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Simple Summary: This study explores the impact of the Encephalitozoon cuniculi on pet rabbits. It examines the link between antibody levels against this parasite and various health parameters in pet rabbits. By dividing 90 pet rabbits into healthy, subclinical, and clinical groups, this study carefully analyzed blood and biochemical changes. Key findings reveal that higher antibody levels in subclinical and clinical rabbits are crucial for diagnosis and management. This study also associates neurological signs and anemia with active infection, emphasizing the need for vigilant clinical and hematologic monitoring. The research found a higher infection rate in mature adult rabbits and a significant prevalence in Bangkok, indicating regional and age-related risk factors. Anemia was significantly linked to E. cuniculi infection, while no connections were found with other parameters. Recognizing anemia and neurological symptoms aids early diagnosis. Encephalitozoon cuniculi can cause serious disease and subclinical infection in rabbits and requires active surveillance to control the infection. This study investigated the association between anti-Encephalitozoon cuniculi antibody status and various health parameters in pet rabbits. A total of 90 rabbits were divided into healthy (N = 30), subclinical (N = 30), and clinical (N = 30) groups based on their anti-Encephalitozoon cuniculi antibody status and clinical presentations. The mean ages of the control (37 ± 40 months) and subclinical groups (38 ± 34 months) were notably lower compared to that of the clinical group (63 ± 38 months, p < 0.01). Serum titers for anti-Encephalitozoon cuniculi antibodies were significantly elevated in rabbits with subclinical and clinical infections compared to those of healthy rabbits (p < 0.05). Neurological signs were predominant in rabbits with active E. cuniculi infection (80.0%), with additional pathological features including urinary dysfunction (10.0%) and cataracts (10.0%). The source of rabbits was not associated with E. cuniculi infection (p = 0.159). Anemia was significantly linked to E. cuniculi infection (p = 0.026); however, no significant associations were found with leukocytosis, thrombocytopenia, or serum biochemistry parameters. Mature adult rabbits were more likely to be infected with E. cuniculi. Recognizing anemia and neurological signs facilitates in early diagnosis of E. cuniculi infection. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi is a microsporidian, domestic rabbits being the main host. The disease can be acute or subclinical, but treatment options are limited and usually with unrewarding results; therefore, diagnosis and prevention of encephalitozoonosis in rabbits are of the utmost importance. This study aims to obtain the first preliminary information of the prevalence of E. cuniculi in the north-western region of Romania. A total of 176 rabbits were clinically examined and 2 mL of blood was sampled from each. An enzyme-linked immunosorbent assay (ELISA) kit by Medicago (Medicago, Uppsala, Sweden) on the resulted blood serum was utilized. Statistical analysis of the results was conducted using the EpiInfo 2000 software (CDC, Atlanta, GA, USA). A total prevalence of 39.2% (69/176) was identified, with statistically significant differences in relation to the rabbits' clinical status, age, season of sampling, breeding system, body condition score and county of origin; the different family farms tested also had a statistically significant difference. This study gives the first preliminary information on this pathogen distribution on Romania's territory, but further studies need to be performed on larger regions to declare the prevalence in the country. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi can cause serious disease and subclinical infection in rabbits and requires active surveillance to control the infection. This study investigated the association between anti-Encephalitozoon cuniculi antibody status and various health parameters in pet rabbits. A total of 90 rabbits were divided into healthy (N = 30), subclinical (N = 30), and clinical (N = 30) groups based on their anti-Encephalitozoon cuniculi antibody status and clinical presentations. The mean ages of the control (37 ± 40 months) and subclinical groups (38 ± 34 months) were notably lower compared to that of the clinical group (63 ± 38 months, p < 0.01). Serum titers for anti-Encephalitozoon cuniculi antibodies were significantly elevated in rabbits with subclinical and clinical infections compared to those of healthy rabbits (p < 0.05). Neurological signs were predominant in rabbits with active E. cuniculi infection (80.0%), with additional pathological features including urinary dysfunction (10.0%) and cataracts (10.0%). The source of rabbits was not associated with E. cuniculi infection (p = 0.159). Anemia was significantly linked to E. cuniculi infection (p = 0.026); however, no significant associations were found with leukocytosis, thrombocytopenia, or serum biochemistry parameters. Mature adult rabbits were more likely to be infected with E. cuniculi. Recognizing anemia and neurological signs facilitates in early diagnosis of E. cuniculi infection.

