Molecular and Cellular Evaluation of Horses With Summer Pasture Associated Asthma Syndrome.

Equine asthma is an airway disease that affects a large number of horses annually leading to considerable economic losses in the horse industry. Despite advances in research in this area, there is still a lack of information on its etiology and molecular characterization in pasture associated asthma. The objective of the current study was to characterize the inflammatory disease of lower airways in horses maintained on pasture through cytologic and immunologic profile during the summer in a tropical environment by analysis of the gene expression of Th1 cytokines (IFN- λ, IL-8), Th2 cytokines (IL-4 and IL-5), and pro-inflammatory cytokines (IL-1, TNF-α) in the bronchoalveolar lavage (BAL) fluid in healthy and asthma horses on pasture. A group 39 of clinically healthy horses maintained on native pasture and supplemented with concentrate was evaluated by BAL analyzed for differential cellular count and assigned into a control and an asthma group. The gene expression of pro-inflammatory cytokines was analyzed in the BAL by reverse time PCR (RT-PCR) (IL-1α (alpha), IL-4, IL-5, IL-8, TNF-α alpha and IFN-λ), using ß-actin as housekeeping gene. Higher gene expression of IL-1, IL-4, IL-5, IL-8, IFN-λ in the BAL of asthma horses was found. Current results indicate an increase in Th2, characterizing an allergic inflammatory reaction due to the significant increase in IL-5 in asthmatic horses (10.3 ± 1.13), when compared to the values ​​obtained in normal horses (3.27 ± 0.46). The only down regulated cytokine in the asthma group was TNF-α, suggesting a chronic antigenic reaction.

Serum proteins and electrophoretic profile in horses undergoing crotalid venom hyperimmunization.

Indisputably, the use of antivenoms for the treatment of snakebite envenoming is beneficial for the victims. However, there are few studies addressing the effect of long-term hyperimmunization in inoculated horses. It is known that the injection of snake venoms and adjuvants leads to local and systemic reactions in horses, but little is known about the response of inflammatory proteins. The aim of this study was to evaluate serum proteins and the electrophoretic profile of horses undergoing crotalid venom hyperimmunization. Twenty horses were divided into two groups: an inoculated group, comprising ten horses that were already being used for production of a Crotalus sp. antivenom, and a control group, comprising ten animals that had never been used for hyperimmunization. All animals were clinically healthy and without laboratory abnormalities. Total protein and albumin concentrations were measured in serum. Serum globulins were obtained by calculation. Plasma fibrinogen estimates were determined by the heat precipitation method. Serum proteinograms were obtained using agarose gel electrophoresis. The results revealed a significant increase in the concentrations of total serum proteins, globulins, and ß-globulins in the inoculated group, exceeding the reference values. There were slight increases in the α-1- and α-2-globulin subfractions in serum-producing horses, with no statistical significance. We also observed that horses used to produce hyperimmune plasma developed hypoalbuminemia, although the decrease in albumin production was not statistically significant. Our findings suggest that the continuous use of horses to produce crotalid antivenom may lead to a chronic inflammatory stimulus, with changes in plasma levels of inflammatory proteins.

Equine Pasture Asthma in Brazil

Background: Summer Pasture Associated Obstructive Pulmonary Disease (SPAOPD), or Equine Pasture Asthma (EPA),has been described as an environmentally-induced respiratory disease that occurs during the warmer and more humidmonths, leading to reversible airway obstruction, persistent and non-specific airway hyper-responsiveness, and chronicneutrophilic airway inflammation. Exacerbation of clinical signs vary according to warm seasons and range from mildto severe episodes of wheezing, coughing, and laboured breathing, being highly debilitating to the equine. This reportdescribed two cases of Equine Pasture Asthma that showed clinical and environmental similarities with Summer PastureAssociated Obstructive Pulmonary Disease.Cases: The patients were crossbreed geldings that have never been stabled and were used for cattle management in a farmin southeastern Brazil. They presented poor performance and a persistent cough for over 3 years. Initially, the respiratorysigns were only observed after exercise but, over the years, it gradually progressed to being observed when the horses wereat rest. Both animals had a history of regular deworming and were previously treated by other veterinarians with antibiotics,clenbuterol, and mucokinetics. Little improvement was noticed by the owner and the signs returned over time as treatmentwas often discontinued. Clinical findings were compatible with the grade 3 mucus classification as well as with score 2for Severe Asthma. BALF cytology was done according to routine procedure. Animal 1 presented slides with free yeast,macrophages, and mucus with Curschmanns spiral and counting of 29.7% of neutrophils (NE), 43.7% of lymphocytes(LP), 25.3% of macrophages (MC) and 1.3% of eosinophils (EO). Animal 2 presented slides with phagocytized yeast,mucus and counting of 27% of NE, 38.5% of LP, 33% of MC and 1.5% of EO...(AU)

Equine Pasture Asthma in Brazil

Background: Summer Pasture Associated Obstructive Pulmonary Disease (SPAOPD), or Equine Pasture Asthma (EPA),has been described as an environmentally-induced respiratory disease that occurs during the warmer and more humidmonths, leading to reversible airway obstruction, persistent and non-specific airway hyper-responsiveness, and chronicneutrophilic airway inflammation. Exacerbation of clinical signs vary according to warm seasons and range from mildto severe episodes of wheezing, coughing, and laboured breathing, being highly debilitating to the equine. This reportdescribed two cases of Equine Pasture Asthma that showed clinical and environmental similarities with Summer PastureAssociated Obstructive Pulmonary Disease.Cases: The patients were crossbreed geldings that have never been stabled and were used for cattle management in a farmin southeastern Brazil. They presented poor performance and a persistent cough for over 3 years. Initially, the respiratorysigns were only observed after exercise but, over the years, it gradually progressed to being observed when the horses wereat rest. Both animals had a history of regular deworming and were previously treated by other veterinarians with antibiotics,clenbuterol, and mucokinetics. Little improvement was noticed by the owner and the signs returned over time as treatmentwas often discontinued. Clinical findings were compatible with the grade 3 mucus classification as well as with score 2for Severe Asthma. BALF cytology was done according to routine procedure. Animal 1 presented slides with free yeast,macrophages, and mucus with Curschmann’s spiral and counting of 29.7% of neutrophils (NE), 43.7% of lymphocytes(LP), 25.3% of macrophages (MC) and 1.3% of eosinophils (EO). Animal 2 presented slides with phagocytized yeast,mucus and counting of 27% of NE, 38.5% of LP, 33% of MC and 1.5% of EO...

