Improvement of the Nursery and Grow-out Phases of the American Oyster Life Cycle

Final Report

Badger Bay Mussel Farms Ltd.
AIMAP-2010-N07

Table of contents

Introduction
Workplan
Results/Discussion
Procedure for Settlement/Epinephrine Treatments
2008 Oyster Stock
2009 Oyster Stock
2010 Oyster Stock
Conclusion/Recommendations

Introduction

Badger Bay Mussel Farms Ltd. (BBMF), with  the financial  assistance of  the Canadian  Centre for Fisheries  Innovation  (CCFI), the National Research Council's Industrial Research Assistance Program (NRC-IRAP) and  the technical assistance of  the  Marine  Institute's (MI) Centre for  Aquaculture and Seafood  Development (CASD) has sought to establish a commercial oyster  aquaculture venture since 2001.  Using this knowledge, BBMF and the Marine Institute designed a three year plan to develop market sized oysters in a timely manner with accepted growth rates and minimal mortality.

The 2010-2011 oyster work was a continuation of work completed in the first year of the three year project and the tracking of the 2008, 2009 and 2010 at-sea oysters.  The current project has permitted the MI and BBMF to maintain some of the 2010 oyster seed at the MI until at sea deployment, track the growth and survivability of the 2008, 2009 and 2010 oysters at the grow-out site in Placentia Bay (Bearberry Cove) and to continue with transfers of eyed larvae, spat and broodstock from the Shippagan Hatchery to MI. This past year also allowed the MI and BBMF staff to deploy two different types of seed collectors at Two Guts Pond in August 2010 anti to refine the techniques used in the oyster rearing cycle to achieve greater success in survivability and quality. There are approximately 60,000 2010 year class oysters remaining at the MI that were maintained over the winter for spring 2011 deployment.

BBMF has an ongoing project with Department of Fisheries and Ocean's Aquaculture Innovation and Market Access Program (DFO-AIMAP) to purchase nursery equipment needed to increase the productivity of the oyster venture and to decrease labour costs.  Funding was presented to BBMF to attain a Quick-tube grader, several different grow-out gear types at various sizes required for oyster maintenance and an Italian brush system used to automatically free gear of biofouling organisms. All of these equipment purchases will be assessed in year 3 of the oyster project (2011-2012).

With the financial assistance of the Canadian Centre for Fisheries Innovation (CCFI) and the National Research Council's Industrial Research Assistance Program (NRC-IRAP) BBMF intends to attempt staged oyster rearing using the three year plan with both Newfoundland and Shippagan  broodstock. This 3 year plan includes gear development for algae growing, hatchery and at sea grow-out, growth and survival trials and site selection for grow-out.

The following report highlights results from April 1, 2010 to March 31, 2011, although the results and discussion of this report will include information from previous oyster projects to establish a clearer understanding of the process and the successes developed over the years.

Workplan

Badger Bay Mussel Farms requested the technical assistance of the Marine Institute to complete the following tasks from April 2010 to March 31, 2011:

1. Transfer of diploid oyster spat from the Shippagan Hatchery to the MI nursery downweller/upweller system in April and May 2010. These animals were held at the MI and fed algae and algae paste until they were deployed in July, 2010.  The number of diploids transferred was approximately 400,000 over two separate transfers.  Transfers did not take place until approval from DFA and DFO was granted.
   
   
2. Transfer of approximately 600,000 eyed oyster larvae from the Shippagan hatchery to the MI in April 2010.
   
   
3. The 2009 diploid oyster spat that were maintained at the MI over the winter were deployed to grow-out gear in July 2010 and some of the 2010 diploid oyster spat were deployed in Placentia Bay in July 2010 when the health status of the oyster spat was confirmed.
   
   
4. Spat collection at Two Guts Pond was attempted using two types of industry spat collectors. Deployment occurred in August 2010 and in October 2010 the collectors were retrieved.
   
   
5. Spawning attempts continued for the Newfoundland and Shippagan broodstock in April and May 2010 using modifications to the 2009 protocols to ensure success of spawning.
   
