCR products were purified and sequenced. All samples (100%) presented sequences that were grouped in the Balantioides coli cluster. The type A0 variant predominated. These sequences were 96% to 99% identical to those deposited in GenBank, including a B. coli sequence that had been obtained from human fecal material in Bolivia. It seems that the culturing system did not select variants, because this variant was also seen in the amplified sequences of fecal samples containing cysts. The isolate sequences in the cultures showed few ambiguities and substitutions, thus generating reliable chromatograms. This was the first study to identify B. coli in captive animals in Brazil, through molecular biology. In addition, it was the first to evaluate a large panel of isolates of the parasite through culturing.

Trypanosoma cruzi I genotype among isolates from patients with chronic Chagas disease followed at the Evandro Chagas National Institute of Infectious Diseases (FIOCRUZ, Brazil).

INTRODUCTION:: Trypanosoma cruzi is the etiologic agent of Chagas disease in humans, mainly in Latin America. Trypanosome stocks were isolated by hemoculture from patients followed at Evandro Chagas National Institute of Infectious Diseases (FIOCRUZ) and studied using different approaches. METHODS:: For species and genotype identification, the stocks were analyzed by parasitological techniques, polymerase chain reaction assays targeted to specific DNA sequences, isoenzyme patterns, besides sequencing of a polymorphic locus of TcSC5D gene (one stock). RESULTS:: The isolates presented typical T. cruzi morphology and usually grew well in routine culture media. Metacyclic trypomastigotes were found in cultures or experimentally infected Triatoma infestans. All isolates were pure T. cruzi cultures, presenting typical 330-bp products from kinetoplast DNA minicircles, and 250 or 200-bp amplicons from the mini-exon non-transcribed spacer. Their genetic type assignment was resolved by their isoenzyme profiles. The finding of TcI in one asymptomatic patient from Paraíba was confirmed by the sequencing assay. TcVI was found in two asymptomatic individuals from Bahia and Rio Grande do Sul. TcII was identified in six patients from Pernambuco, Bahia and Minas Gerais, who presented different clinical forms: cardiac (2), digestive with megaesophagus (1), and indeterminate (3). CONCLUSIONS:: The main T. cruzi genotypes found in Brazilian chronic patients were identified in this work, including TcI, which is less frequent and usually causes asymptomatic disease, unlike that in other American countries. This study emphasizes the importance of T. cruzi genotyping for possible correlations between the parasite and patient' responses to therapeutic treatment or disease clinical manifestations.

Comparison of five parasitological techniques for laboratory diagnosis of Balantidium coli cysts.

Balantidium coli is a protozoon that can cause dysentery in humans, pigs and nonhuman primates, with zoonotic potential. In the literature, there is still little information on the effectiveness of different laboratory techniques for diagnosing this disease. This study compared and evaluated the performance of the Lutz, modified Ritchie, Faust, modified Sheather and direct examination techniques for detecting cysts of this protozoon. Between 2012 and 2014, 1905 fecal samples were collected from captive animals in the state of Rio de Janeiro. Of these, 790 were obtained from the rectum of pigs and 1115 from enclosures occupied by nonhuman primates. B. coli cysts were most evident through direct examination (22.4% of the samples) and the Lutz technique (21%). Fair agreement (Kappa = 0.41; p < 0.05) was observed only between direct examination and Lutz. The flotation techniques (Faust and modified Sheather) did not show good recovery of cysts. A statistically significant difference (p < 0.05) in the frequency of cysts between pigs and nonhuman primates could only be observed through direct examination and the Lutz technique. The most efficient method for diagnosing this parasitosis was seen to an association between direct examination and the spontaneous sedimentation technique.

