Molecular detection of Chlamydophila abortus in post-abortion sheep at oestrus and subsequent lambing

Enzootic abortion of ewes (EAE), resulting from infection with the bacterium Chlamydophila abortus (C. abortus), is a major cause of lamb loss in Europe. The purpose of this study was to assess the potential impact of the shedding of organisms in post-abortion ewes at oestrus and subsequent lambing on the epidemiology of EAE. Using a newly developed C. abortus specific real-time PCR assay, few chlamydial genomes could be detected in vaginal swabs taken from post-abortion ewes at oestrus. At subsequent parturition, all ewes lambed normally with no macroscopic or microbiological evidence of infection. Real-time PCR analysis of placental samples identified very few or no chlamydial genomes, which contrasted significantly with samples taken at the time of abortion, where an average of 2.7x10(7) chlamydial genomes per microgram of total tissue DNA was detected. Few genomes could also be detected from vaginal and cervical tissue samples and lymph nodes taken post-mortem. The results, although not discounting the possibility of a chronic low level persistent infection in post-abortion ewes, suggest that the low levels of chlamydial DNA detected during the periovulation period and at lambing do not significantly impact on the epidemiology of EAE. In terms of flock management, the products of abortion should be considered the major and principal source of infection for transmission to na?ve ewes.     

A retrospective study of cutaneous equine sarcoidosis and its potential infectious aetiological agents

Nine horses from ages 5 to 21 years were diagnosed with cutaneous equine sarcoidosis (ES) over an 18-year period. In addition to skin, the lungs were frequently involved, with other organ systems affected less commonly. A predisposition for thoroughbreds and geldings was noted. Cutaneous lesions and signs included crusts, scales, alopecia and pruritus. These were found at various sites, particularly the legs/thighs/elbows, thorax, neck, face and ventral abdomen. Three horses were euthanized shortly after hospitalization; others survived as long as 12 years. Histopathologic stains, immunohistochemistry and polymerase chain reaction assays on paraffin-embedded cutaneous specimens from eight horses for Mycobacterium spp., Coccidioides immitis, Cryptococcus neoformans, Corynebacterium pseudotuberculosis, and Borrelia burgdorferi were all negative. The aetiology of ES is unlikely microbial and continues to be a diagnosis of exclusion. ES, when limited to the skin, is associated with a good prognosis, with either partial or complete response to glucocorticoid therapy in all the surviving horses.     

Leukocyte emigration in the early stages of laminitis

The mechanisms that initiate the pathophysiologic changes in the digital laminae in equine laminitis are poorly understood. Due to the fact that (1) the horse at risk of laminitis has many similarities clinically to the human sepsis patient and (2) our recent finding of marked laminar proinflammatory cytokine expression at the developmental time point of the black walnut extract (BWE) model of laminitis, we tested the possibility that, similar to organ damage in human sepsis, leukocyte emigration is an early event in laminitis. Using immunoperoxidase methods with an anti-equine CD13 monoclonal antibody that recognizes neutrophils and monocytes, we discovered that, whereas the dermal microvasculature of the skin commonly has a marginal pool of leukocytes, the normal laminar dermal microvasculature has minimal to no perivascular leukocytes. However, increases in leukocyte numbers occurred around the dermal vasculature of both the laminae and the skin in the majority of BWE-treated horses in the developmental stage and at the onset of clinical signs of lameness in the BWE model. These findings indicate that, similar to organ failure in human sepsis, leukocyte emigration is likely to play a significant role in initiating numerous pathophysiologic mechanisms that lead to the development of laminitis.     

