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Background: In rabbits, renal abscesses (pus-filled sores) are rare and diagnosis remains challenging. Therefore, in this study, we aimed to determine the clinical manifestation and diagnostic tests associated with renal abscess identification in rabbits. Case Description: A four-and-a-half-year-old castrated male Lionhead rabbit with a history of poor appetite and abdominal distension was admitted to the animal hospital. Blood analysis, radiography, ultrasonography, and computed tomography scans revealed a kidney abscess found within the renal parenchyma, with severe loss of the cortex and medulla, extending toward the capsule. Consequently, the rabbit underwent nephrectomy. The enlarged right kidney was surgically removed. Histopathological examination of the affected kidney showed severe necrosis and ischemic zones, atrophy of the renal tubules, and prominent heterophils with mixed inflammatory cell infiltrates. Immunohistochemistry and polymerase chain reaction confirmed Encephalitozoon cuniculi and Escherichia coli infections, respectively. Conclusion: This report provides novel insights into the diagnosis of renal abscesses in Lionhead rabbits. [ABSTRACT FROM AUTHOR]

Simple Summary: Encephalitozoonosis is a disease caused by Encephalitozoon cuniculi. It is diagnosed primarily in rabbits, and is less frequently so in other animal species. E. cuniculi is classified as Microsporidia—fungi frequently found in the environment, that are resistant to numerous external factors. This pathogen is diagnosed primarily in rabbits, and is less frequently so in other animal species. The objective of the study was to analyze the prevalence of E. cuniculi infections in guinea pigs with different clinical disorders. The infected animals most frequently exhibited nervous and urinary system symptoms, as well as issues with vision organs, while several animals were recorded as having problems with the respiratory system and thyroid gland dysfunction. The study shows that encephalitozoonosis constitutes a significant problem in rodents kept as domestic animals, which in turn may be a source of infection for humans. Encephalitozoonosis is a disease caused by E. cuniculi. It is diagnosed primarily in rabbits but is less frequently so in other animal species. E. cuniculi is classified among Microsporidia—fungi frequently found in the environment, that are resistant to numerous external factors. Apart from rabbits, rodents form the next group of animals most exposed to infection with these pathogens. The objective of the study was to analyze the prevalence of E. cuniculi infection in guinea pigs with different clinical disorders. The study included 67 animals with E. cuniculi infection confirmed via real-time PCR. The infected animals most frequently exhibited nervous and urinary system symptoms, as well as issues with vision organs, while several animals were also recorded as having problems with the respiratory system and thyroid gland dysfunction. The study shows that encephalitozoonosis constitutes a significant problem in rodents kept as domestic animals, which in turn may be a source of infection for humans. [ABSTRACT FROM AUTHOR]

Simple Summary: Microsporidia are obligatory intracellular fungi that cause infection in a broad variety of eukaryotes. However, there are few epidemiological studies on microsporidia in red pandas in China. Therefore, we conducted an epidemiological survey of microsporidia in red pandas from six zoos in Sichuan Province, China. In 198 fecal samples, Enterocytozoon bieneusi was detected in 24 samples (12.1%), while Encephalitozoon spp. was detected in 8 fecal samples (4.0%). Further analysis revealed the presence of genotypes of E. bieneusi with zoonotic concerns. Interestingly, we also observed that the infection rate of microsporidia was higher in semi-free red pandas with more frequent contact with other animals or humans. This is the first report on the occurrence and genotypes of E. bieneusi and Encephalitozoon spp. in red pandas in Sichuan Province, China, contributing to our understanding of the worldwide distribution and genetic diversity of this pathogen. Enterocytozoon bieneusi and Encephalitozoon spp. are microsporidian pathogens with zoonotic potential that pose significant public health concerns. To ascertain the occurrence and genotypes of E. bieneusi and Encephalitozoon spp., we used nested PCR to amplify the internal transcribed spacer (ITS) gene and DNA sequencing to analyze 198 fecal samples from red pandas from 6 zoos in China. The total rate of microsporidial infection was 15.7% (31/198), with 12.1% (24/198), 1.0% (2/198), 2.0% (4/198) and 1.0% (2/198) for infection rate of E. bieneusi, Encephalitozoon cuniculi, Encephalitozoon intestinalis and Encephalitozoon hellem, respectively. One red panda was detected positive for a mixed infection (E. bieneusi and E. intestinalis). Red pandas living in semi-free conditions are more likely to be infected with microsporidia (χ2 = 6.212, df = 1, p < 0.05). Three known (SC02, D, and PL2) and one novel (SCR1) genotypes of E. bieneusi were found. Three genotypes of E. bieneusi (SC02, D, SCR1) were grouped into group 1 with public health importance, while genotype PL2 formed a separate clade associated with group 2. These findings suggest that red pandas may serve as a host reservoir for zoonotic microsporidia, potentially allowing transmission from red pandas to humans and other animals. [ABSTRACT FROM AUTHOR]

Simple Summary: It is well known that mammals can harbor various pathogens that can affect humans (known as zoonotic pathogens) including viruses, bacteria, fungi and parasites. Microsporidia are a group of pathogens related to fungi and parasites of several animals that can cause diarrhea or systemic infection in humans. Due to the limited knowledge about microsporidia infection in hedgehogs worldwide, this study aimed to analyze the presence and identity of microsporidia in a group of North African hedgehogs from the Canary Islands (Spain). Enterocytozoon bieneusi and Encephalitozoon cuniculi, two zoonotic species of microsporidia, were identified. These results suggest that microsporidia species with zoonotic risk circulate in the archipelago. Microsporidia are unicellular eukaryotic obligate intracellular parasites with a wide range of hosts reported worldwide; however, little is known about the epidemiological data on microsporidia infection in animals from the Canary Islands. Since data on microsporidia infection in hedgehog species are scarce, the aim of this study was to analyze the presence and identity of microsporidia in a group of North African hedgehogs (Atelerix algirus) using microscopic and molecular methods. From December 2020 to September 2021, a total of 36 fecal samples were collected from naturally deceased hedgehogs from Tenerife and Gran Canaria. All samples showed spore-compatible structures (100%; 36/36) under microscopic analysis, of which 61.1% (22/36) were amplified via the nested-polymerase chain reaction (PCR) targeting the partial sequence of the 16S rRNA gene, the internal transcribed spacer (ITS) region, and the partial sequence of the 5.8S rRNA gene. After Sanger sequencing and ITS analysis, Enterocytozoon bieneusi was detected in 47.2% (17/36) of the samples, identifying two novel genotypes (AAE1 and AAE2), followed by the detection of an undetermined species in 8.3% (3/36) and Encephalitozoon cuniculi genotype I in 5.6% (2/36) of the samples. This study constitutes the first report of microsporidia species in Atelerix algirus worldwide, highlighting the high prevalence of zoonotic species. [ABSTRACT FROM AUTHOR]

