Seroprevalence of Encephalitozoon cuniculi and Toxoplasma gondii in domestic rabbits (Oryctolagus cuniculus) in China.

The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites.

Seroprevalence of Encephalitozoon cuniculi in wild rodents, foxes and domestic cats in three sites in the United Kingdom.

Encephalitozoon cuniculi is an obligate intracellular microsporidian that is the causal agent of encephalitozoonosis, an important and emerging disease in both humans and animals. Little is known about its occurrence in wildlife. In this study, serum samples from 793 wild rodents [178 bank voles (BV), 312 field voles (FV) and 303 wood mice (WM)], 96 foxes and 27 domestic cats from three study areas in the UK were tested for the presence of antibodies to E. cuniculi using a direct agglutination test (DAT). Seroprevalence in the wild rodents ranged from 1.00% to 10.67% depending on species (overall 5.31%) and was significantly higher in foxes [49.50% (50/96)]. None of the 27 cats sampled were found to be seropositive. This is the first report of seroprevalence to E. cuniculi in BV, FV, WM, foxes and cats in the UK and provides some evidence that foxes could act as sentinels for the presence of E. cuniculi in rodents. The study demonstrates that wildlife species could be significant reservoirs of infection for both domestic animals and humans.

Seroprevalence of Encephalitozoon cuniculi and Toxoplasma gondii in domestic rabbits (Oryctolagus cuniculus) in China

The breeding of domestic rabbits (Oryctolagus cuniculus) for human consumption has a long tradition in China. Infections that can affect the production of meat or even be transmitted from animals to humans are important to monitor, especially for public health reasons as well as for their impact on animal health. Thus, a total of 1,132 domestic rabbit sera from 4 regions in China were collected for serological screening for Encephalitozoon cuniculi and for Toxoplasma gondii by ELISA and modified agglutination test (MAT), respectively. Antibodies to E. cuniculi were detected in 248/1,132 (21.9%) sera tested while antibodies against T. gondii revealed a seroprevalence of 51/1,132 (4.5%). We believe that the present results are of epidemiological implications and public health importance due to the acknowledged susceptibility of humans to E. cuniculi and T. gondii infections. Therefore, routine screening tests of domestic rabbits are proposed considering the zoonotic potential of these parasites.

Encephalitozoonosis in New Zealand rabbits and potential transmission risk.

Encephalitozoon cuniculi is a small protozoan parasite in the phylum Microspora. It has been shown to naturally infect several host species, including humans. Encephalitozoonosis is routinely diagnosed in vivo by serological examination or post mortem by histopathology. In a conventional rabbit colony, two animals suddenly showed clinical signs (torticollis and asthenia of limbs). Serum samples of these rabbits were seropositive for E. cuniculi after definitive diagnosis (Toxoplasma gondii and Listeria monocytogenes). The animals in the same breeding facility were also clinical examined, and the present study evaluated the prevalence of specific anti-E. cuniculi antibodies using serological testing, both in animals and in people working with animals, after two clinical cases. The rabbits showed no clinical symptoms of the disease. Blood samples were taken for E. cuniculi infection from 50 clinically healthy rabbits. Anti-E. cuniculi antibodies were found in two asymptomatic and two clinically affected animals belonging to the same rabbit colony. Finally, the present study found that the 7.7% (4/52) prevalence of CIA, test positive in rabbits. E. cuniculi spores were detected in the urine of one clinically affected rabbit, and one seropositive animal caretaker after staining with the modified trichrome stain. In conclusion, the presence of seropositive, but apparently healthy rabbits indicates the need for screening examinations to detect the anti-E. cuniculi antibody in rabbits, especially considering the potential zoonotic risk. Therefore, persons should avoid contact with the urine of infected or healthy animals, and always use good personal hygiene when handling animals.

Evaluation of the usefulness of an ELISA and protein electrophoresis in the diagnosis of Encephalitozoon cuniculi infection in rabbits.

