Bilateral bronchoalveolar lavage cytology profiles in a warmblood horse population during a 1-year period.

BACKGROUND: Bronchoalveolar lavage (BAL) cytology results from 1 lung might not be representative of both lungs. OBJECTIVES: To determine whether the lung site sampled would influence the horse's BAL cytology profile, and if a pooled BAL sample would be superior with regard to BAL cytology diagnosis in a cohort of healthy and subclinical asthmatic warmblood horses. ANIMALS: Fifty-nine horses in 2021 and 70 horses in 2022, the follow-up included 53 of the same in each year. METHODS: A cross-sectional study with follow-up included BAL cytology samples from individual lungs and from pooled BAL samples. The BAL samples were enumerated and differential cell count were applied to categorize the horses as control or with airway inflammation (AI). RESULTS: Bronchoalveolar lavage mast cell count was higher in left lung compared to right lung (2021; median 1.6 [range, 0.6-3.3] vs 1.2 [0.7-1.5] P = .009, 2022; median 3.1 [2.1-4.2] vs 2.4 [1.7-3.4], P < .001) and compared to pooled samples (2022; median 2.6 [1.7-3.7], P < .001). Between year 2021 and 2022, 17 of the horses had changes in BAL cytology from control to AI or vice versa. CONCLUSIONS AND CLINICAL IMPORTANCE: Pooled BAL sample was the least reliable for detecting AI, and was not representative of the overall lung condition.

Differential cell analysis in ovine bronchoalveolar lavage fluid: Effect of different sampling methods and seasons.

Background: Bronchoalveolar lavage (BAL) is a method for collecting the cellular and fluid components of the airway surface in the lungs. The assessment of differential cell profiles is potentially valuable in the diagnosis of pulmonary diseases, but there is no information about the normal BAL profiles in the Gezel breed. Aim: This study aimed to characterize the normal cryptologic findings of BAL with Gezel sheep. Methods: Twenty healthy sheep (15 females: 5 males, bodyweight: 55-65 kg) were sedated with xylazine (0.02-0.04 mg/kg IV). Two methods; the transtracheal bronchoalveolar lavage technique and the bronchoscopic bronchoalveolar lavage using a scope were evaluated. Sampling was performed in the summer and winter seasons. Results: Normal value (Mean ± SEM) for total cell, macrophage, lymphocyte, neutrophil, epithelial, and bronchial cells in BAL sampled in summer were (343.75 ± 30.23), ) 24.50 ± 2.62), (2.81 ± 0.51), (1.43 ± 0.88), and (3.12 ± 0.32), respectively. The normal values for the total cell, macrophage, lymphocyte, neutrophil, epithelial, and bronchial cells in BAL sampled in winter were (355.55 ± 37.67), (59.11 ± 4.30), (21.33 ± 3.10), (3.88 ± 1.07), (8.88 ± 3.78), and (6.33 ± 1.44), respectively. Conclusion: No significant change in the percentage of neutrophils was detected between seasons, although the percentages of bronchial and epithelial cells in winter were significantly high (p < 0.05). Except for the mentioned cases, neither the total cell number nor the percentage differential cell populations of BAL changed significantly (p < 0.05) in different sampling methods and seasons. Normal BAL profiles in the Gezel breed were determined and could be used in result interpretations. Also, both sampling methods can be used without significantly affecting the results.

Three-minute nebulization of gentamicin in healthy dogs results in therapeutic concentrations in bronchoalveolar lavage fluid while remaining below the toxic range values in blood.