Simple Summary: Microsporidia are a group of fungal-related pathogens widely distributed in the environment, with some species having a negative impact on animals and public health. The European rabbit (Oryctolagus cuniculus) is considered the natural host of Encephalitozoon cuniculi, a microsporidian pathogen of mammals, including humans. The infection caused by E. cuniculi, encephalitozoonosis, ranges from asymptomatic to severe lesions in rabbits, with clinical signs involving the central nervous system, kidney, and eye being the most common. The majority of reported cases have been in domestic rabbits, while cases in wild rabbits are uncommon. Due to the lack of data on microsporidia in the Canary Islands, the aim of this work was to analyze the prevalence and identity of microsporidia in fecal samples from rabbits in Tenerife. Enterocytozoon bieneusi and Encephalitozoon spp. are microsporidia with zoonotic potential that have been identified in humans, as well as in a large group of wild and domestic animals. Several wildlife species have been studied as reservoirs of zoonotic microsporidia in mainland Spain, including the European rabbit (Oryctolagus cuniculus). Due to a lack of data on microsporidia infection in wildlife on the Canary Islands, the aim of this work was to analyze the prevalence and identify the species of microsporidia in rabbits in Tenerife. Between 2015 and 2017, a total of 50 fecal samples were collected from rabbits in eight municipalities of Tenerife, Canary Islands, Spain. Seven of the fifty samples (14%) were amplified using nested polymerase chain reaction (PCR) targeting the partial sequence of the 16S rRNA gene, the internal transcribed spacer (ITS) region, and the partial sequence of the 5.8S rRNA gene. Sanger sequencing reveals the presence of Encephalitozoon cuniculi genotype I in two samples (4%), and undescribed microsporidia species in five samples (10%). This study constitutes the first molecular detection and genotyping of E. cuniculi in rabbits in Spain. [ABSTRACT FROM AUTHOR]

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]

We identified Encephalitozoon cuniculi genotype II parasites as a cause of extraintestinal microsporidiosis in 2 owners of birds also infected with E. cuniculi. Patients experienced long-lasting nonspecific symptoms; the disease course was more progressive in a patient with diabetes. Our findings suggest direct bird-to-human transmission of this pathogen. [ABSTRACT FROM AUTHOR]

Objective: The aim of this study is to investigate the seroprevalence of Encephalitozoon cuniculi infection in rabbits in facilities with different care conditions in three different provinces. Methods: The sera samples of rabbits that have been hosted in open, semi-open and controlled areas which were obtained from three different enterprises with different environmental possibilities, hygienic situations, maintenance and feeding conditions were evaluated by using the ELISA kits. Results: Consequently, the seropositivity of the rabbits in open, semi-open and controlled shelters were found to be 70%, 40% and 80%, respectively. In this study, in the rabbit under the different care conditions, antibodies anti - E. cuniculi (63.3%) were found in all sera samples. Conclusion: In the context of the data obtained, it was seen that the animals taken into the facility may be infected from the beginning and an infected animal in the facility may have been a source of infection for the many others in the facility. It was also concluded that the septic and aseptic rules which are potentially accepted in the facilties are not enough in battling with the agent and that the zoonotic disease can be a risk for animal keepers and researchers. [ABSTRACT FROM AUTHOR]

Background: Encephalitozoon cuniculi is an important microsporidian parasite with zoonotic potential. The present study highlights the impact of encephalitozoonosis on rabbit health in Egypt. Three rabbit farms in Giza, with a total of 16,400 rabbits were investigated due to occurrence of rabbits displaying clinical signs consistent with encephalitozoonosis. Results: Clinical signs observed during a 4 months observation period in 2018 included vestibular disease, paresis, limb paralysis, cataracts, phacoclastic uveitis, frequent urination, marked decrease in body weight and in some pregnant females, also repeated abortions. The total morbidity rates in adult and young rabbits were 76.7% and 81.5%, respectively. The highest mortality rate was recorded in offspring (12.3%), followed by dams (5.6%), and the lowest recorded mortality rate was in males (0.04%). Post-mortem findings included enteritis, pale enlarged kidneys, congested leptomeninges, focal brain necrosis, and endometrial congestion. Histopathological examination revealed nonsuppurative meningoencephalitis and glial nodules with central necrosis in the brain, vacuolation and necrosis of renal tubular epithelium, and corneal ulceration and ruptured lens capsule with fragmentation of lenticular fibres. E. cuniculi were observed in the brain, retinal ganglion cells, kidneys, and liver. Transmission electron microscopy examination revealed the presence of different developmental stages of E. cuniculi in the brain and kidney. Presence of E. cuniculi was confirmed by conventional polymerase chain reaction using a universal 16S gene for Encephalitozoon spp. followed by sequencing and sequence analysis. Conclusions: The presence of E. cuniculi in rabbits was confirmed at three farms in Egypt. Nervous signs and ocular lesions were the most predominant findings in these farms. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi is a unicellular, spore-forming, obligate intracellular eukaryote belonging to the phylum Microsporidia. It is known to infect mainly immunocompromised and immunocompetent mammals, including humans. The parasite–host relationship has been evaluated using both in vitro cell culturing and animal models. For example, Balb/c and C57BL/6 mouse strains have been used interchangeably, although the latter has been considered more susceptible due to the higher fungal load observed after infection. In the present study, we identified the characteristics of the immune response of C57BL/6 mice treated or not with the immunosuppressant cyclophosphamide (Cy) and challenged with E. cuniculi by intraperitoneal route. After 14 days of infection, serum was collected to analyze Th1, Th2, and Th17 cytokine levels. In addition, peritoneal washes were performed, and the spleen sample was collected for immune cell phenotyping, whereas liver, spleen, kidney, lung, intestine, and central nervous system (CNS) samples were collected for histopathological analysis. Although infected mice displayed a reduced absolute number of macrophages, they showed an M1 profile, an elevated number of CD4+T, CD8+T, B-1, and B-2 lymphocytes, with a predominance of Th1 inflammatory cytokines (interferon [IFN]-γ, tumor necrosis factor [TNF]-α, and interleukin [IL]-2) and Th17. Furthermore, Cy-Infected mice showed a reduced absolute number of macrophages with an M1 profile but a reduced number of CD4+T, CD8+T, B-1, and B-2 lymphocytes, with a predominance of Th1 inflammatory cytokines (IFN-γ, TNF-α, and IL-2) and Th2 (IL-4). This group displayed a higher fungal burden as well and developed more severe encephalitozoonosis, which was associated with a reduced number of T and B lymphocytes and a mixed profile of Th1 and Th2 cytokines. [Display omitted] • E. cuniculi infection determined clinical signs and inflammatory infiltrate in liver. • E. cuniculi infection reduced the macrophage population and Cy modified its profile. • Cy-Infected mice had reduced T and B lymphocytes. • Increased inflammatory cytokines in infected mice. • C57BL/6 mice showed a difference in macrophage population compared to Balb/c mice. [ABSTRACT FROM AUTHOR]

Microsporidia are opportunistic pathogens that infect a wide range of invertebrates and vertebrates. To assess the potential role of dogs in the transmission of these zoonotic pathogens, a total of 282 fecal samples from dogs in the Central Anatolia Region of Turkey were analyzed by utilizing species specific polymerase chain reaction for the four most frequent human microsporidia. Two microsporidia species were recognized in 41 samples (14.5%). Encephalitozoon intestinalis was detected in 35 samples (12.4%) and it was the most common microsporidium. The second microsporidium, E. cuniculi, was identified in six (2.1%) of the samples. Sequence analysis of the intergenic spacer of the ribosomal ribonucleic acid (RNA) internal transcribed spacer (ITS) gene revealed the presence of three E. intestinalis haplotypes closely associated with each other. No polymorphic region was found among the ITS sequences of E. cuniculi isolates and they were characterized as genotype III. This study provides the first data on the zoonotic microsporidia species from dogs in Turkey. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi is an obligate macrophage parasite of vertebrates that commonly infects rodents, monkeys, dogs, birds, and humans. In the present study, we aimed to assess the phagocytosis and intracellular survival of E. cuniculi spores using untreated and lipopolysaccharide (LPS)-activated J774A.1 murine macrophages and assess the macrophage viability. The experimental groups comprised untreated spores, spores killed by heat treatment at 90 °C, and spores killed by treatment with 10% formalin. LPS-activated macrophages significantly increased the phagocytosis of spores and reduced their intracellular growth after 24 and 48 h (P < 0.01); however, after 72 h, we observed an increase in spore replication but no detectable microbicidal activity. These results indicate that LPS activation enhanced E. cuniculi phagocytosis between 24 and 48 h of treatment, but the effect was lost after 72 h, enabling parasitic growth. This study contributes to the understanding of the phagocytosis and survival of E. cuniculi in murine macrophages. [ABSTRACT FROM AUTHOR]