Encephalitozoon cuniculi is a microsporidian, domestic rabbits being the main host. The disease can be acute or subclinical, but treatment options are limited and usually with unrewarding results; therefore, diagnosis and prevention of encephalitozoonosis in rabbits are of the utmost importance. This study aims to obtain the first preliminary information of the prevalence of E. cuniculi in the north-western region of Romania. A total of 176 rabbits were clinically examined and 2 mL of blood was sampled from each. An enzyme-linked immunosorbent assay (ELISA) kit by Medicago (Medicago, Uppsala, Sweden) on the resulted blood serum was utilized. Statistical analysis of the results was conducted using the EpiInfo 2000 software (CDC, Atlanta, GA, USA). A total prevalence of 39.2% (69/176) was identified, with statistically significant differences in relation to the rabbits’ clinical status, age, season of sampling, breeding system, body condition score and county of origin; the different family farms tested also had a statistically significant difference. This study gives the first preliminary information on this pathogen distribution on Romania’s territory, but further studies need to be performed on larger regions to declare the prevalence in the country.

Encephalitozoon cuniculi is a microsporidial parasite that primarily infects domestic rabbits (Oryctolagus cuniculus). It is the causative agent of encephalitozoonosis, a disease with an internationally recognized seroprevalence among rabbits. This study determines the presence, clinical manifestation, and serological status of encephalitozoonosis in pet rabbits in Slovenia using various diagnostic procedures. From 2017 to 2021, 224 pet rabbit sera were collected and tested for encephalitozoonosis with the indirect immunofluorescence assay. Immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies against E. cuniculi were confirmed in 160 (65.6%) cases. Most seropositive rabbits suffered from neurological clinical signs or signs of gastrointestinal disorders such as recurrent hypomotilities, chronic weight loss, cachexia, or anorexia, and fewer showed clinical signs related to the urinary system or phacoclastic uveitis. A quarter of the positively tested rabbits presented without clinical signs. Hematological and biochemical blood analysis confirmed that seropositive animals had elevated globulin and deviated albumin levels in comparison to the normal reference values of non-infected animals. Furthermore, rabbits with neurological clinical signs showed statistically significant higher levels of globulins and total protein. Sixty-eight whole-body radiographs and thirty-two abdominal ultrasound reports were analyzed, looking for changes in the shape or size of the urinary bladder, presence of urinary sludge or uroliths, and any abnormalities related to the kidneys (shape, size, or nephrolites). The results suggest that neurological defects in the urinary bladder caused by E. cuniculi lead to a distended urinary bladder and consequently dysuria, incontinence, urine scalding, and sludgy urine. [ABSTRACT FROM AUTHOR]

Obligately intracellular microsporidia regulate their host cell life cycles, including apoptosis, but this has not been evaluated in phagocytic host cells such as macrophages that can facilitate infection but also can be activated to kill microsporidia. We examined two biologically dissimilar human-infecting microsporidia species, Encephalitozoon cuniculi and Vittaforma corneae, for their effects on staurosporine-induced apoptosis in the human macrophage-differentiated cell line, THP1. Apoptosis was measured after exposure of THP-1 cells to live and dead mature organisms via direct fluorometric measurement of Caspase 3, colorimetric and fluorometric TUNEL assays, and mRNA gene expression profiles using Apoptosis RT2 Profiler PCR Array. Both species of microsporidia modulated the intrinsic apoptosis pathway. In particular, live E. cuniculi spores inhibited staurosporine-induced apoptosis as well as suppressed pro-apoptosis genes and upregulated anti-apoptosis genes more broadly than V. corneae. Exposure to dead spores induced an opposite effect. Vittaforma corneae, however, also induced inflammasome activation via Caspases 1 and 4. Of the 84 apoptosis-related genes assayed, 42 (i.e. 23 pro-apoptosis, nine anti-apoptosis, and 10 regulatory) genes were more affected including those encoding members of the Bcl2 family, caspases and their regulators, and members of the tumour necrosis factor (TNF)/TNF receptor R superfamily.

Encephalitozoon cuniculi is a eukaryote, unicellular, spore-forming, obligate intracellular microorganism of the phylum Microsporidia, with domestic rabbits as its main host. Another important species in which this pathogen has been identified are humans, the infection being therefore called a "zoonosis". The transmission takes place via the horizontal route or the vertical route, and cell-mediated immunity plays the biggest role in the infected hosts' protection. Encephalitozoonosis can manifest itself as an acute infection, with neurological signs, renal signs, and ocular lesions, or as a chronic or subclinical infection, which is usually the case for asymptomatic carriers. The diagnostic techniques usually carried out are histological examination, serological tests, and molecular genetic techniques. The treatment of encephalitozoonosis is usually symptomatic, with unrewarding results, and prevention methods include periodical serological screening, prophylactic administration of fenbendazole, and maintenance of a clean environment. The purpose of this article is to review the current data regarding the pathogenesis, host immunity, clinical signs, diagnostic methods, treatment, and prevention methods of encephalitozoonosis in the domestic rabbit, as well as to analyze the prevalence of this disease in different countries of the world. [ABSTRACT FROM AUTHOR]