Penectomia em equino com parafimose / Partial penectomy in a horse with paraphimosis

Background: In horses, an increase in the volume of penis and foreskin can be caused by trauma. The resultant edema interferes with the retraction of the penis and cause paraphimosis. Surgical intervention through penectomy is indicated in cases wherein an alternative treatment is not feasible. Partial penectomy can prolong the life and reproductive function of many stallions. The present study aims to report on the methodological, functional, and economic feasibility of a successful case of the use of Williams technique for partial penectomy performed in the field in a horse with paraphimosis, preserving urinary, productive, and reproductive functions. Case: A 10-year-old horse weighing 500 kg had clinical signs of paraphimosis due to the formation of granulomatous tissue following trauma to the glans region. A surgical intervention, namely a partial penectomy was decided to be performed to prevent injury from priapism. Due to the limited resources provided by the owner and the impossibility of moving the animal to a surgical center in a veterinary hospital, the procedure was performed in the field, with prior sanitization and preparation of the environment used to perform surgery. The horse was tranquilized with intravenous xylazine hydrochloride and acepromazine, with subsequent induction of anesthesia with ready-to-use (RTU) guaifenesin bolus and maintenance of anesthesia with an intravenous association of RTU guaifenesin, xylazine hydrochloride, and ketamine. The distal third of the penis was amputated using the recommended Williams technique. Although the complications like dehiscence and emergence of granulation tissue occurred after surgery, these were controlled in the daily follow-up of the animal and post-surgical treatment. Discussion: Paraphimosis predisposed the horse to abrasions and edema of the exposed portion of the penis. However, there was no urine retention, which suggested that the urethral ostium and the urethra had no lesions. Because the granulomatous lesions were located in the distal third of the penis and the extent of penile exposure was small, the partial penectomy technique proved effective in solving the permanent exposure of the penis. The anesthetic protocol used was inexpensive, easy to execute, and effective, and no anesthetic complications occurred, proving this protocol to be efficient for the anesthetic induction of animals in the field. In addition to being reliable and widespread, the Williams technique was recommended to prevent possible urethral stenosis and the development of contact dermatitis by urine. This technique makes a rapid recovery of the animal possible, with improvement of its physiological parameters, and due to ease of being able to be done in the field, it is also inexpensive. The edema and the granulation tissue that occurred after surgery were controlled with medication. In general, post-penectomy animals are not used for reproduction. This makes the present report an important contribution, because in cases in which the lesions present a distal disposition and the penile exposure is small, stallions have a chance of maintaining reproductive function after surgery, even with a long period of evolution before surgical treatment. This was demonstrated in the present case, as the animal in this case later impregnated a mare, with the pregnancy confirmed by ultrasound examination. Partial penectomy is a resolution technique for cases of traumatic paraphimosis, in which exuberant granulation tissue is formed in the distal third of the penis, with a long period of evolution. The surgery can be done in the field, with a low surgical cost to the owner, and a good productive and reproductive prognosis for the animal.

Evaluation of acquired passive immunity in mule foals up to 60 days of age.

Ten mule foals were used to investigate serum IgG concentration. Blood was collected from mares and foals on the day following parturition (D1), at 48 and 72 hr after birth (D2 and D3), and on D7, D14, D21, D28, D35, D42, and D60. Serum IgG concentration was determined by zinc sulfate turbidity test and was above 400 mg/dl at all time points, except for one foal on D2. Mean values were above 800 mg/dl from D1 to D7 and at D60. The difference between mares and foals at each collection time was significant, except at D1. For all other days, foal IgG values were lower than the mare IgG values. Mean IgG concentrations were lower from D14 to D42 compared with D1 and D60. The results show that the critical period for the mule foal is at D28, which can be a vulnerable window for infections. The variation pattern in IgG values is similar to that in data published for horse and donkey foals. There are no published studies to date on the subject, and the present contribution enables further understanding of the newborn mule foal.

Association between exercise-induced pulmonary hemorrhage and inflammatory airway disease in polo ponies.

The respiratory system is essential for health and high athletic performance in horses. Respiratory diseases have been recognized as having a major impact on training equine animals and are commonly cited as the second most common cause of wasted training time. Inflammatory airway disease (IAD) is an important cause of poor performance in young racehorses. Exercise-induced pulmonary hemorrhage (EIPH) is considered a major issue for the equine industry because of its high prevalence and association with reduced athletic performance. In Brazil, polo is a growing equestrian sport, but studies on it are still scarce. The aim of this study was to evaluate the occurrence of EIPH, the association between EIPH and IAD, and EIPH influence on the tracheal cytological profile of polo ponies. Thirty-seven horses regularly used for polo were included in this study. Endoscopic examination was performed every 30 to 90 min after practice, and tracheal lavage was performed after 18 to 24 hr. Sixteen animals (43.2%) presented a score of 0 for mucus in endoscopy; twelve animals (32.4%) presented a score for 1 and nine animals (24.3%) presented score 2 of mucus. IAD was characterized by tracheal cytology in 12 animals (32.4%). The occurrence of EIPH in this study was 29.7% (11/37). No significant difference was found in the cell types in tracheal cytology when EIPH-positive and EIPH-negative horses were compared. Polo ponies are affected by IAD and EIPH in relevant proportions, but there was no association between EIPH and tracheal cytological profile.

