I am comparing fish communities at 2 sites (upstream vs downstream) with data collected in two seasons (wet and dry) over several years (2017-2022), with data from the first pair of wet and dry seasons representing the period before a treatment and subsequent data representing periods after the treatment. During each season I sampled each site four times, and recorded abundance of each fish species from each site. I did not conduct sampling during the last dry season due to resource constraints. The community compositions of the two sites over different seasons and periods are visualised in
I am trying to do further analysis using PERMANOVA to look for any spatial-temporal changes in the fish communities, mainly if the two groups are becoming more similar in the years following the treatment. As there are samples with no fishes recorded I have to remove those samples from the dataset, which means I have only three instead of four replicates in some of the site x season x year groups.
My question is, does it still make sense to use PERMANOVA if I have unequal sample size among groups, given the number of replicates from each are small (3-4)? I am planning to run the test separately for wet and dry seasons, but that means I still need to do two-way (site x year) PERMANOVAs for each of the seasons.
I learned from some of the online discussions that unequal sample size would be a problem for two (or more)-way PERMANOVAs and the problem of unequal sample size would be more prominent when the sample sizes are small. Would be grateful for any comments or insight on this.
