Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Encephalitozoon cuniculi”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 55 records · Page 3Linked to original sources

[Experimental Encephalitozoon cuniculi infection in dexamethasone-immunosuppressed mice].

OBJECTIVE: Microsporidian Encephalitozoon cuniculi has been recognized as an opportunistic pathogen in immunosuppressed individuals, such as AIDS patients. The objective of the study was to develop pharmacologically immunosuppressed animals as a model of the natural occurring E. cuniculi infection. METHODS: Distinct groups of adult Balb-C mice were immunosuppressed with different doses of dexamethasone (Dx, 3 or 5 mg/kg/day, intraperitoneal route - IP) and inoculated with E. cuniculi spores by IP route intraperitoneally. Control groups (inoculated animals but non-immunosuppressed and non- inoculated animals but immunosuppressed) were also used. The spores of E. cuniculi were previously cultivated in MDCK cells. The animals were sacrificed and necropsied at 7, 14, 21, 28 and 35 days post-inoculation. Tissue fragments were collected and processed for light microscopy studies, using Gram-chromotrope and hematoxylin-eosin staining techniques. RESULTS: In all immunosuppressed and inoculated inoculated immunosuppressed mice,specially in those that received 5 mg/kg/day of dexamethasone, the most prominent necropsy findings were hepatomegaly and splenomegaly. The experimental inoculation resulted in a disseminated non-lethal infection, characterized by granulomatous lesions in several organs (liver, lungs, kidneys, gut and brain) but notably in the hepatic tissue. Spores of E. cuniculi were only seen in few animals treated with 5 mg/kg/day of Dx at 35 days post-infection. CONCLUSIONS: Microsporidiosis in Dx-immunosuppressed mice provides a useful model for studies of the microsporidial infection, resembling that one naturally occurring in immunodeficient individuals with AIDS.

Animals↗

The infectivity of Encephalitozoon cuniculi in vivo and in vitro.

The infectivity of Encephalitozoon cuniculi grown in cell cultures was determined in cultured cells and in wild and domestic rabbits. The ratio of the total to tissue culture viable count was 1,300 (median of seven determinations). The mean ratio of intact spore count to total count, as determined by electron microscopy was 0.12. Although variation between infectivity experiments was large, the median animal infective dose contained 51 FFU (cell culture focus-forming units) for wild rabbits (Oryctolagus cuniculus) and 40 FFU for domestic rabbits. These two infectivities were not statistically different.

Animals↗

Mapping of repetitive and non-repetitive DNA probes to chromosomes of the microsporidian Encephalitozoon cuniculi.

The molecular karyotype of a murine isolate of Encephalitozoon cuniculi, a microsporidian with a wide range of mammalian hosts, comprises eleven chromosomes ranging in size between 217 and 315 kb. To determine specific chromosomal markers, a partial genomic library was constructed and cloned DNA fragments were hybridized to chromosomal bands separated by pulsed-field gel electrophoresis. Most probes were assigned to single chromosomes, indicating prevalence of low-copy number nucleotide sequences within the very small genome of E. cuniculi (2.9 Mb). A few probes were shown to hybridize to all chromosomes. These repetitive DNA fragments corresponded to either rRNA genes or some non-coding regions whose sequences were characterized by short micro- and minisatellites. The chromosomal locations of beta-tubulin genes and six newly identified protein-encoding genes were determined. Genes encoding dihydrofolate reductase, thymidylate synthase, serine hydroxymethyl transferase, a cdc2 kinase-like protein and helicase ERCC6-like protein were each located on a single chromosome whereas genes for both beta-tubulin and aminopeptidase were on two different chromosomes. The mapping will serve as a reference for further analysis of intraspecific karyotype polymorphism in different isolates from different host species.

Animals↗

Ribosomal DNA sequences of Encephalitozoon hellem and Encephalitozoon cuniculi: species identification and phylogenetic construction.

A segment of ribosomal DNA, about 1,350 base pairs long, was amplified from the microsporidian species Encephalitozoon hellem, isolated from AIDS patients, and Encephalitozoon cuniculi. The amplified DNA segment extends from position 530 in the small ribosomal RNA subunit to position 580 in the large ribosomal RNA subunit. A comparison of sequence data from this region for Encephalitozoon hellem and Encephalitozoon cuniculi shows relatively high sequence similarity, supporting the placement of these two organisms in the same genus. At the same time, sequence differences between the two organisms confirm that they are not the same species. Three separate isolates of E. hellem were sequenced in the highly variable intervening spacer region. The sequence was identical for all three isolates. Within the amplified DNA segment, regions were sequenced which yield highly variable, moderately variable and highly conserved sequence information, appropriate for comparison with other species in the phylum Microspora at all taxonomic levels. We suggest that sequence data from these regions be included in future species descriptions for the purposes of species identification and phylogenetic analysis. Restriction digests of the amplified region are presented and give a rapid method for distinguishing between the two Encephalitozoon species.