Abstract Background Neurological signs, such as head tilt, torticollis, paralysis, and seizures, are common in rabbits. Differential diagnoses include two zoonotic infections caused by the microsporidial fungi Encephalitozoon cuniculi and the apicomplexan protozoa Toxoplasma gondii. Both infections are mainly latent in rabbits but may cause severe or even fatal disease. Although several international studies have reported the seroprevalence of these pathogens in different commercial rabbit populations, similar prevalence studies and risk-factor analyses among family-owned pet rabbits are uncommon and lacking in Scandinavia. We sought to estimate the seroprevalence and possible risk factors for E. cuniculi and T. gondii among Finnish pet rabbits. We used ELISA to measure E. cuniculi IgG seroprevalence of 247 rabbits and modified direct agglutination test for T. gondii seroprevalence of 270 rabbits. Samples were collected as part of the Finnish Pet Rabbit Health Research project. Internet-based questionnaires (n = 231) completed by the rabbit owners were used for risk-factor analysis. Results The apparent seroprevalence of E. cuniculi was 29.2% and true seroprevalence of T. gondii 3.9%. Risk factors were analysed only for E. cuniculi due to the low T. gondii seroprevalence. The final multivariable logistic regression model revealed that rabbits spending the whole summer outdoors had a higher risk of being E. cuniculi seropositive than rabbits with limited outdoor access. Additionally, rabbits living in households with only one or two rabbits had higher risk of being E. cuniculi seropositive than those in multi-rabbit households. Conclusions Nearly one third of Finnish pet rabbits participating in this study had E. cuniculi IgG antibodies, indicating previous exposure to this pathogen. The prevalence is similar to that reported previously in clinically healthy rabbit populations in UK and Korea. While the seroprevalence of T. gondii was low (3.9%), antibodies were detected. Therefore, these zoonotic parasitic infections should be considered as differential diagnoses when treating rabbits.

Encephalitozoon cuniculi is a microsporidian parasite mostly associated with its natural host, the rabbit (Oryctolagus cuniculus). However, other animals can be infected, like other mammals, birds, and even humans. Although it usually causes subclinical infection, it can also lead to encephalitozoonosis, a clinical disease characterized by neurological, ocular, and/or renal signs that can be even fatal, especially in immunocompromised individuals. Therefore, this multidisciplinary review contributes with updated information about the E. cuniculi, deepening in its molecular and genetic characterization, its mechanisms of infection and transmission, and its prevalence among different species and geographic locations, in a One Health perspective. Recent information about the diagnostic and therapeutic approach in the main host species and the prophylaxis and infection control measures currently suggested are also discussed. [ABSTRACT FROM AUTHOR]

• Previous infection with E. cuniculi reduced CD8 T lymphocytes and macrophages viability. • CD8 T lymphocytes from Xid mice showed low proliferation. • High expression of CD69 and Lamp-1 in CD8 T cells from WT mice uninfected. • A higher microbicidal activity of macrophages co-cultures with CD8 T lymphocytes from Xid mice. Encephalitozoon cuniculi is an opportunistic intracellular pathogen that establishes a balanced relationship with immunocompetent individuals depending on the activity of their CD8+ T cells lymphocytes. However, lower resistance to experimental infection with E. cuniculi was found in B-1 deficient mice (Xid), besides increased the number of CD8 T lymphocytes. Here, we evaluated the profile of CD8+ T lymphocytes from Balb/c wild-type (WT) or Balb/c Xid mice (with B-1 cell deficiency) on the microbicidal activity of macrophages challenged with E. cuniculi. Naïve CD8 T lymphocytes from WT or Xid mice uninfected and primed CD8 T lymphocytes from WT or Xid mice infected with E cuniculi were co-cultured with macrophages previously challenged with E. cuniculi. We evaluated macrophages viability and microbicidal activity, and CD8 T lymphocytes viability and presence of activating molecules (CD62L, CD69, and CD107a). Macrophages co-cultured with naïve CD8 T lymphocytes from WT demonstrated high microbicidal activity. Naïve CD8 T lymphocytes obtained from WT mice had a higher expression of CD69 and LAMP-1-activating molecules compared to Xid CD8+ T lymphocytes. Primed CD8 T lymphocytes from Xid mice proliferated more than those from WT mice, however, when the expression of the activating molecule CD69 associated with the expression of CD62L was kept low. In conclusion, naïve CD8+ T lymphocytes from Xid mice, deficient in B-1 cells, they had reduced expression of activation molecules and cytotoxic activity. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi is a microsporidial parasite that primarily infects domestic rabbits (Oryctolagus cuniculus). It is the causative agent of encephalitozoonosis, a disease with an internationally recognized seroprevalence among rabbits. This study determines the presence, clinical manifestation, and serological status of encephalitozoonosis in pet rabbits in Slovenia using various diagnostic procedures. From 2017 to 2021, 224 pet rabbit sera were collected and tested for encephalitozoonosis with the indirect immunofluorescence assay. Immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies against E. cuniculi were confirmed in 160 (65.6%) cases. Most seropositive rabbits suffered from neurological clinical signs or signs of gastrointestinal disorders such as recurrent hypomotilities, chronic weight loss, cachexia, or anorexia, and fewer showed clinical signs related to the urinary system or phacoclastic uveitis. A quarter of the positively tested rabbits presented without clinical signs. Hematological and biochemical blood analysis confirmed that seropositive animals had elevated globulin and deviated albumin levels in comparison to the normal reference values of non-infected animals. Furthermore, rabbits with neurological clinical signs showed statistically significant higher levels of globulins and total protein. Sixty-eight whole-body radiographs and thirty-two abdominal ultrasound reports were analyzed, looking for changes in the shape or size of the urinary bladder, presence of urinary sludge or uroliths, and any abnormalities related to the kidneys (shape, size, or nephrolites). The results suggest that neurological defects in the urinary bladder caused by E. cuniculi lead to a distended urinary bladder and consequently dysuria, incontinence, urine scalding, and sludgy urine.

