Molecular detection of coccidian Apicomplexa Parasites isolated from wild crab-eating and pampas foxes through novel TaqMan™ probes: a contribution to their molecular epidemiology.

Neospora caninum, Toxoplasma gondii and Hammondia spp. are coccidian parasites similar in morphology. Molecular techniques are necessary to detect parasite DNA isolated from stool samples in wild canids because they were reported as definitive hosts of N. caninum life cycle. The objective of this study was to develop a highly sensitive and accurate molecular method for the identification of coccidian Apicomplexa parasites in crab-eating fox (Cerdocyon thous) and pampas fox (Lycalopex gymnocercus). Tissue samples from road-killed animals (pampas fox = 46, crab-eating fox = 55) and feces (pampas fox = 84, crab-eating fox = 2) were collected, and species were diagnosed through molecular assay. PCR was used for the amplification of a fragment of the coccidian Apicomplexa nss-rRNA gene. Additionally, we developed a novel real-time PCR TaqMan™ probe approach to detect T. gondii- Hammondia spp. and N. caninum. This is the first report of N. caninum DNA in pampas fox feces (n = 1), thus it was also detected from pampas fox tissues (n = 1). Meanwhile, T. gondii was found in tissues of pampas (n = 1) and crab-eating (n = 1) foxes and H. triffittae in one crab-eating fox tissue. Despite the low percentage (2.5%) of positive samples, the molecular method developed in this study proved to be highly sensitive and accurate allowing to conduct an extensive monitoring analysis for these parasites in wildlife.

Dual RNA-seq reveals no plastic transcriptional response of the coccidian parasite Eimeria falciformis to host immune defenses.

BACKGROUND: Parasites can either respond to differences in immune defenses that exist between individual hosts plastically or, alternatively, follow a genetically canalized ("hard wired") program of infection. Assuming that large-scale functional plasticity would be discernible in the parasite transcriptome we have performed a dual RNA-seq study of the lifecycle of Eimeria falciformis using infected mice with different immune status as models for coccidian infections. RESULTS: We compared parasite and host transcriptomes (dual transcriptome) between naïve and challenge infected mice, as well as between immune competent and immune deficient ones. Mice with different immune competence show transcriptional differences as well as differences in parasite reproduction (oocyst shedding). Broad gene categories represented by differently abundant host genes indicate enrichments for immune reaction and tissue repair functions. More specifically, TGF-beta, EGF, TNF and IL-1 and IL-6 are examples of functional annotations represented differently depending on host immune status. Much in contrast, parasite transcriptomes were neither different between Coccidia isolated from immune competent and immune deficient mice, nor between those harvested from naïve and challenge infected mice. Instead, parasite transcriptomes have distinct profiles early and late in infection, characterized largely by biosynthesis or motility associated functional gene groups, respectively. Extracellular sporozoite and oocyst stages showed distinct transcriptional profiles and sporozoite transcriptomes were found enriched for species specific genes and likely pathogenicity factors. CONCLUSION: We propose that the niche and host-specific parasite E. falciformis uses a genetically canalized program of infection. This program is likely fixed in an evolutionary process rather than employing phenotypic plasticity to interact with its host. This in turn might limit the potential of the parasite to adapt to new host species or niches, forcing it to coevolve with its host.

Prevalence of helminth ova in soil samples from public places in Shiraz.

To determine the prevalence of helminth eggs in public places and children's playgrounds, 112 soil samples were collected in 26 sites in Shiraz, southern Islamic Republic of Iran, during September 2002-September 2003. Toxocara cati ova were found in 7 (6.3%) samples, 2 had Ascaris lumbricoides ova, 3 had larvae morphologically similar to Strongyloides stercoralis. Coccidia oocysts were also observed in 4 samples. No contamination was observed during the dry season.

Coccidia of turkey: from isolation, characterisation and comparison to molecular phylogeny and molecular diagnostics.

Coccidiosis is a disease caused by apicomplexan parasites of the genus Eimeria, which has a significant economic impact on poultry production. Multiple species infecting the turkey have been described; however, due to the general lack of unambiguous description, their identification and taxonomy is debatable. In this work, a systematic approach was taken to isolate, characterise and compare coccidian species in the turkey. Individual species were tracked according to their unique 18S ribosomal DNA sequence. The single-oocyst isolation technique and passaging of mixed species field isolates in selectively immunised birds enabled the derivation of pure species. Six distinct strains representing five eimerian species that infect the turkey were obtained. It appears highly probable that these species represent all species described in the past with the exception of Eimeria subrotunda. The species were analysed using both traditional methods and DNA sequencing. For each strain the oocyst morphology, prepatent period, gross pathology, pathogenicity, host specificity and endogenous cycle were studied. Antigenic similarity was investigated in multiple cross-immunity experiments. For identification and quantification of each individual species or strain, quantitative real-time PCR markers were also developed. Parallel characterisation of pure strains allowed comprehensive comparison with the original descriptions and assignment of correct species names. The species Eimeria meleagridis, Eimeria dispersa, Eimeria gallopavonis, Eimeria meleagrimitis and Eimeria innocua were identified. Comparison of our data with those of previous studies indicates that Eimeria adenoeides is most probably a synonym for either E. meleagridis or E. gallopavonis, or a description based on a mixture of these species, and thus nomen dubium. The species E. dispersa and E. innocua were also found to infect Bobwhite Quail. Phylogenetic reconstruction based on 18S rDNA and cytochrome c oxidase subunit I gene (COI) sequences showed that these two species form a distinct clade unrelated to other turkey coccidia and point to a polyphyletic origin of the species infecting the turkey.

