The objective of this study was to determine if R. equi IgG purified from commmercially available hyperimmune R. equi plasma and delivered to the lungs of adult ponies would cause a local inflammatory response and if increases in total and R. equi specific IgG occurred post administration. IgG was purified and concentrated from plasma via protein G affinity chromatography. A cross over study was performed. Eight healthy adult ponies were randomly assigned to two groups of four; each pony acted as its own control. Either the IgG product or 0.9% Saline was delivered via a vibrating mesh nebulizer during the first treatment phase. During the second treatment phase ponies recieved the oppostie treatment. A 4 week washout period was allowed between phases. Bronchoalveolar fluid was recovered using a low volume endoscopic technique prior to aerosolization (time 0), and at 1 hr, 6 hrs, and 24 hours post administration. The BAL fluid total IgG concentration and R. equi specific IgG titers were determined via ELISA and cytologic analysis was performed. No clinically significant local inflammatory response was identified in response to IgG treatment. While total IgG concentrations were increased at T1 compared to T0, no significant effects of time were found (P=0.19). However, overall significantly higher concentrations of total IgG were found after administration of saline when compared to IgG administration (P=0.023).  While the R. equi specific titer increased at T1 after IgG administration, no significant difference was identified between treatment or time (P=0.261). Overall the individual response to IgG was variable. It is possible that the protein rich IgG acted as a relatively hypertonic solution and caused fluid influx from the pulmonary parenchyma after treatment thereby diluting the total IgG present when compared to saline administration. This conclusion cannot be verified as BAL dilution correction was not performed. However, it is unknown what titer or level of increased IgG is nessecary to assist with prevention of disease. Future research should focus on the effect of R. equi specific IgG on pulmonary cells to determine if administration of local R. equi specific IgG would alter intrapulmonary immune responses to R. equi. / Master of Science

This research evaluated serum amyloid A (SAA) concentration in synovial fluid of healthy horses as a potential marker for use in the diagnosis and monitoring of horses with septic arthritis. The first study evaluated the effect of arthroscopic lavage of healthy joints on concentrations of systemic and synovial SAA; as well as total protein concentration, nucleated cell count and percentage of neutrophils in synovial fluid. The second study, evaluated the effect of repeated through-and-through joint lavage on SAA in systemic blood and SAA, total protein, nucleated cell count and percentage of neutrophils in synovial fluid from healthy joints. In the first study, middle carpal joints of 6 horses were randomly assigned to one of the following treatments 1) arthrocentesis (controls) or 2) arthroscopic lavage. A washout period of 30 days was allowed in between treatments. Synovial fluid and blood samples were collected at 0, 24, 48, 72, 96 and 120 h. Measurements included SAA in blood and synovial fluid, and total protein, nucleated cell count and percentages of neutrophils in synovial fluid. In the second study, one tarsocrural joint was randomly assigned to receive repeated through-and-through joint lavage at 0, 48 and 96 h in 6 horses. Synovial fluid and blood samples were collected at 0, 24, 48, 72, 96 and 120 h. Measurements included SAA in blood and synovial fluid, and total protein, nucleated cell count and percentages of neutrophils in synovial fluid. For this study, synovial fluid samples collected at time 0 were considered as control values. After arthroscopic lavage and repeated through-and-through joint lavage, systemic and synovial SAA did not increase from baseline values (except for systemic SAA at 24h after arthroscopic lavage and in controls). Total protein values were significantly increased at all time points after arthroscopic and through-and-through joint lavages (except at 96h on both lavage procedures) but not in controls. With both lavage procedures, nucleated cell count significantly increased from baseline values at all time points (except at 96h after through-and-through joint lavage). Percentage of neutrophils was significantly increased after arthroscopic lavage at all time points and only at 24h in controls; however, the percentages of neutrophils were not significantly increased after repeated through-and-through joint lavage. Synovial SAA was not affected by arthroscopic or repeated through-and-through joint lavage; however, synovial total protein and nucleated cell counts were significantly increased. Synovial SAA may be a valuable inflammatory marker that is not affected by procedures as arthroscopic or repeated through-and-through joint lavage in horses. Further validation of synovial SAA as a marker for evaluating the progression of septic joints while treatment is installed is warranted.

