EFFICIENT AND RELIABLE PROTOCOLS FOR THE PRODUCTION OF LIVE FEEDS FOR LARVAL FLORIDA POMPANO

Meghan L. Anderson*, Charles R. Weirich, and Fernando G. Cavalin USDA Agricultural Research Service and Center for Aquaculture and Stock Enhancement
Harbor Branch Oceanographic Institute at Florida Atlantic University 5600 US Highway 1 North Ft. Pierce, FL 34946 USA
BACKGROUND Production of live feed organisms represents the majority of time and labor associated with marine finfish larviculture operations. As a byproduct of establishing a reproduction and larviculture research program at our facility, procedures for the production and enrichment of rotifers and Artemia for the successful rearing of larval Florida pompano have been improved and streamlined. Our live feeds culture protocols are efficient, reliable, and easily support both research and commercial-scale larviculture of this species. ROTIFER PRODUCTION Rotifers are batch cultured using a series of four fiberglass tanks containing 125 L of water. Water temperature and salinity are maintained at 26-27 C and 20-25 g/L, respectively. Each tank is supplied with gentle blown air at four points with a one point source of pure oxygen to maintain DO above 100% saturation. Rotifers in each culture tank are fed concentrated Nannochloropsis paste via peristaltic pump every 4 h. Culture Selco Plus is added at 1030 and 2200 hours. One tank is harvested daily (three days after stocking) at a density of 700-1,000 rotifers/ml. Once they are enumerated, rinsed rotifers are stocked into enrichments buckets and remaining rotifers are restocked into a tank containing sterilized water at a rate of 250 rotifers/ml.
Feeding regime for larval pompano
Overhead view of rotifer culture tank Harvesting rotifers

ARTEMIA DECAPSULATION Artemia cysts are decapsulated on site using standard procedures. Cysts are then stored at 1-3 C in a hypersaline solution until needed.

Larval Florida pompano at 9 days post hatch

S strain rotifers

Rotifer culture area

HATCHING ARTEMIA CYSTS Hatching occurs in two fiberglass tanks containing 175 L of sterilized water. Water temperature and salinity are maintained at 30 C and 28-32 g/L, respectively. Each tank is supplied with vigorous blown air and pure oxygen to maintain DO above 100% saturation. Prior to the addition of cysts, a disinfectant (Sanocare), sodium bicarbonate, and an antifoaming agent are added. Hatching is complete in 12-14 h with typical harvest amounts of 50-90 million 1st instar nauplii.
1st instar Artemia nauplii

Decapsulating Artemia cysts

Stored decapsulated Artemia cysts

Harvesting and rinsing Artemia

Artemia daily schedule

8:30 AM 10:00 AM 11:00 AM 1:00 PM 5:00 PM 6:00 PM 9:00 PM 11:00 PM

Feed larvae

1st

instar nauplii and/or enriched

2nd

instar nauplii from cold storage

Harvest 1st instar nauplii, start enrichment tank, cold store remaining if necessary Harvest enriched 2nd instar nauplii Artemia, cold store Feed larvae, add DC DHA Selco to enrichment tank (T0) Feed larvae Artemia hatching tank started (~16 hours to hatch) Feed larvae Add enrichment (T10)
Removing membranes using oxygen

Microfeeds Enriched 2nd instar nauplii 1st instar nauplii Enriched rotifers

ROTIFER ENRICHMENT AND COLD STORAGE Enrichment occurs in buckets containing a final volume of 15 L (salinity = 28-32 g/L) at a density of 1,500 rotifers/ml. Buckets receive blown air and pure oxygen to maintain DO levels above 100% saturation. The enrichment diet (Ori-Green) is then applied and rotifers are allowed to feed for 3 h at 26-27 C. Once enrichment is completed, rotifers are rinsed to remove residual enrichment diet and are placed into a clean bucket containing a final volume of 15 L. Ice bottles are added as needed to lower the temperature to 10 C over a two hour period. Buckets are then placed into a refrigerated cooler (with ports for blown air) for cold storage up to 24 h at 5-7 C.

Adding DC DHA Selco to Artemia enrichment tank

Adjusting aeration and lights of Artemia hatching tank

Green water Metamorphosis

0
Distributing rotifers to experimental larviculture tanks

12

15

18

21
Adding ice bottles to Artemia prior to cold storage 2nd instar enriched Artemia nauplii

Age (days post hatch)

Daily rotifer schedule

8:30 AM 9:00 AM 9:30 AM 10:00 AM 10:30 AM 1:00 PM 5:00 PM 9:00 PM 10:00 PM

Feed larvae enriched rotifers Assess rotifer populations Harvest tank, prepare enrichment containers, restart tank Enrich rotifers Refill algae containers, AM Selco feeding Harvest, rinse, feed, and cold store rotifers Feed larvae enriched rotifers Feed larvae enriched rotifers PM Selco feeding
Buckets of Artemia in cold storage

ARTEMIA ENRICHMENT AND COLD STORAGE Artemia are rinsed and cold stored (at a density not exceeding 2,500 Artemia/ml) as described for rotifers or they are stocked at a rate not exceeding 300 Artemia/ml into one of two available 175 L enrichment tanks containing sterilized water (temperature = 26 C, salinity = 28-32 g/L). Artemia are enriched for 20 h with the addition of equal amounts of DC DHA Selco at T0 and T10. Harvested enriched 2nd instar nauplii are rinsed thoroughly and cold stored (5-7 C) at a density not exceeding 2,500 Artemia/ml.