MUMBAI, India, July 11 -- Intellectual Property India has published a patent application (202541053295 A) filed by Ramanujam Narayanan; and Dr. Sivaselvakumar Muthusamy, Coimbatore, Tamil Nadu, on June 2, for 'development and validation of a bronchoalveolar lavage technique in mouse model for toxicology risk assessment.'
Inventor(s) include Ramanujam Narayanan; and Dr. Sivaselvakumar Muthusamy.
The application for the patent was published on July 11, under issue no. 28/2025.
According to the abstract released by the Intellectual Property India: "Bronchoalveolar lavage (BAL) technique is a mandatory requirement in regulatory toxicology studies, as per OECD guidelines. However, it requires extreme skill and precision especially in low-cost experiments incorporating minimal sample sizes. Few studies have emphasized the significance of a standard technique of BAL in rodents in publication databases. The importance of this invasive technique cannot however be overemphasized for incorporation of two of the three Rs (Russell and Burch): alternatives to usage of experimental animals. Methods: IAEC approval was obtained. Three animals are selected and weighed. BAL ringer fluid solution is prepared. Methanol solution is the prepared as negative control. One animal is preselected as positive control animal. The second animal is given 1 ppm methanol inhalation in an inhalational chamber. The third mouse is given 1 ppm Aroclor 1232 in a whole body inhalational chamber. Under Ketamine euthanasia IV at 55 mg/kg with Xylazine 1 mg/kg IV at least fifteen minutes prior to the BAL technique, after tracheostomy is secured, the BAL ringer is perfused into the mouse lungs using 21 G tube inserted to a balloon catheter thrice. The aspirated BAL-fluid is incorporated onto 1 ml tubes with PBS prefilled at three times the volume of the BAL fluid yield, centrifuged at 300 g for 5 minutes at 4 C and analyzed for cytology, Aroclor concentrations using UPLC-DAD and inflammatory markers. Lung tissue can be additionally harvested for assessing inflammation using H&E staining. Results: Minimal inflammation was observed for all the three mouse lung samples. BAL-fluid cytology showed Lymphocytosis and granulocytosis in Aroclor treated mouse as compared to both controls. Discussion: A minimally invasive animal BAL technique was developed and validated for mouse inhalational toxicology assessment using Aroclor 1232. The BAL fluid cytology correlated well with histopathology staining for minimal animal respiratory tract inflammation and confirmed to Reduction and Refinement standards of 3Rs by reduction of sample size and animal suffering. This technique can be used for regulatory toxicology risk assessments using mouse models. Conclusion: A minimally-invasive animal BAL technique was developed and validated for mouse inhalational toxicology assessment using Aroclor 1232. The 270 275 280 285 290 BAL fluid cytology correlated well with histopathology staining for minimal animal respiratory tract inflammation and confirmed to Reduction and Refinement standards of 3Rs by reduction of sample size and animal suffering. This technique can be used for regulatory toxicology risk assessments using mouse models. Keywords: Bronchoalveolar lavage, Rodent toxicology, Animal alternatives, Inhalational toxicity, Aroclor 1232."
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