First insight into Encephalitozoon cuniculi infection in laboratory and pet rabbits in Iran

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.     

Molecular detection and genotype identification of E. cuniculi from pet rabbits

Encephalitozoon cuniculi is a microsporidian which is frequently reported from rabbits. This microorganism can either ravage rabbit farms or transmit to humans from pet rabbits. This study aimed to investigate the prevalence and the genotype distribution of E. cuniculi among pet rabbits. In this study urine samples were collected from 50 pet rabbits, aged 2 months to 3 years, admitted to teaching veterinary hospital. Four races Lop, Dutch, Mix, and Angora were screened for E. cuniculi. The clinical symptoms were recorded and total DNA was extracted from urine samples. E. cuniculi was identified using amplification of the small subunit ribosomal RNA (ssu rRNA) gene and its genotypes were characterized using PCR/sequencing of the polar tube protein (PTP) gene. Phylogenetic tree was drawn to confirm the characterized genotypes. Out of 50 samples, 41 (82 %) of rabbits were asymptomatic, while nine (18 %) had at least one of symptoms including head-tilt, circling, and ataxia. A statistical correlation was seen between mean age + SD and symptoms (P-value = 0.039). The presence of E. cuniculi was confirmed in 16/50 (32 %) rabbits and all of them were identified as the genotype I. Our findings represented no consistency between E. cuniculi PCR – positive and the presence of symptoms (P-value = 0.318). Our results showed positive correlation between symptoms and age; however, the lack of correlation between PCR results with age may signify the latent infection in younger rabbits. All identified E. cuniculi were the genotype I, which is reported from rabbits and humans, highlighting the zoonotic concern for this genotype, particularly among subjects who keep pet rabbits.     

Serological survey for antibodies to Encephalitozoon cuniculi and Toxoplasma gondii in pet rabbits in eastern coastal areas of China

Encephalitozoon cuniculi (E. cuniculi) is a microsporidian parasite commonly detected in rabbits and can infect humans and cause encephalitozoonosis. And Toxoplasma gondii is a prevalent parasite distributed worldwide and can infect almost all warm-blooded animals, including humans. The aim of the current study was to investigate the seroprevalence of E. cuniculi and Toxoplasma gondii, and risk factors of infection in pet rabbits reared in eastern coastal areas of China (Tianjin, Shandong, Jiangsu, Zhejiang, Shanghai and Fujian). Total 222 blood samples of pet rabbits were collected from local veterinary hospitals. The seropositivity rates of E. cuniculi were 16.22% (36/222) according to an Enzyme-linked immunosorbent assay (ELISA). Female pet rabbits was significantly higher than that in males (P=0.002), Zhejiang were markedly higher than those in Jiangsu and Shanghai (P=0.017, P=0.022), and cross-breed rabbits were dramatically higher than those in Chinchilla, New Zealand white, Rex (P=0.02, P=0.006, P=0.008). The seroprevalence of T. gondii was 13.06% (29/222) by the method of ELISA. The seroprevalence in Zhejiang was significantly higher than that in Shanghai (P=0.017). No difference in seroprevalence was detected with respect to the gender, age, species, health status, or season. These findings show that E. cuniculi and T. gondii are present and spread in pet rabbits. Therefore, pet rabbits should be considered as an important reservoir of encephalitozoonosis for humans and maybe important implication for public health in eastern coastal areas of China.     

Encephalitozoon cuniculi infection in rabbits

The microsporidian parasite Encephalitozoon cuniculi commonly infects rabbits. Most infections are initially asymptomatic, but for reasons yet to be explained, many rabbits subsequently develop disease as a result of infection with this organism. Three common forms of this disease are recognized, and they can occur individually or in combination. The ocular form is associated with cataracts and when there is extensive damage to the lens, uveitis. The neurological form can vary from a mild change in the rabbit’s behavior to severe vestibular disease. The signs associated with the renal form of the disease are those of chronic progressive renal disease. Definitive diagnosis of encephalitozoonosis in the rabbit is difficult. Animals with encephalitozoonosis are expected to be seropositive, but many apparently healthy rabbits are also seropositive, so this assay is not specific and its results must be considered in the light of other diagnostic findings. The absence of antibody, however, should cause the practitioner to consider other differentials. Drugs proven to be efficacious for E. cuniculi infections include albendazole and fenbendazole. Supportive care and treatment with antiinflammatory medications may also be necessary in some forms of encephalitozoonosis. Lens removal or removal of the lens contents is indicated in some rabbits with E. cuniculi-induced ocular disease.     