Classification of Actinobacillus spp isolates from horses involved in mare reproductive loss syndrome

OBJECTIVE: To identify Actinobacillus spp isolates recovered from fetuses and pericardial fluid from horses affected with mare reproductive loss syndrome (MRLS) and determine whether these bacterial species are the same as those isolated from clinically normal horses. SAMPLE POPULATION: Isolates of actinobacilli recovered from 18 horses with pericarditis and 109 fetuses aborted by mares affected by MRLS. Procedures-Actinobacillus spp isolates were identified to the level of species or subspecies by use of conventional phenotypic tests and biochemical and enzyme test kits. The 16S rRNA gene from selected isolates was amplified, purified, and sequenced. Sequence data were compared with sequence data for actinobacilli in GenBank. RESULTS: Of the 109 isolates obtained from fetuses, 14 were Actinobacillus equuli subsp equuli, 65 were A equuli subsp haemolyticus, 28 were Bisgaard taxon 10-like bacterium, and 2 were Actinobacillus genomospecies 1. Of the 18 isolates from horses with pericarditis, 4 were A equuli subsp equuli, 13 were A equuli subsp haemolyticus, and 1 was Bisgaard taxon 10-like bacterium. Comparisons with published data and GenBank data revealed that the isolates recovered from horses with MRLS were the same as those isolated from the oral cavity or alimentary tract of healthy horses. CONCLUSIONS AND CLINICAL RELEVANCE: Actinobacillus spp isolates recovered from fetuses and pericardial fluid samples of horses affected by MRLS in 2001 to 2003 were identical to Actinobacillus spp found in the oral cavity and alimentary tracts of healthy horses.     

Cortical allograft and endoprosthesis for limb-sparing surgery in dogs with distal radial osteosarcoma: a prospective clinical comparison of two different limb-sparing techniques

OBJECTIVE: To compare surgical and oncologic outcome in dogs with osteosarcoma (OSA) of the distal aspect of the radius treated with limb-sparing surgery, using either a cortical allograft or endoprosthesis, and postoperative chemotherapy; and to evaluate predictive factors for outcome. STUDY DESIGN: Prospective cohort study. ANIMALS: Dogs (n = 20) with spontaneous, non-metastatic OSA of the distal aspect of the radius. METHODS: Dogs were prospectively randomized for limb-sparing surgery with either a cortical allograft (n = 10) or endoprosthesis (10) and full-course adjuvant chemotherapy using single or dual agent protocols of cisplatin, carboplatin, and/or doxorubicin. Surgical (intraoperative findings, postoperative infection, construct failure) and oncologic (local tumor recurrence, metastasis, survival) outcomes were compared. The influence of intraoperative and postoperative variables on surgical and oncologic outcome were evaluated. RESULTS: No clinically significant differences in surgical and oncologic outcome were detected between groups. The percentage of radius replaced by the implant was significantly greater in the endoprosthesis group (60.9% compared with 48.6%, P = .008). Median survival time (MST) for dogs with construct failure, regardless of implant type, was 685 days and significantly greater than MST of dogs without construct failure (322 days, P = .042; hazard ratio [HR] 16.82). Median metastasis-free interval and MST (685 days versus 289 days; P = .034, HR 24.58) were significantly greater in dogs with postoperative infection. Disease-free and overall limb-salvage rates were 70% and 85%, respectively. Overall MST was 430 days. CONCLUSIONS: For dogs with OSA of the distal aspect of the radius, a cortical allograft or endoprosthesis can be used for limb-sparing surgery. Construct failure and postoperative infection significantly improve survival time regardless of implant type. CLINICAL RELEVANCE: An endoprosthesis is an attractive alternative to cortical allografts for limb-salvage of the distal aspect of the radius in dogs because surgical and oncologic outcomes are similar, but the endoprosthesis is an immediately available off-the-shelf implant which is not complicated by the bone harvesting and banking requirements associated with cortical allografts. Mechanisms whereby postoperative infection improves survival time requires further investigation and, if elucidated, may provide the opportunity to improve the outcome of dogs and humans with OSA.     