OBJECTIVE: To determine and compare the concentration of gentamicin in the lower airways and serum of healthy spontaneously breathing dogs after nebulization with 5% undiluted gentamicin during 3 versus 10 minutes. ANIMALS: 10 healthy experimental Beagles. METHODS: This was a prospective crossover study. A standardized bronchoalveolar lavage (BAL) procedure was performed in each dog after 1 week of administration of each of 2 different gentamicin nebulization protocols separated by a 1-week washout period. The 2 protocols consisted of nebulization of 5% undiluted gentamicin (50 mg/mL) twice daily either during 10 minutes per session (± 95 mg; 10-minute protocol) or 3 minutes per session (± 28 mg; 3-minute protocol). BAL fluid (BALF) was obtained under general anesthesia using a bronchoscope within 15 minutes after administration of the last nebulization. Blood was collected within 5 minutes after BALF collection. BALF and serum gentamicin concentrations were determined by particle-enhanced turbidimetric inhibition immunoassay. Concentrations between protocols were compared using a paired t test. RESULTS: Both BALF and serum gentamicin concentrations were higher after the 10-minute protocol compared with the 3-minute protocol (mean ± SD: 2.41 ± 0.87 mg/L vs 1.25 ± 0.31 mg/L, P = .001; and 1.02 ± 0.59 mg/L vs 0.31 ± 0.24 mg/L, P < .0001 in BALF and serum, respectively), while the BALF-to-serum ratio did not differ between the protocols (3.75 [1.37 to 5.75] (median [IQR]) in the 3-minute protocol vs 2.48 [2.02 to 2.67] in the 10-minute protocol; P = .754). CLINICAL RELEVANCE: A 3-minute nebulization of gentamicin seems to achieve sufficient concentrations of gentamicin in the BALF to have good efficacy against aminoglycoside-sensitive bacteria while remaining below the toxic range values in blood.

Cytokine mRNA expression in the bronchoalveolar lavage cells from horses affected by different equine asthma subtypes.

Equine asthma (EA) is a respiratory syndrome associated with the increase of different leukocyte populations in the bronchoalveolar lavage fluid (BALF). Its pathogenetic mechanisms remain unclear. This study aimed to evaluate the associations between the mRNA expression of different cytokines in the BALF, different EA subtypes and lung function. Fifteen horses underwent physical examination, airway endoscopy, BALF cytology and lung function testing (8/15). One horse did not have evidence of EA and was used as healthy reference, while the others were classified as affected by neutrophilic or mixed granulocytic EA. Cells isolated from the residual BALF were used for IL-1ß, IL-2, IFN-γ, IL-4, IL-17A genes expression by quantitative RT-PCR., Cytokine expression was compared between groups, and their correlations with BALF leukocyte and lung function were evaluated. IL-1ß expression was positively correlated with BALF neutrophils count (p=0.038, r=0.56) and with increased expiratory resistance (p=0.047, r=0.76). IFN-γ was correlated with BALF mast cells (p=0.029, r=0.58). IL-4 was higher in horses with mixed granulocytic EA than neutrophilic (p=0.008), positively correlated with BALF mast cells (p=0.028, r=0.59) and inversely with whole-breath (p=0.046, r=-0.76) and expiratory reactance (p=0.003, r=-0.93). Finally, IL-17A was inversely correlated with expiratory reactance (p=0.009, r=-0.92). These results support that multiple immune responses are involved in EA pathogenesis; innate, Th2, and Th17 responses. Innate immunity appeared associated with neutrophilic inflammation, and Th2 response with increased mast cells. The role of Th1 response in EA remains questionable.

Cytological diagnosis of exercise-induced pulmonary haemorrhage: Comparison of tracheal wash and bronchoalveolar lavage in standardbred racehorses.

BACKGROUND: Cytology of airway samples is sensitive for diagnosis of exercise-induced pulmonary haemorrhage (EIPH), but the association between tracheal wash (TW) and bronchoalveolar lavage fluid (BALF) is unknown. The objective of this study was to determine whether diagnosis of EIPH, using haemosiderophages/macrophages (H/M) ratio, differs when based on TW or BALF. METHODS: A prospective cross-sectional study was conducted on 102 standardbred horses in training. TW and BALF were collected concomitantly from all horses at rest (at least 24 hours after their last training or race), and their H/M ratios were calculated. Spearman's correlation, Cohen's kappa and Gwet's coefficient tests were performed to evaluate the association between TW and BALF samples. RESULTS: With BALF, 21 horses met the cytological inclusion criteria for an EIPH diagnosis from individual and/or pooled samples. With TW, 20 horses had occasional (H/M < 10%) haemosiderophages, and nine, one and three horses had small (10%-25%), moderate (25%-50%) and large (>50%) proportions, respectively. Poor correlations and inconsistent concordances between TW and BALF were found for H/M ratio. LIMITATIONS: Limitations include the use of a single staining method and the absence of a total haemosiderin score. CONCLUSION: No association between TW and BALF was found for the cytological diagnosis of EIPH. Based on H/M ratio, BALF remains the sample type of choice for cytological diagnosis of EIPH.