   
6. In order for the oyster industry to grow in Newfoundland, better nursery equipment is required. The pearl nets have a tendency to become fouled at a rapid rate and this was observed in 2010 to a greater extent than previous years. With assistance from AIMAP, different technologies such as, the Italian automated brushes and SEAPA longline baskets with caps were bought and some of the equipment was tested to determine the best nursery equipment to use in Placentia Bay.
   
   
7. The  growth  and  survival of the 2009 year class oysters was determined for both the oysters which were overwintered at MI and the oysters that were overwintered in Placentia Bay waters when the MI overwintered oysters were deployed in July 2010. This gives a better understanding if it’s more feasible to feed the oysters in a controlled environment compared to a natural environment over the winter in Newfoundland.
   
   
8. At sea oyster grow-out gear was evaluated in summer 2010 to March 2011. There were routine samplings in summer and fall to evaluate the performance of the grow-out gear in 2010-2011. Badger Bay Mussel Farms and Merasheen Island Mussel Farm in conjunction with DFO had already  purchased some gear types to accommodate different sized oysters in October 2009 (For example: pearl nets, floating bags, dark sea trays, Australian purses, Oyster Gro Systems,   etc.). Therefore, the results will take into account these gear types.
   
   
9. Some of the commercial grow-out gear consists of plastic parts which may be compromised in Newfoundland's unpredictable ocean environment. Therefore, it was proposed to research, design and build an at sea grow-out cage that would withstand harsh conditions. This cage was not designed or built due to budget constraints and the decision to focus on getting the algal growing system in working condition which cost more than was originally budgeted.
   
   
10. Transfer of approximately 20 broodstock in November/December 2010 from the Shippagan Hatchery in New Brunswick or from another source. This transfer was delayed due to permit issues and did not occur until March 2011. Spawning attempts are being initiated.
    
    
11. Transfer of approximately 500,000 eyed oyster larvae from the Shippagan hatchery to the MI in January/February 2011 was proposed. In actuality, there were approximately one million eyed larvae transferred to the MI on December 20, 2010.  Larval rearing and settlement were tried using the recommendations that were made at the time of the previous project final reports. The collected spat were switched over to an upwelling system where they will grow until at sea deployment.
    
    
12. It was proposed to transfer 150,000 to 200,000 triploid oyster spat from the Shippagan Hatchery to MI for delivery in April 2011. Unfortunately there were no triploid oysters available.
    
    
13. Continuation of tracking the 2008 and 2009 year class oysters that were deployed in July to November 2009.  Survival and growth rates were tracked for the 2010 diploid spat.  The survival and growth rates were determined through regular sampling trips to Placentia Bay.  Sampling included morphometries, mortalities and Sonde casts to monitor oceanographic parameters.
    
    
14. Samples were taken in fall 2010 and glycogen level testing was performed to determine energy reserves of oysters to give an indication of overwintering survival.
    
    
15. Three new upwellers with improvements from the older upwellers were built in preparation for future successful spawnings and larvae settlement.  The extra space was used to sufficiently spread out the oyster spat population.
    
    
16. In a prior project, different algae pastes were evaluated.  A shellfish diet from Reed Mariculture in conjunction with live food is now being used as a result of that project.  No new shellfish diets were found this year, therefore experiments were not completed with diets.

Results/Discussion

Transfer of New Brunswick broodstock, eyed larvae, spat and husbandry

Transfer of 17 conditioned broodstock from the Shippagen Hatchery in New Brunswick to the MI Aquaculture facility occurred on April 12, 2010.  The NB broodstock was already conditioned at the Shippagan hatchery and was held and treated in the same fashion as the NL broodstock (semi­square one metre tanks with aeration and water changes every second day or when required).  Spawning attempts were started as soon as they arrived at MI.