Th. 3e cycle--Sci. pharm.--Paris 5, 1981. N°: 19.

Master of Science / Department of Clinical Sciences / Warren L. Beard / Objectives: To quantify recovery of colored microspheres from cadaver tarsocrural joints via arthroscopic or needle lavage, and to compare recovery for 1-5L of lavage fluid. Study design: Randomized experimental trial. Methods: 8 adult Quarter Horse cadavers had 1.5 million 15μm, colored microspheres injected into each tarsocrural joint. Each joint was randomly assigned to receive lavage with an arthroscope and egress cannula (group A) or three (1 ingress, 2 egress) 14 gauge needles (group N) with 5L 0.9% NaCl. The egress fluid from each liter of lavage was collected separately, and the number of microspheres present in each recovered liter was determined via spectrophotometry. Results: A significant interaction (p<0.01) was present between treatment group and liter. The number of microspheres recovered in the first liter of lavage fluid was significantly higher in the needle group than in the arthroscope group (p<0.01). For both groups the number of microspheres recovered in the first liter of lavage fluid represented a majority of the total microspheres collected, and was significantly different from the subsequent liters collected (p<0.01). The number of microspheres recovered did not differ between liters 2, 3, 4, and 5, within or between treatment groups. Conclusions: In this model, tarsocrural lavage with three 14-gauge needles was more effective at removing colored microspheres from the joint than arthroscopic lavage, suggesting the number or placement of portals present may be more important than portal size and flow rate. No difference in microsphere recovery was seen with lavage volumes greater than 1L.

Afecções do sistema respiratório estão entre as mais frequentes em equinos, sendo uma das principais causas de baixo desempenho em animais atletas. Dentre essas doenças pode-se citar as afecções inflamatórias das vias aéreas, que apresentam uma prevalência considerável e afetam a saúde e vida atlética do animal. Um dos principais meios de atuação terapêutica no tratamento dessas doenças é a administração de medicamentos corticoides, sendo a dexametasona um dos principais fármacos dessa classe utilizados na medicina equina. No tratamento de doenças semelhantes em humanos preconiza-se a administração dos fármacos por via inalatória, otimizando assim o seu efeito terapêutico e diminuindo os efeitos colaterais. Em equinos existem trabalhos que mostram a deposição pulmonar de medicamentos quando administrados por via inalatória, no entanto, não foram encontrados estudos nesse sentido utilizando a dexametasona. O objetivo desse estudo foi avaliar a deposição pulmonar de dexametasona quando administrada por via inalatória em equinos, bem como fatores que possam interferir no seu nível de deposição e a concentração plasmática do fármaco quando administrada por essa via. Para isso foram utilizados seis equinos que foram submetidos à inalação com dexametasona duas vezes, cada uma utilizando um veículo diferente (aquoso ou oleoso) na formulação do fármaco, e quatro animais foram utilizados para o grupo controle, sendo submetidos à inalação somente com o veículo. Após cada inalação foi realizado um lavado broncoalveolar (LAB) e coletas sanguíneas seriadas para quantificação da dexametasona. Após a inalação o fármaco foi identificado nas amostras de LAB dos animais nos grupos tratados. Não houve diferença significativa entre os veículos utilizados e houve diferença entre os animais agitados e os animais calmos, sendo que os primeiros apresentaram uma concentração significativamente maior de dexametasona no LAB. O fármaco não foi identificado no plasma dos animais. A dexametasona quando administrada via nebulização pneumática atinge a região de bronquíolos e alvéolos respiratórios em equinos, principalmente os de comportamento agitado, e não atinge níveis plasmáticos significativos / Disorders of the respiratory system are among the most frequent deseases in horses and one of the main causes of low performance in athletic animals. Inflammatory diseases of the airways should be mentioned due to its considerable prevalence and to affect the health and athletic life of the animal. One of the main ways of therapeutic action of these diseases is the administration of corticosteroid drugs, being dexamethasone one of the main drugs used in equine medicine. In the treatment of similar diseases in humans it is recommended the administration of drugs by inhalation, which optimizes its therapeutic effect and reduces side effects. In horses, there are studies that show the pulmonary deposition of drugs when administered by inhalation, however, there are no studies using dexamethasone. The aim of this study was to evaluate the pulmonary deposition of dexamethasone when administered by inhalation in horses, as well as factors that may interfere with their level of deposition and the plasma concentration of the drug when administered by inhalation. To that, it was used six horses that were submitted to inhalation of dexamethasone twice, each one using a different vehicle (aqueous or oily) in the drug formulation, and four animals were used as control group, being submmited to inhalation just with the vehicle. After each inhalation a bronchoalveolar lavage (BAL) and serial blood collections for quantification of dexamethasone were performed. After the inhalation, the drug was identified in BAL samples from the animals of the treated groups. There was no significant difference between the vehicles used and there was difference between the agitated animals and calm animals, and the first ones had a significantly higher concentration of dexamethasone in the BAL. The drug was not identified in the plasma of animals. Dexamethasone, when administered via nebulization air, reaches the bronchioles region and respiratory alveoli in horses, especially the agitated behavior ones, and it does not affects plasma levels