A European goldfinch (Carduelis carduelis), a canary (Serinus canaria), and a lovebird (Agapornis roseicollis) captive-bred at three different private aviaries in Spain were submitted for necropsy with a history of weakness and ruffled feathers, weight loss associated with glossitis, and respiratory disease, respectively. Microscopically, enterocytes in the jejunum and ileum contained colonies of gram- and Stamp-positive, oval to elliptical microorganisms within parasitophorous vacuoles in the apical cytoplasm. Nested PCR using MSP primers that target microsporidian RNA genes produced amplicons of expected size for Encephalitozoon species, and analysis of forward and reverse DNA sequences confirmed the presence of Encephalitozoon hellem in all cases. The main cause of death of all three birds consisted of concurrent infections. However, intestinal encephalitozoonosis may have contributed to exacerbated catabolism. Encephalitozoonosis (or microsporidiosis) has been rarely described in passerine birds.

Background: Neurological signs, such as head tilt, torticollis, paralysis, and seizures, are common in rabbits. Differential diagnoses include two zoonotic infections caused by the microsporidial fungi Encephalitozoon cuniculi and the apicomplexan protozoa Toxoplasma gondii. Both infections are mainly latent in rabbits but may cause severe or even fatal disease. Although several international studies have reported the seroprevalence of these pathogens in different commercial rabbit populations, similar prevalence studies and risk-factor analyses among family-owned pet rabbits are uncommon and lacking in Scandinavia. We sought to estimate the seroprevalence and possible risk factors for E. cuniculi and T. gondii among Finnish pet rabbits. We used ELISA to measure E. cuniculi IgG seroprevalence of 247 rabbits and modified direct agglutination test for T. gondii seroprevalence of 270 rabbits. Samples were collected as part of the Finnish Pet Rabbit Health Research project. Internet-based questionnaires (n = 231) completed by the rabbit owners were used for risk-factor analysis. Results: The apparent seroprevalence of E. cuniculi was 29.2% and true seroprevalence of T. gondii 3.9%. Risk factors were analysed only for E. cuniculi due to the low T. gondii seroprevalence. The final multivariable logistic regression model revealed that rabbits spending the whole summer outdoors had a higher risk of being E. cuniculi seropositive than rabbits with limited outdoor access. Additionally, rabbits living in households with only one or two rabbits had higher risk of being E. cuniculi seropositive than those in multi-rabbit households. Conclusions: Nearly one third of Finnish pet rabbits participating in this study had E. cuniculi IgG antibodies, indicating previous exposure to this pathogen. The prevalence is similar to that reported previously in clinically healthy rabbit populations in UK and Korea. While the seroprevalence of T. gondii was low (3.9%), antibodies were detected. Therefore, these zoonotic parasitic infections should be considered as differential diagnoses when treating rabbits. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi is a eukaryote, unicellular, spore-forming, obligate intracellular microorganism of the phylum Microsporidia, with domestic rabbits as its main host. Another important species in which this pathogen has been identified are humans, the infection being therefore called a ”zoonosis”. The transmission takes place via the horizontal route or the vertical route, and cell-mediated immunity plays the biggest role in the infected hosts’ protection. Encephalitozoonosis can manifest itself as an acute infection, with neurological signs, renal signs, and ocular lesions, or as a chronic or subclinical infection, which is usually the case for asymptomatic carriers. The diagnostic techniques usually carried out are histological examination, serological tests, and molecular genetic techniques. The treatment of encephalitozoonosis is usually symptomatic, with unrewarding results, and prevention methods include periodical serological screening, prophylactic administration of fenbendazole, and maintenance of a clean environment. The purpose of this article is to review the current data regarding the pathogenesis, host immunity, clinical signs, diagnostic methods, treatment, and prevention methods of encephalitozoonosis in the domestic rabbit, as well as to analyze the prevalence of this disease in different countries of the world.

