Apache don't work after setting the mod_wsgi

Question

#I want to connect Apache with mod_wsgi and create the server which serves the reuslt of python files. I don't use any flameworks.
I use apache=ver.2.4.48 and mod_wsgi=ver.4.9.0.

#Firtst, I confirm that Apache serves the HTMLfiles without mod_wsgi.
Concreatly, when curl localhost:80 or curl localhost:81 is input, I can see the output of the HTMLfile I make.

#Then, I want to connect Apache with mod_wsgi.
mod_wsgi is installed in the virtual environment of conda.The conda environment is constructed with anaconda installed from the resorce cord.

#In this environment, four settings are done as below.
###(1)Setting of WSGIScriptAlias(/usr/local/apache2/conf/extra/httpd-vhosts.conf)

Listen 81
<VirtualHost *:81>
    ServerName wsgiapp.example.com

    DocumentRoot "/usr/local/vhosts"
    <Directory "/usr/local/vhosts">
        AllowOverride All
        Require all granted
    </Directory>

    Alias /vhalias "/usr/local/vhalias"
    <Directory "/usr/local/vhalias">
        AllowOverride All
        Require all granted
    </Directory>

    WSGIScriptAlias /myapp "/usr/local/www/myapp.wsgi"
    <Directory "/usr/local/www">
        AllowOverride All
        Require all granted
    </Directory>

</VirtualHost>

###(2)Setting of Load module(/usr/local/apache2/conf/httpd.conf)

LoadModule wsgi_module　/home/vagrant/anaconda3/envs/gaia/lib/python3.9/site-packages/mod_wsgi/server/mod_wsgi-py39.cpython-39-x86_64-linux-gnu.so

This directory is checked by mod_wsgi-express module-config.
###(3)Uncomment Include conf/extra/httpd-vhosts.conf(/usr/local/apache2/conf/httpd.conf)

# Virtual hosts
Include conf/extra/httpd-vhosts.conf

###(4)Setting of myapp.wsgi(/usr/local/www/myapp.wsgi)

def application(environ, start_response):
    status = '200 OK'
    output = b'Hello World!'

    response_headers = [('Content-type', 'text/plain'),
                        ('Content-Length', str(len(output)))]
    start_response(status, response_headers)

    return [output]

#After I set as above and restart the apache, apache don't react against curl localhost:80,curl localhost:81,curl and localhost:81/myapp. Errors don't even happen.

I confirm that Apache is running by using the command like systemctl status apache2.
The below is the status.

apache2.service - Apache httpd daemon
Loaded: loaded (/usr/lib/systemd/system/apache2.service; enabled; vendor preset: disabled)
Active: active (running) since 木 2021-09-16 16:15:24 JST; 4s ago
Process: 27620 ExecStart=/usr/local/apache2/bin/apachectl -k start (code=exited, status=0/SUCCESS)
Main PID: 27623 (httpd)
Tasks: 5
CGroup: /system.slice/apache2.service
└─27623 /usr/local/apache2/bin/httpd -k start

And I try some other ways of settiing as below.
(1)On httpd-vhosts.conf, Set Both Load module and WSGIScriptAlias
(2)On Both httpd.conf and httpd-vhosts.conf, Set Load module. On httpd-vhosts.conf, set WSGIScriptAlias
In both case, Apache don't reply.

#Can you tell me the right way of setting of mod_wsgi? Thank you in advance.