Equine tracheobronchial wash filtration and its effects on differential cell count / Efeito da filtração do lavado traqueobrônquico de equinos sobre a contagem diferencial de células

Tracheobronchial wash (TBW) is a method to recover cell samples from the airways. The cytology of TBW fluid is an important technique for the diagnosis of pulmonary diseases in horses. Excessive mucus in TBW may cause cell damage and morphological changes that hinder cell type recognition, resulting in a misdiagnosis. The aim of this study was to compare the results of differential cell count in a tracheobronchial wash of filtered and non-filtered samples. Endoscopy and TBW procedures were performed in thirty horses. Each TBW sample was split into two aliquots. Two groups were formed: non-filtrated aliquots (NF) and filtrated aliquots (F). The filtration was performed using a hydrophilic gauze pad. After centrifugation, the differential cell count was performed considering 300 nucleated cells. The filtrated aliquots results presented a significant increase of macrophages count and a significant decrease in neutrophils count comparing to the results of non-filtrated aliquots. These findings were consistent with results of filtered bronchoalveolar wash published studies. Therefore, the filtration of TBW is not an efficient method(AU)

O lavado traqueobrônquico (LTB) é um método usado para recuperar uma amostra de células das vias respiratórias. A citologia do LTB é descrita como uma importante ferramenta no diagnóstico de doença pulmonar em equinos. O excesso de muco geralmente presente no LTB pode alterar a morfologia, dificultando a identificação dos tipos celulares e, por consequência, prejudicando a contagem diferencial de células e a interpretação diagnóstica. O objetivo deste trabalho foi verificar o efeito da filtração do LTB de equinos sobre a contagem diferencial de células na avaliação citológica do lavado traqueobrônquico. Foram utilizados 30 equinos que foram submetidos a coletas de amostras de LTB via endoscópio. De cada amostra obtida foi separada uma primeira alíquota não filtrada (NF). Uma segunda alíquota (F) foi obtida filtrando-se o LTB, utilizando-se uma compressa de gaze hidrófila. As alíquotas F e NF foram submetidas à citocentrifugação para contagem diferencial de 300 células nucleadas. Verificou-se um aumento significativo no percentual de macrófagos e redução significativa de neutrófilos nas amostras filtradas. As alterações observadas foram condizentes com achados de estudos prévios realizados em lavado broncoalveolar. Conclui-se que a filtração do LTB não é uma metodologia aconselhável, pois acarreta em alterações de contagem significativas nas proporções de importantes marcadores celulares de inflamação, podendo gerar possíveis erros de diagnóstico(AU)

Equine tracheobronchial wash filtration and its effects on differential cell count / Efeito da filtração do lavado traqueobrônquico de equinos sobre a contagem diferencial de células

Tracheobronchial wash (TBW) is a method to recover cell samples from the airways. The cytology of TBW fluid is an important technique for the diagnosis of pulmonary diseases in horses. Excessive mucus in TBW may cause cell damage and morphological changes that hinder cell type recognition, resulting in a misdiagnosis. The aim of this study was to compare the results of differential cell count in a tracheobronchial wash of filtered and non-filtered samples. Endoscopy and TBW procedures were performed in thirty horses. Each TBW sample was split into two aliquots. Two groups were formed: non-filtrated aliquots (NF) and filtrated aliquots (F). The filtration was performed using a hydrophilic gauze pad. After centrifugation, the differential cell count was performed considering 300 nucleated cells. The filtrated aliquots results presented a significant increase of macrophages count and a significant decrease in neutrophils count comparing to the results of non-filtrated aliquots. These findings were consistent with results of filtered bronchoalveolar wash published studies. Therefore, the filtration of TBW is not an efficient method

O lavado traqueobrônquico (LTB) é um método usado para recuperar uma amostra de células das vias respiratórias. A citologia do LTB é descrita como uma importante ferramenta no diagnóstico de doença pulmonar em equinos. O excesso de muco geralmente presente no LTB pode alterar a morfologia, dificultando a identificação dos tipos celulares e, por consequência, prejudicando a contagem diferencial de células e a interpretação diagnóstica. O objetivo deste trabalho foi verificar o efeito da filtração do LTB de equinos sobre a contagem diferencial de células na avaliação citológica do lavado traqueobrônquico. Foram utilizados 30 equinos que foram submetidos a coletas de amostras de LTB via endoscópio. De cada amostra obtida foi separada uma primeira alíquota não filtrada (NF). Uma segunda alíquota (F) foi obtida filtrando-se o LTB, utilizando-se uma compressa de gaze hidrófila. As alíquotas F e NF foram submetidas à citocentrifugação para contagem diferencial de 300 células nucleadas. Verificou-se um aumento significativo no percentual de macrófagos e redução significativa de neutrófilos nas amostras filtradas. As alterações observadas foram condizentes com achados de estudos prévios realizados em lavado broncoalveolar. Conclui-se que a filtração do LTB não é uma metodologia aconselhável, pois acarreta em alterações de contagem significativas nas proporções de importantes marcadores celulares de inflamação, podendo gerar possíveis erros de diagnóstico

EQUINE TRACHEOBRONCHIAL WASH FILTRATION AND ITS EFFECTS ON DIFFERENTIAL CELL COUNT

Abstract Tracheobronchial wash (TBW) is a method to recover cell samples from the airways. The cytology of TBW fluid is an important technique for the diagnosis of pulmonary diseases in horses. Excessive mucus in TBW may cause cell damage and morphological changes that hinder cell type recognition, resulting in a misdiagnosis. The aim of this study was to compare the results of differential cell count in a tracheobronchial wash of filtered and non-filtered samples. Endoscopy and TBW procedures were performed in thirty horses. Each TBW sample was split into two aliquots. Two groups were formed: non-filtrated aliquots (NF) and filtrated aliquots (F). The filtration was performed using a hydrophilic gauze pad. After centrifugation, the differential cell count was performed considering 300 nucleated cells. The filtrated aliquots results presented a significant increase of macrophages count and a significant decrease in neutrophils count comparing to the results of non-filtrated aliquots. These findings were consistent with results of filtered bronchoalveolar wash published studies. Therefore, the filtration of TBW is not an efficient method.