Microsporidia are a group of obligate, intracellular, spore‐forming eukaryotic pathogens, which predominantly infects immunocompromised individuals worldwide. Encephalitozoon spp. is one of the most prevalent microsporidia known to infect humans. Host immune system plays a major role in combating pathogens including Encephalitozoon spp. infecting humans. Both innate and adaptive arms of host immune system work together in combating Encephalitozoon infection. Researchers are conducting studies to elucidate the role of both arms of immune system against Encephalitozoon infection. In addition to cell‐mediated adaptive immunity, role of innate immunity is also being highlighted in clearance of Encephalitozoon spp. from host body. Therefore, the current review will give a clear and consolidated update on the role of innate as well as adaptive immunity in protection against Encephalitozoon spp. [ABSTRACT FROM AUTHOR]

Monitoring of infectious diseases is one of the most important pillars of preventive medicine in zoos. Screening for parasitic and bacterial infections is important to keep animals and equipment safe from pathogens that may pose a risk to animal and human health. Zoos usually contain many different animal species living in proximity with people and wild animals. As an epidemiological probe, 188 animals (122 mammals, 65 birds, and one reptile) from a zoo in Slovenia were examined for selected pathogens. Antibodies to Toxoplasma gondii and Neospora caninum were detected by ELISA in 38% (46/122) and 3% (4/122) of mammals, and in 0% (0/64) and 2% (1/57) of birds, respectively; the reptile (0/1) was negative. A statistically significant difference in T. gondii prevalence was found in Carnivora compared to Cetartiodactyla and primate antibodies to Encephalitozoon cuniculi were detected by IFAT in 44% (52/118) of mammals and 20% (11/56) of birds, respectively; the reptile (0/1) was negative. Herbivores had a higher chance of being infected with E. cuniculi compared to omnivores. Antibodies to Chlamydia abortus and Coxiella burnetii were not detected in any of the 74 tested zoo animals. The sera of 39 wild rodents found in the zoo were also examined; they were negative for all three parasites. The parasite T. gondii was detected by PCR in the tissue of two mute swans (Cygnus olor), three eastern house mice (Mus musculus), one yellow-necked field mouse (Apodemus flavicollis), and one striped field mouse (A. agrarius). Positive samples were genotyped by a single multiplex PCR assay using 15 microsatellite markers; one sample from a mute swan was characterized as type II. This micro-epidemiological study offers a better understanding of pathogens in zoo animals and an understanding of the role of zoos in biosurveillance.

• This study provides the first insight into Encephalitozoon cuniculi infection in laboratory and pet rabbits in Iran. • Brains and kidneys of 117 asymptomatic pet and lab rabbits were examined using molecular and histopathological methods. • Results revealed that 3.3% of laboratory rabbits and 59.6% of pet rabbits harboured E. cuniculi in their brains. • FELASA guidelines for monitoring of laboratory animals' health status should be employed in breeding and experimental units. • Special care should be taken in diagnosis of this zoonotic infection in pet rabbits. Encephalitozoon cuniculi infects a wide variety of domestic and wild mammalian species including humans. Although the infection status has been studied in laboratory and pet rabbits worldwide, there is shortage of information regarding the disease in Iran. In the present study, the occurrence of infection in brains of 117 asymptomatic rabbits from six breeding and experimental units with highest population of rabbit colonies in the country (n = 60) as well as pet rabbits of pet stores in two cities (n = 57) were examined by nested-PCR. Histological sections of brains and kidneys were also studied by light microscopy. PCR results revealed that 3.3% of laboratory rabbits (2/60) and 59.6% of pet rabbits (34/57) harboured E. cuniculi in their brains. Histopathology on the other hand showed spores of the parasite in kidney and brain of one and kidney of another pet rabbit. As encephalitozoonosis may interfere with results of experiments performed on laboratory rabbits, routine screenings for identification and culling of infected animals is recommended. Furthermore, infected companion rabbits can transmit E. cuniculi to people in close contact with them, therefore, improving public knowledge of this zoonotic infection is suggested. [ABSTRACT FROM AUTHOR]