<i>Mycoplasma bovis</i> is generally considered the causative pathogen associated with Chronic Pneumonia and Polyarthritis Syndrome (CPPS) in feedlot cattle. However, <i>M. bovis</i> virulence may vary between strains as it is also isolated from asympytomatic cattle. The following study aims to determine the prevalence of <i>M. bovis</i> in the respiratory tract of western Canadian cattle using two sampling methods and at two time points following feedlot entry. Three study groups were sampled. In the first group nasal swabs (NS) and bronchoalveolar lavages (BAL) were taken from 36 clincally healthy cattle at the University of Saskatchewan feedlot at both 14 and 90 days on feed (DOF). In a second experiment, NS were taken from 56 animals upon arrival at a commercial feedlot and one week to three months later upon treatment for respiratory disease. Lung and joint tissue swabs were collected at necropsy from a third group of 19 animals with CPPS clinical pathology originating in 10 different western Canadian feedlots. All samples were selectively cultured for <i>Mycoplasma</i> spp. DNA was extracted from isolated putative <i>Mycoplasma</i> colonies and amplified with universal 16S rRNA gene primers for identification. Amplified Fragment Length Polymorphism (AFLP) was used to genetically differentiate <i>M. bovis</i> positive isolates. More <i>M. bovis</i> was isolated from NS than BAL and <i>M. bovis</i> prevalence increased with DOF in the feedlot in both the University of Saskatchewan and commercial feedlot trials. Three genetically distinct clusters (A, B, and C) were isolated from the necropsy group. Two of these clusters were primarily associated with isolates collected from feedlot cattle and one strain was exclusively found in CPPS-associated mortalities. No significance difference in the prevalence of <i>M. bovis</i> strains was observed between different days on feed or sampling methods. It was concluded that either the difference in disease state is a host dependent outcome, due to a multi-factorial disease complex, or the AFLP assay was not sensitive enough to differentiate strains based on virulence.

<i>Mycoplasma bovis</i> is generally considered the causative pathogen associated with Chronic Pneumonia and Polyarthritis Syndrome (CPPS) in feedlot cattle. However, <i>M. bovis</i> virulence may vary between strains as it is also isolated from asympytomatic cattle. The following study aims to determine the prevalence of <i>M. bovis</i> in the respiratory tract of western Canadian cattle using two sampling methods and at two time points following feedlot entry. Three study groups were sampled. In the first group nasal swabs (NS) and bronchoalveolar lavages (BAL) were taken from 36 clincally healthy cattle at the University of Saskatchewan feedlot at both 14 and 90 days on feed (DOF). In a second experiment, NS were taken from 56 animals upon arrival at a commercial feedlot and one week to three months later upon treatment for respiratory disease. Lung and joint tissue swabs were collected at necropsy from a third group of 19 animals with CPPS clinical pathology originating in 10 different western Canadian feedlots. All samples were selectively cultured for <i>Mycoplasma</i> spp. DNA was extracted from isolated putative <i>Mycoplasma</i> colonies and amplified with universal 16S rRNA gene primers for identification. Amplified Fragment Length Polymorphism (AFLP) was used to genetically differentiate <i>M. bovis</i> positive isolates. More <i>M. bovis</i> was isolated from NS than BAL and <i>M. bovis</i> prevalence increased with DOF in the feedlot in both the University of Saskatchewan and commercial feedlot trials. Three genetically distinct clusters (A, B, and C) were isolated from the necropsy group. Two of these clusters were primarily associated with isolates collected from feedlot cattle and one strain was exclusively found in CPPS-associated mortalities. No significance difference in the prevalence of <i>M. bovis</i> strains was observed between different days on feed or sampling methods. It was concluded that either the difference in disease state is a host dependent outcome, due to a multi-factorial disease complex, or the AFLP assay was not sensitive enough to differentiate strains based on virulence.

