PHYTOHAEMAGGLUTININ’S EFFECT ON NEOSPORA CANINUM ANTIGEN PRODUCTION

The parasite Neospora caninum affects mainly cattle and dogs. This study aimed to evaluate the effect of phytohaemagglutinin (PHE) in antigen production of N. caninum NC-1 strain in gerbils (Meriones unguiculatus) and in vitro. 20 gerbils were used, 10 inoculated intraperitoneally with 1 x 106 tachyzoites and 10 with 1 x 106 tachyzoites plus 300 µL/mL of PHE. 16 bottles of Vero cell culture were inoculated, 8 with 1.5 x 105 tachyzoites and 8 with 1.5 x 105 tachyzoites plus 30 µL/mL of PHE. Serology of gerbils was performed on day 0 and before euthanasia. Tachyzoites present in peritoneal fluid and cell culture bottles were quantified by Neubauer chamber and by real-time PCR (qPCR). PHE has not interfered in the production of tachyzoites of N. caninum in intraperitoneal inoculated gerbils and the effect of PHE in cell culture had a negative impact, considering the qPCR technique as the gold standard

Ecological study of Paracoccidioides brasiliensis in soil: growth ability, conidia production and molecular detection

<p>Abstract</p> <p>Background</p> <p><it>Paracoccidioides brasiliensis </it>ecology is not completely understood, although several pieces of evidence point to the soil as its most probable habitat. The present study aimed to investigate the fungal growth, conidia production and molecular pathogen detection in different soil conditions.</p> <p>Methods</p> <p>Soils samples of clayey, sandy and medium textures were collected from ground surface and the interior of armadillo burrows in a hyperendemic area of Paracoccidioidomycosis. <it>P</it>. <it>brasiliensis </it>was inoculated in soil with controlled humidity and in culture medium containing soil extracts. The molecular detection was carried out by Nested PCR, using panfungal and species specific primers from the ITS-5.8S rDNA region.</p> <p>Results</p> <p>The soil texture does not affect fungus development and the growth is more abundant on/in soil saturated with water. Some soil samples inhibited the development of <it>P. brasiliensis</it>, especially those that contain high values of Exchangeable Aluminum (H+Al) in their composition. Some isolates produced a large number of conidia, mainly in soil-extract agar medium. The molecular detection was positive only in samples collected from armadillo burrows, both in sandy and clayey soil.</p> <p>Conclusion</p> <p><it>P. brasiliensis </it>may grow and produce the infectious conidia in sandy and clayey soil, containing high water content, mainly in wild animal burrows, but without high values of H+Al.</p

Extracellular vesicles in infectious diseases caused by protozoan parasites in buffaloes

Background: Extracellular vesicles (EVs) are small membrane-bound vesicles of growing interest in vetetinary parasitology. The aim of the present report was to provide the first isolation, quantification and protein characterization of EVs from buffalo (Bubalus bubalis) sera infected with Theileria spp. Methods: Infected animals were identified through optical microscopy and PCR. EVs were isolated from buffalo sera by size-exclusion chromatography and characterized using western blotting analysis, nanoparticle tracking analysis and transmission electron microscopy. Subsequently, the proteins from isolated vesicles were characterized by mass spectrometry. Results: EVs from buffalo sera have shown sizes in the 124-140 nm range and 306 proteins were characterized. The protein-protein interaction analysis has evidenced biological processes and molecular function associated with signal transduction, binding, regulation of metabolic processes, transport, catalytic activity and response to acute stress. Five proteins have been shown to be differentially expressed between the control group and that infected with Theileria spp., all acting in the oxidative stress pathway. Conclusions: EVs from buffaloes infected with Theileria spp. were successfully isolated and characterized. This is an advance in the knowledge of host-parasite relationship that contributes to the understanding of host immune response and theileriosis evasion mechanisms. These findings may pave the way for searching new EVs candidate-markers for a better production of safe biological products derived from buffaloes