Resumo O lavado traqueobrônquico (LTB) é um método usado para recuperar uma amostra de células das vias respiratórias. A citologia do LTB é descrita como uma importante ferramenta no diagnóstico de doença pulmonar em equinos. O excesso de muco geralmente presente no LTB pode alterar a morfologia, dificultando a identificação dos tipos celulares e, por consequência, prejudicando a contagem diferencial de células e a interpretação diagnóstica. O objetivo deste trabalho foi verificar o efeito da filtração do LTB de equinos sobre a contagem diferencial de células na avaliação citológica do lavado traqueobrônquico. Foram utilizados 30 equinos que foram submetidos a coletas de amostras de LTB via endoscópio. De cada amostra obtida foi separada uma primeira alíquota não filtrada (NF). Uma segunda alíquota (F) foi obtida filtrando-se o LTB, utilizando-se uma compressa de gaze hidrófila. As alíquotas F e NF foram submetidas à citocentrifugação para contagem diferencial de 300 células nucleadas. Verificou-se um aumento significativo no percentual de macrófagos e redução significativa de neutrófilos nas amostras filtradas. As alterações observadas foram condizentes com achados de estudos prévios realizados em lavado broncoalveolar. Conclui-se que a filtração do LTB não é uma metodologia aconselhável, pois acarreta em alterações de contagem significativas nas proporções de importantes marcadores celulares de inflamação, podendo gerar possíveis erros de diagnóstico.

Electrocardiographic evaluation of healthy dogs treated with ophthalmic solution of timolol maleate 0.5%: preliminary study / Avaliação eletrocardiográfica de cães clinicamente hígidos sob tratamento com solução oftálmica de maleato de timolol 0.5%: estudo preliminar

Timolol maleate 0.5% is a drug recommended for glaucoma treatment in dogs. After administration, it is absorbed to systemic circulation and being an antagonist of beta adrenergic receptors it has important systemic side effects on cardiac electrical conduction. The present study evaluated the modification caused by ophthalmic timolol 0.5% in the electrocardiogram. Six clinically normal dogs were selected to participate in two different ophthalmic treatments: in the first one, a placebo (hypromellose 0.5%) was used and in the second one, timolol 0.5% was administered. Each solution was applied twice a day, for 14 days. The electrocardiographic parameters were measured in times: zero, 10, 60, 120, 240, 360 and 720 min after ocular solution instillation in first, seventh and 14th days of each treatment. The differences found in electrocardiogram were more evident between 120 and 240 min after instillation of timolol 0.5% when compared with placebo treatment. The rhythm was irregular, with sinus arrhythmia and sinus bradycardia moments. The RR and PR intervals lengthen notoriously (p 0.05) from the first day of timolol administration, and are more expressive in the 14th day of treatment. The QT interval demonstrated a few changes, just lengthening noticeably (p 0.05) in the 14th day of timolol application. The QTc interval did not show expressive change. Despite the changes in the electrocardiogram, no clinical manifestation related to beta-adrenoceptor antagonists were observed. One must consider, however, that the animals studied were healthy and thus, clinical signs could result from the changes implied by the use of timolol in animals with pre-existing heart disease. Therefore, cardiac assessment of patients prior to prescription of ophthalmic timolol is recommended.

O maleato de timolol 0,5% é um fármaco recomendado para tratamento de glaucoma em cães. Após instilação, e absorvido para a circulação sistêmica e por ser um antagonista beta-adrenérgico pode promover efeitos colaterais sistêmicos importantes sobre a condução elétrica cardíaca. No presente estudo, foi avaliada a alteração causada pelo timolol 0,5% oftálmico no eletrocardiograma. Foram selecionados seis cães hígidos para participar de dois tratamentos oftálmicos diferentes: no primeiro foi instilado placebo (hipromelose 0,5%) e no segundo utilizou-se timolol 0,5%. O colírio foi instilado a cada 12 horas por 14 dias. Os parâmetros eletrocardiográficos foram mensurados nos tempos: zero, 10, 60, 120, 240, 360 e 720 minutos após instilação da solução ocular nos dias primeiro, sétimo e décimo quarto de cada tratamento. As alterações eletrocardiográficas foram mais evidentes entre 120 e 240 minutos pós-instilação de timolol 0,5% quando comparado com o tratamento placebo. O ritmo foi irregular, com momentos de arritmia sinusal e bradicardia sinusal. Os intervalos RR e PR prolongaram significativamente (p 0,05) desde o primeiro dia de instilação de timolol, sendo o prolongamento mais expressivo no décimo quarto dia de tratamento. O intervalo QT demonstrou pouca variação, apenas prolongando significativamente (p 0,05) no décimo quarto dia de aplicação de timolol. O intervalo QTc não demonstrou alteração significativa (p > 0,05). Apesar das alterações encontradas, não foram observadas manifestações clínicas relacionadas ao timolol nos animais estudados. Deve-se considerar, porém, que os animais em questão eram hígidos e, portanto, as alterações decorrentes do uso do timolol em animais com cardiopatias preexistentes poderiam promover sinais clínicos, sendo recomendada a avaliação cardíaca de pacientes previamente à prescrição do timolol oftálmico.

