1. Please state project objectives and what work was done this quarter to address them: Obj. 1. Document laboratory and field biologyLaboratoryRearing of snails has been successful to date. Mortality has been limited, even among young snails born in captivity in the spring. I have experimented with different rearing set ups to improve rearing efficiency. The most important factors in order of most to least important seem to be: adequate ventilation, container height, appropriate moisture, and availability of hides. (Fig 1.). Development of a standard protocol for rearing the snails and obtaining life stages of interest is the first step in developing robust data regarding both molluscicidal and biological control measures. FieldSnail population activity is currently being monitored biweekly at three locations in central Florida. Sampling is being done using grids throughout groves and in the edge habitats where possible to determine (a) the timing of activity and (b) patterns of activity in space. This information will inform when to apply molluscicides, or biological controls in the future, for optimal management. Nested within this trial is a comparison of two styles of traps- a pyramid and a flat trap (Fig 2a, b). Initially we planned to use flat traps to quantify snail presence as this was shown to be effective in row crops in North Florida, however in reading the genus Bulimulus, we learned that this group of snails is known for their tree climbing tendencies. Additionally, Tedder’s traps in an ongoing Diaprepes trial are consistently covered in them. In site 1, which has been monitored since March 2023, there is a consistent, though low population of B. bonariensis. This site is the only one in which there is abundant within and between row vegetation. Because numbers of snails on traps during sampling have been low, we have checked the vegetation and have found more snails in the weed middle vegetation than near/on trees or microjets so far (Fig 3). In site 2, which has been monitored since early April 2023, snails are consistently present on the traps, though also at a low density. While this site lacks the lush vegetation seen in Site 1, the mossy understory provides a moist habitat in which find more snails than on the traps (Fig. 3). Site 3, which has had the longest history of B. bonariensis activity of the three sites has a much higher snail to trap ratio (Fig 3.). This site, unlike the others, has very little ground cover for snails to seek shelter and moisture, which may factor into the seemingly higher populations than at the other sites. Both trap types, as well as microjects and trees, provide respite from the hot sand. We have insufficient data to provide a trap type comparison at this time, this will occur at a future date. Based on observations at these sites, we will look for additional trapping sites that have similar variations in ground cover to add to these data. Observationally, activity of snails was noticeable in late April 2023, with all life stages visible in groves. We expect that this may be an optimal time to apply management actions. It has also been suggested that the fall population dip, whenever that shall occur, may be a good time to manage to impact the population going into the overwintering period. This information is based on conversations with colleagues working on snails in less tropical environments. At IRREC, B. bonariensis has been causing challenges with an irrigation study which incorporates black plastic mulch for water retention. We took this as an opportunity to gather some preliminary data regarding the influence of this ground cover on snail populations. The data discussed below are extremely preliminary. Findings may change as more data are collected and as environmental conditions change over the course of the year.Snails were collected from a commercial citrus grove. This grove has two treatments implemented as part of an irrigation study: some rows have black plastic mulch, others do not. In two of each row type, five 1x1x1 m areas (quadrats) within each of these rows were selected for sampling. Each quadrat contained the trunk of one citrus tree. The entirety of the tree, ground, and leaf litter in each quadrat was searched for snails, and snails were collected. In the laboratory, the total number of snails and the total mass (weight) of the snails collected in each quadrat was recorded. Our findings are presented in Figures 4 and 5. In short, currently there are no statistically significant findings. This is due in part to several outlier counts. We plan to increase our sampling efforts in this grove to gather more information. Additionally, we plan to examine the underside of the black plastic in order to determine if the plastic is providing a refuge for the snails.Obj 2. Determine factors that influence snail movement/dispersalBulimulus bonariensis, much like other molluscs require moisture to proliferate. With recent rains, we have noted fewer snails on microjets and on our traps, suggesting that when the rainy season begins, snail pressure impacting irrigation dissipates in most groves. This is purely observational and will be followed up with future studies. Obj 3. Field evaluation of baitsUnfortunately, our baits showed up 2 months later than anticipated, so this portion of the work has not moved forward in a grove setting. We attempted to evaluate some baits we had from a previous trial in a semi-field setting using plastic pools with sand substrate and field-collected vegetation for refuge. Unfortunately, our design lacked sufficient moisture to maintain the snails and all perished, though clearly not from ingesting baits. While the commercial baits labelled for field use arrived only recently, we found that some active ingredients were found in snail and slug baits available for residential use. To move this area of study forward, we designed an assay with a vulnerable host (pepper plant), moist soil, and then added the treatments. We are still working on these assays, however none of the materials tested to date killed more than 45% of snails. Obj 4. Determine if arthropod or mammalian predators exist in groves for Bulimulus bonariensisIn observing B. bonariensis under a stereomicroscope, co-PI Quin noticed mites walking on the shells of several from a recent collection. The snails from this collection have mostly died of unknown causes. Out of curiosity, co-PI Quinn has been looking into mites that are pests of snails and found that there is one mite that is a known pest of pet snails which can also occur in open habitats. These mites take up residence in the lungs of the snails where they reproduce. There can be multiple generations of these mites living embedded like ticks in snails’ lungs and at some point, they impact the health of the snails. Mites were found in snails collected in a citrus grove in Fort Pierce on June 29. 5 days after collecting, the colony began to die off, which is unusual, leading to the inspection of snails under the microscope. After finding the mites, snails were dissected to look for their presence in snail lungs. Mites were found in 5 of 6 snails dissected (Fig. 6) This mite is a known threat to both the pet snail and escargot industries, suggesting it may play a role in either weakening the snails or leading to their mortality, particularly in dense populations. Obj. 5. Explore nematodes as a management option for Bulimulus bonariensisPermit paperwork has been submitted to obtain nematodes of interest. 2. Please state what work is anticipated for next quarter: Obj. 1 Document laboratory and field biologyWe will continue work on both the laboratory based life cycle research and the field based population biology. With protocols in place, both elements of this objective will expand to include more elements with the start of our incoming students in August. Obj. 2 Determine factors that influence snail movement/dispersalWe will troubleshoot the methods outlined in our proposal to decipher factors that influence dispersal including land cover, access to moisture, and temperature. Obj 3. Field evaluation of baitsWe will continue the lab assays of baits as described to determine efficacy in the presence of a food resource and in larger arenas where snails can avoid them. Depending on field populations, we may be able to run a field trial of baits, although I expect that will later in the fall.Obj 4. Determine if arthropod or mammalian predators exist in groves for Bulimulus bonariensisWe will continue exploring the role of the mite described previously for its potential role in population regulation. Additionally, we will begin camera trapping as described in our proposal to identify other potential predators in the system. 3. Please state budget status (underspend or overspend, and why): Our project is underspent due to delays in hiring students for the research projects. Staff and PIs have been able to obtain initial data and troubleshoot, but the majority of the research proposed will be led by our incoming students. Additionally, some supplies were backordered, and we have only recently been able to order these materials. 4. Please show all potential commercialization products resulting from this research, and the status of each: It is too early to suggest this yet.