Protected Brush Sampling of the Respiratory Tract of Tursiops truncatus to Determine the Microbial Contamination in Healthy Animals
IAAAM 2012
Steven J.M. Gans1; Eric van Kregten2; Agnes M. Marik2; Nancy Boeve-Epping1; Cornelis E. van Elk3
1Department of Respiratory Diseases, St. Jansdal Hospital, Harderwijk, Netherlands; 2Department of Microbiology, St. Jansdal Hospital, Harderwijk, Netherlands; 3Dolfinarium, Harderwijk, Netherlands

Abstract

Infections of the respiratory tract are a common cause for morbidity in dolphins.1,2 Obviously, identifying the disease-causing microorganism is of great importance in management. Historically, blowhole swabs and exhalation plates have been used as a tool for this purpose. More recently, flexible bronchoscopy with bronchial washings has shown to be helpful.3,4 However, during passage through the pharynx, a bronchoscope will be contaminated with resident microbial flora. Therefore, a bronchial washing is likely to be polluted with microorganisms of the proximal airways. A technique that might overcome this problem is the Protected Brush Specimen (PBS), in which a sterile brush catheter is pushed out of its sealed sheath only after reaching the spot of interest with the bronchoscope.5-7 This technique has been known in human medicine for two decades and was also tested for use in horses.8 The purpose of this study was twofold. First, we wanted to determine whether the PBS showed less contamination with resident upper airway flora compared with bronchial washings. Second, we wanted to investigate whether the conducting airways of healthy dolphins were contaminated with bacterial flora.

Bronchoscopy was performed in five healthy specimens of Tursiops truncatus as part of their health checkup. In all animals, bronchial washings were collected from the lower airways and a protected brush sample was taken from the lower tracheal area just above the carina. The washings and PBS samples were immediately transported to the microbiology laboratory for culturing. We observed that in two out of the five animals the PBS culture remained sterile, even though PBS cultured in broth is a highly sensitive method to find bacterial presence. In the other animals, the PBS showed considerably less species of microorganisms than the washings, leaving the question whether this is the result of partial contamination during the procedure or that it represents resident flora in the lower respiratory tract of these animals. Another observation was the fact that in the bronchial washings several species of water bacteria (species commonly found in an aqueous environment without a host animal) were cultured and in the PBS none. It is very clear that no firm conclusions can be drawn from measurements in only five animals, but our observations indicate that PBS sampling might be a better diagnostic tool than bronchial washings in respiratory infections allowing a more precise and prudent use of antibiotic treatment. Furthermore, our findings suggest that in healthy animals the bacterial contamination of the conducting airways could be low or even absent. Repeating this study in a larger number of animals should be helpful in confirming and clarifying our results. Performing quantitative cultures could be a useful addition.

References

1.  Sweeney JC, Ridgway SH. Common diseases of small cetaceans. J Am Vet Med Assoc. 1975;167:533–540.

2.  Medway W, Schryver HF. Respiratory problems in captive small cetaceans. J Am Vet Med Assoc. 1973;163:571–573.

3.  Tsang KW, Kinoshita R, Rouke N, Yuen Q, Hu W, Lam WK. Bronchoscopy of cetaceans. J Wildlife Dis. 2002;38:224–227.

4.  Hawkins EC, Townsend FI, Lewbart GA, Stamper MA, Thayer VG, Rhinehart HL. Bronchoalveolar lavage in a stranded bottlenose dolphin. Proceedings 27th annual workshop of IAAAM 1996;124.

5.  Wimberley N, Faling LJ, Bartlett JG. A fiberoptic bronchoscopy technique to obtain uncontaminated lower airway secretions for bacterial culture. Am Rev Resp Dis. 1979;119:337–343.

6.  Marquette CH, Ramon P, Courcol R, Wallaert B, Tonnel AB, Voisin C. Bronchoscopic protected catheter brush for the diagnosis of pulmonary infections. Chest. 1988;93:746–750.

7.  Higuchi JH, Coalson JJ, Johanson WG Jr. Bacteriologic diagnosis of nosocomial pneumonia in primates: usefulness of the protected specimen brush. Am Rev Resp Dis. 1982;125:53–57.

8.  Sweeney CR, Sweeney RW, Benson CE. Comparison of bacteria isolated from specimens obtained by use of endoscopic guarded tracheal swabbing and percutaneous tracheal aspiration in horses. J Am Vet Med Assoc. 1990;196:392–393.

  

Speaker Information
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Steven J.M. Gans
Department of Respiratory Diseases
St. Jansdal Hospital
Harderwijk, Netherlands


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