Electrocardiographic evaluation of healthy dogs treated with ophthalmic solution of timolol maleate 0.5%: preliminary study / Avaliação eletrocardiográfica de cães clinicamente hígidos sob tratamento com solução oftálmica de maleato de timolol 0.5%: estudo preliminar

Timolol maleate 0.5% is a drug recommended for glaucoma treatment in dogs. After administration, it is absorbed to systemic circulation and being an antagonist of beta adrenergic receptors it has important systemic side effects on cardiac electrical conduction. The present study evaluated the modification caused by ophthalmic timolol 0.5% in the electrocardiogram. Six clinically normal dogs were selected to participate in two different ophthalmic treatments: in the first one, a placebo (hypromellose 0.5%) was used and in the second one, timolol 0.5% was administered. Each solution was applied twice a day, for 14 days. The electrocardiographic parameters were measured in times: zero, 10, 60, 120, 240, 360 and 720 min after ocular solution instillation in first, seventh and 14th days of each treatment. The differences found in electrocardiogram were more evident between 120 and 240 min after instillation of timolol 0.5% when compared with placebo treatment. The rhythm was irregular, with sinus arrhythmia and sinus bradycardia moments. The RR and PR intervals lengthen notoriously (p 0.05) from the first day of timolol administration, and are more expressive in the 14th day of treatment. The QT interval demonstrated a few changes, just lengthening noticeably (p 0.05) in the 14th day of timolol application. The QTc interval did not show expressive change. Despite the changes in the electrocardiogram, no clinical manifestation related to beta-adrenoceptor antagonists were observed. One must consider, however, that the animals studied were healthy and thus, clinical signs could result from the changes implied by the use of timolol in animals with pre-existing heart disease. Therefore, cardiac assessment of patients prior to prescription of ophthalmic timolol is recommended. (AU) 

O maleato de timolol 0,5% é um fármaco recomendado para tratamento de glaucoma em cães. Após instilação, e absorvido para a circulação sistêmica e por ser um antagonista beta-adrenérgico pode promover efeitos colaterais sistêmicos importantes sobre a condução elétrica cardíaca. No presente estudo, foi avaliada a alteração causada pelo timolol 0,5% oftálmico no eletrocardiograma. Foram selecionados seis cães hígidos para participar de dois tratamentos oftálmicos diferentes: no primeiro foi instilado placebo (hipromelose 0,5%) e no segundo utilizou-se timolol 0,5%. O colírio foi instilado a cada 12 horas por 14 dias. Os parâmetros eletrocardiográficos foram mensurados nos tempos: zero, 10, 60, 120, 240, 360 e 720 minutos após instilação da solução ocular nos dias primeiro, sétimo e décimo quarto de cada tratamento. As alterações eletrocardiográficas foram mais evidentes entre 120 e 240 minutos pós-instilação de timolol 0,5% quando comparado com o tratamento placebo. O ritmo foi irregular, com momentos de arritmia sinusal e bradicardia sinusal. Os intervalos RR e PR prolongaram significativamente (p 0,05) desde o primeiro dia de instilação de timolol, sendo o prolongamento mais expressivo no décimo quarto dia de tratamento. O intervalo QT demonstrou pouca variação, apenas prolongando significativamente (p 0,05) no décimo quarto dia de aplicação de timolol. O intervalo QTc não demonstrou alteração significativa (p > 0,05). Apesar das alterações encontradas, não foram observadas manifestações clínicas relacionadas ao timolol nos animais estudados. Deve-se considerar, porém, que os animais em questão eram hígidos e, portanto, as alterações decorrentes do uso do timolol em animais com cardiopatias preexistentes poderiam promover sinais clínicos, sendo recomendada a avaliação cardíaca de pacientes previamente à prescrição do timolol oftálmico. (AU)

Atividade da fosfatase alcalina no lavado broncoalveolar de equinos de policiamento montado no Estado do Rio de Janeiro / Alkaline phosphatase activity in bronchoalveolar lavage of police horses in Rio de Janeiro State, Brazil

A utilidade da determinação das atividades enzimáticas no trato respiratório posterior como ferramenta diagnóstica já foi demonstrada em várias espécies. Nesse contexto, este trabalho teve por objetivo determinar a atividade da Fosfatase Alcalina (FAL) no lavado broncoalveolar (LBA) de equinos da Polícia Militar do Estado do Rio de Janeiro, comparando animais sadios com portadores assintomáticos de doença inflamatória das vias aéreas (DIVA). Para tal, foram avaliados 28 animais adultos, machos, sem histórico de doença respiratória nos dois meses anteriores ao estudo, com os resultados dos exames físicos e laboratoriais (FAL sanguínea, hematócrito, leucograma, proteína total e fibrinogênio plasmáticos) dentro dos parâmetros fisiológicos. Os equinos foram divididos em dois grupos de acordo com o resultado da citologia broncoalveolar. A determinação da atividade da FAL foi realizada por meio de espectrofotometria a partir de alíquotas do sobrenadante do LBA preservadas em nitrogênio líquido. Para a estimativa do fluido epitelial pulmonar e da atividade da FAL neste, foi realizada a correção da diluição provocada pelo lavado. Os equinos com contagem diferencial de tipos celulares compatível com DIVA apresentaram atividade de FAL no LBA menor, quando comparados aos animais sadios, podendo essa dosagem ser utilizada como complementação do diagnóstico da DIVA.

The use of determining the enzymatic activities in the posterior respiratory tract as a diagnostic tool has already been demonstrated in several species. In this context, this paper aims to determine the activity of alkaline phosphatase (ALP) in the bronchoalveolar lavage (BAL) of horses from the Military Police of the State of Rio de Janeiro, comparing healthy animals with asymptomatic carriers of an inflammatory airway disease (IAD). Twenty-eight adult male animals with no history of respiratory diseases in the last two months prior to the study were studied. Physical exam and blood laboratory test results (ALP, hematocrit, leukogram, total protein and plasma fibrinogen) were within physiological parameters. The equines were separated into two groups according to the results of the bronchoalveolar cytology. The determination of ALP was done by spectrophotometry with aliquots of the supernatant of the BAL preserved in liquid nitrogen. To estimate pulmonary epithelial lining fluid and ALP activity, correction of the dilution caused by the lavage was done. The horses with a cell type differential count compatible with IAD presented a lower ALP activity in BAL when compared to healthy animals, therefore this dosage can be used as a complement in the diagnosis of IAD.