Serological survey of Encephalitozoon cuniculi in farm rabbits in Italy

Rabbit sera (n = 1600) from 40 commercial farms were submitted to a serological screening for Encephalitozoon cuniculi by an enzyme-linked immunosorbent assay (ELISA) and a carbon immunoassay (CIA test). Antibodies anti-Encephalitozoon cuniculi were found in 505/1600 (31.6%) sera analysed, and all the farms (100%) resulted positive. Rabbits older than 4 months showed a significantly higher seropositivity for E. cuniculi (chi-squared test: p < 0.0001) than rabbits under 4 months, E. cuniculi sero-prevalence showed an increasing trend in rabbits within the farm along with the increase in the “number of rabbits on the farm”; however, this trend was not significant (Spearman’s correlation: p = 0.073).The findings of the present study confirm that rabbit is the main reservoir of E. cuniculi; they are of epidemiological relevance and immediate public health importance because of the recognized infectivity in humans by the microsporidium.     

Immunological diagnosis of encephalitozoonosis from post-mortem specimens

Antibody to $\text{Encephalitozoon cuniculi}$ for immunopathological diagnosis was obtained from organs and fluids from rabbit carcasses. By the use of the india-ink immunoreaction antibodies to $\text{E. cuniculi}$ were detected in lungs, kidneys and pleural fluid of seropositive animals. Mean values of the antibody titres in the lungs were $\text{1}\text{10}$ to $\text{1}\text{2}$ of that in serum. Antibody titres from other organs were inconsistent. No titres were found in organs or fluids from seronegative rabbits.     

Variation of the CD4, CD8, and MHC II cell population in granulomas of immunocompetent and immunosuppressed rabbits in Encephalitozoon cuniculi infection

Encephalitozoon cuniculi (E. cuniculi) is a fungi-related, obligate, zoonotic, spore-forming intracellular eukaryotic microorganism. This emerging pathogen causes granulomas in brain and kidneys of infected individuals. The objective of this study was to detect the distribution of CD4, CD8 and MHCII-positive cells within granulomas in these organs in infected immunocompetent (group A) and infected immunosuppressed (group B) New Zealand white rabbits using immunohistochemistry. In brain, labeled CD4 immune cells were mainly located in the periphery of granulomas in group B. Kidneys of groups A and B, displayed CD4-positive in granulomas and were significant different when compared to brain. CD8 immune cells in brain and kidneys were disseminated in the granulomas in groups A and B; however, no significant difference was observed. MHCII-positive cells were more numerous in brain sections of group B and were significantly different when compared to kidney sections. Granulomas were not observed in control animals of group C and D. In conclusion, we identified CD4-positive cells in both the brain and kidneys of immunocompetent and immunosuppressed animals; CD8-positive cells were more numerous in brain of immunosuppressed rabbits and MHCII cells were more predominant in brain of immunocompetent rabbits. Apparently, the immunosuppression stimulated a change in the cellular phenotype of Th1- to Th2-like granulomas in brain and kidneys by an unknown mechanism. These results increase our understanding of CD4, CD8 and MHCII positive cells within the E. cuniculi granuloma microenvironment and will help in future microsporidian granulomas studies of both immunocompetent and immunosuppressed individuals.     

Clinical Update on Testing Modalities for Encephalitozoon cuniculi in Clinically Sick Rabbits

This article describes the clinical presentation of rabbit patients infected with Encephalitozoon cuniculi that present to veterinary hospitals. The difficulty in obtaining a definitive diagnosis with this disease, even in animals with severe clinical signs, is frustrating to veterinarians and owners alike. A detailed review of current diagnostic testing methodologies and recent advances should help with the interpretation of E. cuniculi test results to communicate and inform owners on treatment and prevention of this devastating disease.     