Highlights • Life-long immunosuppression increases the risk of respiratory microsporidiosis. • Symptoms of respiratory microsporidiosis are non-specific. • Encephalitozoon cuniculi should be diagnosed in respiratory samples of transplant patients. Abstract Objectives Encephalitozoon spp. and Enterocytozoon bieneusi are intracellular parasitic fungi from the phylum Microsporidia, which initially localize to the intestine. As opportunistic pathogens, Encephalitozoon spp. in particular can disseminate to the respiratory tract, among other locations. Patients on life-long immunosuppression are at higher risk of such infections, mostly symptomatic. Methods Sputum samples and bronchial washings from 72 renal transplant recipients and 105 patients with various respiratory diseases were screened for Encephalitozoon spp. and E. bieneusi by microscopic examination and genus-specific nested PCR followed by genotyping. Results A total of 8.3% (6/72) of immunosuppressed renal transplant recipients and 1.9% (2/105) of patients with various respiratory diseases, both immunocompetent and immunosuppressed, were positive for respiratory microsporidial infection. All six transplant recipients were Encephalitozoon cuniculi -positive by PCR/sequencing and five of them suffered from respiratory symptoms. The presence of microsporidial spores was also confirmed microscopically in three of the transplant recipients. Of the two immunocompetent patients with various respiratory diseases, one had an E. cuniculi infection, while the second had an E. bieneusi infection. Conclusions Life-long immunosuppression in renal transplant recipients increases the risk of respiratory infection by E. cuniculi. Microsporidia should be screened in respiratory samples of these patients, particularly when they have respiratory symptoms. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi can cause latent disease, especially in lagomorphs and many wild and domestic animals in various countries. This infection is important for veterinary and public health because it is caused by a potentially zoonotic and opportunistic pathogen. The aim of this study was to investigate the relationship between seropositivity of E. cuniculi and renal function markers, which may be useful in predicting the disease in clinically healthy rabbits. In this study, the seropositivity of E. cuniculi infection in clinically healthy rabbits was determined, and necropsy findings were evaluated along with the results of renal function tests. In a laboratory rabbit breeding facility, enzyme-linked immunosorbent assay (ELISA) tests showed that 48 (49.5%) of 97 rabbits were seropositive against E. cuniculi. Blood urea nitrogen (BUN) and creatinine levels were significantly elevated in seropositive animals. Two seropositive rabbits were necropsied to confirm the infection. According to histopathological findings in the kidney, degenerative changes and E. cuniculi spores were identified in the tubule epithelia. Serum creatinine (p<0.001) and BUN (p<0.01) levels were found to have a statistically significant relationship with the serological status of rabbits. Serological and histopathological methods are not routinely used in rabbits to diagnose E. cuniculi infection. The kidneys were one of the most affected organs in encephalitozoonosis in rabbits. As revealed in this study, the testing blood urea nitrogen and creatinine levels would be useful for the evaluation of general health status and renal function of the seropositive rabbits, and clinical interpretation as well. [ABSTRACT FROM AUTHOR]

Background Among patients with hip joint endoprosthesis, periprosthetic osteolysis is the most common complication following primary arthroplasty, and subsequent implant loosening is the leading cause of arthroplasty revision. Causes of stability loss, though not always evident, can be mechanical, allergic, or infectious (bacterial and fungal agents) in nature. Microsporidia, widespread opportunistic fungal pathogens that infect most human tissues, are a potential infectious cause of stability loss. Infections caused by Encephalitozoon species—one of the most common microsporidial pathogens in humans—primarily localize to intestinal and respiratory tracts, but can disseminate to tissues throughout the body. Methods We examined 53 immunocompetent patients, 23 after revision and 30 after primary hip arthroplasty, for infection by Encephalitozoon species. Periprosthetic tissue, urine sediments, and stool samples were tested by microscopic examination and genus-specific nested polymerase chain reaction followed by genotyping. Results Ten patients had Encephalitozoon -positive periprosthetic tissues, 9 (39%) after revision and 1 (3.3%) after primary hip arthroplasty. Among the tissue-positive postrevision patients, 7 had a positive urine sample and 1 had a positive stool sample. Encephalitozoon cuniculi genotype II was identified in 88.8% (16/18) of samples. Two urine samples were positive for a novel Encephalitozoon species. Conclusions Encephalitozoon cuniculi should be considered as a cause of osteolysis in hip periprosthetic tissue, leading to a loss of implant stability. [ABSTRACT FROM AUTHOR]

Highlights • Encephalitozoon cuniculi infection in rabbits with neurological manifestations. • Parasite DNA (genotype I) detected in brain sample. • Brain lesions observed at the early stage of infection. Abstract Encephalitozoon cuniculi , a zoonotic and opportunistic pathogen, can cause latent infection, especially in lagomorphs. Nowadays, this member of the Eukaryotes has drawn significant attention in the fields of veterinary and public health. The purpose of this study was to determine the seroprevalence of infection in a New Zealand rabbit farm that has a clinical history of neurological manifestations including head tilt ataxia, aggressiveness, seizures, and circling and rotational movements around the body length axis, but the general conditions and food intake were normal. Blood samples were taken from 42 breeding rabbits and researched for E. cuniculi antibodies. Out of that, 25 (59%) animals resulted positive against the pathogen. The rabbit was found to be seropositive for E. cuniculi antibodies, but negative for Toxoplasma gondii and Listeria monocytogenes antibodies. Hematological and serum biochemical parameters were measured at reference intervals. No brain tissue impairment was observed the computed tomography (CT) scan. As a result of these histopathological findings, the brain cortex presented severe neuronal degeneration and partial myelin loss, with reactive diffuse gliosis against the parasite spores was observed to the histopathology. These results are possibly related to the early stage of infection because the parasitic infestation comprise long time spreading. E. cuniculi DNA was detected on brain tissues using polymerase chain reaction (PCR), and it partial DNA sequence was identified as E. cuniculi genotype I. [ABSTRACT FROM AUTHOR]

