In this picture you can see the shade cloth overhead and the complicated plumbing for the 6 zones of the structure so that I can program each zone (which are all colour coded!) to come on for a certain time separately! 
This is a rain gauge, one of many sensors I have in the plot to record the conditions the plants are experiencing. the rain is funnelled into a small tipping bucket which holds a specific amount, each tip is recorded and the water drips out of the bottom of the bucket. 
Above are my first male and female betta fish or siamese fighting fish, unfortunately the male died soon after I got him and I replaced him with a new one and also replaced the female for fear that she too was sick with the same problem which killed the male. I got an almost Identical pair and tried to mate them but I couldn't get them to do it, the male would make a bubble nest but then they didn't ever put eggs in it. In the end the female died, I don't know what of, I know the water chemistry is fine and I'm feeding them bloodworms which they love. Anyway I am bored with Bettas, they are not that interesting and have to be kept in separate bowls which is annoying. Below is my original male. 
This is my newer male who I still have. There is also a trap door snail in here too on the bottom. He slides around the glass keeping it clean! 
In a larger bowl I have 2 red capped gold fish and a black moor fish with bulging eyes (see bottom photo): 
Conclusions
The analysis of variance clearly showed that there was a significant difference in the amount of zoospores obtained between the four different treatments (F(3,24)=42.3, p<0.001)>(2,24)=6.75, p=0.005) and between the isolates nested within species (F(6,24)=9.01, p<0.001). The 5mL treatment with SPW or n/s soil extract was identified as being the best treatment, yielding the highest concentration of zoospores to volume of inoculum produced (Table 4). There was no significant difference between the remaining three treatments (Figure 5).
No further analysis of the differences between species and isolates were carried out as these differences were not of interest to the research being undertaken. It may be interesting for future studies to investigate the extent of any significant differences between isolates of the same species; however this did not pertain to the research question for this study.
My aim for this experiment was to optimise the concentration of zoospores produced from a single Petri plate culture. These results show that the best treatment, out of the 4 treatments tested herein, was the 5mL application of SPW or n/s soil extract as this yielded the highest ratio of concentration of zoospores to volume of inoculum obtained. In answer to the research question posed; reducing the volume of SPW or n/s soil extract applied to the broth cultures does increase the concentration to volume ratio of zoospore inoculum obtained.
I will now be able to obtain the concentration of zoospore inoculum that I require for all of the eight Phytophthora spp. that I am working with. Previously P. citricola and P. cambivora had consistently been poor producers of zoospores and I was unable to achieve the desired concentration of inoculum for my plant inoculation experiments. The results of this experiment lead me to conclude that I should treat these species with only 5mL of SPW or n/s soil extract (respectively) rather than 10mL which I had used previously.
I am unable to suggest a biological reason for this stark difference between the 5mL treatment and the 10, 15 and 20mL treatments. The purpose of treating the broth cultures with this SPW or n/s soil extract is to induce sporangia (which release zoospores) production through starvation conditions and so I can only assume that for the species studied herein the reduction of this rinse volume to 5mL enhances the effect of the starvation conditions and promotes greater sporangia or zoospores production. It is unclear whether the changes in zoospore concentration result from greater sporangia production or increased zoospores produced per sporangia. It seems particularly interesting, and unexplained, however that there are no significant differences between the other three treatments so that the relationship between treatment volume and response is not linear. Further research into the factors involved in this response must be undertaken in order to ascertain the nature and cause of the relationship.
Here I am preparing the inoculum to a specified concentration and then dipping the whole small plants in the inoculum. 
This is what I did my statistics term project on:
In the course of my studies of the epidemiology of Phytophthora species on Rhododendrons I need to produce inoculum with which to infect the host plants. Phytophthora spp. are Oomycete plant pathogens, also known as water moulds. The inoculum produced comes from structures called sporangia, borne from the vegetative hyphae of the Phytophthora spp. cultured in vitro. These sporangia release motile zoospores which are the asexual propagules which infect the host tissue.
The process of producing zoospore inoculum involves inoculating a liquid broth media (V8 100) with the Phytophthora isolate, rinsing the 5 day old culture with distilled water to remove the V8 100 broth, and replace it with a volume of either filter sterilised pond water (SPW) or non sterile filtered soil (n/s soil) extract to induce sporangia production by creating starvation stress conditions (Pettitt et al. 2002, Ahonsi et al. 2007).
I am working with 3 isolates of each of 8 species of Phytophthora and I have found that different species produce zoospores at different concentrations. For the inoculations of host plants I must standardise the concentrations of zoospore inoculum produced to 10,000 zoospores mL-1. I have found that two Phytophthora spp. consistently produce half the inoculum concentration of the other species such that I am unable to dilute the inoculum to 10,000 zoospores mL-1(because you cannot increase the concentration, you can only dilute it). I aim to optimise the concentration of zoospores produced from a single Petri plate culture.
One way to try to do this is to adjust the volume of SPW or n/s soil extract applied to induce sporangia production to see what effect this may have on the zoospore inoculum produced. Obviously just decreasing the volume of liquid in the plate will increase the concentration of zoospores in that volume of liquid, however it is possible that either increasing or decreasing the volume of SPW or n/s soil extract applied to the cultures may alter the stress conditions to encourage increased sporangia production thus improving the concentration to volume ratio of inoculum obtained.
References:
Pettitt, T.R. et al., Comparison of serological, culture, and bait methods for detection of Pythium and Phytophthora zoospores in water. Plant Pathology, 2002. 51(6): p. 720-727.
Ahonsi, M.O., Banko T.J., and Hong C., A simple in-vitro `wet-plate' method for mass production of Phytophthora nicotianae zoospores and factors influencing zoospore production.70(3): p. 557-560. Journal of Microbiological Methods, 2007.
Research question: Does reducing the volume of SPW or n/s soil extract applied to induce sporangia production increase the concentration to volume ratio of zoospore inoculum obtained?
Research objective: Determine the volume of SPW or n/s soil extract to use to maximise the number of zoospores produced.
Experimental Design: This experiment will be a nested design with 4 levels of treatment (5, 10, 15, 20 mL; volume of SPW or n/s soil extract) applied to 9 isolates of Phytophthora, 3 nested within each of 3 Phytophthora spp. Each isolate acts as a replicate for treatment so that r = 9. The experimental unit is one isolate growing in a Petri dish. The response variable will be the ratio of the concentration of zoospores to the volume of inoculum yielded from the method described above. MINITAB and MS Excel software will be used to conduct an Analysis of Variance (ANOVA) and a multiple comparisons test to determine the best treatment.
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