Recombinant Iss as a potential vaccine for avian colibacillosis

Avian pathogenic Escherichia coli (APEC) cause colibacillosis, a disease which is responsible for significant losses in poultry. Control of colibacillosis is problematic due to the restricted availability of relevant antimicrobial agents and to the frequent failure of vaccines to protect against the diverse range of APEC serogroups causing disease in birds. Previously, we reported that the increased serum survival gene (iss) is strongly associated with APEC strains, but not with fecal commensal E. coli in birds, making iss and the outer membrane protein it encodes (Iss) candidate targets for colibacillosis control procedures. Preliminary studies in birds showed that their immunization with Iss fusion proteins protected against challenge with two of the more-commonly occurring APEC serogroups (O2 and O78). Here, the potential of an Iss-based vaccine was further examined by assessing its effectiveness against an additional and widely occurring APEC serogroup (O1) and its ability to evoke both a serum and mucosal antibody response in immunized birds. In addition, tissues of selected birds were subjected to histopathologic examination in an effort to better characterize the protective response afforded by immunization with this vaccine. Iss fusion proteins were administered intramuscularly to four groups of 2-wk-old broiler chickens. At 2 wk postimmunization, chickens were challenged with APEC strains of the O1, O2, or O78 serogroups. One week after challenge, chickens were euthanatized, necropsied, any lesions consistent with colibacillosis were scored, and tissues from these birds were taken aseptically. Sera were collected pre-immunization, postimmunization, and post-challenge, and antibody titers to Iss were determined by enzyme-linked immunosorbent assay (ELISA). Also, air sac washings were collected to determine the mucosal antibody response to Iss by ELISA. During the observation period following challenge, 3/12 nonimmunized chickens, 1/12 chickens immunized with 10 microg of GST-Iss, and 1/12 chickens immunized with 50 microg of GST-Iss died when challenged with the O78 strain. No other deaths occurred. Immunized chickens produced a serum and mucosal antibody response to Iss and had significantly lower lesion scores than nonimmunized chickens following challenge, regardless of the challenge strain. This study expands on our previous report of the value of Iss as an immunoprotective antigen and demonstrates that immunization with Iss can provide significant protection of chickens against challenge with three different E. coli strains.     

Detection of Salmonella organisms and assessment of a protocol for removal of contamination in horse stalls at a veterinary teaching hospital

OBJECTIVES: To assess methods of detecting environmental contamination with Salmonella organisms and evaluate a cleaning and disinfection protocol for horse stalls in a veterinary teaching hospital. DESIGN: Original study. SAMPLE POPULATION: 37 horses with diarrhea likely to be caused by Salmonella infection and their stall environments. PROCEDURES: Fecal samples were collected from horses daily during hospitalization; samples were obtained from stall sites after cleaning and application of disinfectants. Fecal and environmental samples were cultured for Salmonella spp and tested via polymerase chain reaction (PCR) assay to detect Salmonella DNA. RESULTS: 1 horse died and 2 were discharged prior to sample collection. Fecal samples from 9 of 34 horses yielded growth of Salmonella organisms on bacteriologic culture, and 23 yielded positive results via PCR assay on > or = 1 occasion. Among environmental samples from 21 stalls, salmonellae were detected at > or = 1 stall site on 6 of 78 occasions, and > or = 1 stall site yielded positive results via PCR assay on 69 of 77 occasions. Salmonella DNA was detected more frequently in samples of stall drains, cracks, and corners. Salmonella spp were cultured from samples of 3 stalls after both initial and second cleaning and disinfection cycles, but no organisms were detected in samples obtained after use of a peroxygen disinfectant. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that stalls in which horses with salmonellosis were housed should only be used to accommodate newly hospitalized horses after samples (collected after 2 cycles of cleaning and disinfection) from drains, cracks, and corners yield negative results on bacteriologic culture.     

Disseminated candidiasis secondary to fungal and bacterial peritonitis in a young dog

Objective – To describe a severe case of bacterial sepsis and disseminated candidiasis in a previously healthy dog.
Case Summary – Fungal sepsis was identified in a 2-year-old dog following intestinal dehiscence 4 days after abdominal surgery. Septic peritonitis was identified at admission and evidence of dehiscence at the previous enterotomy site was found during an exploratory laparotomy. Both gram-positive cocci and Candida albicans were cultured from the abdominal cavity. Candida sp. was also subsequently cultured from a central venous catheter. Euthanasia was performed due to failure to respond to therapy. Fungal organisms, morphologically consistent with Candida spp., were found in the lungs and kidney on postmortem histopathologic examination indicating disseminated candidiasis.
New or Unique Information Provided – Candida peritonitis is a well-recognized entity in humans and contributes to morbidity and mortality in critically ill patients. Abdominal surgery, intestinal perforation, presence of central venous catheters, and administration of broad-spectrum antibiotics are all considered to be suspected risk factors. This report describes the first known case of systemic candidiasis occurring secondary to Candida peritonitis and bacterial sepsis in a critically ill dog.     

