I am describing a new species of ant. Molecular and Morphological data suggests that there are 2 separate species. In fact we are very certain they are separate based on behavioral information. I would like to statistically analyze the measurements I have taken from my samples to support what I have found morphologically and molecularly.
However, I am having trouble with where to begin with my statistical analysis, or what sort of analysis I should do.
Issue: I have a data set of n=42 Colonies, with n=172 individuals measured; 9 measurements and 5 indices for each worker (indices are standard practice). Broken down further morphologically; I have n=13 colonies and n=46 workers from the described species; and n=29 colonies and n=127 workers from the new species. The biggest issue is the sampling error for the measurements. This is from how uncommon the described species is, and its confusion with the undescribed species. I have some colonies sampled that had 2-3 workers measured, while most had 5 workers measured, a few with 5+ workers measured , and even some that were measured as the described species but should have been measured as the undescribed. We did not have behavioral information for those colonies that were misplaced.
I have two questions I want to answer: Are the measurements from each colony are similar (Variance in a colony)? Show that they are separate species from these measurements. When I measured total length of the ants, it strongly suggested they are separate species.
My preliminary data showed that they were separate, but I only used a Z-test. Here are two characters we reported on a poster. A z-test was "helpful" but not really informative or useful without an ANOVA, which I can't do since the measurements are dependent of the colony and the sampling issue.
I had started to do a principal components analysis of the measurements (using PNG) with 4 principles resolving ~90-95% of the variance; as suggested by a colleague. The scatter plot did show the expected grouping by species(Red= Described, Grey- Undescribed, Yellow= Should have been measured as undescribed). But I am not certain of what to do with that data or if it really resolves my issue of dependent variables.
I had considered finding the averages of colonies, since each worker sampled came from the same mother and has the same genetic information as its sisters. But the sampling issue arises. Are the measurements I took of workers from a colony replication for that colony? Should the type specimen of the described species I measured hold more weight than other specimen of that species I measured (I measured a paratype and 3 holotypes)?
Please let me know if I should provide more information. Or there is something wrong with my train of thought/reasoning. Thank you.