Fractionation of the reference inoculum of epizootic rabbit enteropathy in discontinuous sucrose gradient identifies aetiological agents in high density fractions

Epizootic rabbit enteropathy (ERE) is a major cause of economic loss in intensive rabbit production. Since its first recognition in 1997, much work has been done to determine the pathogenic mechanisms of the disease and to identify the aetiological agent(s). Unfortunately, the quest for aetiology has only met with limited success despite the ability to reproduce the syndrome by inoculation of intestinal contents from field cases. These intestinal inocula contain a huge number of microorganisms which could all be involved in the aetiology of ERE. To decrease the number of putative agents, the French reference inoculum TEC3 was fractionated on a discontinuous sucrose gradient so that seven fractions (supernatant, 10%, 20%, 30%, 40%, 50% and pellet) were obtained. Specific-pathogen-free rabbits were inoculated with three out of these seven fractions (supernatant, 30%, and pellet). The objectives were: (1) to characterise the seven fractions by bacteriological examination; (2) to verify whether the aetiological agent was present in the fractions by inoculation of rabbits; (3) to assign the aetiological agent of ERE to a morphological group of pathogens; (4) to identify a fraction which could replace the reference inoculum TEC3 in applications such as cell cultures or egg inoculation. The results strongly suggest that ERE is a bacterial disease and does not have a viral or parasitic aetiology.     

ULTRASONOGRAPHIC INTESTINAL HYPERECHOIC MUCOSAL STRIATIONS IN DOGS ARE ASSOCIATED WITH LACTEAL DILATION

In this retrospective study, the medical records of 23 dogs with the sonographic feature of small intestinal hyperechoic mucosal striations and an endoscopic or surgical intestinal biopsy were reviewed. Histopathologic lacteal dilation was present in 96% of dogs with mucosal striations. Sonographic findings associated with mucosal striations included: mild jejunal wall thickening (96%), mild duodenal wall thickening (78%), mucosal speckles (70%), and abdominal effusion (87%). The mucosal striations were diffuse (70%) or multifocal (30%) and did not cause loss of wall layering, except in one dog with a severe mural lipogranuloma. Mesenteric lymphadenopathy was identified in 9% of dogs. Thirteen dogs with endoscopic biopsies had mild to moderate villus lacteal dilation and the nine dogs with surgical biopsies had moderate to severe dilation. Inflammatory infiltrates were mild (61%) or moderate (30%) with variable numbers and combinations of cells, including eosinophils (65%), plasma cells (61%), lymphocytes (57%), and neutrophils (30%); one dog had disseminated villus histiocytic sarcoma. The biochemistry changes and clinical signs were consistent with protein-losing enteropathy in 78% of dogs. Hyperechoic mucosal striations in dogs are associated with lacteal dilation and are frequently associated with mucosal inflammation and protein losing enteropathy.     

Epizootic rabbit enteropathy: comparison of PCR-based RAPD fingerprints from virulent and non-virulent samples

Epizootic rabbit enteropathy is a gastrointestinal disorder of unknown aetiology of farmed rabbits characterised by inanition and mortality. Genomic analyses of virulent and non-virulent samples of inocula were performed using random amplification of polymorphic DNA (RAPD). Differences in bacterial DNA composition were found between inocula, but specific sequences were not linked with field cases of epizootic rabbit enteropathy.     

Comparison of a commercial ELISA with an indirect fluorescent antibody test to detect antibodies to Lawsonia intracellularis in experimentally challenged pigs

Objective The ability of a new commercial ELISA to detect pigs with subclinical proliferative enteropathy (PE) was compared with the traditional indirect fluorescent antibody test (IFAT). Methods Serum samples were selected from pigs with known Lawsonia intracellularis infection status and clinical signs of PE, but the sample population consisted predominantly of pigs subclinically affected by PE. Results Significant association and agreement were shown between the ELISA and IFAT assays. ELISA results correlated well with the duration of L. intracellularis shedding as detected by polymerase chain reaction. Conclusion ELISA can be successfully used to monitor L. intracellularis infection in pigs.     

Attenuation of virulence of Lawsonia intracellularis after in vitro passages and its effects on the experimental reproduction of porcine proliferative enteropathy

Non-pathogenic Lawsonia intracellularis variants have been obtained through multiple passages in cell culture but there is no information regarding the number of passages necessary to attenuate a pathogenic isolate. The present study evaluated the susceptibility of pigs to L. intracellularis after 10, 20 and 40 passages in vitro. Three groups (six animals/group) were inoculated with pure culture of L. intracellularis on passage 10, 20 or 40 and one group with placebo. The animals were monitored for clinical signs, fecal shedding and serological IgG response during 28 days post-inoculation. Gross and histologic lesions and the level of infection based on the amount of L. intracellularis-specific antigen in the intestinal mucosa identified by immunohistochemistry were evaluated in two animals from each group on days 14, 21 and 28. Animals inoculated with passages 10 and 20 demonstrated proliferative lesions typical of porcine proliferative enteropathy associated with the presence of Lawsonia-specific antigen in the intestinal mucosa. Passage 40-inoculated pigs did not show proliferative lesions or presence of Lawsonia antigen at any time point throughout the study. Similar patterns of the fecal shedding were observed in passage 10 and 20-infected pigs but those infected with passage 40 shed for a short period. Serological IgG responses in passage 10 and 20-inoculated pigs were detected from day 14 post-infection but not at all in passage 40-inoculated animals. These results demonstrate attenuation of the virulence properties of L. intracellularis between 20 and 40 cell passages in vitro. This information will be valuable for design of future experimental models and for studying the mechanisms involved in the attenuation of L. intracellularis virulence.     

Equine proliferative enteropathy – a review of recent developments

Equine proliferative enteropathy (EPE) is a disease of foals caused by the obligate intracellular organism Lawsonia intracellularis. This emerging disease affects mainly weanling foals and causes fever, lethargy, peripheral oedema, diarrhoea, colic and weight loss. The diagnosis of EPE may be challenging and relies on the presence of hypoproteinaemia, thickening of segments of the small intestinal wall observed upon abdominal ultrasonography, positive serology and molecular detection of L. intracellularis in faeces. Although the clinical entity, diagnostic approach and treatment of EPE are well established and described, the epidemiology for this disease has remained largely unaddressed. This article focuses on new developments in the field of EPE, including epidemiology, pathophysiology, clinical signs, diagnosis, treatment and prevention. The Summary is available in Chinese – see Supporting information.