OBJECTIVE-To evaluate the usefulness of an antibody detection ELISA and protein electrophoresis (PE) for diagnosing Encephalitozoon cuniculi (ECUN) infection in pet rabbits. ANIMALS-203 pet rabbits. PROCEDURES-Serum and plasma samples from pet rabbits were submitted from veterinary clinics within the United States. Participating veterinarians completed a questionnaire that was used to classify rabbits as clinically normal (n=33), suspected of having an ECUN infection (103), or clinically abnormal but not suspected of having an ECUN infection (67). An ELISA for detection of serum or plasma IgG against ECUN was developed by use of commercially available reagents. Results of the ELISA and PE were used to detect ECUN infection. RESULTS-A high seroprevalence of antibody against ECUN was detected in all 3 groups of rabbits. In rabbits suspected of having an ECUN infection, the mean IgG titer was 1.7 times as high as the values in the other rabbit groups. Rabbits suspected of having an ECUN infection and those that were simply clinically abnormal had a higher concentration of gamma-globulins than clinically normal rabbits. This increase in globulins concentration was accompanied by a decrease in the albumin-to-globulin ratio. Results of the ELISA and PE were significantly different between clinically normal rabbits and those suspected of having an ECUN infection. CONCLUSIONS AND CLINICAL RELEVANCE-The combination of an ELISA and PE may aid in the diagnosis of ECUN infection in pet rabbits. IMPACT FOR HUMAN MEDICINE-Because ECUN is a potential zoonotic agent, diagnostic methods for pet rabbits need to be improved to protect human health.

Urinary protein:creatinine ratio in rabbits in relation to their serological status to Encephalitozoon cuniculi.

The concentrations of protein and creatinine were measured in urine samples from 74 healthy domestic pet rabbits, 54 of them seronegative to Encephalitozoon cuniculi and 20 seropositive. The calculated reference range for the urinary protein:creatinine ratio (UPC) of E cuniculi-seronegative rabbits was 0.11 to 0.40. There was no significant variation in the UPC due to the bodyweight, breed, sex, neutered status or husbandry of the rabbits. Seroconversion to E cuniculi was not found to be associated with clinical renal disease because none of the seropositive rabbits had azotaemia or proteinuria.

Analysis of cerebrospinal fluid in healthy rabbits and rabbits with clinically suspected encephalitozoonosis.

Samples of uncontaminated cerebrospinal fluid (csf) were collected from the cisterna magna of 20 healthy laboratory rabbits and 21 pet rabbits with vestibular disease and/or paresis due to clinically suspected encephalitozoonosis. In the healthy rabbits' csf the leucocyte count was

Immune responsiveness associated with experimental Encephalitozoon intestinalis infection in immunocompetent rats.

PURPOSE: Microsporidial infections have been recognized as an increasingly important infection in immunocompromized patients, particularly those infected with HIV/AIDS. This study was designed to study immune responses associated with experimental Encephalitozoon intestinalis infection in immunecompetent rats. MATERIALS AND METHODS: Thirty-four rats in 3 groups, A (Control), B (Intraperitoneal) and C (Oral) were given injections of 0.5 ml of 2 x 10(6) of purified spores of Encephalitotozoon intestinalis spores and were observed for serum specific IgG for 21 days using both Direct and Indirect ELISA. RESULTS: In indirect ELISA, specific lgG were detected on days 7, 14 and 21 for the group B rats and on day 21 for group C and in direct ELISA method, specific lgG were detected in-group B rats on days 7 and 21, for group C rats on day 21 only, while in the control rats, specific lgG were not detected. There was no significant difference between the direct and indirect methods (df=1, X(2), P>0.05). E. intestinalis was observed in stool samples of rats in 1/12 (08.33%) on days 14 and 21 in group B and in 4/10 (33.33%), 3/10 (25.00%) and 2/10 (16.67%) on days 7, 14 and 21 respectively in group C. In-group, A which is the control rats, no microsporidia were observed on days 0, 7, 14 and 21. CONCLUSIONS: There were no changes in the T-lymphocyte counts of rats prior to and after inoculation with spores. Extensive lesions were observed along the intestinal walls especially on the middle and lower sections of group C rats only.

Immunohistochemical identification of Encephalitozoon cuniculi in phacoclastic uveitis in four rabbits.

