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

Abstract

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

APPLICATION OF 3-HYDROXYBUTYRATE MEASUREMENT AND PLASMA PROTEIN ELECTROPHORESIS IN THE DIAGNOSIS OF ASPERGILLOSIS IN AFRICAN PENGUINS (SPHENISCUS DEMERSUS)

“…Blood samples were drawn from penguin flipper vein, then heparinized in lithium tubes and centrifuged. If free from hemolysis and lipemia, [3][4][5][6] plasma specimens were refrigerated until analysis within 24 hours of submission to the laboratory. 10 Otherwise, they were frozen at -80°C.…”

“…7 Galactomannan antigen was measured in blood according to the manufacturers' recommendations (Bio-Rad Clinical Diagnostics, Hercules, CA, 94547, U.S.A.). 3,7,8,13 EPH results were obtained from the SPIFE 3000 ® electrophoresis analyzer (Helena Labs, Beaumont, TX, 77707, U.S.A.). 5 They were subcategorized as abnormal on the basis of at least a 10%-change (in absolute concentration) compared to the reference intervals of at least one protein fraction.…”

“…7 Veterinary practitioners currently depend on plasma protein electrophoresis (EPH) to address the diagnosis of aspergillosis and initiate preemptive antifungal treatment. 3,8 Quantitation of serum proteins has been shown to provide a reflection of acute phase responses through the demonstration of change in albumin, a negative acute phase protein, and four globulin fractions which are often increased with different disease processes. 12 The diagnostic and prognostic application of data obtained from commercial semi-automated methods of protein fractionation has been well described in veterinary medicine.…”

Application of mass spectrometry to elucidate the pathophysiology of Encephalitozoon cuniculi infection in rabbits

“…As the humoral response is theoretically only indicative of contact—current, recent or past—with the pathogen, including IgM titers which can persist over several months [6,28], it appeared relevant to focus not only on the description of antigenic microsporidian proteins but to comprehensively study all the changes in host responses in the pathophysiology of E . cuniculi infections in rabbits [6,8,29]. To address this, we conducted a direct differential comparison of proteomes via the iTRAQ ® protocol.…”

Questions around Encephalitozoon cuniculi in rabbits

“…In addition, the use of PCR technology allows the clinician to confirm the presence or absence of E cuniculi DNA in various organs and body fluids. Regardless, seropositivity still does not confirm that clinical signs are due to E cuniculi (Künzel and others 2008), which is where information like that presented in papers such as those by Meyer-Breckwoldt (1996), Ewringmann and Göbel (1999), Harcourt-Brown and Holloway (2003), Keeble and Shaw (2006), Cray and others (2009) and Csokai and others (2009), as well as the paper by Hein and others (2014), which is summarised on p 350 of this issue of Veterinary Record , becomes important. All of these papers attempt to explain the relevance of seropositivity in terms of broader epidemiological constructs.…”