Abstract Microsporidia are obligate intracellurar unicellular parasite of wide range of vertebrates. Although ingestion or inhalation of microsporidian spores is the main route of infection, assumed vertical transmission was described in some mammals. The present study was focused on proof of vertical transmission in mice under experimental conditions. Mice were infected with E. cuniculi genotype II intraperitoneally after mating, or perorally followed by mating in acute or chronic phase of infection. Fetuses were delivered by Caesarean section or mice were kept up to the parturition. Some of cubs were immediately after birth transferred to uninfected surrogate mothers. Group of cubs was immunosuppressed. All cubs were examined using polymerase chain reaction for the presence of Encephalitozoon after birth or in their age of 3 or 6 weeks, respectively. All fetuses delivered by Caesarean section, which were intraperitoneally or perorally infected were negative as well as all neonatal mice and youngsters tested in age of 6 weeks. Only immunosuppressed cubs and cubs of immunodeficient mice in age of 21 days were positive for Encephalitozoon cuniculi genotype II. Present results provided the evidence that transplacental transmission of Encephalitozoon cuniculi in mice occurs, but the mechanism of these transport is still unknown. Graphical abstract Image Highlights • Encephalitozoon cuniculi is vertically transmitted in mice. • The load of spores was very low in newborn cubs. • The infection developed in cubs born to infected mothers after immunosupression. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi was first recognized as the disease agent in rabbits in 1922. The genotype of E. cuniculi isolated from laboratory rabbits with the neurologic disease was described as genotype I. In the eye, this parasite causes damage to the lens, causing phacoclastic uveitis and cataracts. Intraocular infection often occurs in cases of transplacental transmission. There has been no report on the molecular diagnosis of the parasite in Turkey. The current study is the first report on the detection of E. cuniculi spores using the molecular method in Turkey. In our previous study, a rabbit breeding facility was determined seropositive for E. cuniculi infection monitored for five years in terms of clinical symptoms. An autopsy was performed for a definite diagnosis of the infection. Samples were stained according to the hematoxylin-eosin (H&E) staining after tissue processing procedure and histopathologic analysis was performed. In addition to, the samples for DNA extraction were also taken during the autopsy. ECUNF and ECUNR species-specific primer pairs were used for amplification and genotyping of E. cuniculi. The animals were observed no clinical symptoms except ocular lesion (n=9). Therefore, one of these rabbits was used in the autopsy to definite diagnosis and determination of the damage to the eye. As histopathological, the lesions in the eye were found in the initial or middle stage of progressive infection. The DNA sequence showed that E. cuniculi examined in the present study were genotype I. Possible cause of the visible white mass in the rabbit's eye may be the parasite infection. Therefore, clinicians may consider E. cuniculi as one of the possible causes of ocular lesions in rabbits during daily inspection or ophthalmological examination. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To describe the presentation of 15 rabbits with naturally occurring cataracts referred for phacoemulsification surgery, the procedure in 13 cases and the follow‐up in 12. Animals included: Fifteen rabbits (30 eyes), nine of which stopped following visual cues in association with cataract progression. Materials and Methods: Rabbits underwent preoperative ophthalmic and ocular ultrasound examination. Results: Thirteen rabbits (22 eyes) had mature cataracts. Ten were bilateral and three unilateral. Two rabbits had an anterior chamber abscess. The cataract in one of these was incipient. One rabbit had bilateral immature cataracts. One rabbit had a subluxated lens, and one had a retinal detachment. Thirteen rabbits (22 eyes) underwent phacoemulsification. Eighteen, 13.5‐mm capsular tension rings (CTRs) and seventeen, 13‐mm IOLs (Acrivet®, Berlin, Germany) were fitted including one 41D 60V‐model, and three 49D and thirteen 58D 20S‐models. Intraoperative complications included one unilateral posterior‐capsular tear, one lens subluxation, and one expulsive choroidal hemorrhage. One rabbit died during anesthetic recovery. Nine cases were PCR‐tested for Encephalitozoon cuniculi, and only three were positive. The median follow‐up time was 12 months (4‐24 months). Rabbits that were not following visual cues preoperatively did so postoperatively, and surgery resulted in a clear visual axis for the follow‐up period in every case except in two, due to reasons other than the surgery. Conclusions: Phacoemulsification with CTR and IOL implantation offers good long‐term results and can improve the quality of life of pet rabbits. Retinal detachment, lens luxation, expulsive choroidal hemorrhage, and anesthetic death are potential complications. [ABSTRACT FROM AUTHOR]

Two 3 year-old, healthy, client-owned Lop rabbits presented with bilateral cataracts. After performing a physical examination, bloodwork, ocular ultrasonography and electroretinography, both animals were deemed good surgical candidates for phacoemulsification. Bilateral cataract surgery was performed and both rabbits regained vision in both eyes. Both animals developed post-operative ocular hypertension and one animal developed corneal ulcers immediately after surgery. Both surgical complications resolved with medical management. This case series describes phacoemulsification of bilateral cataracts in 2 companion rabbits and the use of an intraocular lens in 1 rabbit. Surgical treatment of cataracts can be considered as a treatment option whenever a healthy rabbit is visually impaired due to cataracts. [ABSTRACT FROM AUTHOR]

The expression of tumor necrosis factor (TNF) -α, interleukin (IL) −4 and IL-10, as well as apoptosis and nitric oxide (NO) levels were measured in the brain and kidneys of immunocompetent and immunosuppressed New Zealand White rabbits infected with Encephalitozoon cuniculi . All of the animals had clinical signs histopathological lesions compatible with encephalitozoonosis and were E. cuniculi -positive by using a carbon immunoassay test. Encephalitozoon cuniculi infection promoted the expression of TNF-α and NO production in the kidneys of infected rabbits, and a synergic effect was observed in animal treated with dexamethasone. The IL-4 expression was similar in the brain and kidneys of infected rabbits, regardless of their immunologic status. The IL-10 mRNA expression in the brain of infected immunosuppressed rabbits was elevated when compared with positive controls. Apoptosis of granuloma mononuclear-like cells was detected in immunocompetent E. cuniculi –infected rabbits, but it was more evident in infected-immunosuppressed animals. Nitric oxide levels were elevated both in immunocompetent and immunosuppressed infected animals, but it was more apparent in the kidneys. These data suggest that modulation of the immune response by E. cuniculi could contribute to the survival of the parasite within phagocytic cells in granulomas via an as yet undetermined mechanism. [ABSTRACT FROM AUTHOR]

Background: Encephalitozoon cuniculi is an opportunistic microsporidian parasite that can affect a number of different species of mammalian animals and humans. The parasite can pose also threat for rabbits even though it causes several sporadic and asymptomatic infections. Infection of eyes is common and clinical symptom of ocular infection may include uveitis and cataracts. We found out subacute findings in naturally infected animals and show here a first described eye lesions as well as central nervous system and kidneys in Turkey. Methods: The rabbits (n:171) of breeding units were observed to daily clinical examination for infection of E. cuniculi during three years. The eyes of five rabbits (2.9%) showed white intraocular masses or cataracts in the breeding units during daily examinations. The infection was described clinicopathologically in collected organ samples in the animals. During observation, macroscopically, corneal lesions and opacity and impaired lens were taken into attention as well as hyperemia in central nervous system and kidney. Histopathologically, parasitophorous vacuoles pertaining to E. cuniculi were detected in all three tissues during different routine Haematoxylin-Eosin and Gram stainings. Results: Degenerative and necrotic changes in epithelium of cornea and lens and also neurons and tubules were predominantly observed in addition to nonpurulent interstitiel nephritis and encephalitis. Conclusion: The results from study lead to subacute findings especially in eye during natural E. cuniculi infections following asymptomatic and latent changes among breeding colony. The lesions indicated sub-acute stage of E. cuniculi infection in eye lesions of rabbit in Turkey. [ABSTRACT FROM AUTHOR]

This study revises our understanding of the effectiveness of cell-mediated adaptive immunity and treatment against microsporidia using molecular detection and quantification of microsporidia in immunocompetent C57Bl/6 and immunodeficient CD4−/− and CD8−/− mice for the first time. We demonstrate an intense dissemination of microsporidia into most organs within the first weeks post-infection in all strains of mice, followed by a chronic infection characterized by microsporidia persistence in CD4−/− and C57Bl/6 mice and a lethal outcome for CD8−/− mice. Albendazole application reduces microsporidia burden in C57Bl/6 and CD4−/− mice, whereas CD8−/− mice experience only a temporary effect of the treatment. Surprisingly, treated CD8−/− mice survived the entire experimental duration despite enormous microsporidia burden. On the basis of our results, we conclude that microsporidia survive despite the presence of immune mechanisms and treatments that are currently considered to be effective and therefore that CD8 T lymphocytes represent a major, but not sole effector mechanism controlling microsporidiosis. Furthermore, the survival of mice does not correspond to spore burden, which provides new insight into latent microsporidiosis from an epidemiological point of view. [ABSTRACT FROM AUTHOR]

