I am looking for some advice on an appropriate statistical analysis in R

*Experimental question* : We are interested in how communities from different streams may vary in their response to experimental temperature. Specifically we are interested to test for differences in the slope and intercept of the relationship between temperature and oxygen consumption between the different stream communities.

*Outline of experiment*

There were four streams and from each we took samples which had a characteristic community of organisms. These four different communities are labelled “S1,.. S4.

Each ‘Stream’ was incubated together in the same water bath at six temperatures in succession, giving covariates (~ 5,10,15,20,25,and 30°C) to the factor “Temp”.

However this also meant six repeated measures were made on the same ‘Stream’ over time. As time between incubations varied ‘Time’ is given a categorical label a,....f.

This procedure was repeated four times using different samples from the same streams, thus giving ‘Replicates’ labelled 1,...4. Each ‘Replicate’ had different temperature values but always six values in total.

The response variable measured in all 96 incubations, was the rate at which the communities consumed oxygen over a given time in incubation: ‘Rate’

Some incubation’s were excluded (5 from 96)

*Models*

Ideally we would do an ANCOVA to test for differences in slope or intercepts for the different streams. However as there were repeated measures and unequal n and unbalanced design, I have used a linear mixed effect model (from nlme package in R) in the form:

model <- lme (Rate ~ Temp* Stream, random = ~ Time|Replicate)

*Question* : Do these models appropriately account for the temporal / spatial pseudoreplication in the experimental design ?

Many thanks in advance

Daniel

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