Rabbit Clinical Pathology

With rabbit patients, as in other species, analyzing blood and urine samples can be useful and informative, although interpretation of the results is sometimes challenging. This article summarizes the interpretation of laboratory results from rabbits. Hematological parameters can yield information about the red blood cell population and leukocyte response to stress and pathogens. Biochemistry evaluation can be used to investigate liver, kidney, and other organ function, and urinalysis results may yield additional information about kidney function and electrolyte imbalances. Serological tests are available for several pathogens of rabbits, including Encephalitozoon cuniculi, although the significance of positive results and antibody titers is not clear. Serum protein electrophoresis aids the understanding of protein disorders and the immune response to acute and chronic inflammation.     

Comparison of enzyme-linked immunosorbent assay and immunofluorescence test for determination of anti-Encephalitozoon cuniculi antibodies in sera from rabbits with different clinical and histopathological presentations

Encephalitozoon cuniculi is a microsporidian that infects rabbits resulting in a varied pathology of the nervous, renal, and ocular tissues. Serology based studies using different methods have been conducted indicating a high seroprevalence worldwide in pet rabbits. While qualitative results are not especially helpful in the diagnosis of this infection, quantitative titers can have increased predictive value. In the current study, results of enzyme-linked immunosorbent assay (ELISA) and immunofluorescence test methods were compared using banked samples from rabbits with confirmed infection and clinically normal rabbits. Pearson's analysis found good and significant correlation (R = 0.56, P = 0.0002). Bland Altman analysis demonstrated the methods were similar but not equivalent. The sensitivity ranged from 0.79 to 0.84 and the specificity was 1.00 for both assays. Anti-E. cuniculi IgM titers were also determined by ELISA to be an independent measure from IgG (R = 0.31, P = 0.06) with a sensitivity of 0.24 and specificity of 1.00. In total, these results confirm an agreement in the detection of IgG antibody by two different methods conducted at laboratories in the United States and Austria that are involved in research and clinical diagnostic testing of this agent.     

Acute Phase Protein Levels in Rabbits with Suspected Encephalitozoon cuniculi Infection

The objective of this study was to evaluate the application of acute phase protein assays for C-reactive protein (CRP), haptoglobin (HP), and serum amyloid A (SAA) in the diagnosis of Encephalitozoon cuniculi (ECUN) infection in pet rabbits. Serum samples from 48 pet rabbits were submitted from veterinary clinics within the United States. Participating veterinarians completed a questionnaire that was used to classify rabbits as either non-ECUN suspect (n = 19) or suspected of having ECUN infection (n = 29). A previously described enzyme-linked immunosorbent assay diagnostic test was used to detect immunoglobulin G (IgG) titers against ECUN. Samples were additionally tested for levels of CRP, HP, and SAA. A nearly 10-fold mean increase in CRP levels was observed in the ECUN-suspect group. This increase was significant (P < 0.05). There was no significant difference in HP or SAA levels between the clinical groups. These data support the use of CRP as an adjunct test in the diagnosis of ECUN infection in pet rabbits.     

Cross-sectional survey of pet ownership,veterinary service utilisation,and pet-related expenditures in New Zealand

ABSTRACT

Aims: To describe the demographics of and predictors for pet ownership, reasons for visiting a veterinarian, and pet-related expenditure in pet owners in New Zealand.

Methods: A cross-sectional online survey was conducted from 18–22 June 2015 using a permission-based panel of New Zealand residents aged ≥18 years. Questions included demographics of respondents, number of pets and reasons for owning or not owning pets, number of visits and reasons for visiting a veterinarian, and pet-related expenditure.

Results: Of the 1,572 respondents who completed the survey, 1,013 (64.4%) owned ≥1 pet. Of these, 443 owned dogs, 696 cats, 32 horses, 103 birds, 55 rabbits, and 159 owned fish. Companionship was the most common reason for getting dogs, cats, and birds; horses were mostly owned as a hobby, rabbits to provide fun for children, and fish as a source of relaxation. The majority of dog, cat, and rabbit owners considered their pets to be family members; horse owners almost equally considered their horses a hobby or a family member. The odds of pet ownership increased for respondents from a rural region, having a higher household income, having children and being female. Overall, 711/1,013 (70.2%) pet-owning respondents had taken ≥1 animal to the veterinarian in the previous year, with the most common reasons being for vaccination or annual check-ups or health issues. Respondents who considered their pets trusted companions, had a higher income, and owned dogs or cats compared with other species, were most likely to have taken their pet to a veterinarian. The greatest pet-related expenditure for all species was food. The median yearly veterinary expenditure was $200–499 by dog owners, $100–199 by cat owners, and <$100 by horse, bird, rabbit, and fish owners. The best source of information for pet-related issues was considered to be veterinarians by 724/1,001 (72.3%) owners, and the internet by 509/1,001 (50.8%) owners.