The present study was conducted to evaluate the methanolic extracts from several plant leaves widely used in traditional medicine to cure digestive tract disorders and in the self-medication of wild animals such as non-human primates, namely Archidendron fagifolium , Diospyros sumatrana , Shorea sumatrana, and Piper betle leaves, with regard to their antimicrosporidial activity against Encephalitozoon cuniculi in immunocompetent BALB/c mice determined using molecular detection of microsporidial DNA (qPCR) in various tissues and body fluids of infected, treated mice. Of the plant extracts tested, Diospyros sumatrana provided the most promising results, reducing spore shedding by 88% compared to untreated controls. Moreover, total burden per 1 g of tissue in the D. sumatrana extract-treated group reached 87% reduction compared to untreated controls, which was comparable to the effect of the standard drug, Albendazole. This data represents the baseline necessary for further research focused on determining the structure, activity and modes of action of the active compounds, mainly of D. sumatrana , enabling subsequent development of antimicrosporidial remedies. [ABSTRACT FROM AUTHOR]

Diagnosis of Encephalitozoon cuniculi infection in rabbits remains a major veterinary issue. ELISA or immunofluorescence assays are the current reference standards of serological tests. However, these conventional techniques suffer from a lack of accuracy for distinguishing active from past infections, as a positive serostatus is common in clinically normal rabbits. In this study, we assessed the diagnostic performance of Western blot (WB) to detect both anti- E. cuniculi immunoglobulin G (IgG) and immunoglobulin M (IgM) in comparison with ELISA and to address the intensity of the immune response through a quantitative approach. Positive WB results were highly correlated with the E. cuniculi-related diseased status ( P < 0.0001). Although it was more labor intensive and less standardized, quantitative WB provided detailed comparable analysis regarding the humoral response and diagnostic performance similar to ELISA testing with statistically higher sensitivity (88.4 vs. 76.1% for IgG detection and 84.3 vs. 70.4% for IgM, P < 0.01). Several specific WB bands were shown to be significantly associated with concomitant clinical signs, like the one located at 50 kDa (OR = 8.2, [2.4-27.7], P = 0.0008) for IgG and (OR = 27.9, [4.2-187.9], P = 0.0006) for IgM. Therefore, the quantitative WB may have application in veterinary diagnostic laboratories to increase the accuracy of the clinical diagnosis of E. cuniculi infection. In addition, this tool may help to further understand the development and function of the humoral immune response to this infectious agent. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi is an opportunist intracellular pathogen of mammals. The adaptive immune response is essential to eliminate E. cuniculi , but evidence is mounting that the response initiated by the innate immune response may ultimately define whether or not the parasite can survive. B-1 cells may act as antigen-presenting cells or differentiate into phagocytes, playing different roles in many infection models. However, the role of these cells in the dynamics of Encephalitozoon sp. infections is still unknown. To investigate the role of B-1 cells in E. cuniculi infection, BALB/c and BALB/c XID (B-1 cells deficient) mice were infected with E. cuniculi spores. Cytometric analyses of peritoneal cells showed that B-1 cells and macrophages increased significantly in infected BALB/c mice compared to uninfected controls. Despite the increase in the number of CD4 + and CD8 + lymphocytes in XID mice, these animals were more susceptible to infection as evidenced histologically with more prominent inflammatory lesions and parasite burden. Pro-inflammatory cytokines increased in both infected BALB/c and BALB/c XID mice. To confirm B-1 cells role in encephalitozoonosis, we adoptively transferred B-1 cells to BALB/c XID mice and this group showed few symptoms and microscopic lesions, associated with an increased in cytokines. Together, these results suggest that B-1 cells may increase the resistance of BALB/c mice to encephalitozoonosis, evidencing for the first time the important role of B-1 lymphocytes in the control of microsporidia infection. [ABSTRACT FROM AUTHOR]

Serological prevalence of E. cuniculi infection was assessed in 183 rabbits from central Italy. In seropositive deceased rabbits, histopathological lesions were also evaluated. Sera from 118 rabbits from 6 intensive farms, 10 rabbits from 6 family farms, 16 rabbits from a zoo, 30 rabbits from 5 research laboratories and 9 pet rabbits from 9 different owners, were tested by an enzyme-linked immunosorbent assay. Data were statistically analysed. Tissue samples from brain and kidney of 10 deceased rabbits were formalin-fixed and subsequently analysed by histopathology and immunohistochemistry. Anti -E. cuniculi antibodies were found in 129/183 (70.5%) analysed sera. At statistical analysis, E. cuniculi seropositivity was significantly higher ( p < 0.05) in industrial and zoo rabbits. At histology, different degrees of pathological lesions were found in serological positive (9) deceased animals. In three rabbits deceased after showing neurological signs, the severity of the lesions was interpreted as a likely cause for their death. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi (E. cuniculi) is an intracellular zoonotic parasite that is recognized as a common parasite of rabbits. Infected rabbits show neurological and renal disorder. The polar tube protein 1 of the spore is revealed to be antigenicity. The present study aimed to produce a recombinant polar tube protein 1 (rPTP1) for which the codon usage of a native ptp1 gene was optimized and synthesized for protein production in Escherichia coli (E. coli). Both a native ptp1 gene amplified by PCR technique and an optimized ptp1 gene were ligated into a pQE30 expression vector and transformed into an E. coli strain M15. The rPTP1 production was induced by 1M isopropyl--Dthiogalactoside. A protein band with a molecular weight of about 45 kDa was found in the extracted protein of both E. coli transformed with optimized ptp1 and native ptp1 plasmids. SDS-PAGE and western blot analysis using an antihistidine antibody revealed a notably higher protein yield of the E. coli containing an optimized ptp1 plasmid than that of the E. coli containing a native ptp1 plasmid. Biological function of the recombinant protein was evaluated by western blotting using E. cuniculi-positive and negative rabbit sera. Result showed that the biological function of the optimized rPTP1 protein expressed from E. coli could differentiate between E. cuniculi-negative and positive rabbit sera. Therefore, rPTP1 can be used for the development of a diagnostic test for E. cuniculi infection in the future. [ABSTRACT FROM AUTHOR]

Levels of interferon (IFN)-γ and interleukin (IL)-10 were measured in the serum of immunocompetent and immunosuppressed New Zealand White rabbits naturally infected with Encephalitozoon cuniculi . IFN-γ levels were elevated in infected rabbits, and a synergic effect was observed in animals treated with the immunosuppressive agent dexamethasone (Dex). The role of IL-10 in infected rabbits remains unclear, as IL-10 levels were similar to those of negative controls. Dex appeared to exhibit a proinflammatory effect, as IFN-γ levels were elevated in infected immunosuppressed rabbits. Similarly, Dex exhibited a synergic effect in infected immunosuppressed rabbits, as evidenced by the elevation in IFN-γ production. These data indicate that the immune response to this glucocorticoid should be considered in the design of future animal model studies of immunosuppression. [ABSTRACT FROM AUTHOR]

This study aimed to investigate pathology of oral E. cuniculi infection during pregnancy in pregnant mice and embryos. A total of 40 pregnant mice at first day of gestation were divided into two groups, first group were infected orally by E. cuniculi sores of 107 spores/ mice, second group left without any treatment. At 18th days of gestation all pregnant mice were euthanized. Gross pathology finding in pregnant mice of infected group included congestion of liver and lung, the embryos lesions consisted from enlargement of head and abdomen. Histological lesions in pregnant mice of infected group consisted of hepatic non-suppurative granulomatous lesions with E. cuniculi spores aggregation with lymphocytic infiltration, the lungs lesions consisted of infiltration of lymphocytes with E. cuniculi spores, kidney lesions composed from degenerative and necrotic changes in renal tubules, brain lesions consisted from lymphocytic infiltration with increase in number of glial cells, while intestine tissue sections showed hyperplasia of lymphatic tissue with present of parasitic vacuoles at tips of villi, the placenta exhibited E. cuniculi spores with hyperplasia of trophoblast in chorionic villi, while histological lesions in embryos showed lymphocytic infiltration around alveoli with hyperplasia of lymphatic tissue around bronchioles with absent the normal architecture of hepatic cords and vacuolation of hepatocytes with hyperplasia of lymphocytes in white pulp of spleen. This study provides insight into the pathology of E. cuniculi infection in pregnant mice and their embryos, also supports the hypothesis of intrauterine transmission of E. cuniculi infection to embryos during pregnancy period. [ABSTRACT FROM AUTHOR]