Acute-phase protein concentration in bronchoalveolar lavage fluid from healthy horses / Proteínas de fase aguda do lavado broncoalveolar de equinos hígidos

Bronchoalveolar lavage fluid (BALF) was analyzed to obtain information on leakage of acute-phase proteins from the blood into the respiratory lumen and about local synthesis. Ceruloplasmin, transferrin, albumin, α1-antitripsin, immunoglobulin G heavy, immunoglobulin G light, immunoglobulin A, haptoglobin, acidic glycoprotein, and P23 were measured in BALF from 30 horses without inflammatory disease by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). In serum, the same proteins were identified except for α1-antitrypsin. In conclusion, this study demonstrated that polyacrylamide gel electrophoresis (SDS-PAGE) can be used for the determination of acute-phase proteins in BALF samples from horses. In healthy horses, the values are very low, but they can be compared with reference values to assist in the diagnosis of animals with respiratory diseases.(AU)

O líquido obtido através da lavagem broncoalveolar (LBA) foi analisado para obter informações sobre as proteínas da fase aguda. Ceruloplasmina, transferrina, albumina, α1-antitripsina, imunoglobulina G pesada, imunoglobulina G leve, imunoglobulina A, haptoglobina, glicoproteína ácida e P23 foram medidas nos LBA de 30 cavalos sem doença inflamatória por eletroforese em gel de poliacrilamida com dodecilsulfato de sódio (SDS-PAGE). No soro, as mesmas proteínas foram identificadas, exceto a α1-antitripsina. Em conclusão, este estudo demonstra que a eletroforese em gel de poliacrilamida (SDS-PAGE) pode ser usada para a determinação de proteínas de fase aguda em amostras de LBA em cavalos. Em cavalos saudáveis, os valores são muito baixos, no entanto, podem ser comparados e auxiliar no diagnóstico de animais com doenças respiratórias.(AU)

Efeitos da estabulação sobre o trato respiratório de equinos em dois tipos diferentes de camas e a pasto / Stabling effects on equine respiratory tract using two different types of stable beds and pasture

As afecções do sistema respiratório são a segunda causa de queda de desempenho e afastamento dos equinos do esporte ou trabalho, as quais estão atrás somente das desordens musculoesqueléticas. A estabulação e a alimentação à base de fenos predispõem os equinos à inalação de grande número de agentes irritantes às vias aéreas. No intuito de avaliar a influência dos tipos de cama utilizados em baias, estudaram-se três grupos de cinco equinos cada, durante 45 dias. Dois grupos foram mantidos em cocheiras com forrações diferentes à base de madeira (maravalha de pínus autoclavada-MA e serragem-SE) e o terceiro grupo permaneceu no pasto. Foram realizadas avaliações quinzenais, que incluíram exame físico, hemograma, hemogasometria e lavado broncoalveolar. Ao exame físico, notou-se a presença de tosse e espirros em animais do grupo SE. No lavado broncoalveolar, a contagem de células nucleadas totais apresentou maiores valores nos animais a pasto, seguido do grupo SE e, por último, o MA. Concluiu-se que as duas camas testadas neste estudo não apresentaram diferença com relação à capacidade irritativa das vias aéreas superiores no período analisado.(AU)