Reproduction de : Thèse d'exercice : Médecine vétérinaire : Toulouse 3 : 2007. / Titre provenant de l'écran titre. Bibliogr. p. 57-63.

Several tests are available for evaluation of respiratory disorders but most of them are invasive and associated with some risk or patient discomfort. Examples include bronchoscopy (bronchoalveolar lavage, BAL) [1], venopuncture [2] and sputum induction [3]. Noninvasive sampling of nongaseous substances contained in expired air, collected as exhaled breath condensate (EBC) has been used to detect inflammatory markers and by-products including nitric oxide and arachidonic acid metabolites and proteins [4]. Nerve growth factor (NGF) is a protein that has been implicated in neurogenic airway inflammation and this pilot study aimed to develop a non-invasive approach for evaluation of allergic airway inflammatory disease by measuring and comparing levels of NGF in the induced sputum and EBC of ten (10) asthmatics and ten (10) non-asthmatics.Though twenty (20) subjects were sampled, an unexpected event due to a defective NGF kit inadvertently resulted in an unsuccessful analysis of fifteen (15) sets of specimen (6 non-asthmatics and 9 asthmatics), limiting the study.This study is significant because occupational lung diseases are the number one work-related illness in the United States and occupational asthma is the most common form [9]. Toluene diisocyanate (TDI) is the commonest cause of occupational asthma and workers exposed to TDI vapor may develop inflammatory conditions including asthma, rhinitis and nasal irritation [7].Results: NGF was detected and measured only in sputum, with a mean NGF level of 210 (210-210, range 0) in asthmatics and 164 (7-280, range 273) in non-asthmatics. Nonetheless, we failed to reject the null hypothesis (number 3).Conclusion: This limited study did not have adequate power (power 11%) due to the small sample size and thus lacks internal validity. Further studies are needed using a larger sample size.

Bronchoalveolar lavage (BAL) is a minimally invasive technique utilized in human and veterinary medicine to sample the lower generation bronchi and alveolar spaces. The basic technique for BAL involves infusion of sterile saline into the lower airways and re-aspiration of the fluid (bronchoalveolar lavage fluid; BALF). Certain aspects of BAL technique and BALF processing affect sample quality, and sample quality is important to ensure meaningful cytology. Aspiration techniques for retrieval of BALF have not been critically evaluated in companion animal medicine. This research project compared three aspiration techniques for retrieval of BALF in dogs [manual aspiration without tubing (MA), manual aspiration through polyethylene tubing (MAPT) and suction pump aspiration (SPA)] and their effect on sample quality in healthy dogs and dogs with respiratory tract disease. SPA consistently retrieved a higher proportion of BALF than MA and MAPT. In addition, SPA yielded improved sample quality compared to MA and MAPT. The improved BALF retrieval and cellularity scores in SPA samples did not significantly increase the diagnostic rate achieved from BALF cytology in dogs with pulmonary disease. The results indicated that both MA and SPA are suitable for BAL in dogs with respiratory tract disease. Yet, for the purpose of creating a standardized BAL technique in dogs, SPA is recommended for BALF retrieval due to the improved sample quality parameters. / Ontario Veterinary College Pet Trust