This study was conducted to evaluate the efficacy of oral administration of fenbendazole (20 mg/kg body weight) prior to and after experimental infection of immunosuppressed rabbits with Encephalitozoon cuniculi. A total of thirty rabbits were divided into five groups: NN (non-immunosuppressed; non-infected), IN (immunosuppressed; non-infected), IPI (immunosuppressed; protectedinfected), ITI (immunosuppressed; treated-infected), and II (immunosuppressed; infected) groups. Fenbendazole was administered as a prophylactic for seven successive days before infection with E. cuniculi and as a treatment for four weeks initiated on the 28th day post-challenge (PC). Experimental rabbits were infected with intraperitoneal injection of 2 ? 105 E. cuniculi spores. Parameters evaluated were body weight, detection of spores in urine, serum antibody assay, hematological, biochemical and histopathological changes. The IPI and ITI groups showed a significant better final bwt than the II group. Spores were detected in urine of all infected rabbits from the 28th day PC until the end of the study. The IPI group showed the least values of antibodies (IgG) compared to the ITI and II groups. Concerning histopathological changes, the intensity of the lesions was marked particularly in the II rabbits and to a lesser extent in the ITI rabbits. Noticeable improvement was found in the IPI rabbits. It could be concluded that fenbendazole was effective to some extent in protection of rabbits against E. cuniculi infection, while when administered as a therapeutic no significant effects were observed. [ABSTRACT FROM AUTHOR]

Several enteric microsporidia species have been detected in humans and other vertebrates and their identifications at the genotype level are currently being elucidated. As advanced methods, reagents, and disposal kits for detecting and identifying pathogens become commercially available, it is important to test them in settings other than in laboratories with 'state-of-the-art' equipment and well-trained staff members. In the present study, we sought to detect microsporidia DNA preserved and extracted from FTA (fast technology analysis) cards spotted with human fecal suspensions obtained from Cameroonian volunteers living in the capital city of Yaoundé to preclude the need for employing spore-concentrating protocols. Further, we tested whether amplicon nucleotide sequencing approaches could be used on small aliquots taken from the cards to elucidate the diversity of microsporidia species and strains infecting native residents. Of 196 samples analyzed, 12 (6.1%) were positive for microsporidia DNA; Enterocytozoon bieneusi (Type IV and KIN-1), Encephalitozoon cuniculi, and Encephalitozoon intestinalis were identified. These data demonstrate the utility of the FTA cards in identifying genotypes of microsporidia DNA in human fecal samples that may be applied to field testing for prevalence studies. [ABSTRACT FROM AUTHOR]

Background: The microsporidian Encephalitozoon cuniculi is an obligate intracellular eukaryotic pathogen with a small nuclear genome (2.9 Mbp) consisting of 11 chromosomes. Although each chromosome end is known to contain a single rDNA unit, the incomplete assembly of subtelomeric regions following sequencing of the genome identified only 3 of the 22 expected rDNA units. While chromosome end assembly remains a difficult process in most eukaryotic genomes, it is of significant importance for pathogens because these regions encode factors important for virulence and host evasion. Results: Here we report the first complete assembly of E. cuniculi chromosome ends, and describe a novel mosaic structure of segmental duplications (EXT repeats) in these regions. EXT repeats range in size between 3.5 and 23.8 kbp and contain four multigene families encoding membrane associated proteins. Twenty-one recombination sites were identified in the sub-terminal region of E. cuniculi chromosomes. Our analysis suggests that these sites contribute to the diversity of chromosome ends organization through Double Strand Break repair mechanisms. The region containing EXT repeats at chromosome extremities can be differentiated based on gene composition, GC content, recombination sites density and chromosome landscape. Conclusion: Together this study provides the complete structure of the chromosome ends of E. cuniculi GB-M1, and identifies important factors, which could play a major role in parasite diversity and host-parasite interactions. Comparison with other eukaryotic genomes suggests that terminal regions could be distinguished precisely based on gene content, genetic instability and base composition biais. The diversity of processes assciated with chromosome extremities and their biological consequences, as they are presented in the present study, emphasize the fact that great effort will be necessary in the future to characterize more carefully these regions during whole genome sequencing efforts. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi spores cause severe granulomatous inflammation in the brain where mononuclear cells and macrophages infiltrate. Here, we orally infected New Zealand white rabbits with 1 × 106 E. cuniculi viable spores to study the recruitment and localization of macrophages in brain granulomas. At day 30 post-infection, the positive phenotype markers iNOS (M1) and Arg-1 (M2) were located in the periphery and center of granulomas, respectively. Live intracytoplasmic spores were found only in positive Arg-1 cells. This is the first work to describe the recruitment and distribution of M1 and M2 macrophages in the brain granulomas of rabbits infected with E. cuniculi. [Display omitted] • All infected brain sections contained immunolabels for M1 and M2 cells. • M1 cells were peripherally distributed inside the granuloma. • M2 cells localized at the center of the granuloma. • M2 cells were unable to destroy the spores. [ABSTRACT FROM AUTHOR]

Background: Microsporidia species are obligatory intracellular agents that can infect I all major animal groups including mammals, birds, fishes and insects. Whereas worldwide human infection reports are increasing, the cognition of sources of infection particularly zoonotic transmission could be helpful. We aimed to detect zoonotic microsporidia spore in fecal samples from some animals with close -- contact to human. Methods: Overall, 142 fecal samples were collected from animals with closed-contact to human, during 2012-2013. Trichrome -- blue staining were performed and DNA was I then extracted from samples, identified positive, microscopically. Nested PCR was also I carried out with primers targeting SSU rRNA gene and PCR products were sequenced. Results: From 142 stool samples, microsporidia spores have been observed microscopically in 15 (10.56%) samples. En. cuniculi was found in the faces of 3 (15%) small white mice and 1 (10%) laboratory rabbits(totally 2.81%). Moreover, E. bieneusi was detected in 3 (10%) samples of sheep, 2 (5.12%) cattle, 1 (10%) rabbit, 3 (11.53%) cats and 2 (11.76%) ownership dogs (totally 7.74%). Phylogenetic analysis showed interesting data. This is the first study in Iran, which identified E. bieneusi and En. Cuniculi in fecal samples of laboratory animals with close -- contact to human as well as domesticated animal and analyzed them in phylogenetic tree. Conclusion: E. bieneusi is the most prevalent microsporidia species in animals. Our results can also alert us about potentially zoonotic transmission of microsporidiosis. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Encephalitozoon cuniculi is a microsporidian parasite commonly found in rabbits that can infect humans, causing encephalitozoonosis. Our objective in this study was to evaluate the seroprevalence of this parasite in rabbits and humans in China Methods: Overall, 300 serum samples each from clinically healthy rabbit and human were collected from three regions of China (Sichuan Province, Chongqing Municipality and Jilin Province) from January to September 2013 and tested for anti- E. Cuniculi antibodies using an ELISA. Results: An overall seroprevalence of E. cuniculi was recorded as 56/300 (18.76%) and 29/300 (9.76%) in rabbit and human sera, respectively. The seropositivity of rabbit samples collected from Jilin province was 41%, which was significantly higher (P<0.01) than Sichuan Province (9%) and Chongqing Municipality (6%). Three breeds of rabbit were used in the present study and antibody detection in Rex Rabbit was significantly (P<0.01) higher than Japanese White and New Zealand Rabbit. In human, Jilin province was more prevalent (18%) followed by Sichuan Province (6%) and Chongqing Municipality (5%). Conclusions The E. cuniculi was present and widespread among healthy rabbits and humans in China [ABSTRACT FROM AUTHOR]

