Integration of Animal Behavioral Assessment and Convolutional Neural Network to Study Wasabi-Alcohol Taste-Smell Interaction

Summary

This article describes a set of methods for measuring the suppressive ability of sniffing alcoholic beverages on the wasabi-elicited stinging sensation.

Abstract

The commercial wasabi pastes commonly used for food preparation contain a homologous compound of chemosensory isothiocyanates (ITCs) that elicit an irritating sensation upon consumption. The impact of sniffing dietary alcoholic beverages on the sensation of wasabi spiciness has never been studied. While most sensory evaluation studies focus on individual food and beverages separately, there is a lack of research on the olfactory study of sniffing liquor while consuming wasabi. Here, a methodology is developed that combines the use of an animal behavioral study and a convolutional neural network to analyze the facial expressions of mice when they simultaneously sniff liquor and consume wasabi. The results demonstrate that the trained and validated deep learning model recognizes 29% of the images depicting co-treatment of wasabi and alcohol belonging to the class of the wasabi-negative liquor-positive group without the need for prior training materials filtering. Statistical analysis of mouse grimace scale scores obtained from the selected video frame images reveals a significant difference (P < 0.01) between the presence and absence of liquor. This finding suggests that dietary alcoholic beverages might have a diminishing effect on the wasabi-elicited reactions in mice. This combinatory methodology holds potential for individual ITC compound screening and sensory analyses of spirit components in the future. However, further study is required to investigate the underlying mechanism of alcohol-induced suppression of wasabi pungency.

Introduction

Wasabia japonica, commonly known as wasabi, has gained recognition in food preparation^1,2. The intense sensory experience it elicits upon consumption, characterized by tearing up, sneezing, or coughing, is well-known. This distinctive pungency of wasabi can be attributed to a homologous compound of chemosensory isothiocyanates (ITCs). They are volatile organosulfur phytochemicals that can be categorized into ω-alkenyl and ω-methylthioalkyl isothiocyanates³. Among these compounds, allyl isothiocyanate (AITC) is the most predominated natural ITC produc....

Protocol

In this study, two 7-week-old ICR male mice weighing between 17-25 g were utilized for the animal behavioral assessment. All housing and experimental procedures were approved by the Hong Kong Baptist University Committee on the Use of Human and Animal Subjects in Teaching and Research. The animal room was maintained at a temperature of 25 °C and a room humidity of 40%-70% on a 12-h light-dark cycle.

1. Cage design

1. Prepare acrylonitrile butadiene styrene bricks.......

Discussion

The proposed method for studying taste-smell interaction in this work is based on the original method of behavioral coding for facial expression of pain in mice, which was developed by Langford et al.¹⁶. Several recently published articles have introduced CNN for automatic mouse face tracking and subsequent MGS scoring^21,26,27,28. Applying CNNs offers an advantage over t.......

Materials

Name	Company	Catalog Number	Comments
Absolute ethanol (EtOH)	VWR Chemicals BDH	CAS# 64-17-5
Acrylonitrile butadiene styrene bricks	Jiahuifeng Flagship Store	https://shop.paizi10.com/jiahuifeng/chanpin.html
Acrylonitrile butadiene styrene plates	Jiahuifeng Flagship Store	https://shop.paizi10.com/jiahuifeng/chanpin.html
Allyl isothiocyanate (AITC)	Sigma-Aldrich	CAS# 57-06-7
Anhydrous dimethyl sulfoxide	Sigma-Aldrich	CAS# 67-68-5
Chinese spirit	Yanghe Qingci	https://www.chinayanghe.com/article/45551.html
Commercial wasabi	S&B FOODS INC.	https://www.sbfoods-worldwide.com
Formic acid (FA)	VWR Chemicals BDH	CAS# 64-18-6
GraphPad Prism 5	GraphPad	https://www.graphpad.com
HPLC-grade acetonitrile (ACN)	VWR Chemicals BDH	CAS# 75-05-8
HPLC-grade methanol (MeOH)	VWR Chemicals BDH	CAS# 67-56-1
Microsoft Excel 2016	Microsoft	https://www.microsoft.com
Microsoft PowerPoint 2016	Microsoft	https://www.microsoft.com
Milli-Q water system	Millipore	https://www.merckmillipore.com
Mouse: ICR	Laboratory Animal Services Centre (The Chinese University of Hong Kong, Hong Kong, China)	N/A
Peanut butter	Skippy	https://www.peanutbutter.com/peanut-butter/creamy
Python v.3.10	Python Software Foundation	https://www.python.org
Transparent acrylic plates	Taobao Store	https://item.taobao.com/item.htm?_u=32l3b7k63381&id=60996545797 0&spm=a1z09.2.0.0.77572e8dFPM EHU