Amputation and Carboplatin for Treatment of Dogs With Osteosarcoma: 48 Cases (1991 to 1993)

Forty-eight dogs with histologically confirmed appendicular osteosarcoma (OSA) entered a prospective clinical trial evaluating treatment with amputation and up to 4 doses of carboplatin given every 21 days. The median disease-free interval (DFI) was 257 days, with 31.2% of the dogs disease-free at 1 year. The median survival time was 321 days, with 35.4% of the dogs alive at 1 year. Dogs with proximal humeral OSA had shorter DFI (P = .016) and survival (P = .037) times than dogs with OSA at other locations. Dogs with lower body weights (40 kg) had longer DFI (P = .0056) and survival (P = .007) times than larger dogs. Survival times for dogs that received carboplatin were statistically longer than those previously reported for amputation alone (P < .001). DFI and survival times are similar to those previously reported for 2 to 4 doses of cisplatin. Carboplatin appears to be a well-tolerated chemotherapeutic drug that can be given safely every 21 days at a dose of 300 mg/m². Neutropenia was the dose-limiting toxicity in this study. J Vet Intern Med 1996_;10:76–81. Copyright © 1996 by the American College of Veterinary Internal Medicine .     

Perspectives on emerging zoonotic disease research and capacity building in Canada

Zoonoses are fundamental determinants of community health. Preventing, identifying and managing these infections must be a central public health focus. Most current zoonoses research focuses on the interface of the pathogen and the clinically ill person, emphasizing microbial detection, mechanisms of pathogenicity and clinical intervention strategies, rather than examining the causes of emergence, persistence and spread of new zoonoses. There are gaps in the understanding of the animal determinants of emergence and the capacity to train highly qualified individuals; these are major obstacles to preventing new disease threats. The ability to predict the emergence of zoonoses and their resulting public health and societal impacts are hindered when insufficient effort is devoted to understanding zoonotic disease epidemiology, and when zoonoses are not examined in a manner that yields fundamental insight into their origin and spread. Emerging infectious disease research should rest on four pillars: enhanced communications across disciplinary and agency boundaries; the assessment and development of surveillance and disease detection tools; the examination of linkages between animal health determinants of human health outcomes; and finally, cross-disciplinary training and research. A national strategy to predict, prevent and manage emerging diseases must have a prominent and explicit role for veterinary and biological researchers. An integrated health approach would provide decision makers with a firmer foundation from which to build evidence-based disease prevention and control plans that involve complex human/animal/environmental systems, and would serve as the foundation to train and support the new cadre of individuals ultimately needed to maintain and apply research capacity in this area.     

The prevention of attachment and the detachment effects of a novel combination of fipronil, amitraz and (S)-methoprene for Rhipicephalus sanguineus and Dermacentor variabilis on dogs

A novel combination of fipronil, amitraz and (S)-methoprene (CERTIFECT?, Merial Limited, GA, USA) was evaluated for the prevention of attachment of ticks and its ability to cause detachment of ticks. For the two prevention of attachment studies, 20 purpose-bred beagles were allocated each to two equal groups based on pretreatment tick counts (treated and untreated). Each dog was exposed to 50 adult Rhipicephalus sanguineus and Dermacentor variabilis weekly starting 24h after treatment. In study 1 infestations with R. sanguineus were discontinued after Day 7 but continued to Day 28 for D. variabilis in both studies. Counts of ticks by species were made 2, 4 and 24h after exposure to ticks. Ticks not attaching to dogs were evaluated for viability. For the evaluation of detachment study, 16 purpose-bred beagles were allocated each to two equal groups based on pretreatment tick counts (treated and untreated). Each dog was infested with 50 unfed R. sanguineus and D. variabilis adults on Day -2. Ticks were thumb counted without removal on all dogs on Day -1, and at 4, 12, and 24h after treatment. Ticks were counted and removed at 48 h after treatment. Dogs treated with the novel combination had significantly (p<0.05) lower total numbers of attached R. sanguineus and D. variabilis than untreated controls at 4h through Day 7. For R. sanguineus, percent reduction of attachment at 24h after infestation through Day 29 ranged from 94.5% to 100%. For D. variabilis, the percent reduction of attachment at 24h through Day 22 was above 98.0%. These studies demonstrate that novel combination can disrupt attachment of R. sanguineus and D. variabilis for up to 28 days following treatment. Of those ticks that are exposed to the treatment, even if they do not attach to the dog and remain in the environment, greater than 90% (p<0.05) die within 24h for 2-3 weeks following treatment. Also, for those dogs infested with ticks at the time of treatment, the novel combination causes significant detachment (p<.05) starting at 12h and reaching 98.9% by 48 h after treatment. This product provides an effective means for controlling ticks infesting dogs and limiting the spread of tick transmitted diseases. Additionally, the mortality of ticks exposed to CERTIFECT will reduce infestation of the dog's environment.     