Background: As a zoonotic pathogen, Encephalitozoon cuniculi is a cause of serious disease in animals and people. The present study was to evaluate the health status examination of this seropositive animal care worker in our previous study. Methods: Blood samples were taken from five workers. CIA test was applied to detect antibodies against E. cuniculi in blood serum. The indirect immunofluorescence antibody test was used as confirmation test. Seropositive worker had a complete medical examination. Results: Only one worker was found to be seropositive according to the results of the serological test. Sera positive to E. cuniculi was confirmed with IFAT and spores were detected in the urine sample of the worker. The worker was treated with albendazole. Conclusion: Rabbits should be examined routinely for the presence of anti-E. cuniculi antibody. People working with laboratory animal should avoid contact with urine and faeces of infected or pay attention to personal hygiene. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi is an obligate intracellular microsporidian parasite that infects a wide range of mammalian hosts. The present study investigated the prevalence of E. cuniculi in different animal hosts from different provinces of Egypt (Alexandria, Behera, and Assuit) using serological (IFAT and ELISA) and molecular (PCR) assays. A total of 324 serum and 274 urine samples were collected from seven different species of animals (cattle, buffaloes, sheep, goat, rabbit, dog, and rat). The results of serological examination confirmed the occurrence of antibodies against E. cuniculi in 38.9 % (126 out of 324) of the examined animals. The significant ( P < 0.01) highest positivity was observed in goats (67 %) followed by buffaloes, rabbits, dogs, rat, and cattle (46.42, 41, 40, 36.2, and 28.1 %, respectively), while the least was recorded in sheep (9 %). Behera province showed the highest ( P < 0.01) infection rate (40.68 %) followed by Alexandria and Assuit (39.2 and 22.73 %, respectively). The infection rate was significantly higher ( P < 0.01) in females (45.34 %) than that in males (30.47 %). Positive cases were observed in all age categories. The highest infection rate (64.66 %) was recorded in the age group 1-5 years and the least was recorded in the age group <1 year (34.85 %).On the other hand, only five positive out of 274 urine samples (1.82 %) were detected by PCR. Our study provides a wide database on prevalence and epidemiology of an ignored parasite ( E. cuniculi) for the first time in Egypt. [ABSTRACT FROM AUTHOR]

• Intravenous infection of E. cuniculi determined interstitial pneumonia with a predominance of lymphocytic infiltrate. • Inflammatory infiltrate was characterized by the predominance of CD8 and CD4 T lymphocytes. • Immunosuppression aggravated pneumonia and increased fungal burden. Opportunistic fungal pneumonia is a cause of concern in immunocompromised patients due to its high morbidity and mortality rates. One such opportunistic agent affecting immunocompromised patients is the microsporidia called Encephalitozoon cuniculi. This study aimed to evaluate pneumonia caused by E. cuniculi in mice treated with the immunosuppressive agent cyclophosphamide (Cy). This study also aimed to describe the immune cells associated with the microsporidial pneumonia. C57BL/6 mice were infected intravenously with E. cuniculi spores and treated with Cy (75 mg/kg/week, intraperitoneally). Thirty days post-infection, the fungal burden (qPCR), histopathological lesions, cytokine production, and the phenotype of the immune cells in the lung parenchyma were evaluated. Histologically, interstitial pneumonia with lymphocytic infiltrate was observed in the infected animals. The infiltrate mainly consisted of CD8+ and CD4+ T lymphocytes, with reduced populations of B lymphocytes and macrophages. The production of tumor necrosis factor-alpha (TNF-α) was significant in the animals of the infected groups. Also, the fungal burden was higher in the Cy-treated animals, which was confirmed by the immunohistochemical observation of spores. These results demonstrated that E. cuniculi infection of C57BL/6 mice caused lymphocytic interstitial pneumonia (characterized by a predominant lymphocytic infiltrate), which was aggravated by Cy-induced immunosuppression. Thus, these results can be used to understand the different pathological, immunological, and therapeutic aspects of lymphocytic interstitial pneumonia. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi is a parasite that can infect a variety of animals, including immune-compromised humans. Rabbits are prone to encephalitozoonosis but the subclinical course and high serum prevalence make it difficult to diagnose with certainty in live animals. Different approaches have been used to support the presence of an active infection in symptomatic rabbits: antibodies titers, IgM and IgG serum evaluation, electrophoresis, PCR in urine, and cerebrospinal fluid analysis. In this study, 76 symptomatic rabbits were divided in three groups based on their Ig results: IgM+ IgG+, IgM-IgG+ and IgM-IgG-. By statistical analysis total serum protein was proved not to be effective in predicting an antibody response, in contrast the γ-globulin ranges from serum electrophoresis can predict whether the patient is positive for IgM and/or IgG. In fact, a significant quantitative correlation between percentage of γ-globulins and positive IgM and IgG serum for E. cuniculi was established in symptomatic rabbit. These values can be useful to perform a screening both for symptomatic and asymptomatic rabbits. [ABSTRACT FROM AUTHOR]

The breeding of domestic rabbit (Oryctolagus cuniculus) for human consumption has a long tradition mainly in European and Asian countries. 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 animals health. This study aimed to collect sera from rabbits bred in different conditions and test the presence of Toxoplasma gondii and Encephalitozoon cuniculi antibodies. Whether infections were active or latent was assessed by determining the occurrence of IgM or IgM together with IgG antibodies which indicated active infection whereas latent infection was characterized by finding IgG antibodies only. An ELISA test was performed with 1883 sera samples collected throughout the Czech and Slovak Republics. The seroprevalence of T. gondii in 902 samples from 6 commercial farms (CF) was very low with only 4 rabbits (0.4%) being positive. In total 99 (10.1%) individuals out of 981 samples from 29 household farms (HF) were positive for T. gondii antibodies. Only 2 (50%) of the T. gondii positive CF rabbits had active infections while the rest were latently infected. The serological results showed that 35 (35.4%) rabbits from the T. gondii positive HF group suffered from active infection. Out of CF samples 185 (20.5%) were positive for E. cuniculi. Antibodies of E. cuniculi were detected in 497 (50.7%) HF rabbits. Active E. cuniculi infections were determined in 85.9% of CF and 56.3% of HF rabbits; respectively. Interestingly, the E. cuniculi positive rabbits were significantly more often positive for anti-T. gondii antibodies in comparison to E. cuniculi negative individuals. Prevalence of T. gondii in CF rabbits was negligible. According to our results meat of HF rabbits still poses a risk of T. gondii infection. Nevertheless, the risk is on its lowest level in 20 years which is apparently caused due to changes in feeding practices. The occurrence of E. cuniculi antibodies was significantly lower in rabbits from commercial farms, apparently because of better hygiene conditions. [ABSTRACT FROM AUTHOR]

Abstract: The sequence encoding SWP1 was cloned from the genome of Encephalitozoon cuniculi. Recombinant SWP1 (rSWP1) was expressed in Escherichia coli and used to detect E. cuniculi infections in farmed foxes and dogs with an indirect enzyme-linked immunosorbent assay (ELISA) in the present study. The sera of foxes infected with E. cuniculi could be distinguished from the sera of foxes infected with Toxoplasma gondii, Neospora caninum, and Cryptosporidium parvum using the ELISA. In total, 198 fox samples collected in Liaoning were used to determine the prevalence of antibodies against this disease. The results showed that 16.7% of the fox serum samples were positive according to the ELISA using rSWP1, which agreed with the ELISA results based on recombinant PTP2 (rPTP2). The sensitivity and specificity of the ELISA based on rSWP1 suggest that this could be an alternative method for the diagnosis of E. cuniculi infections in foxes. In addition, 298 dog samples collected in Beijing, Shanghai, and Hunan were also detected in this study, of which six dog samples (2%) were positive according to the ELISA using rSWP1. To the best of our knowledge, this is the first study to demonstrate the serological prevalence of E. cuniculi infections in dogs and foxes in China. [Copyright &y& Elsevier]

