I recommend Fragstat which can calculate many landscape metrics including some indicators of fragmentation. It can be accessed at the link below:

https://www.umass.edu/landeco/research/fragstats/fragstats.html

Thank you, Timothy! I'm not familiar with this software but I'm pretty comfortable with GIS. How can I prepare my data to input in FRAGSTAT?

Satellite images helped by arc GIS

You can determine your variables and their scales including vegetation size, road situation, water resources and other factors both before and after the highway. In my opinion Gıs and Maxent systems can be used

Thanks for the innovative topic. With best wishes for the success of your research findings.

I think

ArcGIS is a good tool to be used for this kind of analysis especially if you add remote sensing software such as envi to monitor the detection through time for raster images

Fragstat is a very good tool you should use. Please visit YouTube for some simple tutorials on how Fragstat operates.

There exists additional very powerful tools for potential connectivity analysis. Conefor Sensinode is a very powerful tool.

Recommended. The suggestions may be adopted for the analysis.

If you use raster maps, also Idrisi (now called TerrSets) cal calculates some fragmentation indexes.