Attachment E_FINAL.pdf

Florida Fish and Wildlife Conservation Commission

Eelgrass Propagation and Restoration Standard Operating Procedures (SOPs)

BACKGROUND

In 2021, a hybrid species of eelgrass (V. spiralis and V. denseserrulata; hereafter, ‘the hybrid’) was reported for the first time in Florida (Gorham et al. 2021). Both parent species are non-native to North America and their respective natural ranges do not overlap in the wild, suggesting that the hybrid is non-native to Florida and its origin and mode of introduction might be related to the aquarium pet trade.

Populations of the hybrid have also been found in Japan, Hungary, and Australia (Wasekura et al. 2016);

however, little is known about its distribution in Florida, its competitive potential with native Vallisneria spp., or its effect on oligohaline macrophyte communities and freshwater ecosystems. Limiting the spread of the hybrid is therefore essential to the management of eelgrass habitat in Florida.

To minimize the spread of the hybrid, the Florida Fish and Wildlife Conservation Commission (FWC) developed a set of standard operating procedures (below) to guide the agency’s eelgrass restoration activities, specifically brood stock collection, eelgrass propagation, and transplantation. The agency will continue to research the statewide distribution of the hybrid and its sympatry with native eelgrass. As such, this will be a living document, which will be updated based on new scientific information and methods and changing management goals and needs.

STANDARD OPERATING PROCEDURES FOR FWC VALLISNERIA RESTORATION

1. V. spiralis, V. denseserrulata, or a hybrid of these species, or any other non-native Vallisneria (eelgrass) species will NOT be used for propagation or planting by the FWC or its contractors.

2. Waterbodies with known hybrid populations are NOT eligible to supply donor material for direct transplanting or nursery out-planting activities conducted by FWC or its contractors.

3. FWC will not purchase eelgrass from nursery facilities found to contain non-native eelgrass.

4. Within waterbodies where the hybrid has not yet been detected, all donor beds intended for direct transplanting must be screened for hybrid presence using genetic methods and following the collection protocol (“Methods for Transplanting Projects”) in the document, Detection and Mitigation Protocols for Hybrid Eelgrass in Restoration.

5. All plants intended for nursery out-planting must be screened for hybrid presence using genetic methods and following the collection protocol, “Methods for Nursery Out-Planting Projects” in the document, Detection and Mitigation Protocols for Hybrid Eelgrass in Restoration.

6. Only native Vallisneria (Les et al. 2008) will be propagated in FWC nursery or hatchery locations.

a. Any non-native eelgrass will be eradicated from FWC nursery or hatchery locations.

b. All eelgrass plants to be used for nursery brood stock must be genetically screened for hybrid status prior to entering an FWC nursery or hatchery location for grow out.

Note: waterbodies with known hybrid presence are not eligible for nursery brood stock collection.

7. Recommendation: For genetic screening for the hybrid and genetic authentication of native eelgrass plants, FWC recommends the rapid PCR assay developed by the FWC Fish and Wildlife Research Institute (Tringali et al. 2023).

GUIDANCE FOR FWC CONTRACTED NURSERY GROW-OUT

1. Each eelgrass plant for transplanting should possess at least four (4) leaves measuring at least six

(6) inches in length from an intact root mass.

2. If eelgrass is propagated in liners, pots, “mechanical planting units” (MPU), or “burritos”, the propagation unit should contain a minimum of one (1) individual plant for "MPUs"; two (2) individual eelgrass plants for 2" liners or pots; three (3) individual eelgrass plants for "burritos";

and four (4) individual eelgrass plants for 4" liners or pots.

3. If eelgrass is propagated in liners, pots, “mechanical planting units”, or “burritos”, the enclosing material containing the soil matrix and plants must be biodegradable (e.g., peat or burlap). The biodegradable portion may be grown within a non-biodegradable unit (e.g., 2” or 4” plastic liner);

however, the plants shall only be planted in their biodegradable wrapping.

4. The recommended soil matrix in which propagated plants are grown is 75% sand and 25%peat.

5. All propagated eelgrass plants should be potted and grown for at least two (2) months prior to out-planting.

6. Plants should be free of any noxious plant material, specifically any plant species listed by the

Florida Invasive Species Council (formerly known as FLEPPC) as a Category I or II invasive species, and/or exotic wildlife species (e.g., non-native Pomacea snails).

GUIDANCE FOR FWC CONTRACTING OF EELGRASS RESTORATION ACTIVITIES

1. All vendors contracted for eelgrass propagation and out-planting should possess a Certificate of Nursery Registration (DACS-08004), which shall be made available upon request by the Project Manager.

2. Contractors shall provide location[s] of plant origin or collection and documentation of genetic authentication for the native species to the Project Manager prior to propagation.

3. All plants proposed for out-planting should be available for inspection by the FWC Project Manager/Contract Manager at least two (2) weeks prior to out-planting. FWC may sample a portion of the plants grown in the nursery facilities prior to out-planting (“Methods for Hybrid Detection in Existing Nursery Stock” in Detection and Mitigation Protocols for Hybrid Eelgrass in Restoration).

4. All contracted out-planting operations should be coordinated and conducted under the direct daily supervision of designated FWC personnel.

5. The Project Manager and/or appropriate FWC staff overseeing the contractor should maintain a count of the number of eelgrass plants meeting specification requirements that are successfully installed. The Contractor and FWC Contract Manager should agree in writing on the number of plants successfully supplied and installed at the end of each day in accordance with the “Contract Management Daily Report: Aquatic Revegetation” (Attachment).

REFERENCES

Gorham, S. B., S. Seyoum, B. T. Furman, K. M. Darnell, L. K. Reynolds, and M. D. Tringali. 2021.

Molecular detection of a non-native hybrid eelgrass, Vallisneria spiralis Linnaeus (1753) × V.

denseserrulata Makino (1921), in the southeastern United States. Aquatic Botany 175:103445.

Les, D. H., S. W. L. Jacobs, N. P. Tippery, L. Chen, M. L. Moody, and M. Wilstermann-Hildebrand.

2008. Systematics of Vallisneria (Hydrocharitaceae). Systematic Botany 33:49-65.

Tringali, M. D., S. B. Gorham, S. Seyoum, B. T. Furman, C. Puchulutegui, M. S. Bass, B. T. Furman, and C. Mallison. 2023. A PCR assay for the detection of introduced Vallisneria spiralis, V.

denseserrulata and their hybrids. Conservation Genetics Resources https://doi.org/10.1007/s12686-023-01311-9

Wasekura, H., S. Horie, S. Fujii, and M. Maki. 2016. Molecular identification of alien species of Vallisneria (Hydrocharitaceae) species in Japan with a special emphasis on the commercially traded accessions and the discovery of hybrid between nonindigenous V. spiralis and native V.

denseserrulata. Aquatic Botany 128:1-6.