Atividade da fosfatase alcalina no lavado broncoalveolar de equinos de policiamento montado no Estado do Rio de Janeiro / Alkaline phosphatase activity in bronchoalveolar lavage of police horses in Rio de Janeiro State, Brazil

A utilidade da determinação das atividades enzimáticas no trato respiratório posterior como ferramenta diagnóstica já foi demonstrada em várias espécies. Nesse contexto, este trabalho teve por objetivo determinar a atividade da Fosfatase Alcalina (FAL) no lavado broncoalveolar (LBA) de equinos da Polícia Militar do Estado do Rio de Janeiro, comparando animais sadios com portadores assintomáticos de doença inflamatória das vias aéreas (DIVA). Para tal, foram avaliados 28 animais adultos, machos, sem histórico de doença respiratória nos dois meses anteriores ao estudo, com os resultados dos exames físicos e laboratoriais (FAL sanguínea, hematócrito, leucograma, proteína total e fibrinogênio plasmáticos) dentro dos parâmetros fisiológicos. Os equinos foram divididos em dois grupos de acordo com o resultado da citologia broncoalveolar. A determinação da atividade da FAL foi realizada por meio de espectrofotometria a partir de alíquotas do sobrenadante do LBA preservadas em nitrogênio líquido. Para a estimativa do fluido epitelial pulmonar e da atividade da FAL neste, foi realizada a correção da diluição provocada pelo lavado. Os equinos com contagem diferencial de tipos celulares compatível com DIVA apresentaram atividade de FAL no LBA menor, quando comparados aos animais sadios, podendo essa dosagem ser utilizada como complementação do diagnóstico da DIVA.(AU)

The use of determining the enzymatic activities in the posterior respiratory tract as a diagnostic tool has already been demonstrated in several species. In this context, this paper aims to determine the activity of alkaline phosphatase (ALP) in the bronchoalveolar lavage (BAL) of horses from the Military Police of the State of Rio de Janeiro, comparing healthy animals with asymptomatic carriers of an inflammatory airway disease (IAD). Twenty-eight adult male animals with no history of respiratory diseases in the last two months prior to the study were studied. Physical exam and blood laboratory test results (ALP, hematocrit, leukogram, total protein and plasma fibrinogen) were within physiological parameters. The equines were separated into two groups according to the results of the bronchoalveolar cytology. The determination of ALP was done by spectrophotometry with aliquots of the supernatant of the BAL preserved in liquid nitrogen. To estimate pulmonary epithelial lining fluid and ALP activity, correction of the dilution caused by the lavage was done. The horses with a cell type differential count compatible with IAD presented a lower ALP activity in BAL when compared to healthy animals, therefore this dosage can be used as a complement in the diagnosis of IAD.(AU)

Comparison between linear smear concentration and cytocentrifugation techniques to evaluate equine bronchoalveolar lavage cytology / Comparação entre as técnicas de esfregaço linear e citocentrifugação para avaliação citológica do lavado broncoalveolar equino

The bronchoalveolar lavage (BAL) is a sensitive method to diagnose diseases of the distal portion of the lower respiratory tract and has been broadly used by numerous researchers. Cytocentrifugation is the choice cytological preparation technique, but demands specific and costly equipment. Therefore, the present paper intends to verify the applicability of the linear smear technique to evaluate BAL samples. For this, BAL samples of 30 equines were used and the cytological preparations were done by cytocentrifugation and linear smear techniques. All glass microscope slides were fixed and stained with Giemsa for the differential cell count. Regarding the effect of the preparation technique on differential counts, no significant difference in any cell type was found. The linear smear is a reliable alternative and can be recommended as a substitution to cytocentrifugation.(AU)

O lavado broncoalveolar (LBA) é um método sensível para diagnosticar doenças do trato respiratório posterior e vem sendo utilizado por diversos pesquisadores. A citocentrifugação, técnica de escolha para processar amostras citológicas de LBA, exige equipamentos específicos e caros. Por isso, este trabalho verificou a aplicabilidade da técnica de esfregaço linear para avaliação citológica do LBA. Foram utilizadas amostras de LBA de 30 equinos adultos. As preparações citológicas foram realizadas tanto por citocentrifugação quanto por esfregaço linear. Todas as lâminas foram fixadas e coradas com Giemsa para realização da contagem celular diferencial. Não foram encontradas alterações morfológicas significativas e nem diferenças estatísticas entre nenhum dos tipos celulares processados pelos dois métodos, o que permite afirmar que o método de esfregaço linear é uma alternativa segura para avaliação morfológica celular do LBA de equinos, podendo ser utilizado no lugar da citocentrifugação quando esta não estiver disponível.(AU)

Comparison between linear smear concentration and cytocentrifugation techniques to evaluate equine bronchoalveolar lavage cytology / Comparação entre as técnicas de esfregaço linear e citocentrifugação para avaliação citológica do lavado broncoalveolar equino

The bronchoalveolar lavage (BAL) is a sensitive method to diagnose diseases of the distal portion of the lower respiratory tract and has been broadly used by numerous researchers. Cytocentrifugation is the choice cytological preparation technique, but demands specific and costly equipment. Therefore, the present paper intends to verify the applicability of the linear smear technique to evaluate BAL samples. For this, BAL samples of 30 equines were used and the cytological preparations were done by cytocentrifugation and linear smear techniques. All glass microscope slides were fixed and stained with Giemsa for the differential cell count. Regarding the effect of the preparation technique on differential counts, no significant difference in any cell type was found. The linear smear is a reliable alternative and can be recommended as a substitution to cytocentrifugation.