Occurrence of anti-Neospora caninum antibodiesin dogs in rural areas in Minas Gerais, Brazil / Ocorrência de anticorpos anti-Neospora caninum em cães de áreas rurais em Minas Gerais, Brasil

The aim of this study was to determine the frequency of anti-Neospora caninum antibodies and risk factors associated with seropositivity in 240 dogs from rural areas of the Lavras, Belo Horizonte and Nanuque regions, state of Minas Gerais, Brazil. The indirect fluorescence antibody test (IFAT) with a cutoff of 50 was used to assess the frequency of seropositive dogs. The risk factor analysis was performed using the Chi-square test (χ2) and multiple logistic regression. The frequency of seropositivity for N. caninum was 15% of the dogs (36/240). Presence of dogs without defined breed (p = 0.018; OR = 5.9) and presence of cattle on the farm (p = 0.053, OR = 4.3) were associated with N. caninum seropositivity. Dogs in the Nanuque region had higher seropositivity for N. caninum (29.9%) than did those in the Lavras (6.2%) and Belo Horizonte (2.2%) regions (p < 0.05). The presence of seropositive dogs in rural areas of Minas Gerais demonstrates the potential risk of horizontal transmission of N. caninum to cattle, especially from dogs without defined breed, which were four times more likely to be seropositive than purebred dogs were.

O objetivo deste estudo foi determinar a frequência de anticorpos anti-Neospora caninum e fatores de risco associados à soropositividade, em 240 cães de áreas rurais, nas regiões de Lavras, Belo Horizonte e Nanuque, Minas Gerais, Brasil. Foi utilizada a reação de imunofluorescência indireta (RIFI), com um ponto de corte igual a 50, para avaliar a frequência de cães soropositivos. A análise dos fatores de risco foi realizada pelo teste de Qui-quadrado (χ2) e regressão logística múltipla. A frequência de cães soropositivos para N. caninum foi de 15% (36/240). A presença de cães sem raça definida (SRD) (p = 0,018; OR = 5,9) e bovinos na propriedade (p = 0,053; OR = 4,3) foi associada à soropositividade pelo N. caninum. A frequência de cães soropositivos foi mais elevada na região de Nanuque (29,9%), quando comparados aos das regiões de Lavras (6,2%) e Belo Horizonte (2,2%) (p < 0,05). A presença de cães soropositivos, em áreas rurais de Minas Gerais, demonstra o risco potencial da transmissão horizontal de N. caninum para os bovinos, especialmente os cães SRD, que apresentaram quatro vezes mais a chance de serem soropositivos em relação aos cães com raça definida.

Experimental coccidiosis in goats. 1. Experimental model. Effects of parasitism on the feeding behaviour and the growth of animals; intestinal lesions.

Experimentally infecting goats with three species of coccidia (Eimeria ninakohlyakimovae, E. arloingi, E. christenseni) and placing them in metabolism cage helped us to obtain a parasitic development which was significant and continuous. This development is not necessarily accompanied by clinical symptoms. However slight constipation and not diarrhoea was observed. Feed consumption is reduced and a subsequent reduction in the rate of growth is observed. As far as the feeding behaviour is concerned, the reduction in the rate of growth is compensated by the animals choosing those parts of their ration which are most easily digested.

Eimeria minasensis n.sp. (Apicomplexa: Eimeriidae) in the domestic goat Capra hircus, from Brazil

Eimeria minasensis n.sp. is described in the domestic goat Capra hircus from Brazil. Oocysts ellipsoidal are 35 x 24.5 (32-37.7 x 20.9-27.9) µm. Sporocysts elongate-ellipsoid are 15.2 x 9 (12.3-18.4 x 7.8-10.2) µm, with a Stieda body at the narrow end. Oocyst wall smooth and bilayered; outer layer about 1.2 (0.8-1.6) µm and colorless; inner layer about 0.5 (0.4-0.8) µm and dark-brown. Micropyle, a mound-shaped micropylar cap 1,6 x 8,9 (0,8-2 x 7-10,2) easily dislodged; one or more oocyst polar granules present. Oocyst residuum absent. Sporocyst residuum present, composed of many scattered granules. Sporozoites elongate, lying lengthwise, "head to tail" in the sporocysts; one or two refractile globules are usually visible. Sporulation time was 120 hr at 27ºC, prepatent period, 19 to 20 days and patent period 15 to 25 days. Gamonts, gametes and oocysts present in cecum and colon. Prevalence was 12.8 per cent (6/47) in goats from Minas Gerais, Brazil.

