Equine bullous pemphigoid IgG autoantibodies target linear epitopes in the NC16A ectodomain of collagen XVII (BP180, BPAG2)

Bullous pemphigoid (BP) is an autoimmune subepithelial blistering dermatosis of humans, dogs, cats and pigs. It is characterized by skin-fixed and circulating IgG autoantibodies that target one or both BP antigens. An immunological homologue of BP in humans was diagnosed in two horses with cutaneous and mucosal ulcerations as well as microscopic subepithelial vesiculation. Immunological investigations revealed similar findings for both the horses. Direct immunofluorescence demonstrated the presence of IgG deposited linearly at the dermoepidermal junction in mucosal and skin biopsy specimens. Indirect immunofluorescence testing confirmed the existence of circulating basement membrane-specific IgG autoantibodies. Using intact and salt-split epithelial substrates, serum IgG were shown to target antigens situated not only at the basal, but also at the lateral and apical aspects of stratum basale keratinocytes. Immunoblotting and ELISA corroborated that the IgG from affected horses, but not those from normal controls, exhibited high immunoreactivity against the NC16A extracellular domain of type XVII collagen (BPAG2, BP180). Equine BP could be proposed, therefore, as another spontaneous model of this most common basement membrane autoimmune dermatosis of humans.     

Temporal patterns and quantification of excretion of Mycobacterium avium subsp paratuberculosis in sheep with Johne's disease

OBJECTIVES: To determine the frequency of excretion of Mycobacterium avium subsp paratuberculosis in Merino sheep with Johne's disease and to quantify excretion in a group of Merino sheep. DESIGN: A pen and laboratory experiment. PROCEDURE: Seven sheep selected from an affected flock on the basis of acid-fast bacilli in the sheep's faeces were housed and total daily faecal output was collected, weighed and subjected to culture for M avium subsp paratuberculosis. An end-point titration method was used to enumerate viable M avium subsp paratuberculosis in a 15 day pooled sample from five sheep that had acid-fast bacilli in their faeces while housed. RESULTS: Four sheep with subclinical multibacillary Johne's disease excreted M avium subsp paratuberculosis each day for 11 days of cultural observation. A further three sheep were intermittent excreters but lacked other evidence of infection with M avium subsp paratuberculosis. The average number of viable bacteria excreted was 1.09 x 10(8) per gram of faeces while total daily excretion was 8.36 x 10(10) viable M avium subsp paratuberculosis per sheep. Examination of faecal smears stained with Ziehl Neelsen was an unreliable means of assessing daily excretion in individual animals except in those with severe lesions. CONCLUSION: Excretion of M avium subsp paratuberculosis in Merino sheep with multibacillary Johne's disease occurred daily, proving that environmental contamination can be continuous on farms with endemic ovine Johne's disease. Faecal culture is a useful method for detecting infection as it does not appear to be affected by the timing of collection of a sample from sheep with multibacillary disease however, to maximise the sensitivity of disease surveillance using faecal culture, sampling rates should be adjusted to take account of the proportions of multibacillary and paucibacillary cases.     

Aplasia of the gallbladder in a dog

A young, female Maltese dog was presented with intermittent vomiting of bile. Biochemical evidence of persistent mild hepatopathy had been present for 11 months. Exploratory celiotomy was performed. Absence of the gallbladder with malformation of the quadrate lobe of the liver was identified. There was histological evidence of bile duct proliferation and portal fibrosis.     

Lateral musculature in the whitemouth croaker (Micropogonias furnieri): its characterization with respect to different gonadal conditions

Histochemical and ultrastructural studies were performed on the lateral musculature from individual female whitemouth croaker, Micropogonias furnieri, at the anterior, medium and posterior regions. Based upon histochemical myosin-ATPase (m-ATPase) determination, diverse types of red, pink and white fibres were discerned. Red muscle had abundant mitochondria and stained intensely for aerobic enzymes, white muscle scarcely stained for the same enzymes and pink muscle responded in an intermediate manner. Both white and pink muscle had few mitochondria. The relative proportion of red muscle increased towards the caudal region; pink muscle diminished towards this region and white muscle modified its proportion only in the anterior region. m-ATPase activity showed differences in relation to the gonadal condition along the body, particularly in the white fibres at the anterior and medium regions.     

Bilateral aural hyalohyphomycosis in sable antelope (Hippotragus niger).

Three juvenile sable antelope (Hippotragus niger) were diagnosed with bilateral aural hyalohyphomycosis based on histopathology. All three animals were suspected to be immunodeficient based on low IgG levels determined using the zinc sulfate turbidity test. The serum and hepatic copper levels of one animal were below the bovine reference range. Clinical signs in the three animals included bilateral ventral deviation of the pinnae with multifocal subcutaneous aural tumefaction and poor body condition. Numerous septate, nonpigmented fungal hyphae were found within the auricular cartilage, dermis, and subcutaneous granulomas. No significant fungal agents were isolated by culture, and no signs of systemic fungal dissemination were identified except for a concurrent fungal rhinitis in one animal.     