Prior to the transfer of the eyed larvae, four conical tanks were set up with aeration and the appropriate temperature.  There was to be a transfer of approximately 500,000 eyed oyster larvae from the Shippagen hatchery to the MI in December 2009 or January/February 2010. Unfortunately, this did not occur due to the same delays as the broodstock transfer.  Two eyed larvae transfers took place on April 12 (210,000 larvae) and 14 (400,000 larvae), 2010.  The project team developed a new way of larval rearing and settlement because this was a major bottleneck in the oyster rearing protocols followed in the past. The larval rearing attempts in the past were successful to the point of settlement and development in upwellers.  There were problems with the settlement plates that were used and the timing of transfer into an upwelling system.  In this project, the usage of epinephrine in our settlement process was investigated. It was determined earlier that the upwelling system was used too early and aeration was too turbulent for the sensitive settling larvae.  Therefore, for this project when the oysters were prepared to settle (300 microns) they were placed in a downwelling system, epinephrine was added and they were filtered every day for 2 weeks.   The collected spat were switched over to an upwelling system where they grew until at sea deployment.

Procedure for Settlement/Epinephrine Treatments

Epinephrine treatment:  Dissolved 0.1832g of epinephrine in a small amount of hydrochloric acid, then diluted the mixture in 10L of filtered seawater.  This mixture was used to treat two million larvae. The exposure time was 90 minutes.

Presence of broken shells was probably due to removing the spat from the sides of the upweller.  It was discovered from the Shippagan Hatchery that they used paraffin wax to coat the sides of the upweller so the spat would not settle on the sides of the tank. This technique was used during the next settlement opportunity.

An attempt was made this year to receive larvae at an earlier time than in previous transfers to allow the project team a head start on production. Approximately one million eyed larvae were transferred to the MI from the Shippagan hatchery on December 20, 2010.  Larval rearing and settlement were tried with modifications to the existing protocols.  New conical tanks were used and most of the larvae were swimming within the first couple of hours after introduction to the system.  Larvae were sieved after each water change and the animals greater than 300 microns were placed in the downwelling system. Epinephrine treatments were completed every second day until al of the oysters had settled.  The collected spat were switched over to an upwelling system where they will grow until at sea deployment in spring/summer 2011.  For this settlement the paraffin wax treatment was used on the upwellers and worked well. There was no spat settlement on the sides of the upwellers, but there was some settlement on the airlines, pumps, etc. Approximately five thousand animals successfullysettled and continue to grow as spat.  It was discovered from the hatchery in New Brunswick that they use a settlement substrate in conjunction with the epinephrine treatment.  This   technique will be used at the next available opportunity.

83,000 1mm spat were transferred from New Brunswick on March 31, 2010 and a second transfer of 311,000 spat   occurred on May 18, 2010.  These were held in the upweller systems until at sea deployment in July.  In February 2011 the spat that were left from the 2010 year class were graded and spread out over 4 tanks instead of the 2 tanks they were held in. The majority of the animals are now between 800 microns to 5 mm but approximately 50 of them are 15 mm or greater. A larger upweller cylinder has been constructed from larger pipe - 14 inch diameter instead of 6 inch.  A larger cylinder will give the animals additional surface area in the tank to grow and will enable more area for settlement.

All of the oysters were fed algae in accordance with their size and were supplemented with Shellfish Diet purchased from Reed Mariculture.

Newfoundland and New Brunswick Broodstock Husbandry/Spawning

Transfer of 12 Newfoundland broodstock from the DFO laboratory to a quarantined system at the
Aquaculture Facility at the MI was completed on November 13, 2009.  These oysters, as well as the 17 New Brunswick broodstock, were held in quarantined tanks with aeration and frequent water changes.

Conditioning began when the broodstock were acclimatized to the MI facility after their transfer.  Food in the form of three live algal species (Tetraselmis sp., Chaetoceros sp. and T-Iso) and an artificial diet (Shellfish Diet 1800 from Reed Mariculture) were added daily.  Temperature was controlled by heating units and as temperature was increased additional heaters were added and the   photoperiod was controlled at 16 hours day light: 8 hours darkness.

Gamete production in early February would enable the production cycle to be completed for a June ocean transfer. This would give the new oyster spat the opportunity to take advantage of higher water temperatures and food availability. Spawning of the Newfoundland broodstock was attempted on March 10, 2010 and March 29, 2010. Attempts to induce spawning consisted of thermal cycling with a temperature differential of approximately 11-12°C and addition of serotonin which is a hormone proven to trigger release of gametes. No spawning occurred in the Newfoundland broodstock.

