# Acoustic Detection of Vaccine Reactions in Hens for Assessing Anti-Inflammatory Product Efficacy
###### Abstract  
Acoustic studies on poultry show that chicken vocalizations can be a real-time indicator of the health conditions of the birds and can improve animal welfare and farm management. In this study, hens vaccinated against infectious laryngotracheitis (ILT) were acoustically recorded for 3 days before vaccine administration (pre-reaction period) and also from vaccination onwards, with the first 5 days being identified as the \"reaction period\" and the 5 following days as \"post reaction\". The raw audio was pre-processed to isolate hen calls and the 13 Mel-frequency cepstral coefficients; then, the spectral centroid and the number of vocalizations were extracted to build the acoustic dataset. The experiment was carried out on the same farm but in two different houses. The hens from one house were assigned to the control group, without administration of the anti-inflammatory product, and the other formed the treatment group. Both acoustic data sets were recorded and processed in the same way. The control group was used to acoustically model the animal reaction to the vaccine and we automatically detected the hens' vaccine reactions and side effects through acoustics. From Scikit-Learn algorithms, Gaussian Naive Bayes was the best performing model, with a balanced accuracy of 80% for modeling the reactions and non-reactions caused by ILT in the control group. Furthermore, the importance of algorithm permutation highlighted that the centroid and MFCC4 were the most important features in acoustically detecting the ILT vaccine reaction. The fitted Gaussian Naive Bayes model allowed us to evaluate the treatment group to determine if the vocalizations after vaccine administration were detected as non-reactions, due to the anti-inflammatory product's effectiveness. Of the sample, 99% of vocalizations were classified as non-reactions, due to the anti-inflammatory properties of the product, which reduced vaccine reactions and side effects. The non-invasive detection of hens' responses to vaccination to prevent respiratory problems in hens described in this paper is an innovative method of measuring and detecting avian welfare. and avian influenza [3]. The following different methods are available for diagnosing ILT: (i) a histopathological examination of the formation of syncytial cells and tracheal epithelial cells, with the development of pathogenic intranuclear inclusion bodies, necrosis, and hemorrhage [2], (ii) a commercial Enzyme-Linked Immunosorbent Assay (ELISA) for the detection of antibodies against the ILT virus [1] (iii) DNA detection methods for the ILT virus. Molecular techniques for ILTV detection include Polymerase Chain Reaction (PCR), real-time PCR, nested PCR, multiplex PCR, in situ hybridization, and PCR followed by Restriction Fragment Length Polymorphism (RFLP) [4]. The symptoms of larynxopteracheitis, like other diseases, influence the vocalization and behaviour of animals by modifying their calls due to coughing [5]. ILT vaccination generates stress in animals due to the process itself: the hens are manually removed one by one from the cage to administer an eye drop vaccine. Additionally, side effects such as breathing problems may develop in the following days. This combination of stress and side effects can be recorded as a biological response, using hen calls. New bioacoustic precision livestock farming (PLF) techniques offer the possibility of automatically tracking and monitoring the behavior of poultry through its calls [6].