Detection of Dirofilaria immitis using microscopic, serological and molecular techniques among dogs in Cabo Frio, RJ, Brazil / Detecção de Dirofilaria immitis utilizando técnicas microscópicas, imunológicas e moleculares em cães de Cabo Frio, RJ

Abstract Heartworm disease is a health problem for dogs and cats, especially in tropical and subtropical coastal regions of the world. Some studies have compared the efficacy of the diagnostic techniques used to detect this parasitosis. Therefore, the aim of this study was to compare parasitological optical microscopy (POM), serological and molecular techniques for diagnosing canine heartworm infection. Samples were collected between July 2015 and April 2016 from 103 dogs in Cabo Frio, RJ, Brazil. The wet fresh blood, thick smears, thin smears and modified Knott's test were used to detect microfilariae. ELISA (Snap™ 4Dx ® IDEXX) was used to detect antigens and the polymerase chain reaction (PCR) was used to detect DNA and enable sequencing for species differentiation and confirmation. 19.4% of samples were positive according to microscopy. Through PCR, 15.5% of the total were positive. Using ELISA, the positivity rate was 29.1%. Occult heartworm infection was detected in 11.6% of the samples. ELISA sensitivity was shown to be higher than PCR or microscopy (P = 0.001). Sequencing of samples confirmed the presence of Dirofilaria immitis and Acanthocheilonema reconditum . ELISA was more effective for serological diagnosis canine heartworm and should be used in clinical and epidemiological studies.

Resumo A dirofilariose é um problema de saúde para cães e gatos, especialmente nas regiões costeiras tropicais e subtropicais do mundo. Alguns estudos compararam a eficácia das técnicas de diagnóstico usadas para detectar esta parasitose. Portanto, o objetivo deste estudo foi comparar a microscopia óptica (OM), técnicas sorológicas e moleculares para o diagnóstico de infecção por Dirofilaria immitis . Foram coletadas, entre julho de 2015 e abril de 2016, amostras de 103 cães em Cabo Frio, RJ, Brasil. O exame direto, distensão espessa, distensão delgada e o teste de Knott modificado foram usados para detectar microfilárias. O ELISA (Snap ™ 4Dx ® IDEXX) foi usado para detectar antígenos e a reação em cadeia da polimerase (PCR) foi usada para detectar DNA e o sequenciamento para diferenciação e confirmação de espécie. Das amostras, 19,4% foram positivas de acordo com a microscopia. Por PCR, 15,5% do total foram positivos. Utilizando o ELISA, a taxa de positividade foi de 29,1%. Dirofilariose oculta foi detectada em 11,6% das amostras. A sensibilidade ao ELISA mostrou-se superior à PCR ou microscopia (P = 0,001). O sequenciamento das amostras confirmou a presença de Dirofilaria immitis e Acanthocheilonema reconditum . O ELISA foi mais eficaz no diagnóstico sorológico de dirofilariose canina e deve ser usado em estudos clínicos e epidemiológicos.

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.

Hospital food handlers in Niterói, RJ, Brazil: intestinal parasitism.

A survey for intestinal parasites was carried out with food handlers from two private and three public hospitals in Niterói City, RJ, Brazil. The aim of this research was to verify the enteroparasites prevalence in this professional group. The investigation was divided in two phases. The first phase consisted of interviews with the participants; coproparasitological exams using Lutz, Faust et al. and Baermann techniques; under fingernail material analysis, using Mello et al. modified method; and educational lectures to food handlers. In the second phase, coproparasitological exams were repeated. Positive results were observed in 14.2% (17/120) and 17.1% (12/70) of the individuals in the first and second phases respectively. The most frequent parasite was Entamoeba coli, detected in 48.5% (16/33) of the samples with positive results. Under fingernail residues were observed in 19.2% (23/120) of the food handlers. E. coli cysts were found in one fingernail residue, likewise they were detected in the feces of the same food handler. Such data showed a potential transmission risk of intestinal parasites by food handling, indicating the need of adopting a diagnosis/orientation procedure as a bi-annual routine activity in hospitals, in order to improve the food service quality and population health condition.