The first spawning attempt was made with the New Brunswick broodstock on April 27, 2010 using the thermal cycling and serotonin addition protocols, but no spawning occurred at that time.

A second attempt was made on May 12, 2010. The oysters were held overnight in the refrigerator at 4°C.  They were added to the spawning tank in a cold cycle to begin the thermal eyeing.  After three thermal cycles spawning did not occur for this attempt.

The third attempt was made on May 13, 2010 with refrigerated oysters that were left at 4°C overnight. The cycle was started with warm water and after two cycles the broodstock were moved to a naturally lit area and serotonin was added; spawning was successful. Cell division was observed and the larvae were kept in a container for the initial stages of blastula, gastrula and trocophores.  The larvae were fed daily with cultured and artificial Nannochloropsis.

In November 2010, re-conditioning of the existing broodstock began using some revised techniques. Heated, unfiltered seawater is being used to make sure that as many nutrients are getting to the broodstock as possible. The Newfoundland and New Brunswick Broodstock are currently at l6°C and at a 15 hour photoperiod. Using an experimental design, the hope is to condition these animals using artificial and live feed and controlling some of the environmental factors that trigger gamete development. One such environmental factor is salinity and it is being decreased gradually to simulate spring runoff and the influx of freshwater.

In March 2011, 20 new conditioned broodstock were transferred from the Shippagan hatchery in order to perfect the spawning, fertilizing and larval rearing techniques.

2008 Oyster Stock

July 2009 deployment

The first at sea deployment of oysters occurred on July 23, 2009.  The oysters were graded into four size groups before deployment. The deployment set-ups were pearl nets (2 mm to 6 mm) for all size groups on the single line, but some of the largest oysters were also placed in floating bags (9 to 12 mm) on the double line. The number of oysters deployed initially is an approximation because some of them were very small and it was difficult to get an accurate count.  The counts that were done in September 2009 provided   more accurate total numbers. The thermograph that was deployed at the oyster site on July 23, 2009 was not collected during the 2010 field trip due to difficulty in finding it.

September 2009 on site assessment

On September 22, 2009 all of the 2008 stock that was initially deployed in the pearl nets was transferred to the floating bag set -ups. An additional 10 floating bags were deployed with the 2008 stock.  The 4 grades were kept separate by marking the bags with colored electrical ties.

To track the growth and mortality of the 2008 stock a three pearl net set-up was used with 50 oysters in each net. Two Australian purses (12 mm and 17 mm) were also stocked with 100 of the 2008 stock in each purse. Individual measurements were taken and recorded on each oyster in each pearl net and Australian purse.  These set-ups will remain throughout the grow-out cycle and measurements will be taken on each sampling trip. The large grade had greater morphometries values than the largest grade. This is probably due to the small sample size being used.

November 2009 on site assessment

On November 5, 2009 all of the 2008 stock that were deployed in the floating bag set-ups in September, 2009 were spread out over 24 floating bags.  The four different grades were kept separate by marking the bags with colored electrical ties so they could be tracked over time.

Measurements were also taken on 50 random oysters from each grade with the averages. Onemortality was recorded in the 17 mm Australian purse set-up.

June 20 I 0 on site assessment

A sampling trip to Bearberry Cove occurred on June 23, 2010 and the oysters from the mortality nets and the different oyster equipment were sampled. All of the 2008 stock that was deployed in the floating bag set-ups in November 2009 was sampled.

There were no significant differences in the morphometries of all of the gear types which is interesting especially when observing the pearl nets.  It was believed that the bottom net may have less growth and more mortality as time progressed, but this is not occurring.  Therefore, it may be possible to sink the oyster grow-out equipment lower than originally thought without any ill effects. This will continue to be tracked in the upcoming sampling trips.  This will ultimately affect how the oyster farm is managed and is an important piece of information to have for this particular site in going forward. Measurements were taken on 50 random oysters from each grade that was deployed.