Respiratory system affections in horses are the second cause of decrease in performance and removal from the sport or farm work, which are behind only the musculoskeletal disorders. Stabling and hay feeding predisposes horses' airways to inhalation of large numbers of irritating agents. To evaluate the influence of bed types used in bays, 3 groups of 5 horses each were studied for 45 days. Two groups were kept in stables with different wood beds - based rafters (autoclaved pine - MA and sawdust - SE) and the third group remained in the pasture. Biweekly evaluations with physical examination, hemogram, hemogasometry and bronchoalveolar lavage were performed. On physical examination, the presence of cough and sneezing was observed in animals of the SE group. In the bronchoalveolar lavage, the total nucleated cell count presented higher values in the animals in the pasture, followed by the SE group and finally the MA. It was concluded that the different beds used presented no difference with respect to the irritative capacity of the upper airways.(AU)

Montelukast, Leukotriene Inhibitor, Reduces LPS-Induced Acute Lung Inflammation and Human Neutrophil Activation / El montelukast, un inhibidor de leucotrienos, reduce la inflamación pulmonar aguda inducida por LPS y la activación de neutrófilos humanos

Objectives: Some pro-inflammatory lipids derived from 1 lipooxygenase enzyme are potent neutrophil chemoattractant, a cell centrally involved in acute respiratory distress syndrome (ARDS); a syndrome lacking effective treatment. Considering the beneficial effects of the leukotriene receptor inhibitor, montelukast, on other lung diseases, whether montelukast attenuates inflammation in a mouse model of ARDS, and whether it reduces LPS stimulated activation of human neutrophils was investigated. Methods: Thirty-five C57Bl/6 mice were distributed into control (PBS) + 24h, LPS + 24h (10 μg/mouse), control + 48 h, LPS+48 h, and LPS 48 h+Montelukast (10 mg/kg). In addition, human neutrophils were incubated with LPS ( 1μg/mL) and treated with montelukast (10 μM). Results: Oral-tracheal administration of montelukast significantly attenuated total cells (P < .05), macrophages (P < .05), neutrophils (P < .01), lymphocytes (P < .001) and total protein levels in BAL (P < .05), as well as IL-6 (P < .05), CXCL1/KC (P < .05), IL-17 (P < .05) and TNF-alfa (P < .05). Furthermore, montelukast reduced neutrophils (P < .001), lymphocytes (P < .01) and macrophages (P < .01) in the lung parenchyma. In addition, montelukast restored BAL VEGF levels (P < .05). LTB4 receptor expression (P < .001) as well as NF-κB (P <. 001), a downstream target of LPS, were also reduced in lung parenchymal leukocytes. Furthermore, montelukast reduced IL-8 (P < .001) production by LPS-treated human neutrophils. Conclusion: In conclusion, montelukast efficiently attenuated both LPS-induced lung inflammation in a mouse model of ARDS and in LPS challenged human neutrophils

Objetivos: Algunos lípidos proinflamatorios derivados de la enzima lipooxigenasa 1 son potentes quimioatrayentes de neutrófilos, un tipo celular con una implicación principal en el síndrome de distrés respiratorio agudo (SDRA), para el que no hay tratamiento efectivo. Considerando los efectos beneficiosos del inhibidor de los receptores de leucotrienos montelukast en otras enfermedades pulmonares, se investigó si este fármaco era capaz de atenuar la inflamación en un modelo de ratón de SDRA y de reducir la activación de los neutrófilos humanos inducida por LPS. Métodos: Se utilizaron 35 ratones C57BL/6 distribuidos en los siguientes grupos: control (PBS) + 24 h, LPS+(24 h [10 μg/ratón]), control + 48 h y LPS 48 h + montelukast (10 mg/kg). Por otro lado, se incubaron neutrófilos humanos con LPS (1 μg/ml) y se trataron con montelukast (10 μM). Resultados: La administración orotraqueal de montelukast redujo el número total de células (p < 0,05), de macrófagos (p < 0,05), de neutrófilos (p < 0,01), de linfocitos (p < 0,001) y los niveles totales de proteína en el lavado broncoalveolar (p < 0,05), así como de IL-6 (p < 0,05), CXCL1/KC (p < 0,05), IL-17 (p < 0,05) y TNF-alfa (p < 0,05). Además, el montelukast redujo los neutrófilos (p < 0,001), los linfocitos (p < 0,01) y los macrófagos (p < 0,01) en el parénquima pulmonar. Asimismo, restauró los niveles de VEGF en el lavado broncoalveolar (p < 0,05) y disminuyó la expresión del receptor LTB4 (p < 0,001) y de NF-κB (p < 0,001), una diana downstream del LPS, en los leucocitos del parénquima pulmonar. Por último, redujo la producción de IL-8 por parte de los neutrófilos humanos tratados con LPS. Conclusión: En conclusión, el montelukast atenuó de manera eficaz tanto la inflamación pulmonar inducida por LPS en un modelo de ratón de SDRA como en neutrófilos humanos estimulados con LPS

