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PubMed2003

Intraspecies genotype variability of the microsporidian parasite Encephalitozoon hellem.

Seven isolates of Encephalitozoon hellem from human immunodeficiency virus-positive patients were genotyped through a series of markers: the internal transcribed spacer (ITS) of ribosomal DNA, the polar tube protein (PTP) gene, and two intergenic spacers (IGS-TH and IGS-HZ) whose polymorphism is newly reported. The genome markers were all analyzed at three levels: PCR amplification followed by polyacrylamide gel electrophoresis, single-strand conformation analysis (SSCA), and DNA sequencing. The polymorphisms detected involve insertions/deletions and point mutations. SSCA can distinguish any pair of sequences, even those differing by a single base pair. The different isolates studied fit into the previously described ITS genotype 1A, except one which seems to be a 2A derivative variant (2D). When PTP and the new markers IGS-TH and IGS-HZ were analyzed, most of the isolates displayed different genotypes, demonstrating that E. hellem has a strong intraspecies variability. A set of markers such as those used here may be very useful in genotyping of clinical samples and in the assessment of epidemiological relationships among E. hellem strains.

Animals↗
PubMed1989

Prevalence of antibodies to Encephalitozoon cuniculi in stray dogs as determined by an ELISA.

An ELISA, using Encephalitozoon cuniculi spores as antigen, was used to determine the prevalence of specific anti-E cuniculi IgG in a group of stray dogs. In a preliminary survey 51 of 248 sera were classified as positive with titres of 1:400 to 1:3200. The 18 sera with titres of 1:400 were reclassified as negative when no IgG binding to the spores could be detected by comparison with a standard curve of anti-E cuniculi IgG. The remaining 33 sera (13.3 per cent) were classified as low, moderate or strong positives. Comparison of total IgG and specific IgG showed that specific IgG was greatly increased in the moderately and strongly positive sera. E cuniculi may be of importance as one cause of fading puppy syndrome when transmitted transplacentally, and as a complicating infection in human immunodeficiency diseases.

Animals↗
PubMed2001

Prevention and treatment of Encephalitozoon cuniculi infection in rabbits with fenbendazole.

The efficacy of fenbendazole for preventing an experimental infection of Encephalitozoon cuniculi and for eliminating the spores from the central nervous system of naturally infected rabbits was investigated. Fenbendazole (20 mg/kg bodyweight daily) was administered from seven days before until two or 21 days after rabbits had been infected orally with 10(6) spores of E. cuniculi. Both regimens were effective in preventing the establishment of the parasites, as demonstrated by negative parasitic-specific serology and by the failure to isolate the parasite from brain tissue. In naturally infected, seropositive rabbits, parasites were successfully isolated from seven of nine untreated animals, but not from the brain tissue of eight animals treated with fenbendazole-medicated pellets for four weeks.

Animals↗
PubMed2003

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↗
PubMed2006

Seroprevalence of antibodies to Encephalitozoon cuniculi in domestic rabbits in the United Kingdom.

Serum samples from 97 clinically healthy domestic rabbits were tested for antibodies to Encephalitozoon cuniculi by an indirect elisa technique. Fifty (52 per cent) of them were seropositive. The samples were taken as part of a routine health screen or before general anaesthesia at 22 veterinary practices in England, Scotland and Wales, and the veterinary surgeons were asked to complete a questionnaire to provide information concerning the animal's husbandry, diet, vaccination, health status and any preventive medicine routines. None of these factors was found to be associated with the serological status of the rabbits.

Animals↗
PubMed1992

Hepatic lesions in rabbits infected with Encephalitozoon cuniculi administered per rectum.

Microsporidia have been recognized recently as opportunistic pathogens in acquired immunodeficiency syndrome patients. In an attempt to develop an animal model of enteric microsporidiosis, adult (5 to 6 months old) male Flemish Giant rabbits from a closed New York colony were administered 5 x 10(3), 5 x 10(5), and 5 x 10(7) Encephalitozoon cuniculi per rectum. Rabbits given 5 x 10(5) and 5 x 10(7) E. cuniculi had moderate granulomatous periportal infiltrates, characterized by the presence of numerous macrophages, epithelioid cells and a few multinucleated giant cells, lymphocytes, and plasma cells. Inflammatory cells also were seen infiltrating the tunica adventitia and tunica media of hepatic portal veins and branches of the hepatic artery. This study demonstrates that administration of E. cuniculi per rectum to rabbits results in infection that is characterized by high frequency and severity of hepatic lesions.