Evidence for surfactant contributing to the gastric mucosal barrier of the horse

This study was undertaken to determine the hydrophobicity of the luminal surface of the equine stomach and to elucidate the ultrastructure of the lining imparting that property. Gastric and duodenal mucosal samples from 5 horses were collected immediately after euthanasia and subjected to surface contact angle measurement using a goniometer. Gastric mucosal samples from 4 horses and a foal were examined by electron microscopy following a fixation procedure known to preserve phospholipids and oligolamellar structures. Contact angles for the equine gastric glandular mucosal surface (mean +/- s.e. 78.0 +/- 11.0 degrees) were greater than for the duodenum (33.4 +/- 8.7 degrees), (P = 0.003). The contact angles for gastric squamous tissue (50.4 +/- 4.5 degrees) tended to be greater than for duodenum (P = 0.15). Electron microscopy revealed the existence of surfactant as abundant osmiophilic phospholipid material within both squamous and glandular gastric mucosae. These results indicate the hydrophobic nature of the equine gastric mucosae. We propose that the water-repellent nature of the stomach contributes to the 'gastric mucosal barrier' and is imparted by surface-active phospholipid adsorbed to the surface. Phospholipids may also be utilised as a physical barrier to back-diffusion of acid by lining intracellular canaliculi and oxyntic ducts where other defence mechanisms are absent.     

Inhibition of in vitro Histophilus somni biofilm production by recombinant Hsp60 antibodies

Histophilus somni is an opportunistic pathogen causing respiratory, genitourinary and generalized infections in cattle. An important virulence factor is its ability to produce a biofilm. The aim of this work was to confirm that H. somni Hsp60 (Gro-EL) is a constituent of the biofilm produced by this bacterium in vitro and to check whether or not the presence of a specific antibody within the culture medium can inhibit biofilm production. Biofilm production by H. somni cultured in vitro was confirmed by crystalline violet staining. The presence of Hsp60 in the biofilm was confirmed by using specific antibodies produced in a mouse and goat hyperimmunized with H. somni recombinant Hsp60 (rHsp60). Large complexes of biofilm stained with Hsp60 antibodies were microscopically detected. This indicates that the Hsp60 protein is a common constituent of the biofilm produced by H. somni in vitro. In a second experiment, mouse serum containing anti-H. somni rHsp60 antibodies was added to an H. somni culture. It was found that the presence of anti-rHsp60 antibodies in the culture medium inhibited biofilm production in vitro. Only small biofilm particles were seen in the presence of the specific antibody, whereas in control cultures (without specific antiserum) large biofilm complexes were produced. The results indicate that antibodies specific to Hsp60 may be useful for preventing H. somni biofilm formation in vitro. If this also occurs in vivo, it may be helpful for eradicating H. somni infection in cattle through the elimination of carriers. Further in vivo studies are needed to confirm this idea.     

Use of a mixed sericea lespedeza and grass pasture system for control of gastrointestinal nematodes in lambs and kids

Because of a high prevalence of anthelmintic resistance and consumer demand for chemical free meat products, management tools to minimize the need for deworming are needed. The objective was to examine the effectiveness of grazing sericea lespedeza (SL) in a mixed grass or a pure forage system for control of gastrointestinal nematodes (GIN); in other words pasture systems included grass, grass plus SL, or SL alone (Experiments 2 and 3). Selective use of copper oxide wire particles (COWP) based on the FAMACHA(?) system was used to aid in GIN control. In Experiment 1, lambs co-grazed bermudagrass (BG; n=21) or SL in a mixed grass pasture (SLM; n=22) with dams for 14 days. In Experiment 2, lambs grazed BG (n=14), SLM (n=13), or pure SL (SLP; n=13) pastures for 56 days. In Experiment 3, doe kids grazed BG (n=12), SLM (n=13), or SLP (n=13) for 84 days. Animals were fed a 16% crude protein supplement based on NRC requirements and estimated forage quality of pastures, so that 454, 389, and 200 g/lamb (Experiment 2), or 454, 300, and 150 g of supplement/goat (Experiment 3) was fed to BG, SLM, and SLP, respectively. Animals were dewormed with COWP if FAMACHA(?) was >3. Coprocultures were conducted to identify GIN genus. In Experiment 1, FEC were reduced in lambs grazing SLM compared with BG pastures. In Experiment 2, FEC were reduced in SLP compared with BG lambs on all days, and reduced in SLM compared with BG lambs on day 56. Initially, Haemonchus contortus was the predominant nematode, but the population shifted to other species in the SL groups by the end of the study. The mean number of dewormings/lamb was 0.71, 0.20, and 0.21±0.13 for BG, SLM, and SLP groups, respectively. In goats in Experiment 3, Trichostrongylus spp. was the predominant nematode in May and June and H. contortus in July. There was little meaningful effect of forage treatments on GIN infection in kids. Because H. contortus was not the predominant nematode in kids, the integrated approaches used may not have been effective in controlling GIN. Grazing SL was effective for GIN control in lambs in early summer, but the effect was delayed in lambs grazing a mixed SL/grass pasture unless lambs initially grazed with dams. An integrated approach used that included SL grazing and COWP for deworming was effective in lambs.