O lavado broncoalveolar (LBA) é um método sensível para diagnosticar doenças do trato respiratório posterior e vem sendo utilizado por diversos pesquisadores. A citocentrifugação, técnica de escolha para processar amostras citológicas de LBA, exige equipamentos específicos e caros. Por isso, este trabalho verificou a aplicabilidade da técnica de esfregaço linear para avaliação citológica do LBA. Foram utilizadas amostras de LBA de 30 equinos adultos. As preparações citológicas foram realizadas tanto por citocentrifugação quanto por esfregaço linear. Todas as lâminas foram fixadas e coradas com Giemsa para realização da contagem celular diferencial. Não foram encontradas alterações morfológicas significativas e nem diferenças estatísticas entre nenhum dos tipos celulares processados pelos dois métodos, o que permite afirmar que o método de esfregaço linear é uma alternativa segura para avaliação morfológica celular do LBA de equinos, podendo ser utilizado no lugar da citocentrifugação quando esta não estiver disponível.

Análise do líquido broncoalveolar de equinos portadores de doença inflamatória das vias aéreas / Evaluation of bronchoalveolar fluid of horses with inflammatory airway disease

Objetivou-se caracterizar o perfil citológico broncoalveoar de equinos de policiamento portadores assintomáticos deDoença Inflamatória de Vias Aéreas (DIVA). Utilizaram-se 17 equinos adultos, machos e fêmeas, com idade entre 11 e24 anos. Os animais que constituíram o grupo controle (oito) apresentaram-se normais ao exame físico, à endoscopia eà mensuração da diferença máxima de pressão intrapleural. No lavado broncoalveolar a contagem deveria ser de até 4%,0,7%, 3,3% para neutrófilos, eosinófilos e células epiteliais e entre 17,0 e 56,7%, 29 e 75,7%, 0,7 e 13,7% para linfócitos,macrófagos e mastócitos, respectivamente. O grupo considerado doente (nove animais) apresentou achados compatíveiscom afecções do trato respiratório posterior, pelo menos à endoscopia, além de uma contagem de neutrófilos maiorque 5% e menor que 25%. O quadro assintomático de doença inflamatória das vias aéreas caracterizou-se por infiltradoneutrofílico, discreta redução no número de macrófagos, aumento no número de macrófagos espumosos, discretoinfiltrado eosinofílico e aumento marcante no número de células epiteliais. Em face do caráter assintomático dessaenfermidade nos equinos de policiamento, a utilização do LBA como método de diagnóstico e acompanhamento éfundamental, pois permite reconhecer, tratar e determinar uma utilização mais racional desses animais, de modo amelhorar a qualidade de vida e prolongar a utilização dos mesmos em serviço.(AU)

The aim of this study was to characterize the cytology profile of bronchoalveolar lavage from police horses withasymptomatic Inflammatory Airway Disease (IAD). Seventeen adult horses, males and females, with ages ranging from11 to 24 years old were utilized. The control group (eight horses) presented vital signs within the physiologic range,normal pulmonary percussion and auscultation, no abnormalities at respiratory endoscopy and at the measurement ofthe maximal intra-pleural pressure difference. The cell count of bronchoalveolar lavage (BAL) should be up to 4%, 0.7%,3.3% to neutrophil, eosinophil and epithelial cell, and ranging of 17.0 to 56.7%, 29 to 75.7%, 0.7 to 13.7% to lymphocytes,macrophages and mast cell, respectively. The diseased group (nine animals) presented findings compatible with lowerairway disease, at least during endoscopy and showed a neutrophil counts above 5% and below 25%. AsymptomaticIAD was characterized by neutrophilic infiltrate, discrete reduction in macrophage count with increased number offoamy macrophages and discrete eosinophilic infiltrate and marked increase in the number of epithelial cells. Due to theasymptomatic character of IAD in police horses, BAL is the preferred diagnostic and follow-up method since it allowsrecognition of the disease, evaluation of treatment and helps the implementation of a graded and rational work regimethat aims at improving welfare of those animals as well as extending their usage as patrolling horses.(AU)

Análise do líquido broncoalveolar de equinos portadoresde doença inflamatória das vias aéreas / Evaluation of bronchoalveolar fluid of horses with inflammatory airway disease

Objetivou-se caracterizar o perfil citológico broncoalveoar de equinos de policiamento portadores assintomáticos deDoença Inflamatória de Vias Aéreas (DIVA). Utilizaram-se 17 equinos adultos, machos e fêmeas, com idade entre 11 e24 anos. Os animais que constituíram o grupo controle (oito) apresentaram-se normais ao exame físico, à endoscopia eà mensuração da diferença máxima de pressão intrapleural. No lavado broncoalveolar a contagem deveria ser de até 4%,0,7%, 3,3% para neutrófilos, eosinófilos e células epiteliais e entre 17,0 e 56,7%, 29 e 75,7%, 0,7 e 13,7% para linfócitos,macrófagos e mastócitos, respectivamente. O grupo considerado doente (nove animais) apresentou achados compatíveiscom afecções do trato respiratório posterior, pelo menos à endoscopia, além de uma contagem de neutrófilos maiorque 5% e menor que 25%. O quadro assintomático de doença inflamatória das vias aéreas caracterizou-se por infiltradoneutrofílico, discreta redução no número de macrófagos, aumento no número de macrófagos espumosos, discretoinfiltrado eosinofílico e aumento marcante no número de células epiteliais. Em face do caráter assintomático dessaenfermidade nos equinos de policiamento, a utilização do LBA como método de diagnóstico e acompanhamento éfundamental, pois permite reconhecer, tratar e determinar uma utilização mais racional desses animais, de modo amelhorar a qualidade de vida e prolongar a utilização dos mesmos em serviço.

The aim of this study was to characterize the cytology profile of bronchoalveolar lavage from police horses withasymptomatic Inflammatory Airway Disease (IAD). Seventeen adult horses, males and females, with ages ranging from11 to 24 years old were utilized. The control group (eight horses) presented vital signs within the physiologic range,normal pulmonary percussion and auscultation, no abnormalities at respiratory endoscopy and at the measurement ofthe maximal intra-pleural pressure difference. The cell count of bronchoalveolar lavage (BAL) should be up to 4%, 0.7%,3.3% to neutrophil, eosinophil and epithelial cell, and ranging of 17.0 to 56.7%, 29 to 75.7%, 0.7 to 13.7% to lymphocytes,macrophages and mast cell, respectively. The diseased group (nine animals) presented findings compatible with lowerairway disease, at least during endoscopy and showed a neutrophil counts above 5% and below 25%. AsymptomaticIAD was characterized by neutrophilic infiltrate, discrete reduction in macrophage count with increased number offoamy macrophages and discrete eosinophilic infiltrate and marked increase in the number of epithelial cells. Due to theasymptomatic character of IAD in police horses, BAL is the preferred diagnostic and follow-up method since it allowsrecognition of the disease, evaluation of treatment and helps the implementation of a graded and rational work regimethat aims at improving welfare of those animals as well as extending their usage as patrolling horses.