Animals↗

Fractionation of sporogonial stages of the microsporidian Encephalitozoon cuniculi by Percoll gradients.

Microsporidia are obligate intracellular parasites that are increasingly recognized as a cause of opportunistic infections in immunocompromised individuals. Encephalitozoon cuniculi has been identified in humans with AIDS and infects a wide range of mammalian hosts. Little is known about the metabolic processes that regulate growth and replication of microsporidia. Examination of the individual stages of development will facilitate such studies and reveal possible targets for drug therapy. The purpose of this study was to fractionate and purify stages of the microsporidian life cycle. Encephalitozoon cuniculi were cultured in RK-13 cells. The tissue supernatants containing multiple parasite stages, empty microsporidial husks and host cell debris were collected, washed, and subjected to differential centrifugation in 80% stock isotonic Percoll. Transmission electron microscopy and SDS-polyacrylamide gel electrophoresis were used to compare the content and purity of each fraction. Mature spores formed a band at a density of approximately 1.138 g/ml. Sporoblasts were found at densities between 1.102 g/ml and 1.119 g/ml. A mixture of sporonts, sporoblasts, microsporidial husks, and cell debris remained at the top of the gradient and additional centrifugation in 30% and 50% Percoll resulted in separation of these stages. These results represent the first step toward fractionating stages of microsporidia infecting humans.

Animals↗

Humoral immune response to natural infection with Encephalitozoon cuniculi in rabbits.

Sera from 4 generations of a family of rabbits having a natural Encephalitozoon cuniculi infection were investigated. Antibodies were demonstrated in a litter of newborn rabbits and in another litter 11 days old. Histoloigcal examination of the brains and kidneys of these animals failed to reveal lesions attributable to the disease. Sera from a further litter of 2 rabbits, taken at weekly intervals from 4 to 42 weeks of age, revealed a low initial antibody titre which regressed to an undetectable level. After an interval, titres rose and then plateaued to the end of the period of observation. Encephalitozoon cuniculi infections in the dams of all the litters were confirmed by antibody assay and by histology. Parallel titration of samples treated and not treated with 2-mercaptoethanol showed that the india-ink immunoreaction demonstrates only immunoglobulin G.

Animals↗

Intraocular microsporidiosis due to Encephalitozoon cuniculi in a patient with idiopathic CD4+ T-lymphocytopenia.

Encephalitozoon cuniculi was documented to cause disseminated microsporidial infection including an iris tumor and endophthalmitis in an adolescent with idiopathic CD4+ T-lymphocytopenia. The diagnosis was established by microscopic, serologic and molecular methods. E. cuniculi (rabbit strain) was identified from the iris tumor, as well as from urine, conjunctival, corneal, and nasal swabs. Treatment with oral albendazole led to rapid improvement. This case raises the possibility of disseminated microsporidial infection in the context of idiopathic CD4+ T-lymphocytopenia and possibly advanced human immunodeficiency virus (HIV) infection, and above all the possibility of intraocular infection with E. cuniculi in humans.

AIDS-Related Opportunistic Infections↗

Direct ABC immunohistochemistry to Encephalitozoon cuniculi.

Among the sera from 9 rabbits spontaneously infected with Encephalitozoon cuniculi, a serum which revealed a high titer for E. cuniculi by indirect protein A-gold (IPAG) immunohistochemistry and reacted with the outer layer of the shell of E. cuniculi spores on immunoelectron microscopical examination, was biotinylated. The biotinylated rabbit anti-E. cuniculi IgG reacted immunohistochemically with E. cuniculi, but not with other protozoa tested, namely Neospora caninum, Toxoplasma gondii and sarcocystis. The direct avidin-biotin-peroxidase complex (ABC) immunohistochemistry using biotinylated rabbit anti-E. cuniculi IgG in this study is a useful tool for the diagnosis and study of encephalitozoonosis.

Animals↗

The first finding of antibodies to Encephalitozoon cuniculi in cows in Slovakia.