Many reports have been published on the suspected vertical transmission of Encephalitozoon cuniculi ; however, prior to 2003, these reports were based on circumstantial evidence, such as histopathological, immunohistochemical, or serological diagnosis of the infection. In 2003, vertical transmission of the parasite was confirmed by detection of E. cuniculi DNA in fetuses with the nested polymerase chain reaction (PCR) technique. However, the passage of the parasite to eyes of fetus during the intrauterine stage still requires verification. In the current study, natively infected with parasite spores female rabbits were mated with non-infected males. All resulting offspring that died before ten postpartum days were investigated using molecular techniques to confirm the intrauterine transmission of the parasite to the offspring' eyes. In total, 119 DNA samples from rabbit offspring tissues were collected from blood, kidney, brain, eye (both eyes were used as single samples), lung, placenta, liver and heart were used for PCR. Parasitic DNA in the eyes of offspring was detected (54%) 6 of 11 naturally seropositive mother rabbits. PCR results were found to be positive for the eyes of 63% (19/30) of the offsprings from seropositive rabbits. Therefore, mother rabbits naturally infected with E. cuniculi showed the molecular presence of the parasite in their offspring' eyes. Sequence analysis confirmed the partial DNA sequence data of E. cuniculi and blast analysis identified the agent as genotype I. These results confirm transmission of E. cuniculi to rabbit offspring' eyes in the intrauterine period. This is the first molecular evidence to show ocular transmission of the infection via an intrauterine route in rabbits. [ABSTRACT FROM AUTHOR]

A 47-day-old black-footed ferret (Mustela nigripes) kit was found dead in June 2016. Histologic examination revealed pyogranulomatous tubulointerstitial nephritis, pneumonia, and encephalitis, with intralesional microsporidia. Transmission electron microscopic examination showed microsporidia with ultrastructural characteristics consistent with Encephalitozoon spp. Polymerase chain reaction (PCR) and direct sequencing confirmed the presence of Encephalitozoon cuniculi genotype II. This organism has been reported in other Carnivora (i.e., canids, felids, mustelids, procyonids, otariids). In humans, it is generally described as an opportunistic pathogen in immunocompromised individuals. The source of infection in the quarantine facility remains unknown, although two groups of frozen feeder rat kidneys tested positive for E. cuniculi genotype II via PCR. Feeding whole prey to various zoo taxa carries some potential disease transmission risk. [ABSTRACT FROM AUTHOR]

Problems with parasitic infections are common in zoological gardens and circuses. In some animals it can lead to several disorders such as systemic disease, reproductive disorders (abortions and neonatal mortality), and even to death if severe illness is untreated. Thus, the aim of this study was to evaluate the prevalence of three common parasites in 74 animals from three zoos, and four circuses in Southern Italy. Antibodies to Toxoplasma gondii, Neospora caninum, and Encephalitozoon cuniculi were detected in 51%, 12%, and 20% of animals, respectively. Co‐infections of T. gondii and N. caninum were reported in seven animals (9%) and co‐infection of T. gondii and E. cuniculi in one animal. T. gondii, N. caninum and E. cuniculi seroprevalence differed in type of diet (P ≤ 0.0001; P ≤ 0.037 and P ≤ 0.004, respectively). T. gondii and E. cuniculi seroprevalence also differed in animal families (P ≤ 0.0001) and according to type of housing (P ≤ 0.003), respectively. Statistical differences were not found in other characteristics (gender, age, country of birth, origin, and contact with cats or dogs). This is the first serological study focusing on protozoan and microsporidian parasites in zoo and circus animals from Southern Italy and the first detection of antibodies to E. cuniculi in camels in Europe. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii, Neospora caninum, and Encephalitozoon cuniculi are important infectious agents, with T. gondii and E. cuniculi having zoonotic potential. There are two main clonal lineages (types I and II) of T. gondii in Europe, but little is known about genotypes of T. gondii in wild animals. The aim of our study was molecular detection of these three pathogens in tissues of wild red foxes (Vulpes vulpes) from the Czech Republic. Using PCR (B1 gene), we detected T. gondii in 10% of the animals that we tested (n=100); N. caninum and E. cuniculi were not detected. The T. gondii samples were genotyped by single multiplex PCR assay with 15 microsatellite markers. Five samples were successfully genotyped as genotype II, a unique finding for T. gondii isolated from red foxes from the Czech Republic. [ABSTRACT FROM AUTHOR]