Administration, Rectal↗
PubMed2003

Encephalitozoon cuniculi placentitis and abortion in a quarterhorse mare.

Encephalitozoon cuniculi is a microsporidial parasite, which has rarely been reported to cause placentitis in animals. A late-term aborted fetus and placenta from a Quarterhorse were presented to the Livestock Disease Diagnostic Center, University of Kentucky, for diagnostic examination. There was a necrotizing placentitis, with distension of many chorionic epithelial cells by intracytoplasmic vacuoles containing 1-2-microm-diameter, elongated, gram-positive organisms. The organisms were identified as E. cuniculi by electron microscopy and by polymerase chain reaction using primers to microsporidial ribosomal DNA. Joints of the fetus were swollen, with gross and microscopic lesions of synovitis; however, E. cuniculi DNA was not detected.

Abortion, Veterinary↗
PubMed2002

An indirect ELISA for detection of Encephalitozoon cuniculi infection in farmed blue foxes (Alopex lagopus).

Infection with the intracellular microsporidium Encephalitozoon cuniculi can cause serious disease, encephalitozoonosis, in the blue fox (Alopex lagopus). The disease diagnosis is based on clinical signs and pathological findings, and detection of E. cuniculi or circulating antibodies directed against the parasite. Indirect immunofluorescence (IFAT) and carbon immunoassay (CIA) are the most commonly used serological methods for diagnosis in this species. In the present study, an indirect ELISA (enzyme linked immunosorbent assay) was established and evaluated against IFAT by testing of 205 field samples from blue foxes. There was high agreement between the results of the ELISA and CIA (kappa=0.99), and the ELISA and IFAT (kappa=0.958). There was no significant statistical difference between the tests (p>0.05). It was concluded that the ELISA could be used to identify seropositive farmed blue foxes. The advantage of the ELISA lies in the potential of screening large numbers of animals with the goal of eradicating E. cuniculi infection in the farms.

Animals↗
PubMed2000

Comparison of assays for antibodies to Encephalitozoon cuniculi in rabbits.

Two indirect immunofluorescence (IIF) assays, two enzyme-linked immunosorbent assays (ELISAs) and the carbon immunoassay (CIA) for determination of antibodies to Encephalitozoon cuniculi were compared using 210 sera of rabbits, 135 of which originated from seven infected colonies, while 75 originated from four uninfected colonies. There was no evidence of a difference between the different assays with respect to the number of positive sera. There was a clear correlation between the quantitative response measured by IIF and CIA and the other assays, and between both IIF tests, while no such correlation was found in the quantitative response measured by ELISAs, which might be explained by the less quantitative nature of the ELISA. Therefore quantitative determination of antibodies to E. cuniculi should be performed by IIF and not by ELISA. The nosographic sensitivities N1 and specificities N2 of the assays were > or = 0.94 and > or = 0.97 respectively. Small differences in N1 and N2 between the assays, although not statistically significant, were responsible for differences in the calculated predictive values of a positive test and of a negative test. As expected, the magnitude of these differences depended on the fraction of positive sera sampled from a given colony. There was strong evidence of such a difference between the fraction of positive sera found in different colonies, but the sample size from some colonies was too small to allow any conclusion, whether this was due to differences in the prevalences of the infection in the colonies or something else. We conclude that any of the assays will be suitable for the routine health monitoring of laboratory rabbit colonies for E. cuniculi infection, as recommended by the Federation of European Laboratory Animal Science Associations.

Animals↗
PubMed2003

In utero transmission of Encephalitozoon cuniculi strain type I in rabbits.

Pregnant rabbits were serologically diagnosed as having been infected with Encephalitozoon cuniculi. At necropsy at 28 days of gestation, does, placentas and fetuses were found to be infected with E. cuniculi strain type I as evidenced by using the nested-polymerase chain reaction (PCR) technique, thereby confirming vertical transplacental transmission.

Animals↗
PubMed1991

The incidence of Encephalitozoon cuniculi in a commercial barrier-maintained rabbit breeding colony.