The presence of antibodies against Encephalitozoon cuniculi in cows was observed by the method of indirect immunofluorescence (IFAT). The animals reacting at the titre 1:64 and more were considered positive. Of the total number of 55 sera examined, 20 were positive at the titre 1:64, from which four were positive at the titre 1:256 also. Our results indicate that it is very important to carry out screening examinations in large-scale breeding of cattle due to the persistent high percentage of bovine abortions and disorders in the reproductive cycle of unknown etiology.

Abortion, Veterinary↗

Mechanisms of resistance to the intracellular protozoan Encephalitozoon cuniculi in mice.

Mechanisms of resistance to the obligate intracellular protozoan Encephalitozoon cuniculi were studied in BALB/c mice. Resistance to lethal disease was T cell-dependent because transfer of T-enriched, but not T-depleted, spleen cells from sensitized BALB/c donors would protect infected BALB/c-nu mice. A modified focus-forming assay was utilized to measure effects on E. cuniculi infectivity in vitro. The results show that antibodies exert an opsonization effect and may block parasite entry into nonphagocytic cells. No cytotoxic T cells were demonstrated. Supernatants from E. cuniculi-sensitized spleen cells incubated with E. cuniculi in vitro could induce adherent PEC to kill E. cuniculi.

Animals↗

Can Encephalitozoon cuniculi cross the placenta?

Laboratory rabbits and mice were challenged with Encephalitozoon cuniculi at three different stages of gestation to determine whether the parasite crosses the placenta. No transplacental transmission could be demonstrated.

Animals↗

The identification of rRNA maturation sites in the microsporidian Encephalitozoon cuniculi argues against the full excision of presumed ITS1 sequence.

In Encephalitozoon cuniculi like in other microsporidia, the primary transcript for SSU and LSU rRNAs includes only one internal transcribed spacer (ITS1) which separates SSU rRNA from the 5.8S region associated with LSU rRNA. The extraction of total RNA from E. cuniculi-infected MRC5 cells using a hot phenol/chloroform procedure enabled us to perform primer extension and S1 nuclease protection experiments in the aim of identifying rRNA maturation sites. Our data support a simple processing (four cleavage sites) with elimination of only nine nucleotides between SSU and LSU rRNA regions. Most of the presumed ITS1 sequence characterized by strain-dependent polymorphism therefore remains linked to SSU rRNA 3' end. A new secondary structure for the sixth domain of E. cuniculi LSU rRNA is proposed following the identification of its 3' terminus.

Animals↗

PCR detection of Neospora caninum, Toxoplasma gondii and Encephalitozoon cuniculi in brains of wild carnivores.

Neospora caninum, Toxoplasma gondii and Encephalitozoon cuniculi are important pathogens with affinity to the central nervous system of many animals. 240 brains of wild carnivores were examined by PCR-based diagnosis. The presence of N. caninum DNA was confirmed in 4.61% (7/152) red foxes (Vulpes vulpes). DNA of T. gondii was found in 4.92% (3/61) martens (Martes sp.) and in 1.32% (2/152) red foxes. DNA of E. cuniculi was determined in 3.28% (2/61) martens and in one examined European otter (Lutra lutra). There were no co-infections found. These results provide the first evidence of E. cuniculi in the European otter, the first report of N. caninum in foxes in the Czech Republic and confirm the presence of T. gondii in wild carnivores in the Czech Republic.

Animals↗

Two Encephalitozoon cuniculi strains of human origin are infectious to rabbits.

The microsporidian Encephalitozoon cuniculi can infect a wide variety of mammals including man. In this study, E. cuniculi isolates of animal origin were compared with 6 isolates obtained from HIV-infected patients. Based on results of Western blot analysis, random amplified polymorphic DNA (RAPD) and the sequence of the rDNA internal transcribed spacer (ITS) the isolates were classified into 3 groups with the repeated element 5'GTTT-3' in the ITS being a reliable genetic marker. Five isolates from Swiss patients were found to be homologous to isolates from Swiss rabbits (strain I). The sixth isolate from a patient from Mexico differed by all methods and could be attributed to E. cuniculi strain III that has been described from 2 dogs from the USA. All of these isolates were distinguished from isolates from blue foxes from Norway (strain II). Intraspecific nucleotide divergence of the SSU rRNA gene of E. cuniculi belonging to the 3 strains was in the same low range (0.00-0.15%) as was found for the corresponding sequence of 2 E. hellem isolates. Groups of 2 rabbits were infected by oral inoculation of 10(7) E. cuniculi spores (2 isolates from strain I of human and rabbit origin, 1 from strain III) as shown by antibody responses and the re-isolation of the parasites from brain material. The results provide further evidence that per oral transmission of the parasite between various hosts is feasible.