Leukotriene B4 in equine asthma syndrome: what do we know so far? / Leucotrieno B4 na síndrome asmática equina: o que sabemos até o momento?

The term "equine asthma syndrome" (EAS) was recently proposed due to the resemblance of the equine disease to human asthma. Leukotrienes cause constriction of the bronchi, especially in the lower airways and increase mucus secretion in the respiratory system. Leukotriene B4 (LTB4) has been discovered as a strong chemotactic factor, which plays a role in neutrophil migration. The immunologic background of EAS remains not fully elucidated despite many studies on the pathogenesis. This study aimed to evaluate the LTB4 concentration in the bronchoalveolar lavage fluid (BALF) of horses with and without pulmonary inflammatory disease. Thirty-five mixed breed horses were studied and LTB4 was determined by using specific ELISA Kit. The horses were grouped by 2 different criteria for statistical analysis of data: 1) according to the values for BALF citology and 2) according to the detection of LTB4 in BALF. There was significant difference of effect of age on the LTB4 detection in equine BALF. Younger animals were the majority where it was possible to detect LTB4 values in LBA. In conclusion, there was an effect of age on the detection of LTB4 in equine BALF, where LTB4 levels were more easily detected in younger animals than older animals and the results of this study raise the possibility of considering future studies with the objective of establishing the real role and the best moment to detect LTB4 in BALF of the equine asthma syndrome.(AU)

Recentemente, o termo "síndrome da asma equina" (SAE) foi proposto devido à semelhança da doença equina à asma humana. Os leucotrienos causam constrição dos brônquios, especialmente nas vias aéreas posteriores e aumentam a secreção de muco no sistema respiratório. O leucotrieno B4 (LTB4) foi descoberto como um forte fator quimiotático, que desempenha um papel na migração de neutrófilos. O fundo imunológico do SAE permanece não completamente elucidado apesar de muitos estudos sobre a patogênese. Este estudo teve como objetivo avaliar a concentração de LTB4 no lavado broncoalveolar (LBA) de equinos com e sem doença inflamatória pulmonar. Trinta e cinco cavalos de raças mistas foram estudados e o LTB4 foi determinado usando o kit ELISA específico. Os animais foram agrupados por dois critérios diferentes para análise estatística dos dados: 1) de acordo com os valores para citologia do LBA e 2) de acordo com a detecção do LTB4 no LBA. Houve diferença significativa do efeito da idade na detecção do LTB4 no LBA equino. Os animais mais jovens foram a maioria onde foi possível detectar os valores de LTB4 no LBA. Em conclusão, houve um efeito da idade na detecção de LTB4 em LBA equino, onde os níveis de LTB4 foram mais facilmente detectados em animais jovens do que em animais mais velhos e foi possível detectar a concentração de LTB4 no LBA equino e os resultados deste estudo levantam a possibilidade de considerar futuros estudos com o objetivo de estabelecer o real papel e o melhor momento para detectar LTB4 no LBA da síndrome asmática equina.(AU)