Conclusions: Among survey respondents, pet ownership was common and pets filled a variety of roles in the household. Pet owners reported spending considerable amounts of money on their pets each year, but some of them may be underutilising veterinary services despite veterinarians being considered as valuable sources of information about pet-related issues.     

Inhibited development of Obeliscoides cuniculi in rabbits: the effects of active and passive immunization and resumption of larval development

Rabbits were either actively or passively immunized against Obeliscoides cuniculi and challenged with 5 000 infective larvae (IL). Worm development was inhibited in actively immunized animals and worm egg production was impaired by administration of immune serum. Inhibited fourth stage larvae (L₄) from high-level infections (200 000 IL) and normal L₄ from low-level infections (4 000 IL) were transferred to actively immunized and normal rabbits. In both instances, large numbers of L₄ developed to maturity; however, a significant number of inhibited L₄ failed to resume development. Inhibition of larvae resulted in the development of fewer adult male than female worms, and the change in sex ratio was further accentuated in actively immunized recipients. Worm egg production was also reduced in actively immunized rabbits. These data indicate that host immune responses contribute to inhibition of O. cuniculi in rabbits and that worm egg production is also responsive to immunologic control.     

Obesity in pet rabbits (Oryctolagus cuniculus): A narrative review

IntroductionObesity reduces both the longevity and the quality of life of pet animals. For pet rabbits, there is a dearth of research focusing on obesity. This paper provides an overview of the available information on obesity in rabbits and discusses approaches to its prevention and treatment.MethodThe prevalence of being overweight/obesity in pet rabbits varies from 5.9% to 35%, depending on study origin and assessment method. Six potential risk factors for rabbit obesity are identified, namely age, breed, sex and neutered status, diet, housing, and owner characteristics.ConclusionObesity in rabbits has myriad consequences, including osteoarthritis, pododermatitis, hepatic lipidosis, atherosclerosis, insulin resistance, and increased risk of postanesthetic complications. Obesity in pet rabbits can be practically diagnosed with body condition scoring, but validation is needed. Prevention of obesity should be prioritized before treatment, but both involve facilitating more exercise, limiting concentrated calorie intake, and offering plenty of hay to pet rabbits.     

Seroprevalence of antibodies to Encephalitozoon cuniculi and Encephalitozoon intestinalis in humans and animals

The presence of antibodies against Encephalitozoon cuniculi (E. cuniculi) and Encephalitozoon intestinalis (E. intestinalis) was examined in 215 samples from humans and in 488 samples from five different species of domestic and companion animals in Slovakia. The 215 human samples and samples from 90 swine, 123 non-infected cattle (cattle), 24 cattle infected with bovine leukosis virus (BLV-positive cattle), 140 sheep and 111 dogs were examined by the enzyme-linked immunosorbent assay (ELISA). Samples with serum titres 1:200 or higher were considered as positive. Specific anti-E. cuniculi antibodies were found in humans (0.9%), swine (52%), cattle (2%), sheep (9%) and dogs (15%) except for the BLV-positive cattle at the titre of 1:200. The titre of 1:400 was detected only in humans (0.5%). The presence of specific anti-E. intestinalis antibodies at the titre of 1:200 was confirmed in humans (6%), swine (51%), cattle (11%), BLV-positive cattle (13%) and dogs (6%) but not in sheep. The anti-E. intestinalis antibodies reached the 1:400 in humans (1%), swine (4%) and BLV-positive cattle (17%). The presence of specific anti-E. intestinalis antibodies at the titre of 1:600 was observed only in one swine (1%). Significant differences were observed in animals at titres 1:200 and 1:400 (chi-squared test: p < 0.0001) for both pathogens and in humans only for E. cuniculi at the titre of 1:400 (chi-squared test: p < 0.0075).