Animals↗

Immunologic and clinicopathologic evaluation of adult dogs inoculated with Encephalitozoon cuniculi.

Beagle dogs inoculated intravenously with 1.75 x 10(9) viable Encephalitozoon cuniculi spores at 12.5 months of age were monitored for 18 months to assess infection in the adult dog. Parameters monitored included packed cell volume, total and differential leukocyte counts, and humoral and cellular responses to infection. Immunoglobulin M and G antibodies directed against the parasite tegument were present throughout the 18 months. Peripheral blood monocytes treated with lymphokines, either antigen specific (E. cuniculi) or nonspecific (concanavalin A), killed E. cuniculi spores in vitro. Pretreatment of E. cuniculi spores with normal dog serum or infected dog serum enhanced the killing of the parasite (normal dog serum less than infected dog serum) by canine monocytes. Histologic examination of selected tissues revealed microfocal plasma cell and lymphocyte aggregates at the renal corticomedullary junction and in the medullary interstitium. The results of these experiments suggest that the adult dog is able to mount an effective defense to infection, to minimize host tissue damage, and to eliminate the parasite through complex interactions between monocytes-macrophages and lymphocytes.

Animals↗

Prevalence of antibodies to Encephalitozoon cuniculi in horses in the Israel.

Infection with the intracellular microsporidium Encephalitozoon cuniculi can cause a serious disease--encephalitozoonosis in various animals and people. Several species of mammals, including the horse, were seem to be potential sources of encephalitozoonosis for animal as well as human hosts. The disease diagnosis is based on clinical signs, pathological findings, and the detection of E. cuniculi or circulating antibodies directed against the parasite. This study investigates the seroconversion to E. cuniculi in horses admitted to the Veterinary Teaching Hospital of the Hebrew University of Jerusalem and 3 different private horse-riding farms across Israel. Antibodies to E. cuniculi were determined using the IFA test in the sera from 102 horses. Of 72 asymptomatic horses, 60% were seropositive and 19% of the positive samples showed a titter of 1:512. Of 30 horses with various clinical signs, 80% were seropositive and 68% of the positive samples showed a titer of 1:512. High titers were associated with colic and neurological signs. This could prove to be interesting if the high percentages of prevalence of antibodies level in horses are an indication of health risk in humans.

Animals↗

Encephalitozoon cuniculi in pet rabbits.

The results of a serological test for Encephalitozoon cuniculi in 125 pet rabbits are reviewed, together with follow-up studies of clinical cases. Blood samples were taken from 38 asymptomatic rabbits and 87 rabbits showing neurological, renal or ocular signs suggestive of encephalitozoonosis. In the asymptomatic group, six of 26 (23 per cent) apparently healthy rabbits, sampled as part of a health screen, were seropositive; of the remaining 12 asymptomatic rabbits, sampled because they lived with seropositive companions, eight (66 per cent) were seropositive. Fifty-eight of the rabbits with clinical disease showed neurological signs, including head tilt, seizures, ataxia and swaying; three of them also showed renal signs and two showed ocular signs, and these five rabbits were all seropositive. Head tilt was the most common neurological sign in 21 of 23 (91 per cent) of the seropositive cases. All nine rabbits with ocular lesions were seropositive. In follow-up studies of clinical cases, the rabbits showed variable responses to treatment with albendazole, fenbendazole, antibiotics or corticosteroids, and some cases recovered without treatment.

Adrenal Cortex Hormones↗

Monoclonal antibodies for specific detection of Encephalitozoon cuniculi.

Seven species-specific monoclonal antibodies (MAbs) were produced against Encephalitozoon cuniculi and characterized. The MAbs were immunoglobulin G, and when used for indirect microimmunofluorescence microscopy and Western immunoblot assays, they detected E. cuniculi originating from clinical samples. They did not cross-react with other Encephalitozoon species (E. intestinalis and E. hellem) or with a collection of gram-negative bacteria, yeast, and other parasites. The MAbs reacted primarily with 121-, 56-, 45-, 43-, and 41-kDa protein epitopes of E. cuniculi. These epitopes were demonstrated to be E. cuniculi species specific by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We developed MAbs to strains of E. cuniculi that can be used successfully to distinguish E. cuniculi from other microsporidial species in cultures established from clinical specimens. These MAbs may provide a specific, simple, rapid, and low-cost tool for the identification and diagnosis of infections due to microsporidia.

Animals↗