Use of volumetric capnography to identify pulmonary dysfunction in horses with and without clinically apparent recurrent airway obstruction

OBJECTIVE: To investigate whether volumetric capnography indices could be used to differentiate between horses without recurrent airway obstruction (RAO) and horses with RAO that were in clinical remission or that had clinically apparent RAO. ANIMALS: 70 adult Swiss Warmblood horses (20 used for pleasure riding and 50 used for dressage or show jumping). PROCEDURE: Horses were allocated to 4 groups on the basis of history, clinical signs, results of endoscopy, and cytologic findings (group 1, 21 healthy horses; group 2, 22 horses with RAO that were in remission; group 3, 16 horses with mild RAO; group 4, 11 horses with exacerbated RAO). Expiratory volume and CO2 curves were recorded by use of a computerized ultrasonic spirometer. Volumetric capnograms were plotted, and derived indices were calculated. RESULTS: Dead-space volume (VD) was calculated by use of the Bohr equation (VD(Bohr)) and for physiologic VD (VD(phys)). Ratios for VD(Bohr) to expiratory tidal volume (VT) and VD(phys) to V(T) as well as an index of effective CO2 elimination were significantly different among groups of horses. Age and use of the horses also significantly affected volumetric capnography indices. CONCLUSIONS AND CLINICAL RELEVANCE: Ratios of VD(Bohr) to VT and VD(phys) to VT as well as an index of effective CO2 elimination were sufficiently sensitive measures to distinguish between healthy horses and horses with RAO in remission. To optimize the ability of volumetric capnography indices to differentiate among horses in heterogeneous populations, it is important to account for effects of age and specific use of the horses.     

Muscle reorganisation through local injection of stem cells in the diaphragm of mdx mice

Background

The diaphragm is the major respiratory muscle affected by Duchenne muscular dystrophy (DMD) and is responsible for causing 80% of deaths. The use of mechanical forces that act on the body or intermittent pressure on the airways improves the quality of life of patients but does not prevent the progression of respiratory failure. Thus, diseases that require tissue repair, such as DMD, represent a group of pathologies that have great potential for cell therapy. The application of stem cells directly into the diaphragm instead of systemic application can reduce cell migration to other affected areas and increase the chances of muscle reorganisation. The mdx mouse is a suitable animal model for this research because its diaphragmatic phenotype is similar to human DMD. Therefore, the aim of this study was to assess the potential cell implantation in the diaphragm muscle after the xenotransplantation of stem cells.

Methods

A total of 9 mice, including 3 control BALB/Cmice, 3 5-month-old mdx mice without stem cell injections and 3 mdx mice injected with stem cells, were used. The animals injected with stem cells underwent laparoscopy so that stem cells from GFP-labelled rabbit olfactory epithelium could be locally injected into the diaphragm muscle. After 8 days, all animals were euthanised, and the diaphragm muscle was dissected and subjected to histological and immunohistochemical analyses.

Results

Both the fresh diaphragm tissue and immunohistochemical analyses showed immunopositive GFP labelling of some of the cells and immunonegativity of myoblast bundles. In the histological analysis, we observed a reduction in the inflammatory infiltrate as well as the presence of a few peripheral nuclei and myoblast bundles.

Conclusion

We were able to implant stem cells into the diaphragm via local injection, which promoted moderate muscle reorganisation. The presence of myoblast bundles cannot be attributed to stem cell incorporation because there was no immunopositive labelling in this structure. It is believed that the formation of the bundles may have been stimulated by cellular signalling mechanisms that have not yet been elucidated.     

Grid keratotomy as a treatment for superficial nonhealing corneal ulcers in 10 horses

OBJECTIVE: To describe the clinical symptoms of 10 cases of superficial nonhealing corneal ulcers in horses and to evaluate the results of grid keratotomy in these patients. DESIGN: Retrospective study. ANIMALS: Ten horses with superficial nonhealing corneal ulceration in one eye. PROCEDURE: The signalment, history and clinical symptoms are reported of 10 patients with superficial nonhealing corneal ulcers during the period from August 2003 to February 2005. Grid keratotomy was performed in all cases. In addition, the surgical procedure of grid keratotomy and response to therapy are described. RESULTS: Horses generally responded well to grid keratotomy (eight cases healed after one grid keratotomy and one horse healed after a second grid keratotomy; one case was not available for follow-up). Only 2/10 had discomfort after treatment and only 2/9 had some degree of scarring after treatment. The healing time, which was known in seven cases, averaged 8.4 days (+/- SD 4). CONCLUSIONS AND CLINICAL RELEVANCE: Grid keratotomy is an appropriate option for treatment of superficial nonhealing corneal ulcers in horses. The procedure is simple to perform. It generally induces a rapid and uncomplicated healing of the cornea. Only in a limited number of cases does a small amount of scarring occur.     