Estimativa do volume de fluido epitelial pulmonar recuperado no lavado broncoalveolar no equino / Estimation of volume of epithelial lining fluid recovered by bronchoalveolar lavage in equine

O lavado broncoalveolar (LBA ) é uma técnica que permite coletar o fluido epitelial pulmonar (FEP) e avaliar a integridade do trato respiratório posterior. A instilação de solução isotônica para a sua realização resulta numa diluição significativa do FEP. A ureia é comumente utilizada como marcador de diluição, em virtude do seu livre trânsito pelas membranas biológicas. Contudo, a metodologia previamente estabelecida que faz uso dela é de difícil reprodução, principalmente pela utilização de reagentes e equipamentos de difícil obtenção. O objetivo deste trabalho foi descrever uma metodologia para este cálculo utilizando um kit comercial de determinação de ureia (ureia UV LabTest®). Foram obtidas amostras pareadas de sangue e de lavado broncoalveolar de dezoito equinos adultos. A partir de concentrações conhecidas de ureia no LBA de equinos, foi estabelecida uma equação de regressão linear e sua curva-padrão, utilizadas para cálculo das concentrações nas amostras de LBA. O volume estimado de FEP teve média de 0,52±0,12mL. O percentual médio obtido de FEP em relação ao volume recuperado de LBA foi de 0,39±0,08%. Estes resultados estão compatíveis com os descritos, provando que a metodologia desenvolvida permitiu obter resultados confiáveis, podendo ser empregada para estimar o volume de FEP e, consequentemente, calcular a concentração ou atividade de componentes desejados.(AU)

Bronchoalveolar lavage (BAL ) is a widespread technique that allows collection of pulmonary epithelial lining fluid (PELF) and evaluation of integrity of the lower airways. The instillation of isotonic solution needed for collection results in significant dilution of the PELF . Due to its ability to freely cross biologic membranes urea is commonly used as a marker of PELF dilution. Nevertheless, the previously established methodology is difficult to be reproduced, especially since it utilizes reagents and equipments that are difficult to obtain. The objective of this study was to describe a methodology for measuring urea utilizing a commercial kit (Ureia UVL abTest®). Paired samples of blood and BALF were obtained from 18 adult horses. A standard curve of known urea dilutions in equine BALF provided a linear regression equation that allowed calculation of unknown urea concentrations in BALF samples. The mean estimated PELF volume was 0.52±0.12mL. The mean PELF percentage in relation to recovery BALF was 0.39±0.08%. These results are compatible with the ones previously described, demonstrating that the methodology presented here produced reliable results and can be utilized to estimate the PELF volume and consequently calculate concentration or activity of components or substances of interest.(AU)

Estimativa do volume de fluido epitelial pulmonar recuperado no lavado broncoalveolar no equino / Estimation of volume of epithelial lining fluid recovered by bronchoalveolar lavage in equine

O lavado broncoalveolar (LBA ) é uma técnica que permite coletar o fluido epitelial pulmonar (FEP) e avaliar a integridade do trato respiratório posterior. A instilação de solução isotônica para a sua realização resulta numa diluição significativa do FEP. A ureia é comumente utilizada como marcador de diluição, em virtude do seu livre trânsito pelas membranas biológicas. Contudo, a metodologia previamente estabelecida que faz uso dela é de difícil reprodução, principalmente pela utilização de reagentes e equipamentos de difícil obtenção. O objetivo deste trabalho foi descrever uma metodologia para este cálculo utilizando um kit comercial de determinação de ureia (ureia UV LabTest®). Foram obtidas amostras pareadas de sangue e de lavado broncoalveolar de dezoito equinos adultos. A partir de concentrações conhecidas de ureia no LBA de equinos, foi estabelecida uma equação de regressão linear e sua curva-padrão, utilizadas para cálculo das concentrações nas amostras de LBA. O volume estimado de FEP teve média de 0,52±0,12mL. O percentual médio obtido de FEP em relação ao volume recuperado de LBA foi de 0,39±0,08%. Estes resultados estão compatíveis com os descritos, provando que a metodologia desenvolvida permitiu obter resultados confiáveis, podendo ser empregada para estimar o volume de FEP e, consequentemente, calcular a concentração ou atividade de componentes desejados.

Bronchoalveolar lavage (BAL ) is a widespread technique that allows collection of pulmonary epithelial lining fluid (PELF) and evaluation of integrity of the lower airways. The instillation of isotonic solution needed for collection results in significant dilution of the PELF . Due to its ability to freely cross biologic membranes urea is commonly used as a marker of PELF dilution. Nevertheless, the previously established methodology is difficult to be reproduced, especially since it utilizes reagents and equipments that are difficult to obtain. The objective of this study was to describe a methodology for measuring urea utilizing a commercial kit (Ureia UVL abTest®). Paired samples of blood and BALF were obtained from 18 adult horses. A standard curve of known urea dilutions in equine BALF provided a linear regression equation that allowed calculation of unknown urea concentrations in BALF samples. The mean estimated PELF volume was 0.52±0.12mL. The mean PELF percentage in relation to recovery BALF was 0.39±0.08%. These results are compatible with the ones previously described, demonstrating that the methodology presented here produced reliable results and can be utilized to estimate the PELF volume and consequently calculate concentration or activity of components or substances of interest.