**2. Materials and methods**

#### **2.1 Preparation and storage of experimental diets**

Half-washed basin of fresh red chilli pepper (*Capsicum annum*) was procured from a local market in Nigeria. It was washed and sun-dried for 4 days. The dried samples were then pulverised using electric blender to obtain a fine powder. The pepper powder was then stored in air-tight rubber container from which pepper diets were prepared.

Capsaicin (95% pure) was obtained from Wuxi Gorunjie natural-Pharma Co. Ltd., Jiangsu China.

#### **2.2 Animal treatment**

Thirty (30) male mice of CD-1 strain weighing between 22 and 34 g were used for the study. They were kept in a well-ventilated room under room temperature (25 ± 2°C), humidity of 8 ± 5% and 12/12 h light/dark cycle and allowed 1 week for acclimatisation to the research environment before the experiments. They were randomly assigned into three groups, namely; control group that received normal rodent

**175**

*The Effects of Consumption of* Capsicum *on Some Neurobehavioural Parameters*

their beddings, feed and water were hygienically handled and changed daily.

chow, pepper group that were fed 20% chilli pepper diet and capsaicin group that were given 10% capsaicin diet. Each group comprised 10 mice. Each mouse was allowed drinking water *ad libitum*. This treatment was done for 28 days and within this period,

Approval for this study was obtained from the Faculty Animal Ethics Committee of Faculty of Basic Medical Sciences, University of Calabar with Protocol number

The elevated plus maze apparatus designed according to the description of Lister

) with 15 cm high walls radiating from a central square (5 × 5 cm).

) with 0.25 cm high edges and two closed

[14], and the test protocol adapted by Okon et al. [15] were used in this present study. This apparatus is used to assess the anxiety and fear levels of the mice. The test is based on the inborn aversion of rodents to open or bright illuminated spaces.

The open arms contain a slight edge (4 mm high) to prevent the mice from slipping

Prior to the test, the plus maze arms, surfaces and closed sides were cleaned with methylated spirit to eliminate olfactory clues and to remove faecal boll and urine. The mice were placed in the central square of the plus maze such that the mice faced an open arm away from the experimenter upon placement. Immediately after placement, a stop watch was started and the mice were allowed to explore the apparatus for 5 min. The test sessions were recorded and videotaped. Behaviours scored included open arm entry, open arm entry duration; head dip, rearing and

The beam walking assesses fine motor coordination and balance [17]. This test examines the ability of the mice to remain upright and to walk on an elevated and relatively narrow beam [17]. The beam has a length of 120 cm, a width of 0.6 cm and is suspended about 60 above some foam pads. The beam is marked at 5 cm and 1 cm intervals. It is composed of wood and is coated with black paint. The mouse was placed on one end of the beam. The trial was started after the mouse has secured its grip on the beam and lasted for approximately 5 min. The tests were videotaped for

This test used by Ramabadran et al. [18] assesses the basic pain response in mice

to thermal stimuli. This test measures spinally driven aspects of pain. Here, the

scoring. The parameters scored included the number of foot slips and falls.

*DOI: http://dx.doi.org/10.5772/intechopen.91744*

**2.3 Approval for animal use**

**2.4 Behavioural protocol**

*2.4.1 Assessment of anxiety levels*

The maze has two open arms (45 × 5 cm2

*2.4.1.1 The elevated plus maze*

and falling off the edge [16].

stretch attend posture frequencies.

*2.4.3 Assessment of pain perception*

*2.4.3.1 Tail immersion test*

*2.4.2.1 Beam walking*

*2.4.2 Assessment of motor coordination*

014PY20314.

arms (40 × 5 cm2

chow, pepper group that were fed 20% chilli pepper diet and capsaicin group that were given 10% capsaicin diet. Each group comprised 10 mice. Each mouse was allowed drinking water *ad libitum*. This treatment was done for 28 days and within this period, their beddings, feed and water were hygienically handled and changed daily.