Pharmacokinetic of florfenicol in pulmonary epithelial lining fluid of swine and effects of anesthetic agent on drug plasma disposition kinetics / Farmacoqu'imica de florfenicol em fluido de epitélio pulmonar de suínos e efeitos de agente anestésico em cinética de droga em plasma

The primary objective of the current study was to compare the pharmacokinetic (PK) of florfenicol (FFL) in pulmonary epithelial lining fluid and the plasma in swine. The second objectives were to evaluate the effect of anesthesia with ketamine and propofol on the PK of FFL in plasma. Bronchoaveolar lavage was utilized for quantification of PELF volume and the urea dilution method was used to determine the concentration of FFL in PELF. FFL was administered intramuscularly (IM) to swine in a single dose of 20mg/kg body weight. The main PK parameters of FFL in plasma and PELF were as follows: the area under the concentration-time curve, maximal drug concentration, elimination half-life and mean residence time were 69.45±4.36 vs 85.03±9.26µg·hr/ml, 4.65±0.34 vs 5.94±0.86µg/ml, 9.87±1.70 vs 10.69±1.60hr and 12.75±0.35 vs 14.46±1.26hr, respectively. There was no statistically significant difference between the PK profiles of FFL for the anesthetized and unanesthetized pigs. This study suggest that (i) FFL penetrated rapidly into the pulmonary and the drug concentration decay faster in plasma than in the pulmonary, (ii) the PK profile of FFL in swine was not interfered after administration of anesthetic agent.(AU)

O objetivo primário desse estudo foi comparar a farmacocinética de florfenicol (FFL) em fluido epitelial pulmonar à farmacocinética (PK) de FFL em plasma suíno. O segundo objetivo foi avaliar o efeito de anestesia com ketamina e propofol no PK de FFL em plasma. Lavagem broncoalveolar foi utilizada para quantificar volume de fluido epitelial pulmonar (PELF) e método de diluição de uréia para determinar FFL em PELF. Injeção de FFL foi administrada intramuscular a suínos em dose única de 20mg/kg de peso corporal. Os principais parâmetros de PK em FFL em plasma e PELF foram os seguintes: a área sob a curva de concentração-tempo, concentração máxima da droga, eliminação de meia-vida e média de tempo de permanência foram 69,45±4,36 vs 85,03±9,26µg·hr/ml, 4,65±0,34 vs 5,94±0,86µg/ml, 9,87±1,70 vs 10,69±1,60hr e 12,75±0,35 vs 14,46±1,26hr, respectivamente. Não houve diferença estatisticamente significante entre os perfis de PK de FFL para os porcos anestesiados e não anestesiados. Esse estudo sugere que (i) FFL penetrou rapidamente no pulmão e concentração da droga sofre queda mais veloz em plasma que líquido pulmonar, (ii) o perfil de PK de FFL em suínos não modificou após administração de agente anestésico.(AU)

Expression of the protein serum amyloid A in response to Aspergillus fumigatus in murine models of allergic airway inflammation / Expresión de la proteína amiloide A sérica como respuesta a Aspergillus fumigatus en modelos murinos de inflamación alérgica de las vías respiratorias

Background. Serum amyloid A (SAA) is an acute phase protein that is elevated in blood during inflammation. The role of this protein in allergic diseases of airways remains unclear. Aims. The objective of this study was to evaluate the SAA in blood, lung and bronchial cells in a murine model of bronchial hypersensitivity to Aspergillus fumigatus. Methods. To achieve this purpose, different groups of 5-month-old mice were housed in cages containing hay bedding that was contaminated with A. fumigatus and were kept in an isolation room for 16 days to allow for the induction of allergic airway inflammation. Subsequently, the mice were then exposed once again to Aspergillus spores at 0, 2, 8, 24 and 72 h, and they were bled to acquire serum and sacrificed to obtain bronchoalveolar lavage fluid (BALF) or lung tissues for analysis. SAA levels were measured in lung, serum and BALF by dot blot assay and RT-PCR (reverse transcription polymerase chain reaction). Results. The results indicated that SAA protein levels increased in both serum and lung within 2–24 h after mice were exposed to Aspergillus spores. Moreover, the SAA mRNA expression levels in the lungs and BALF cells demonstrated the same trend that was observed for the protein levels through the dot blot assay; in particular, SAA mRNA levels increased within the first hour after mice were exposed to A. fumigatus. Conclusions. In this allergic airway model, we conclude that A. fumigatus can induce an acute inflammatory response in the airways through the stimulation of the SAA protein, increasing its levels in serum, lung tissue and BALF samples during the early hours of exposure of mice that have been sensitised for this fungus (AU)