October 2010 sampling

The 2008 stock that was deployed in the floating bag set-ups in November 2009 was sampled. The site manager removed the oysters from the Australian purses because of fouling and placed them in two pearl nets but did not take into account the mesh sizes that they were transferred from. The oyster measurements will be combined for these nets.  The decision was made to remove the width from the morphometries because the industry standard is to take account of the length and especially the height (cupping). Mortalities: 1 in top pearl net, 0 in the middle pearl net, 1 mortality in the bottom pearl net and no mortalities in the combined Australian purses. Measurements were taken on 50 random oysters from each grade that was deployed with the averages.

The oysters doubled their size in the first two months in the small and medium grades and the larger grades almost doubled their size.  After the initial growth spurt there was very little growth from November 2009 to June 2010. This is probably due to low food levels or a stressful event that may have occurred.  Then from June to October 2010 the length and height increased again due to the increased levels of nutrients in the water column.  All four grades grew similarly and are all between 33-40 mm lengths and 12- 14 mm heights.

2009 Oyster Stock

September 2009 deployment

On September 22, 2009 the largest of the 2009 oyster stock was deployed in pearl nets (34 pearl net set­ ups with 2 nets per set-up, for a total of 68 pearl nets) on the s ingle line.  The pearl nets were stocked with 12 g of oysters (approximately 936 oysters/net).  The total amount deployed was approximately 63,648 oysters.

November 2009 deployment/sampling

On November 5, 2009 the second lot of 2009 oyster stock was deployed in pearl nets (19 pearl net set­ ups with 2 nets per set-up, for a total of 38 pearl nets) on the single line.  The pearl nets were stocked with 10 g of oysters (approximately 900 oysters/net).  The total amount deployed was approximately 34,200 oysters.

During the November sampling trip, it was noticed that there was a lot of fouling on the pearl nets.   To help remedy this situation 30 periwinkles per pearl net were added.  An instrument was deployed to record tidal cycle which can then be used to help calculate flushing rates and flux (food values).

June 2010 gear and site assessment

The 2009 oysters that were deployed in 3mm pearl nets during September and November of 2009 were assessed. Measurements were unable to be taken because of the excessive amount of fouling and growth of clams, mussels and scallops within the nets.  The September pearl nets were observed to have more fouling than the November deployment.  It seems that the oysters in the pearl nets were deployed at the time for settlement of other animals, this compromised the efficient growth of the oysters.  The oysters were still very small and handling was minimized.  The decision was made to wait until the sampling trip in July in order to have the time to pick through every net to retrieve the oysters.

July 2010 deployment
Prior to the deployment of the remaining 2009 oysters that were held at the MI over the past year a grading and approximate count was completed.  The animals were still small and could be deployed in 3mm oyster gear.  The number of animals deployed was approximately 19,000.

The 2009 runts that were held at the Marine Institute for a year were deployed into three 3 stacks of
3mm oyster baskets and were tied to a main single line that was previously put out. 

The oysters that were deployed in pearl nets were assessed again and separation of the oysters from the other fouling animals was attempted.  Grading of the animals and picking was tried but both attempts failed due to the vast number of clams that were present and attached to the nets.  It was decided and communicated to the site manager that the pearl nets would be dealt with in some manner during the fall sampling.

October 2010 sampling

The Bearberry Cove site was visited on October 5, 2010 and we were informed by the site manager that he had to remove all of the 2009 stock pearl nets due to the extensive fouling and the oyster mortality. Measurements were taken on 50 random oysters.  The oysters increased their size by approximately three-fold from July to October  probably due to increased  food  in the water.

2010 Oyster Stock

2010 diploid spat and eyed larvae were transferred to the Marine Institute from the Shippagan hatchery in March, April and May 2010. These were held in the upweller systems until at sea deployment in July.

On July 14, 2010 the 2010 stock was graded and three sizes emerged: l mm (small grade), 3 mm (medium grade) and 6 mm (large grade).  The approximately 75,000 1 mm animals were held at the Marine Institute aquaculture facility over the winter and will be deployed in spring/summer 2011.