A 2-mo-old Vancouver Island marmot (Marmota vancouverensis), housed at a quarantined breeding facility, presented for acute obtundation and vestibular ataxia. Physical examination revealed poor growth compared with littermates, poor nutritional condition, and mild dehydration. The animal's condition deteriorated over 24 hr, and it was euthanized following the development of generalized seizures. No gross abnormalities were observed upon postmortem evaluation. Histologic evaluation revealed severe, multifocal, granulomatous and lymphoplasmacytic meningoencephalomyelitis and interstitial nephritis, with intralesional, intracytoplasmic spore-filled, parasitophorous vacuoles and segmental, multi-organ, fibrinoid vasculitis (disseminated encephalitozoonosis). The etiologic agent was evident by hematoxylin and eosin and Gram-chromotrope stains, and confirmed as Encephalitozoon cuniculi by polymerase chain reaction on brain tissue. Sequencing of the internal transcribed spacer region of the rRNA gene showed 100% homology with E. cuniculi strain IV, which is a newly described genotype. This is the first report of encephalitozoonosis in this critically endangered species. [ABSTRACT FROM AUTHOR]

Microsporidia are intracellular pathogens that cause severe disease in immunocompromised humans and animals. We recently demonstrated that XID mice are more susceptible to Encephalitozoon cuniculi infection by intraperitoneal route, evidencing the role of B-1 cells in resistance against infection. The present study investigated the resistance and susceptibility against E. cuniculi oral infection, including the role of B-1 cells. BALB/c and BALB/c XID (B-1 cells deficient) mice were orally infected with E. cuniculi spores. No clinical symptoms were observed in infected animals; histopathology showed lymphoplasmocytic enteritis with degeneration of the apexes of the villi in all infected groups. Higher parasite burden was observed in infected BALB/c XID mice. In the spleen and peritoneum, all infected mice showed a decrease of lymphocytes, including CD8 + T cells, mostly in infected BALB/c XID mice. Adoptive transfer of B-1 cells (XID + B-1) was associated with a lower parasite burden. Pro-inflammatory cytokines (IFN-γ, TNF-α and IL-6) increased mostly in infected XID + B1 mice. Together, the present results showed that BALB/c XID mice infected by the oral route were more susceptible to encephalitozoonosis than BALB/c mice, demonstrating the B-1 cells importance in the control of the immune response against oral E. cuniculi infection. [ABSTRACT FROM AUTHOR]

[b]Introduction and objectives.[/b][i] Encephalitozoon cuniculi[/i] is an obligate intracellular parasite infecting especially domestic rabbits; however, spontaneous infections have been documented in other mammalian species such as dogs, cats, rabbits, horses, cows and sheep all over the world. Encephalitozoonosis is a chronic and latent disease leading to renal failure, encephalitis, disorders of brain and urinary tract, and may lead to death. There are limited reports on encephalitozoonosis in wildlife, which is why the aim of this study was to detect the prevalence of antibodies to [i]E. cuniculi[/i] in European hares. [b]Materials and methods. [/b]Samples of blood sera from 701 wild hares from the Czech Republic (n = 245), the Slovak Republic (n = 211) and Austria (n = 245) were examined by indirect immunofluorescence antibody test (IFAT); samples with titer ≥ 40 were marked as positive. [b]Results[/b]. The total seroprevalence of E. cuniculi antibodies was 1.42% with titres in the range 40–640. Antibodies to [i]E. cuniculi[/i] were detected in 2.9% (7/245), 0.8% (2/245) and 0.47% (1/211) hares from the Czech Republic, Austria and the Slovak Republic, respectively. [b]Conclusions[/b]. This is the first detection of antibodies to [i]E. cuniculi[/i] in hares from Europe showing that hares could be exposed to[i] E. cuniculi[/i] infection, however with a low rate.

Encephalitozoonosis is a common infectious disease widely spread among rabbits. Its causative agent, Encephalitozoon cuniculi, is considered as a zoonotic and emerging pathogen capable of infecting both immunocompetent and immunocompromised hosts, including humans. In rabbits, clinical signs include neurological, kidney and ocular disease. The aim of this study was to detect E. cuniculi in ocular structures in immunocompetent rabbits after experimental oral infection using immunohistochemistry. In infected animals, E. cuniculi spores were present in periocular connective tissue, sclera, cornea, choroidea, iris, retina and lens, as a round to ovoid organism reacting with a specific anti-E. cuniculi monoclonal antibody as early as 2 weeks after infection. There were no signs of inflammatory lesions in any of the ocular tissues examined at 2, 4, 6 and 8 weeks after infection. In the present study, E. cuniculi was also detected in the lenses of adult rabbits, which indicates that ways of lens infection other than intrauterine and haematogenic are possible.