White piedra and pediculosis capitis in the same patient

Piedra branca é micose superficial causada pelo gênero Trichosporon e caracterizada por nódulos aderidos à haste do pelo. Pediculose capitis é causada pelo Pediculus humanus var. capitis pertencente à subordem Anoplura. Enquanto que a pediculose é enfermidade comum, relatos clínicos de piedra branca são raros. Técnicas de biologia molecular identificaram o agente de piedra branca do presente relato como T.inkin. Os autores apresentam associação de ambas as infestações no mesmo paciente para salientar seus aspectos clínicos distintos.White piedra is a superficial mycosis caused by the genus Trichosporon. It is characterized by nodules on the hair shaft. Pediculosis capitis is caused by Pediculus humanus var. capitis of the suborder Anoplura. Whereas pediculosis is a common infestation, clinical reports of white piedra are rare. Molecular biology procedures identified T. inkin as the agent of white piedra in this case report. The authors present associations between the two diseases in the same patient in order to highlight their clinical differences

Phytohaemagglutinin's effect on Neospora caninum antigen production

The parasite Neospora caninum affects mainly cattle and dogs. This study aimed to evaluate the effect of phytohaemagglutinin (PHE) in antigen production of N. caninum NC-1 strain in gerbils (Meriones unguiculatus) and in vitro. 20 gerbils were used, 10 inoculated intraperitoneally with 1 × 106 tachyzoites and 10 with 1 × 106 tachyzoites plus 300 μL/mL of PHE. 16 bottles of Vero cell culture were inoculated, 8 with 1.5 × 105 tachyzoites and 8 with 1.5 × 105 tachyzoites plus 30 μL/mL of PHE. Serology of gerbils was performed on day 0 and before euthanasia. Tachyzoites present in peritoneal fluid and cell culture bottles were quantified by Neubauer chamber and by real-time PCR (qPCR). PHE has not interfered in the production of tachyzoites of N. caninum in intraperitoneal inoculated gerbils and the effect of PHE in cell culture had a negative impact, considering the qPCR technique as the gold standard

Genotyping of Toxoplasma gondii and Sarcocystis spp. in road-killed wild mammals from the Central Western Region of the State of São Paulo, Brazil

Abstract INTRODUCTION: Road-killed wild animals host zoonotic pathogens such as Toxoplasma gondii, offering a new opportunity for the epidemiological study of these infectious organisms. METHODS This investigation aimed to determine the presence of T. gondii and other apicomplexan parasites in tissue samples of 64 road-killed wild animals, using polymerase chain reaction (PCR). Positive samples were then typed by PCR-restriction fragment length polymorphism (RFLP) using 7 markers: SAG1, 5′-3′SAG2, SAG3, BTUB, c29-6, PK1, and Apico. PCR-RFLP targeting 18S ribosomal RNA (rRNA) genes was also performed on all samples to detect other apicomplexan parasites. RESULTS T. gondii DNA was detected in 16 tissue samples from 8 individual animals, as follows: 1 Cerdocyon thous (crab-eating fox), 1 Didelphis albiventris (white-eared opossum), 1 Lutreolina crassicaudata (lutrine opossum), 2 Myrmecophaga tridactyla (giant anteater), 1 Procyon cancrivorus (crab-eating raccoon), and 2 Sphiggurus spinosus (Paraguay hairy dwarf porcupine). Seven different T. gondii genotypes were identified, 6 of which were novel. Typing by 18S rRNA verified these 16 T. gondii-infected samples, and identified 1 Sarcocystis spp.-infected animal [Dasypus novemcinctus (nine-banded armadillo)]. The amplified T. gondii (GenBank accession No. L37415.1) and Sarcocystis spp. 18S rRNA products were confirmed by sequencing. CONCLUSIONS Our results indicate that T. gondii is commonly present in wild mammals, which act as sources of infection for humans and animals, including other wild species. The approach employed herein proved useful for detecting T. gondii and Sarcocystis spp. in the environment and identifying their natural reservoirs, contributing to our understanding of host-parasite interactions