Abstract: Encephalitozoon is an obligate intracellular microsporidian parasite that infects a wide range of mammalian hosts. In this study, we used nested PCR to investigate the presence of Encephalitozoon infection in Rodentia and Soricomorpha in Japan. We attempted to amplify and sequence Encephalitozoon-specific DNA from brain and viscera samples of 180 animals collected between 2008 and 2010. Forty-three samples (23.9%) from the orders Rodentia and Soricomorpha were positive for Encephalitozoon. This study is the first report of Encephalitozoon infection in Rodentia and Soricomorpha in Japan, and our findings suggest that these hosts may play a role in the spread of microsporidian spores in the environment. [Copyright &y& Elsevier]

Microsporidia are obligate intracellular pathogens capable of infecting a wide variety of hosts ranging from invertebrates to vertebrates. The infection process requires a step of prior adherence of Microsporidia to the surface of host cells. A few studies demonstrated the involvement of proteins containing a ricin-B lectin (RBL) domain in parasite infection. In this study Anncalia algerae and Encephalitozoon cuniculi genomes were screened by bioinformatic analysis to identify proteins with an extracellular prediction and possessing RBL-type carbohydrate-binding domains, being both potentially relevant factors contributing to host cell adherence. Three proteins named Aa RBLL-1 and Aa RBLL-2 from A. algerae and Ec RBLL-1 from E. cuniculi , were selected and comparative analysis of sequences suggested their belonging to a multigenic family, with a conserved structural RBL domain despite a significant amino acid sequence divergence. The production of recombinant proteins and antibodies against the three proteins allowed their subcellular localization on the spore wall and/or the polar tube. Adherence inhibition assays based on pre-treatments with recombinant proteins or antibodies highlighted the significant decrease of the proliferation of both E. cuniculi and A. algerae , strongly suggesting that these proteins are involved in the infection process. [Display omitted] • RBLL proteins are localized on the polar tube and spore wall. • Aa RBLL-1, Aa RBLL-2 and EcRBLL-1 decreased significantly the host cell infection. • Ec RBLL-1, Aa RBLL-1 and Aa RBLL-2 could contribute to the anchoring of the polar tube. [ABSTRACT FROM AUTHOR]

Encephalitozoon cuniculi , an intracellular pathogen, lives in a balanced relationship with immunocompetent individuals based on the activity of T lymphocytes. We previously highlighted the greater susceptibility of B-1 cell-deficient mice (XID mice) to encephalitozoonosis. This study aimed to develop a model of disseminated and severe encephalitozoonosis in mice with combined immunodeficiency to elucidate the role of B cells. To address this objective, cyclophosphamide (Cy)-treated BALB/c and XID mice were inoculated with E. cuniculi , followed by the evaluation of the immune response and histopathological lesions. Immunosuppressed BALB/c mice manifested no clinical signs with an increase in the populations of T lymphocytes and macrophages in the spleen. Immunosuppressed and infected XID mice revealed elevated T cells, macrophages populations, and pro-inflammatory cytokines levels (IFN-γ, TNF-α, and IL-6) with the presence of abdominal effusion and lesions in multiple organs. These clinical characteristics are associated with extensive and severe encephalitozoonosis. The symptoms and lesion size were reduced, whereas B-2 and CD4+ T cells populations were increased in the spleen by transferring B-2 cells adoptive to XID mice. Moreover, B-1 cells adoptive transfer upregulated the peritoneal populations of B-2 cells and macrophages but not T lymphocytes and decreased the symptoms. Herein, we speculated the consistency in the development of severe and disseminated encephalitozoonosis in mice with genetic deficiency of Bruton's tyrosine kinase (Btk) associated with Cy immunosuppression develop with that of the models with T cell deficiency. Taken together, these data emphasized the crucial role of B cells in the protective immune response against encephalitozoonosis. [Display omitted] • Deficiency of B-1 and B-2 cells in XID mice and Cy treatment caused a more severe and widespread encephalitozoonosis. • Tansfer of B-2 cells decreased all components of immunity in the peritoneal cavity and downregulated cytokines. • Increase in T lymphocytes, macrophages and pro-inflammatory cytokines was demonstrated in Xid mice with encephalitozoonosis. [ABSTRACT FROM AUTHOR]

The aim of the study was to carry out a serological survey of Encephalitozoon cuniculi in commercially reared rabbit does (Oryctolagus cuniculi) in Veneto region (Northern Italy). Two hundred and sixty blood samples from 13 farms were examined by a carbon immunoassay (CIA test) to detect the presence of antibodies anti-E. cuniculi. All sampled rabbit does were clinically healthy. Seropositivity against E. cuniculi was found in 196/260 (75.4%) sera and in all the sampled farms (100%). Logistic regression analysis showed that the size of the farm had no statistically significant effect on E. cuniculi positivity; whereas rabbits of the hybrid X showed a higher seropositivity (p < 0.01) than rabbits belonging to other commercial breeds. Moreover, the age seemed to influence the seropositivity (p < 0.05). This serological survey showed a high prevalence of E. cuniculi infection suggesting that this parasite may be endemic in industrial rabbitries in Northern Italy. [ABSTRACT FROM AUTHOR]

Abstract: Faecal samples were collected from 377 horses on 23 farms with varying management systems in the Czech Republic. Microsporidia were found on 16 farms and the overall prevalence of Enterocytozoon bieneusi and Encephalitozoon cuniculi was 17.3% (66/377) and 6.9% (26/377), respectively. The prevalence of E. cuniculi in horses over 3years of age was significantly higher (10.0%) compared to younger horses (4.0%). No significant differences in prevalence were observed among stallions, geldings, and mares for both microsporidia. Significantly higher infection rates of E. bieneusi and E. cuniculi were recorded in horses kept in stables than those on pasture. Two genotypes of E. cuniculi (I and II) and 15 genotypes of E. bieneusi including six previously described and nine novel genotypes were detected. The most common genotype detected was E. bieneusi genotype D identified in 51.5% (34/66) of positive horses. The identification of E. bieneusi genotypes D, EpbA, G and WL15, which were previously reported in pigs, humans, racoons and horses, indicates that horses could be a potential source of zoonotic infection in humans. [Copyright &y& Elsevier]

Abstract: Between 2000 and 2008, 95 rabbits with suspected encephalitozoonosis and neurological symptoms were treated at the Clinic of Small Animal Medicine, Ludwig Maximilian University Munich. Standard treatment consisted of oxytetracycline (from 2000 to 2003; n =50) or fenbendazole and oxytetracycline (from 2004 to 2008; n =45), and the rabbits were randomly assigned to treatment groups with or without dexamethasone. Each therapeutic regime was given for 10days, with fluids, B vitamins and nutritional support added as needed. Therapeutic success was evaluated by assessing the survival rate on day 10, the neurological score of the surviving rabbits and Kaplan–Meier curves for long-term survival past 10days. Inclusion of fenbendazole in the treatment protocol was associated with increased survival rates on day 10 (p =0.043), better neurological scores (p =0.008), and improved long-term survival (p =0.025) based on the results of univariate analyses. Treatment with dexamethasone showed no effect on neurological score or on short- or long-term survival. The study did not provide any evidence that dexamethasone is an effective component of the treatment scheme. [Copyright &y& Elsevier]