Microsporidia are obligate intracellular, spore-forming fungi. A wide range of vertebrate and invertebrate hosts can be infected; however, cases of infected turtles or tortoises have not yet been described. This is the first description of 4 cases in tortoises (Testudo spp), which showed general signs of illness as well as intestinal and respiratory signs until death occurred or they were euthanized. Granulomatous and necrotizing inflammation was visible in the lung and liver. Fungal organisms were present in heart blood, lung, liver, and intestine. Because of their morphology, staining properties (eg, positive in periodic acid–Schiff and silver reaction), and electron microscopic appearance, they were classified as microsporidia. [ABSTRACT FROM AUTHOR]

A European goldfinch ( Carduelis carduelis ), a canary ( Serinus canaria ), and a lovebird ( Agapornis roseicollis ) captive-bred at three different private aviaries in Spain were submitted for necropsy with a history of weakness and ruffled feathers, weight loss associated with glossitis, and respiratory disease, respectively. Microscopically, enterocytes in the jejunum and ileum contained colonies of gram- and Stamp-positive, oval to elliptical microorganisms within parasitophorous vacuoles in the apical cytoplasm. Nested PCR using MSP primers that target microsporidian RNA genes produced amplicons of expected size for Encephalitozoon species, and analysis of forward and reverse DNA sequences confirmed the presence of Encephalitozoon hellem in all cases. The main cause of death of all three birds consisted of concurrent infections. However, intestinal encephalitozoonosis may have contributed to exacerbated catabolism. Encephalitozoonosis (or microsporidiosis) has been rarely described in passerine birds. [ABSTRACT FROM AUTHOR]

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 (κ = 0.99), and the ELISA and IFAT (κ = 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.

OBJETIVO: O microsporídio Encephalitozoon cuniculi tem sido reconhecido como um patógeno oportunista em indivíduos imunossuprimidos, tais como pacientes com Aids. O objetivo do trabalho foi desenvolver animais farmacologicamente imunossuprimidos como modelo da infecção natural pelo E. cuniculi. MÉTODOS: Foram usados grupos distintos de camundongos Balb-C adultos, imunossuprimidos com diferentes doses de dexametasona (Dx, 3 ou 5 mg/kg/dia por via intraperitoneal ¾ IP) e inoculados com esporos de E. cuniculi por via IP. Também foram usados grupos controle (animais inoculados, mas nãoimunossuprimidos, e animais imunossuprimidos, mas não inoculados). Os esporos de E. cuniculi foram previamente cultivados em células MDCK. Os animais foram sacrificados e submetidos à necropsia aos 7, 14, 21, 28 e 35 dias pós-inoculação. Fragmentos teciduais foram coletados e processados para análise por microscopia de luz, utilizando-se as técnicas de coloração de Gram -chromotrope e de hematoxilina-eosina. RESULTADOS: Em todos os animais imunossuprimidos e inoculados, porém especialmente naqueles que receberam 5 mg/kg/dia de Dx, os achados de necropsia mais proeminentes foram hepato e esplenomegalia. A inoculação experimental resultou em uma infecção disseminada e não-letal, caracterizada por lesões granulomatosas em diversos órgãos (fígado, pulmões, rins, intestino, encéfalo), porém mais notadamente no tecido hepático. Esporos de E. cuniculi foram observados em poucos animais tratados com 5 mg/kg/dia de Dx aos 35 dias pós-infecção. CONCLUSÕES: Microsporidiose em camundongos imunossuprimidos com Dx fornece um modelo útil para estudos da infecção por microsporídios, assemelhando-se àquela naturalmente observada em indivíduos imunodeficientes com Aids.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: Distint 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 immunosupressed 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.

Clinical signs of neurological disease, such as head tilt, hind limb paresis or paralysis, seizures, and muscle weakness, are commonly encountered in pet rabbits, and in the authors׳ experience, less often in rodent species. Moreover, localisation of neurological lesions and establishment of a definitive diagnosis can be challenging for any of the exotic small mammal species. In many rabbit and rodent cases, distinguishing neurological disease from musculoskeletal disease is difficult. The parasitic disease encephalitozoonosis is commonly diagnosed in pet rabbits; in both rabbits and rodents, bacterial infections are also a common underlying cause of neurological disease. Other causes of neurological diseases that adversely affect pet rabbits and rodents include toxins, trauma, metabolic and degenerative disorders, viral infections, neoplasia, and hereditary abnormalities. [ABSTRACT FROM AUTHOR]