July 2010 deployment

On July 16, 2010 the medium and large grades were deployed at the Bearberry Cove site in 3 mm and 6 mm oyster baskets.  The medium grade oysters went into 14 sets of three-3 mm baskets and the large grade went into 3 sets of three-6 mm baskets. No oysters were placed in pearl nets due to the fouling problems that were experienced in the previous year.

October 20 I 0 sampling

A CASD technician and a BBMF employee visited the Bearberry Cove site on October 5, 2010 and sampled the 2010 medium and large grades of oyster stock. Measurements were  taken on 50 random  oysters. The oysters showed only a minimal increase in size from July to October.

This 2010 oyster stock will be tracked in the upcoming year for mortality, growth and morphometries.

Algal growing system

Algal growing system specifications were researched and a Seasalter Continuous Algal Production System was sourced at Bay Shellfish in Florida, USA.  An eight bag system was ordered on March 2, 2010 and arrived in late March 2010.  The system was set-up in an appropriate area at the MI facility in June 2011.  The following lists the items purchased to set up the eight bag system:

1. 100 custom poly bags
2. Eight custom fitted poly shaped bases
3. High temperature glass injection ports for air, water and harvesting
4. Water, harvest and delivery lines
5. Cages to hold the bag structure
6. Lights

The electrical setup for the algal bag system was costlier than was expected. The decision was made by the client,  MI  and  the  funders to allocate the funding that was supposed to be used for researching, designing, building and testing an at sea grow-out cage to the algal bag system. This was a better use of the funds at this point in time for the development of our rearing techniques.

New Upwellers

Three new upwellers with modifications were built in April 2010 to house the extra oyster spat. They are now being used for the spat that were transferred from New Brunswick in March 2011and the 2010 stock that are being held at MI until spring deployment.

Wild Oyster Spat Collection

A larval monitoring program was initiated by providing the monitoring equipment to Mr. Richard Dollimount (Fisheries Officer, Stephenville, NL).  No larval monitoring was completed at Two Guts Pond because of Mr. Dollimount's employment situation.

In June and July 2010, equipment to build oyster spat collectors was purchased and two types of collectors were built: Chinese hat collectors and modified corrugated weeping tile collectors. These collectors were limed and cured, then 10 of each collector was deployed at Two Guts Pond in August 2010.  Collectors were placed in areas where adults were found previously and where historical records showed that oysters were transferred in the 50's.  Larval monitoring was completed at this time and a thermograph was deployed.  Microscopic observations did show that our plankton samples did contain larvae, but it was not determined conclusively if these larvae were clam or oyster larvae because they are similar in appearance.

A field trip to Two Guts Pond in October was carried out to collect the deployed thermograph and the spat collectors and to observe the area.  This area was damaged by Hurricane Igor earlier in October and some of the collectors were displaced onto the land by high winds and seas.  The collectors that were in the water were damaged as well and moved to different areas.   These were retrieved but no spat collection was observed.

Oyster Nursery and Grow-out Equipment

BBMF has an ongoing project with DFO's Aquaculture Innovation and Market Access Program (AIMAP) to purchase nursery equipment needed to increase the productivity of the oyster venture and to decrease labour costs.  $35,000 was presented to BBMF to attain a Quick-tube grader, several different grow-out gear types at various sizes required for oyster maintenance and an Italian brush system used to automatically free gear of biofouling organisms. All of these equipment purchases will be assessed in year 3 of the oyster project (2011-2012).

Glycogen Testing

Glycogen level testing was performed to determine energy reserves of oysters to give an indication of overwintering survival. The 2008 oyster stock was brought back to the MI from the October 2010 field trip to Bearberry Cove, processed and frozen in MI's -80°C ultra low temperature freezer.  The samples were then used in a glucose assay kit to determine a baseline glucose concentration.  This baseline will be used for comparisons in the following years.

Conclusion/Recommendations

New techniques in larval rearing and settlement, such as the epinephrine treatments and wax coating on the upwellers, were tested and evaluated and both were successful.  After speaking with the hatchery manager at the Shippagan hatchery, more techniques were discovered such as the use of specific cultch in conjunction with the wax coating and broodstock handling for conditioning. These new techniques will be tested in March and April 2011 with the new broodstock, larvae and spat.

