AQUACULTURAL ENGINEERING

Development of an open hardware 3-D printed conveyor device for continuous cryopreservation of non-batched samples
Liu Y, Eskridge M, Guitreau A, Beckham J, Chesnut M, Torres L, Tiersch TR and Monroe WT
A great challenge among communities participating in germplasm repository development is to obtain suitable cryopreservation equipment and devices. Commercial programmable freezers are costly and thus unaffordable to many users. Self-made devices have substantial variability among users, resulting in few opportunities for standardization across communities. The development of open hardware with the increasing accessibility of three-dimensional (3-D) printing offers rapid prototyping and easy fabrication of devices by users around the world at low cost. The present study explored the feasibility of developing operational prototypes of 3-D printed motorized cryopreservation devices for continuous freezing of non-batched samples. A controlled cooling conveyor device (CCCD) was designed and fabricated to cryopreserve sperm samples in straws that were loaded onto chain links suspended over liquid nitrogen held in a Styrofoam box. Cooling rates of 5 to 34 °C/min for 0.5-ml French straws were produced by adjusting the height of conveyor chains, slopes, and liquid nitrogen mass. The plunge temperature (-47 °C to -61 °C) was controlled by adjustment of conveyor speed. The cooling curves from the CCCD were comparable to a commercial programmable freezer. There were no significant differences in post-thaw motility of sperm from ornamental (Koi) common carp () among samples frozen with the CCCD and those frozen with a commercial programmable freezer. The post-thaw sperm motility was consistent among samples frozen in the CCCD across a 15-min time span. The CCCD prototypes in the present study proved to be feasible and functional as low-cost, customizable, portable, and yet standardizable options for freezing of individual (non-batched) samples. Additional design alternatives are proposed to facilitate further adaptation and development by diverse user communities.
3-D Printed Customizable Vitrification Devices for Preservation of Genetic Resources of Aquatic Species
Tiersch CJ, Liu Y, Tiersch TR and Monroe WT
Sperm vitrification as an alternative approach to conventional cryopreservation (equilibrium freezing) allows quick and low-cost sample preservation and is suitable for small-bodied aquatic species with miniscule testis, fieldwork at remote locations, and small-scale freezing for research purposes. The goal of this present study was to develop operational prototypes of 3-dimensional (3-D) printed vitrification devices with innovative components that can provide comprehensive functionalities for practical repository development for aquatic species. The design featured an elongated loop to suspend a thin film of sperm sample in cryoprotectant, a retractable sleeve to protect the vitrified samples and allow permanent labeling, a handle to facilitate processing and storage, and a shaft with annular grooves to guide positioning of the protective retractable sleeve. To span a wide range of sample capacities and configurations, a total of 39 different configurations (3 loop lengths ×13 loop heights) were fabricated by 3-D printing with the thermoplastics polylactic acid (PLA) and acrylonitrile butadiene styrene (ABS). A total of 86 devices were fabricated with ABS filament with a print failure rate of 9%, and 97 devices were fabricated with PLA filament with a failure rate of 20%. Major types of printing failures included disconnected loops, insufficient build surface adhesion, stringing, and inconsistent extrusion. The sample volume capacity ranged from 1-47 μL and had linear relationships to the loop lengths and layer numbers. Vitrified samples were observed in 10-mm and 15-mm loops fabricated with PLA and ABS but not in 20-mm loops. This study demonstrated the feasibility of development of standardized low-cost ($0.05 material cost) devices fabricated by 3-D printing with practical functions including vitrification, volume control, labeling, protection, and storage within conventional systems. These prototypes can be further developed, standardized, and used to assist development of germplasm repositories to protect the genetic resources of aquatic species by user groups such as breeders, hatcheries, aquariums, and researchers.
Framework for evaluating external and internal parameters associated with Sea Based Container Culture (SBCC): Towards understanding rearing success in European lobsters ()
Halswell P, Daniels CL and Johanning L
Sea Based Container Culture (SBCC) is a mariculture technique that relies on the natural maintenance of environmental conditions, such as Dissolved Oxygen (DO) concentration and feed availability. This paper discusses a framework to evaluate the rearing success of European Lobsters () in SBCC based on temporal and spatial variations of external parameters, including current velocity, wave velocity, turbulent fluctuations and dissolved oxygen concentrations. The temporal variations considered annual changes to the environment and the effect of biofouling growth, and the spatial variations considered the geographical location (case study of Falmouth bay, Cornwall) and vertical position in the water column. The internal parameters of the containers were modelled using transfer functions derived from previous experimental data. The internal parameters were compared to rearing limitations selected from available literature, which included foraging and mobility behaviours, and DO consumption. The time that internal parameters exceeded the rearing limitations was quantified, allowing rearing success to be predicted. This paper uses a case study of external parameters measured in Cornish waters, UK, to demonstrate the framework methodology. The framework showed that in situ measurements of current, wave and turbulence could be used to predict the internal parameters of SBCC containers, which can be used to predict theoretical rearing success based on rearing limitations. The framework indicated that DO concentrations within the containers should not affect rearing success; however, the foraging and mobility limits were exceeded by 0 to 30% of the time (depending on vertical position in the water column and assessment method). The paper aims to demonstrate the generic framework methodology and understands its limitations in predicting rearing success. The framework provides a tool to optimise the SBCC design for spatial and temporal varying conditions related to a geographical location or (vice versa) identify suitable mariculture sites based on SBCC design and environmental conditions. Additionally, the framework can optimise the vertical position of the SBCC in the water column and identify, from parameters considered, those that are most likely to affect rearing success.
Design and operation of a low-cost and compact autonomous buoy system for use in coastal aquaculture and water quality monitoring
Schmidt W, Raymond D, Parish D, Ashton IGC, Miller PI, Campos CJA and Shutler JD
The need to ensure future food security and issues of varying estuarine water quality is driving the expansion of aquaculture into near-shore coastal waters. It is prudent to fully evaluate new or proposed aquaculture sites, prior to any substantial financial investment in infrastructure and staffing. Measurements of water temperature, salinity and dissolved oxygen can be used to gain insight into the physical, chemical and biological water quality conditions within a farm site, towards identifying its suitability for farming, both for the stock species of interest and for assessing the potential risk from harmful or toxic algae. The latter can cause closure of shellfish harvesting. Unfortunately, commercial scientific monitoring systems can be cost prohibitive for small organisations and companies to purchase and operate. Here we describe the design, construction and deployment of a low cost (<£ 5000) monitoring buoy suitable for use within a near-shore aquaculture farm or bathing waters. The mooring includes a suite of sensors designed for supporting and understanding variations in near-shore physical, chemical and biological water quality. The system has been designed so that it can be operated and maintained by non-scientific staff, whilst still providing good quality scientific data. Data collected from two deployments totalling 14 months, one in a coastal bay location, another in an estuary, have illustrated the robust design and provided insight into the suitability of these sites for aquaculture and the potential occurrence of a toxin causing algae ( spp.). The instruments maintained good accuracy during the deployments when compared to independent in situ measurements (e.g. RMSE 0.13-0.16 °C, bias 0.03-0.08 °C) enabling stratification and biological features to be identified, along with confirming that the waters were suitable for mussel ( spp.) and lobster () aquaculture, whilst sites showed conditions agreeable for spp.