Antecedentes. La proteína amiloide A sérica (AAS) es un reactante de fase aguda cuyos valores sanguíneos aumentan durante los procesos inflamatorios agudos. Todavía no se ha dilucidado el papel que desempeña en las enfermedades alérgicas de las vías respiratorias. Objetivos. El objetivo del presente estudio fue examinar los valores de AAS en sangre, tejido pulmonar y células bronquiales en un modelo murino de hipersensibilidad bronquial frente a Aspergillus fumigatus. Métodos. Diferentes grupos de ratones de 5 meses de vida fueron alojados en jaulas cuyos lechos de paja estaban contaminados por A. fumigatus y se mantuvieron en una sala de aislamiento durante 16 días para permitir la inducción de inflamación alérgica de las vías respiratorias. Tras este período de inducción, a las 0, 2, 8, 24 y 72 h los animales se expusieron de nuevo a esporas de Aspergillus. En cada tiempo de reexposición se obtuvieron muestras sanguíneas de los animales y, acto seguido, fueron sacrificados para obtener líquido de lavado broncoalveolar y muestras de tejido pulmonar. La concentración de AAS se analizó mediante técnica de hibridación del ADN (Southern) y reacción en cadena de la polimerasa-retrotranscriptasa en muestras de suero, tejido pulmonar y células de líquido de lavado broncoalveolar. Resultados. Los resultados del presente estudio demuestran que, al cabo de 2-24 h de la exposición a A. fumigatus aumentaron los valores de proteína AAS en muestras de suero y tejido pulmonar. Además, en células de líquido de lavado broncoalveolar y muestras de tejido pulmonar los niveles de expresión de ARNm de AAS demostraron la misma tendencia, y, en particular, aumentaron al cabo de la primera hora de exposición a las esporas de A. fumigatus. Conclusiones. En este modelo murino de alergia de las vías respiratorias, concluimos que A. fumigatus puede inducir una respuesta inflamatoria aguda de las vías respiratorias a través de la estimulación de la proteína AAS, aumentando su concentración sérica en muestras de tejido pulmonar y de líquido de lavado broncoalveolar durante las primeras horas de exposición de ratones sensibilizados frente a este hongo (AU)

Bronchoalveolar lavage (BAL) in tapirs (Tapirus terrestris) with Mycobacterium tuberculosis / Lavado broncoalveolar (LBA) em antas (Tapirus terrestris) com Mycobacterium tuberculosis

This paper describes the in vivo Bronchoalveolar lavage (BAL) technique by endoscopy in tapirs (Tapirus terrestris) with clinical signs of tuberculosis. The technique was performed in two tapirs, male and female, from Curitiba Zoo, Paraná, Brazil. A flexible endoscope and a polyethylene catheter were used after the chemical restraint of the animals. For BAL technique, 60mL of saline 0.9% were infused with a polyethylene catheter, introduced by the endoscope's working channel, and 15mL of BAL were recovered, analyzed and submitted to cytocentrifugation. Slides were stained by Papanicolaou, periodic acid-Schiff (PAS) and Ziehl-Neelsen methods contained high quantity of inflammatory cells on light microscopy (macrophages 27.5%, lymphocytes 0.5%, neutrophis 67% and eosinophis 5%). BAL samples were submitted to culture, bacilloscopy and PCR and were negative for both animals. Based on this study, it was concluded that the bronchoalveolar lavage technique in tapirs is feasible, simple, noninvasive, practical and fast, providing an important clinical information in vivo regarding the functional status of the lower respiratory tract...