A greater realization of the importance of visiting an oyster hatchery came to the forefront when important techniques were learned after spawning and settlement attempts.  A visit is being organized with Shiappagan hatchery for the spring 2011.

In future oyster projects,  the concerns that Newfoundland provincial officials have regarding additional health testing, new oyster grow-out sites and closing the oyster  rearing loop so that all activities can take place in Newfoundland and Labrador will continue to be addressed.

The 2008 oyster deployments in Placentia Bay continue to be a great success and favourable overwintering results of the 2010 stock are expected at Bearberry Cove in spring 2011 are expected.

The following workplan is recommended for the next project:

1. Year 3 will be a repetition of the life cycle of the American oyster in the hatchery, nursery and grow-out area. There will be a fine tuning of protocols and changes to procedures to allow better efficiency.
   
   
2. Selection of a more appropriate nursery site has become an issue for BBMF.  The current site is not located in close proximity to any roads, wharves or infrastructure required for nursery rearing.  It is an excellent site for grow out but it is not ideal because of the logistics required to get to the site for maintenance and nursery equipment tending.  Site selection is a top priority this coming year and some appropriately located areas will be investigated.  This will entail visits to applicable areas, placing water quality equipment on the sites and applying for experimental licenses if required.
   
   
3. Transfer of diploid oyster spat from the Shippagen Hatchery to the MI nursery downweller/upweller system in March/April 2011. These animals will be held at the MI and fed algae (for first 6 weeks)/algae paste (to deployment) until they reach approximately 3 to 5 mm. The number of diploids will be approximately 500,000 depending on the amount available at the time of transfer. All transfers will be pending approval from DFA and DFO.
   
   
4. Transfer of approximately 20 broodstock from the Shippagan Hatchery in New Brunswick or from another source. The NB broodstock will be conditioned using the standard protocols that were previously mentioned in Year 1 and 2.  Spawning attempts will be made when conditioning is sufficient.
   
   
5. If there are approximately 65 mm. oysters available from the deployments in Placentia Bay the project team will use some of these as continuing Newfoundland broodstock.  A Newfoundland Broodstock Program may be implemented depending on genetics, growth and survivability over time.
   
   
6. Spawning attempts will continue for the Newfoundland and Shippagan broodstock. Modifications to 2009 and 2010 protocols may have to be made to ensure success of spawning.
   
   
7. Oyster spat collection at Two Guts Pond will be attempted again using two industry standard collectors.  More intensive larval monitoring will be attempted before deployment of the collectors.  Divers will also be employed to look for more adult oysters at Two Guts Pond and an experimental license will be in place to transfer some adults back to the Marine Institute if available.
   
   
8. At sea oyster grow-out gear will be evaluated in summer 2011to March 2012. There will be routine sampling to evaluate the performance of the grow-out gear in 2011-2012.
   
   
9. Transfer of approximately 1,000,000 eyed oyster larvae from the Shippagan hatchery to the MI in January/February 2012.  Larval rearing and settlement will be tried as in other projects while using the recommendations that were made at the time of the project final reports. The collected spat will then be switched over to an upwelling system where they will grow until at sea deployment.
   
   
10. Continuation of tracking the 2008, 2009 and 2010 and newly deployed 2011 year class oysters that were deployed in Placentia Bay. Survival and growth rates will also be tracked for the 2010 and 2011 diploid and triploid spat.  The survival and growth rates will be tracked through regular sampling trips to Placentia Bay to March 2012.  Sampling will occur at 1 month after deployment and 3 months after deployment. Sampling will include morphometries, mortalities and Sonde casts to monitor oceanographic parameters.
    
    
11. Glycogen level testing or some other test to determine energy reserves of oysters will be used annually in November to give an indication of overwintering survival.
    
    
12. Conduct a technical mission to visit the Shippagan Hatchery in New Brunswick to learn the techniques used in the rearing of Eastern oysters.  If the Shippagan hatchery is unavailable then a different oyster hatchery will be pursued.