A monoclonal antibody against a canine CD45 homologue: analysis of tissue distribution, biochemical properties and in vitro immunological activity

This report describes the characterisation of a monoclonal antibody (mAb), AB6, which recognises specifically a cluster of canine leukocyte surface molecules. The immunogen used for obtaining the AB6 mAb was a lysate of canine peripheral blood mononuclear cells (PBMC). This novel mAb belongs to the IgG2a isotype, and reacted in Western blot with four different canine leukocyte glycoproteins with apparent molecular weights of 180, 190, 205 and 220 kDa. The AB6 mAb recognised the majority of canine peripheral blood leukocytes as determined by flow cytometry (97%). It also exhibited a broad reactivity pattern against lymphoid and myeloid cells, inhibited the proliferation of mitogen-stimulated canine PBMC and did not recognise human PBMC and murine splenocytes. The biochemical properties, cell and tissue specificity, and in vitro biological activity of the AB6 mAb indicate that it recognises a canine CD45 homologue. The mAb could become a valuable diagnostic and research tool for the evaluation of immune functions in dogs.     

The fibronectin binding protein ShdA is not a prerequisite for long term faecal shedding of Salmonella typhimurium in pigs

Porcine carcasses contaminated with Salmonella typhimurium pose significant public health problems. Prolonged faecal shedding of Salmonella in pigs contributes to the contamination level of carcasses. Although the mechanism of prolonged faecal shedding is not yet clarified, the CS54 Island, and more specifically the shdA gene encoding a fibronectin binding autotransporter protein, was identified as an important locus for intestinal colonization and persistence of Salmonella typhimurium in mice. The aim of this study was to assess the contribution of ShdA in faecal shedding of Salmonella typhimurium in pigs. Pigs were orally inoculated with a Salmonella typhimurium wild type field strain or its isogenic shdA mutant strain. For the first few days after inoculation, the shdA mutant strain was excreted more, the diarrhoea was more pronounced and higher numbers of internal organs were infected. No effect on long-term shedding was found. In a porcine ileal loop model, the wild type strain and shdA mutant strain did not show any differences in the induction of neutrophil influx into the intestinal wall and lumen. In conclusion, we have shown that a Salmonella typhimurium deletion mutant in shdA is more virulent during the first days after inoculation and is not significantly impaired in persistence or prolonged shedding in pigs.     

Synovial fluid changes in induced infectious arthritis in calves

The objective was to develop an experimental model of septic arthritis in calves and to evaluate the effect of treatment on cytologic and bacteriologic variables of synovial fluid. The right tarsus of 7 healthy Holstein bull calves were inoculated with 10(8) colony-forming units of viable Escherichia coli of a pap-positive strain (day 1). On day 2, joint lavage was performed and antibiotic treatment was instituted. Cytologic examinations, bacterial cultures, and pap factor determinations by polymerase chain reaction (PCR) were performed on synovial fluid samples that were collected daily until day 4, then every 4 days until day 24. Results of physical examination, the severity of lameness, and swelling were recorded. Clinical signs of septic arthritis appeared on day 2 and persisted until day 9 for all calves. Bacterial cultures from all calves were positive for E. coli on day 2, and remained positive until day 3 for 1 calf and until day 4 for 5 calves. In addition, PCR results were positive for all calves, with 6 positive through day 3 and 1 positive through day 4, after which a positive result was again obtained on day 24. Synovial fluid neutrophil counts and white blood cell counts were significantly increased on days 2-4; however, synovial total protein concentrations were increased (P < .05) throughout the experiment in comparison to day 1. Results of all bacterial cultures were negative on day 8, although clinicopathologic signs of inflammation persisted until day 20. This model successfully induced acute septic arthritis in calves. Rapid recovery occurred within 1 week when an appropriate treatment was instituted early in the course of the disease.     

C-reactive protein measurement in canine saliva.

An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 microg/ml and 0.0091 microg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7-9.9% and 8.5-16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R2 = 0.76) and clinical studies (R2 = 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments.     

Ultrastructural identification of small granule prolactin (PRL) cells in goat adenohypophysis by the colloidal-gold method

We describe ultrastructural characteristics of PRL cells with small secretory granules, immunostained with ovine antiserum, in adult Murciano-Granadina milking goats during anoestrus, the last third of gestation and lactation. This cell subtype is considered to comprise a stable population that decreases numerically during gestation and lactation, and that may change during these stages to show large granules.     

A quantitative assessment of the validity of animal-health surveys using stochastic modelling

This paper presents a stochastic simulation model to evaluate the efficacy of regional or national surveys aimed at identifying infection in populations of animals. The process of evaluation involves specification or calculation of cluster-level test sensitivity and specificity, which are derived from two probability distributions of the number of individual-level positive tests expected from non-infected and infected clusters, respectively. Probability distributions for the number of positive clusters expected in a situation of freedom from infection and under various levels of cluster prevalence are specified and used to determine survey properties (the survey being considered a diagnostic system), and ROC curves are drawn. Likelihood ratios allow investigators to state the extent to which a survey result is more likely to be observed if the region or country is infected at a given prevalence than if it is free from infection. The result of a survey carried out to investigate the presence of porcine reproductive and respiratory syndrome (PRRS) in Switzerland is used to illustrate this approach. The model can be adapted to a wide range of survey designs.