Between 1982 and 1987 sera from 4952 New Zealand White rabbits (Oryctolagus cuniculus) obtained from a single commercial supplier were tested for the presence of antibodies to Encephalitozoon cuniculi. A commercially available carbon immunoassay test kit was used. Initially 32.9% of the rabbits were seropositive with the number progressively decreasing to 2.3% by 1987. The reason for the significant decline in the incidence of infection was most likely due to a selection process for breeding stock instituted by the supplier based upon productivity, posture and weight of each animal.

Animals↗
PubMed1993

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↗
PubMed2001

Isolation of Encephalitozoon cuniculi using primary tissue culture techniques from a rabbit in a colony showing encephalitozoonosis.

Encephalitozoon spores were isolated in a primary tissue culture of the kidneys from an encephalitozoonosis-suspected rabbit in a municipal zoo in Hokkaido. The isolated spores were morphologically characteristic of microsporidial ones in chromotrope stain, and proven to be E. cuniculi by a polymerase chain reaction (PCR) with a species-specific primer set and by direct DNA sequencing of the PCR products.

Animals↗
PubMed2005

Humoral intestinal immunity against Encephalitozoon cuniculi (Microsporidia) infection in mice.

Three strains of mice, BALB/c, IL-12 knock-out (KO) and INF-gamma knock-out, were chosen as an experimental model for the study of intestinal immunity induction against Encephalitozoon cuniculi Levaditi, Nicolau et Schoen, 1923 infection. Mice were infected perorally with 10(7) spores and re-infected with the same dose 70 days after the first infection. The anti-E. cuniculi IgA, IgG and IgM responses in sera and extracts of stool samples were determined by ELISA. Results have shown specific antibody production in the sera and intestinal secretions of all three strains of mice induced orally by E. ciniculi spores. BALB/c mice developed a stronger humoral immune response than IL-12 KO mice. The lowest antibody response developed in INF-gamma KO mice that succumbed to the infection within 28 days post infection.

Animals↗
PubMed2004

Immune response in mice infected by Encephalitozoon cuniculi and suppressed by dexamethasone.

Several indicators of immune response were observed in immunocompetent mice of the ICR line and those suppressed by dexamethasone upon their experimental infection with the microsporidia of Encephalitozoon cuniculi. The mice were infected by one-shot intraperitoneal administration of 5 x 10(7) pathogenic spores. On Days 7, 14, 28 and 42 after infection, peripheral blood leukocyte phagocytic activity was determined and compared, including phagocytic index and the blastogenic response in spleen cells to mitogenic activation by concanavalin A and phytohaemagglutinin. The results point to the fact that E. cuniculi itself can cause a significant decrease in phagocytic activity of phagocytic leukocytes in the early stages of infection as well as a remarkable decrease in the proliferative response of spleen cells to T-cellular mitogens.

Animals↗
PubMed2006

First report on Encephalitozoon intestinalis infection of swine in Europe.

Encephalitozoon intestinalis infection of sows is reported from a pig farm in Slovakia. Spores were detected by direct microscopic visualisation in the faeces of 25 out of 27 sows (92.6%). This finding was also supported serologically by the presence of specific anti-E. intestinalis antibodies and by a species-specific polymerase chain reaction (PCR). This is the first report on E. intestinalis infection of swine in Europe.

Animals↗
PubMed2005

Experimental meningoencephalomyelitis by Encephalitozoon cuniculi in cyclophosphamide-immunosuppressed mice.

Encephalitozoonosis is an increasingly important opportunistic protozoan infection in immunocompromised individuals. This study aims to examine the development of an experimental Encephalitozoon cuniculi infection in the central nervous system of immunosuppressed mice. Adult Balb-C mice were inoculated intraperitoneally with E. cuniculi spores, treated with cyclophosphamide during the experimental period and killed from 15 to 75 days post-inoculation. Tissue samples were collected and processed for light and transmission electron microscopy investigation. Multifocal granulomas were seen in all organs. A lymphocytic, diffuse, non-suppurative menigoencephalomyelitis was observed, with neuronal degeneration and necrosis, macrophagic infiltration and reactive astrocytosis. E. cuniculi spores were seen in the microgranulomas or occurred unassociated with inflammatory reaction. The parasites were rarely seen in Hematoxylin-Eosin stained sections, but were Gram-Chromotrope positive. Proliferative forms and spores were found in parasitophorous vacuoles within neural cells and macrophages. Experimental encephalitozoonosis in immunosuppressed mice provides an useful model for the study of brain lesions associated with these protozoans in man.

Animals↗