Histological and morphometric lesions in the pre-clinical,developmental phase of insulin-induced laminitis in Standardbred horses

Lamellar pathology in experimentally-induced equine laminitis associated with euglycaemic hyperinsulinaemia is substantial by the acute, clinical phase (～48 h post-induction). However, lamellar pathology of the developmental, pre-clinical phase requires evaluation. The aim of this study was to analyse lamellar lesions both qualitatively and quantitatively, 6, 12 and 24 h after the commencement of hyperinsulinaemia. Histological and histomorphometrical analyses of lamellar pathology at each time-point included assessment of lamellar length and width, epidermal cell proliferation and death, basement membrane (BM) pathology and leucocyte infiltration. Archived lamellar tissue from control horses and those with acute, insulin-induced laminitis (48 h) was also assessed for cellular proliferative activity by counting the number of cells showing positive nuclear immuno labelling for TPX2.Decreased secondary epidermal lamellar (SEL) width and increased histomorphological evidence of SEL epidermal basal (and supra-basal) cell death occurred early in disease progression (6 h). Increased cellular proliferation in SELs, infiltration of the dermis with small numbers of leucocytes and BM damage occurred later (24 and 48 h). Some lesions, such as narrowing of the SELs, were progressive over this time period (6–48 h). Cellular pathology preceded leucocyte infiltration and BM pathology, indicating that the latter changes may be secondary or downstream events in hyperinsulinaemic laminitis.     

Abdominal aortic aneurysm associated with systemic fungal infection in a German shepherd dog

A 2 yr old spayed female German shepherd presented with a chief complaint of acute onset paraparesis and weight loss. At presentation, the dog was pyrexic, nonambulatory, and had generalized muscle wasting. Neurolocalization was consistent with a thoracolumbar spinal cord lesion. An abdominal ultrasound was performed and revealed a focal dilation (4 cm) of the terminal aorta with evidence of blood stasis consistent with an aortic aneurysm. The dog was euthanized shortly after admission to the hospital and a post mortem examination was performed. Fungal organisms were identified in the aortic aneurysm as well as from the thoracic vertebrae, mesenteric lymph nodes, axillary lymph nodes, spleen, kidneys, liver, lungs, and heart. Although the morphology was consistent with Candida spp., immunohistochemistry and PCR could not definitively identify the causative organism. Mycotic aortic aneurysms are a rare finding in humans and have not been previously reported in the dog. To the authors' knowledge, this is the first known report of an aortic aneurysm associated with systemic fungal infection in a dog.     

Equine atopic skin disease and response to allergen-specific immunotherapy: a retrospective study at the University of California-Davis (1991-2008)

This retrospective study reports on the clinical presentation of equine atopic skin disease and evaluates response to treatment with allergen-specific immunotherapy (ASIT) based on intradermal testing and/or serum testing. Computerized medical records from January 1991 to December 2008 yielded 54 horses included in the study. Presenting clinical signs (CS) included urticaria (n=28), pruritus (n=8) or both (n=18). Forty-one of 54 horses received ASIT, and response to ASIT (n=32) was evaluated via telephone survey. Eighty-four per cent (n=27) of owners reported that ASIT reduced their horse's CS; 59% (n=19) were able to manage CS by ASIT alone. Three horses (9%) were managed with ASIT in combination with doxepin and discontinued use of corticosteroids. There was no statistical significance between type of test performed and reported success of ASIT (χ(2) analysis, P=0.53). Ninety-three per cent (n=30) of owners reported use of antipruritic medications prior to starting ASIT; 57% (n=17) of these owners reported discontinuing those medications due to success of ASIT. Adverse effects were limited to swelling at the injection site, seen in 16% (n=5). Seventy-five per cent (n=24) of owners elected to discontinue ASIT after 6 months to 8 years (mean 2.2 years): 15 due to resolution of CS, six due to persistent CS, two because the horse was sold, and one due to cost. Ten owners reported no recurrence of CS after discontinuing ASIT; five had recurrence within a median of 2 years of discontinuing ASIT (range 1-12 years). Allergen-specific immunotherapy is a safe and effective way to manage equine atopic skin disease.