PURPOSE: Encephalitozoon cuniculi is a microsporidium with a wide range of mammalian hosts. In rabbits it can be responsible for cataract and lens-induced uveitis (LIU). The aim of this study was to provide specific immunohistochemical demonstration and localization of E. cuniculi within the eye, in rabbits with LIU. MATERIAL AND METHODS: Four rabbits were presented with a white mass in the eye and iris discoloration. Complete ophthalmic examinations were performed and a presumptive diagnosis of LIU was made in all cases. Initial therapy with a topical steroid, atropine and systemic enrofloxacin was instituted while serologic (IFA or ICA tests) and cytologic lab results were pending. The final outcome in all cases was enucleation. Routine histology and immunohistochemistry (ABC method) with an antiserum anti-Encephalitozoon cuniculi were performed. RESULTS: Indirect immunofluorescence performed on one rabbit serum expressed a titer of 1 : 32; carbon immunoassay on the serum of the other three rabbits expressed a titer of 1 : 5120 in one, and a titer of 1 : 2560 in the other two cases. Histologically, an intraocular, locally extensive pyogranulomatous infiltration that partially filled the posterior chamber, encasing a wide anterior lens capsule break, was detected in all cases. Immunohistochemically, spores reacting with anti-Encephalitozoon cuniculi antiserum were present in all specimens, occasionally within macrophages and lens epithelial cells. CONCLUSION: Detection of E. cuniculi in rabbits with phacoclastic uveitis has been investigated in the past with different methods. Based on our results, we suggest that immunohistochemistry should be regarded as a useful tool both for specific demonstration of E. cuniculi and for its localization within tissues.

Developmental expression of two spore wall proteins during maturation of the microsporidian Encephalitozoon intestinalis.

Microsporidia are intracellular eukaryotes that infect many animals and cause opportunistic infections in AIDS patients. The disease is transmitted via environmentally resistant spores. Two spore wall constituents from the microsporidian Encephalitozoon intestinalis were characterized. Spore wall protein 1 (SWP1), a 50-kDa glycoprotein recognized by monoclonal antibody (MAb) 11B2, was detected in developing sporonts and at low levels on the surfaces of mature spores. In contrast, SWP2, a 150-kDa glycoprotein recognized by MAb 7G7, was detected on fully formed sporonts and was more abundant on mature spores than SWP1. Nevertheless, the SWPs appeared to be complexed on the surfaces of mature spores. SWP1 and SWP2 are similar at the DNA and protein levels and have 10 conserved cysteines in the N-terminal domain, suggesting similar secondary structures. The C-terminal domain of SWP2 has a unique region containing 50 repeating 12- or 15-amino-acid units that lacks homology to known protein motifs. Antibodies from mice infected with E. intestinalis recognized SWP1 and SWP2. The characterization of two immunogenic SWPs from E. intestinalis will allow the study of exospore structure and function and may lead to the development of useful tools in the diagnosis and treatment of microsporidiosis.

Catecholamines and encephalitozoonosis in rabbits.

Twenty four rabbits (Oryctolagus cuniculus f. domestica) were used to detect specific anti-Encephalitozoon cuniculi antibodies. To identify microsporidian infection, a haemolytic test in agar gel was carried out. Blood samples of animals with and without spontaneous encephalitozoonosis were evaluated, and compared for the presence of epinephrine (EPI), norepinephrine (NE), and dopamine (DA). Rabbits infected spontaneously with E. cuniculi had significantly lower levels of catecholamines than healthy animals. This decrease in catecholamines is of special interest because of their role as factors modifying the immune response. These neuromediators also have different influences on the function of immune cells.

Isolates of Encephalitozoon cuniculi from farmed blue foxes (Alopex lagopus) from Norway differ from isolates from Swiss domestic rabbits (Oryctolagus cuniculus).

Encephalitozoon cuniculi has a wide host range among mammals, but whether it represents a homogeneous species is a subject of controversy. We have isolated, cultivated (in human MRC-5 cells) and, for the first time, characterized by immunological and molecular biological methods four isolates of E. cuniculi from Norwegian blue foxes with a history of encephalitozoonosis. The isolates were compared with nine isolates from domestic rabbits from Switzerland. Two E. cuniculi subtypes were identified according to their host species. A 5'-GTTT-3' tetranucleotide repeat was present twice in the rDNA intergenic spacer in all isolates from foxes as opposed to three times in all isolates from rabbits. Furthermore, random amplified polymorphic DNA analysis showed one polymorphic band among the subtypes, and Western-blot analysis using serum from an infected fox discriminated between the two subtypes on the basis of their banding patterns in the ranges of 31-33 and 38-40 kDa. The 5'-GTTT-3' tetranucleotide repeat is a valuable genetic marker for these two subtypes of E. cuniculi and will be of use in continued studies on the molecular epidemiology of this parasite.

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.