Alternate Aquaculture Vaccine Delivery System

Sponsors	Uidaho CNR [U of I][1]
Team Name	Fish-Vac
Duration	Summer 2020 - Fall 2020
Faculty Adviser	Dr. Matthew Swenson
Mentor	Max Johnson
Client	Dr. Ken Cain
Team Members	Hasan Alajmi Khari Amos Cody Gibson Ben McBride

Problem Definition

Project Goal: The goal of this project is to develop an alternate aquaculture vaccine delivery system to enhance antigen uptake and protection in juvenile salmonids.

Background

Fish vaccines used for aquaculture species are typically delivered by injection, immersion, or oral (feed) delivery. Of these methods, injection vaccination is by far the most effective and provides the greatest level of protection from disease. This is primarily due to direct delivery of antigens to immune cells rather than relying on uptake across the integument (skin) such as for immersion vaccination, or the need for intact antigens to pass through the harsh environment of the stomach during oral vaccination. Immersion and oral vaccines are the methods of choice for mass vaccination of small fish (1-15g) because of the ease of delivery when compared to injecting large numbers of small fish. However, if a mass vaccine delivery method could be developed that “injected” a vaccine into the integument in a new way and provide greater disease protection than immersion or oral vaccination, it would revolutionize the aquaculture industry.

Deliverables

The Device Must:

• Create a puncture or break in the integument(skin) of small fish without causing injury.
• Be designed to see repeated use.
• Be designed in a way that allows the transfer of used and new vaccine solution in and out of the system.
• Meet or exceed all IACUC regulations for animal research.

Specifications

Product Specifications:

• Must be sized for bench top testing version (1L of fluid and 240g of fish)
• Must be upscaled to hold 30 gal and be 27.4kg of fish at a time
• Must be functional for 1g-5g fish
• Must create puncture or integument(skin) of 85% of the fish
• Must allow for proper cleaning
• Must allow for all consumables to be easily changed
• Must be made of non-porous material i.e. polished stainless steel and UHMW
• Must have full operational capabilities in environments with both fresh and salt water around 15°C
• Must be capable of doing all necessary fluid changes
• The components shall comply with the Animal care and use protocol standards

Value Proposition Statement

Delivering vaccinations to humans 1 by 1 or by appointment is easy due to the construct of our society and freedom of communication. When giving vaccinations to hundreds of fish, scientists use the immersion vaccination method. This method submerges the animal into a water and vaccine bath, where the treatment is absorbed through the skin. The largest issue in utilizing this method, is the time it takes for the skin to uptake the medicine. Our project is constructed to assist in this method and cut down the immersion time. Our model works as a funnel for fish to to pass through, be harmlessly pierced by hypodermic needles, and to be briefly treated in a vaccinated bath. Our product will enhance the vaccine-skin uptake and make the immersion treatment method more effective and efficient to fish.

Design Considerations

When developing the device the following need to be considered:

• Materials such as wood, galvanized steel, and carbon steels cannot be used.
• If electrical components are used they must be sealed and water tight.
• All components of the system should be light and small enough for one person to safely handle.
• All components need to be easily accessed in order to be disinfected between uses.
• The first prototype should be able to handle 240 grams of fish per vaccine dip.
• Holding tank drains will be compatible with hoses for easy emptying and refilling of tanks.

Project Learning

Initial Design Concepts

Initial Design Idea:

Figure	Description
	Bed of Needles: Requires water to be drained from the tank r a large platform to be lowered into the tank Would require 1000's of needles A bed of needles isn't a readily available item

Micro-Needle Roller

Micro-Needle Rollers:

Figure	Description
	This product is a small cylindrical roller with exposed microneedles Commonly used to pierce the skin of humans to promote collagen development This product is adjustable, easily replaceable and relatively cheap

Design Concepts Using Micro-Needle Roller

Designs Implementing Use of Micro-Needle Rollers:

Sketches	Description
	Abrasion Column: This tank has an open base that allows fish to fall through when raised. Micro needle rollers pierce the fish skin. Treatment Bowl: This bowl allows the vaccine and fish to interact with one another for given amount of time. When time is reached, bowl drains vaccine and carries fish to be released into neutral environment. Solid Bowl: This bowl allows all sections and variables to be controlled and kept together. Compact and simple design with no expensive mechanical instruments.
	Abrasion and Overflow Plate: Fish will be dumped on the abrasion plate, piercing the fish. The overflow plate will catch the fish falling through the space between the roller and plate, then slid into vaccine bowl. Solid Bowl: This bowl allows all sections and variables to be controlled and kept together. Treatment Bowl: This bowl allows the vaccine and fish to interact with one another for given amount of time. When time is reached, bowl drains vaccine and carries fish to be released into neutral environment.
	Chute: For this design the fish along with water will be dumped into a pyramid shaped chute with a slot at the bottom. Positioned at the bottom of this slot will be one needled roller and one smooth roller. The fish will be forced between the rollers, then will fall onto an inclined plane of slotted UHMW which will remove the excess water before dropping them into the vaccine tank.
	Tube: For this design fish as well as rearing water will be poured into the top of the tube, which will flow down to a cone which will direct the fish into a stream that will flow over the roller which is mounted in a second tube. The rearing water will then be drained off and the fish will be deposited into the vaccination tank.
	Chute: This is an open top chute design. Fish will be poured into the top of the chute via net. A small set of spray nozzles will be placed near the top of the chute to assist the fish as they slide down to the rollers. A small sump pump will be placed in the vaccination tank below the chute so there is no need to add or remove rearing water. Two rollers will be placed at the chute exit. The top roller will be smooth and will adjust up and down for a range of fish sizes. The lower roller will be the needle roller. Cart: To make the system portable a cart will be used to transport the chute and vaccination tank. The chute will bolt to the cart and will accommodate a variety of tank sizes. Tanks will be removable to allow fast changeovers of vaccination fluid. A second net in this process will be placed in the tank prior to pouring fish into the chute to save time with netting.

Additional Research

Research:

Topic	Description
Pump:	12 V sprayer pump should be great for our application. They are relatively cheap, and they meet the necessary criteria:Self Priming, Modular, Must have at least 5 feet of vacuum, Pump more than 1 gpm, Standard sizes for fittings, Can be hard mounted A 120 V to 12 V may have to be used to accommodate the labs power source
Transportation of Fish:	Verbal conversation with Mr. Ford. He worked in Ken Cain’s lab for a year while attending the university of Idaho. He has an associates in Aquaculture for the College of Southern Idaho. Fish are moved from one place to another primarily using nets and tanks Other facilities that Mr.Ford worked (Evaqua, CSI fish hatchery, Pahsimeroi fish hatchery) also use the same methods of moving fish from one location to another For moving fish long distances to restock ponds, he used a fish stocking unit which was a large truck with a tank. When arriving at the location, the fish were evacuated through pipes that used gravity for draining Most of his experience of moving fish was when the fish were larger (6 inches and greater) Link to IDFG video for fish transportation :https://youtube/F4SkXAG6mfg
Fish Testing in Ken Cain's Lab:	Verbal conversation with Mr. Vega. He worked in Ken Cain’s lab for a semester. He has an associates in Aquaculture for the College of Southern Idaho. In Ken Cain's lab, tests are done in what are called challenge rooms Tanks in challenge rooms are around the size of an ice cream tub or 5 gallon bucket A stand for the apparatus is needed keep it upright inside of the small tanks Stand heights can vary to supports different water levels

Initial Prototype

Progression of Initial Prototype:

Figure	Description
	3D Model: Funnel Material-Acrylic Roller Mounts-Stainless Steel Weight of Funnel with Rollers-1 lb. 8 oz. Top Opening Dimension-(10x10) in. Bottom Opening Dimension-(1x1) in. Stand Width-(5x5) in.
	Cardboard Model: The cardboard model was created to validate the 3D model. This model was used for a meeting with our client to better explain our design and give a 3D visual aid of actual size.
	Initial Prototype Pre-Assembly: The acrylic funnel has been cut, prepped, and glued together. The roller mounts have been cut out of stainless steel and are in the process of being welded 3 different lengths of needle rollers have been disassembled and are ready to be placed in the roller mount when finished
	Initial Prototype 1st Iteration: This iteration uses a derma roller with the same needle spacing as used on human skin The fish in the picture have been anesthetized for testing Water was added to help the fish flow down the chute It was determined after multiple tests that the fish skin was not being punctured due to the high needle density Using this same iteration, multiple needle configurations were tested (.5, 1, 1.5mm) lengths, spacing needle packs, and clipping needles on needle packs with increased needle pack spacing
	Initial Prototype Final Iteration: This iteration uses a new roller that we designed and 3D printed The rollers are a two piece design that use 25 gauge hypodermic needles There are 4 needles per roller and both rollers have needles unlike the first iteration To decrease needle density we increased roller diameter which required a new stainless steel mount to be cut and welded Testing of this iteration proved that a lower density of needles would allow the skin of fish to be punctured validating our proof of concept The larger needled roller design is going to be implemented on the larger prototype

Final Prototype

Progression of Final Prototype:

Figure	Description
	3D Model Design 1: The size of the funnel has been increased to handle more fish Implements a pump and nozzles to help move fish through the chute Can be carried by one person The frame is adjustable for a range of tank sizes Chute separates from the frame to make the system easier to transport
	3D Model Design 2: Chute has been enlarged to accommodate new rollers which are larger in diameter Dividers have been placed in the chute to direct fish into one of the 3 roller assemblies New rollers are motor driven and are geared so each roller assembly will operate independently The frame has been enlarged to accommodate larger chute The hoses added are not to scale but give a representation of hose placement in final product Adjustable leg pins have been moved from horizontal plane to vertical plane
	Final Prototype Manufacturing: Picture 1) Fitting and tack welding of chute body Picture 2) Chute body fit up with first divider in place to be tack welded Picture 3) Chute and dividers in place and tack welded with 1.5 inch stich pattern Picture 4) Chute with roller guides and frame completely welded and ready for final assembly
	Final Prototype Assembled: Picture 1) Gear side of chute with rollers and hardware installed Picture 2) Motor side of chute with motors, pump, and wiring installed Picture 3) Top view of chute with pump and spray ball in view Picture 4) Isometric view (motor side) of final prototype assembly

Validation

Skin Puncturing Tests

Designs Progression of Initial Prototype:

Figure	Description
	Test 1: Fish carcasses we acquired from Dr.Ken Cains lab for testing skin thickness A 22 gauge hypodermic needle was used in the left 2 pictures to determine skin thickness Skin thickness was found to be ~.25mm The body weight of the fish (~3 grams) was used to determine the force required to puncture the skin The photo on the right was a test to determine if a fish would be able to slide into a needle on the same incline as the initial prototype A .5mm roller was rolled over a fish carcass to determine if the needles would puncture the skin and it was found that the skin could not be punctured without an excessive amount of force.
	Test 2: Water was used to assist fish as they went down the chute The weight of the fish was not great enough to puncture skin To puncture the skin of the fish, rollers were spaced closer together and turned by hand, but resulted in to large of a force and caused damage to the fish Needle length and density was changed, but all results were the same as the first test which caused too much damage to the fish
	Test 3: Lower needle density over a larger diameter roller resulted in punctures to the skin of the fish without causing excess damage No additional force was needed to puncture the skin Water was also used in testing of the new roller along with manually spinning the rollers to help guide fish through the chutes exit The needles used in this test are 25 gauge hypodermic needles and protrude from the roller .030"

Fish Cycle Validation

This is the cycle or flow of fish that will be seen during our testing of the initial prototype:

Team Members

	Cody Gibson Major: Mechanical Engineering Hometown: Burley, Idaho Responsibility: Wiki Page, Documentation Email: gibs7646@vandals.uidaho.edu		Ben McBride Major: Mechanical Engineering Hometown: Idaho Falls, Idaho Responsibility: Project Manager Email: mcbr5602@vandals.uidaho.edu
	Hasan Alajmi Major: Mechanical Engineering Hometown: Kuwait Responsibility: Solid Modeling, Budget Email: alaj9683@vandals.uidaho.edu		Khari Amos Major: Material Science Engineering Hometown: Idaho Falls, Idaho Responsibility:Communications Email: amos8304@vandals.uidaho.edu

Additional Documentation

Budget

• File:Fish Vac Budget.pdf

Client Interview

• File:Fish Vac CI 6 18.pdf
• File:Fish Vac CI 6 23.pdf

Design Validation Plan

• File:Fish Vacc. Design Validation Plan.pdf

Manufacturing Plan

• File:Team Fish Vac. Manufacturing Plan Rev.1.pdf

Meeting Minutes

• File:Fish Vac MM 6 24.pdf
• File:Fish Vac MM 6 30.pdf
• File:Fish Vac MM 7 2.pdf
• File:Fish Vac MM 7 7.pdf
• File:Fish Vac MM 7 9.pdf
• File:Fish Vac MM 7 14.pdf
• File:Fish Vac MM 7 16.pdf
• File:Fish Vac MM 7 21.pdf
• File:Fish Vac MM 7 23.pdf
• File:Fish Vac MM 7 28.pdf
• File:Fish Vac MM 7 30.pdf
• File:Fish Vac MM 8 4.pdf
• File:Fish Vac MM 9 2.pdf
• File:Fish Vac MM 9 9.pdf
• File:Fish Vac MM 9 16.pdf
• File:Fish Vac MM 9 23.pdf
• File:Fish Vac MM 9 30.pdf
• File:Fish Vac MM 10 7.pdf
• File:Fish Vac MM 11 11.pdf
• File:Fish Vac MM 11 18.pdf

Presentations

• File:Fish Vac Concept Design Review.pdf
• File:Engineering Release Review Presentation.pdf
• File:Team FishVac Snapshot 4.pdf
• File:Technical Presentation Fish Vac.pdf

Product Requirements

• File:Fish Vac PR.pdf

Project Schedule

• File:Fish Vac Schedule.pdf

Team Contract

• File:Fish Vac Team Contract.pdf