In vitro susceptibility testing of meticillin-resistant and meticillin-susceptible staphylococci to mupirocin and novobiocin

Antimicrobials effective against meticillin-resistant staphylococci are limited. Mupirocin is a topical antimicrobial used to treat bacterial skin infections. Novobiocin is an oral antimicrobial approved for treatment of staphylococcal upper respiratory infections in dogs. This study reports the in vitro activity of mupirocin and novobiocin on meticillin-susceptible (MSS) and resistant staphylococci (MRS) from healthy dogs and dogs with superficial pyoderma. Staphylococci were isolated from skin swabs at four sites on healthy dogs and from lesions on dogs with superficial pyoderma. Staphylococci were identified by morphology and by catalase and coagulase testing. Speciation and susceptibility testing were performed by the Dade Microscan (W. Sacramento, CA, USA). Meticillin resistance was confirmed by an oxacillin screen plate. Novobiocin and mupirocin susceptibilities were tested by disc diffusion. Staphylococci were cultured from 61 healthy dogs (17 MRS and 44 MSS) and 30 dogs with pyoderma (15 MRS and 15 MSS), with higher proportions of MRS isolates in dogs with pyoderma (P=0.038; χ(2) test). For mupirocin, 79.5% (35 of 44) MSS and 82.3% (14 of 17) MRS isolates from healthy dogs, and 100% (15 of 15) MSS and 86.6% (13 of 15) MRS isolates from dogs with pyoderma were susceptible (MSS, P=0.094; MRS, P=1.0; Fisher's exact test). For novobiocin, 95.4% (42 of 44) MSS and 52.9% (nine of 17) MRS isolates from healthy dogs and 93.3% (14 of 15) MSS and 80% (12 of 15) MRS isolates from dogs with pyoderma were susceptible (MSS, P=1.0; MRS, P=0.148; Fisher's exact test).     

Live Brucella abortus rough vaccine strain RB51 stimulates enhanced innate immune response in vitro compared to rough vaccine strain RB51SOD and virulent smooth strain 2308 in murine bone marrow-derived dendritic cells

Brucella spp. are Gram-negative, coccobacillary, facultative intracellular pathogens. B. abortus strain 2308 is a pathogenic strain affecting cattle and humans. Rough B. abortus strain RB51, which lacks the O-side chain of lipopolysaccharide (LPS), is the live attenuated USDA approved vaccine for cattle in the United States. Strain RB51SOD, which overexpresses Cu–Zn superoxide dismutase (SOD), has been shown to confer better protection than strain RB51 in a murine model. Protection against brucellosis is mediated by a strong CD4+ Th₁ and CD8+ Tc₁ adaptive immune response. In order to stimulate a robust adaptive response, a solid innate immune response, including that mediated by dendritic cells, is essential. As dendritic cells (DCs) are highly susceptible to Brucella infection, it is possible that pathogenic strains could limit the innate and thereby adaptive immune response. By contrast, vaccine strains could limit or bolster the innate and subsequent adaptive immune response. Identifying how Brucella vaccines stimulate innate and adaptive immunity is critical for enhancing vaccine efficacy. The ability of rough vaccine strains RB51 and RB51SOD to stimulate DC function has not been characterized. We report that live rough vaccine strain RB51 induced significantly better (p ≤ 0.05) DC maturation and function compared to either strain RB51SOD or smooth virulent strain 2308, based on costimulatory marker expression and cytokine production.