O presente trabalho descreve a técnica de lavado broncoalveolar (LBA) por endoscopia em antas (Tapirus terrestris) in vivo com sinais clínicos de tuberculose. A técnica foi realizada em duas antas, um macho e uma fêmea, provenientes do Zoológico de Curitiba, Paraná, Brasil, utilizando-se endoscópio flexível e sonda de polietileno, após a contenção química desses animais. Para o LBA, 60mL de solução fisiológica 0,9% foram infundidos com auxílio de cateter de polietileno, introduzido pelo canal de trabalho do endoscópio, e, aproximadamente, 15mL de LBA foram recuperados, acondicionados, analisados e submetidos à citocentrifugação. As lâminas foram coradas pelas técnicas de Papanicolau, ácido periódico de Schiff (PAS) e Ziehl- Neelsen, método que contém altas quantidades de células inflamatórias em microscopia (macrófagos 27,5%, linfócitos 0,5%, neutrófilos 67% e eosinófilos 5%). Amostras de LBA foram submetidas a cultura, baciloscopia e PCR e foram negativas em ambos os animais. Concluiu-se, baseado no presente trabalho, que técnica de lavado broncoalveolar é simples, não invasiva, funcional e rápida. Pode fornecer ao clínico importantes informações acerca do estado de funcionamento do aparelho respiratório in vivo...

Quantitative measurement of influenza virus replication using consecutive bronchoalveolar lavage in the lower respiratory tract of a ferret model

The ferret is an established animal model of influenza virus infection. Although viral replication in the upper respiratory tract is usually measured with consecutively collected nasal washes, daily evaluation of viral replication in the lung is limited because a large numbers of ferrets need to be sacrificed at consecutive time points. To overcome this limitation, we performed a virus quantification assay using bronchoalveolar lavage (BAL) fluid. This non-invasive BAL technique allows consecutive quantification of virus replication in the lungs of living ferrets. Our method can be used for the longitudinal evaluation of virus tropism in the lower respiratory tract.

Lesiones pulmonares y lavado broncoalveolar en ratones expuestos a la inhalación de base de cocaina (Basuco) / Respiratory pathologic manifestations of cocaine inhalation in mice

Desde 1970 por su menor costo, se ha popularizado el uso de formas intermedias, menos purificadas de cocaína (base de cocaína), especialmente por inhalación de cigarrillos; de acuerdo a la región recibe diversos nombres: en USA se conoce como Crack, en nuestro medio como basuco. Sólo existen informes aislados de manifestaciones patológicas respiratorias asociadas con fumar base de cocaína, por lo que se desarrolló el presente estudio para tratar de caracterizar mejor las lesiones que en el pulmón origina la inhalación de basuco. Se utilizaron dos grupos de 22 ratones blancos; uno se expuso a basuco y el otro sirvió como control. La dosis de basuco se calculó de acuerdo con la cantidad promedio utilizada por un consumidor habitual. Se sacrificaron paulatinamente durante un año; se realizó lavado broncoalveolar y los pulmones fueron extraídos, insuflados en formol para ser procesados completamente. Los hallazgos histopatológicos y los recuentos celulares de los lavados broncoalveolares de los controles fueron normales. En el grupo expuesto la bronquiolitis linfoplasmocitaria fué el hallazgo más común (44 por ciento), seguido porla neumonitis intersticial (22 por ciento), inflamación linfocítica perivascular (22 por ciento) y bronquiolitis folicular (17 por ciento). También se observó hemorragia alveolar, bronquiolitis obliterante y neumonía descamativa. La distribución citológica diferencial de los lavados broncoalveolares se correlacionó con los hallazgos histopatológicos. Se demuestra con claridad que la inhalación de basuco induce cambios histopatológicos en el pulmón de ratón así como en el recuento citológico diferencial del lavado broncoalveolar, lo que justifica la realización de investigaciones en grupos humanos expuestos