Contaminaçäo por enteroparasitas em hortaliças comercializadas nas cidades de Niterói e Rio de Janeiro, Brasil / Intestinal parasites contamination from vegetables comercialized in Niterói and Rio de Janeiro cities, Brazil

O objetivo deste estudo foi avaliar a contaminaçäo por enteroparasitas em hortaliças consumidas cruas comercializadas nas cidades de Niterói e Rio de Janeiro. Foram estudadas 128 amostras de hortaliças - alface (Lactuca sativa) e agriäo (Nasturtium officinale) provenientes do comércio (supermercados, feiras-livre e quitandas) e de restaurantes tipo self-services. Apenas 6,2 por cento das amostras apresentaram presença de estruturas parasitárias com morfologia semelhante as de espécies parasitas de animais. Foi encontrado presença de contaminantes como ácaros, ovos de ácaros, insetos, larvas de nematóides e protozoários ciliados em quase todas as amostras (96,1 por cento), inclusive nas de restaurantes. Este alto percentual sugere a presença de risco de infecçäo, pois associado a esses agentes poderiam existir estruturas parasitárias infectantes para o homem

Helmintos e protozoários em fezes de javalis (Sus scrofa scrofa) criados em cativeiro / Helminths and protozoa in wild boars (Sus scrofa scrofa) feces raised in captivity

Faecal samples from 79 wild boars (Sus scrofa scrofa) were examined by sedimentation method, zinc sulfate flotation method, and centrifugal flotation in sugar solution for endoparasites research. The results showed that 97.5 por cento of the samples were positive for helminths and/or protozoa as follows: strongilides (70.9 por cento), Ascaris suum (46.9 por cento), Trichuris suis (29.1 por cento), Metastrongylus sp. (12.6 por cento), Strongyloides ransomi (3.8 por cento), Balantidium coli (38.0 por cento), Entamoeba spp. (15.2 por cento), Giardia spp. (1.3 por cento), Blastocystis sp. (12.6 por cento). The frequency of Entamoeba was higher in young animals. Coccidian oocysts were observed in 59.5 por cento samples and five species of Eimeria and one Isospora were recovered: Eimeria scabra (31.9 por cento), E. deblieck (31.9 por cento), E. perminuta (23.4 por cento), E. cerdonis (17.0 por cento), E. scrofae (12.8 por cento) and Isospora suis (12.8 por cento).

Cryptosporidiosis en pacientes con sida: acciones de la enfermera / Cryptosporidiosis in AIDS patients. Nurse actions

Cryptosporidium spp es una coccidia intestinal con una distribución mundial, que ha sido recientemente reconocida como una importante causa de cuadros diarreicos, malabsorción y pérdida de peso, y que puede llegar a constituir una amenaza para la vida de personas con compromiso inmunológico, como los pacientes con síndrome de inmunodeficiencia adquirida (SIDA). El propósito de este estudio fue reflejar las características de esta parasitosis, en un grupo de 24 pacientes seropositivos al virus de la inmunodeficiencia humana (VIH), que estuvieron ingresados en el Servicio de Medicina del Instituto "Pedro Kourí" (IPK), durante los años 1995 y 1996. Para el análisis de los datos se utilizó la prueba de comparación de proporciones; se encontró que todos los individuos parasitados por esta coccidia estaban comprendidos en el grupo IV de la infección por el VIH. La diarrea y la pérdida de peso fueron las alteraciones clínicas más frecuentemente asociadas con esta parasitosis en los 24 pacientes estudiados (100 porciento). Se revisó el papel fundamental que desempeña la enfermera en la prevención frente a esta problemática en los diferentes niveles de atención en salud

Hallazgo de Eimeria tenella en células epiteliales de la bolsa de Fabricio / Findings of Eimeria tenella in epithelial cells of the bursa of Fabricius

En el presente trabajo se describen observaciones realizadas durante un experimento diseñado para estudiar el comportamiento de los leucocitos polimorfonucleares en infecciones con E. tenella en pollos tratados con 5-FU como agente granulocitopénico. Ciento veinte pollitos de engorda fueron asignados en 4 grupos con 30 pollitos cada uno: 1) testigo blanco, 2) tratado con 200 mg/kg de peso de 5-fluorouracilo (5-FU), 3) infectado oralmente con 500 ooquistes esporulados de Eimera tenella y 4) infectado con E tenella después del tratamiento con 5-FU. Se administró E. tenella a 5 pollos de cada grupo a los días: 2, 4, 6, 8, 10 y 12 días posinoculación con 5-FU (17 días de edad). Siete días posinfección con E. Tenella se tomaron muestras de tejidos de varios órganos, para su estudio histológico. Se observaron coccidias en diferentes estado de desarrollo en 25 y 24 pollos en los grupos 3 y 4, respectivamente. Se observó hiperplasia epitelial moderada a severa, infiltrado mononuclear en el subepitelio, y algunos quistes intraepiteliales, que fueron asociados con la presencia del parásito. Este hallazgo muestra que bajo ciertas circunstancias E. tenella es capaz no sólo de invadir el epitelio de la bolsa de Fabricio, sino tambien de desarrollarse en él