an analysis of costs, safety, and legality
What it Costs
Mastitis costs the dairy industry billions of dollars every year in both treatment of infected dairy cattle and in loss of sellable milk. Through our extensive human centered design outreach, a farmer told us that even the smallest improvement in current management of the disease would be worth an investment from the farmer’s perspective because of just how costly the occurrence of mastitis is.
A typical case of mastitis costs approximately $300. A standard medicine for mastitis such as Spectramast , costs between $6-$8 per treatment. Treatment must be given daily for a course of about a week, so the cost of the medicine itself is between $42-$56. Additionally, while a cow is being treated for a mastitis, as well as a few days post-treatment, the milk produced by that cow cannot be sold. The current price of milk is approximately $16.50/cwt . Since a healthy cow produces around 100 lb of milk a day, over a course of 9 days without saleable milk there would be approximately $150 in losses. Also, once a cow recovers, they usually have permanent defects due to scarring which leads to a 10% less milk production for the rest of their lactation lifespan.
Bacteriocins also are better than the presently available medicines for mastitis. Currently, the most common treatment for mastitis is the use of intramammary antibiotic tubes. Unfortunately, these treatments often lack specificity and are given without knowledge of the type of bacteria actually causing the infection [3,4]. The specificity that our bacteriocin treatment offers leads to a smaller risk of eventual development of resistance, as well as more importantly the ability to specifically target the type of bacteria causing the disease without altering other bacterial ecology. Since the price range of the bacteriocins is likely to be in the range of other current antibiotics, it is clear that there would be a reason to purchase bacteriocins.
Current milking shells on the market cost $40-$50, but are simply solid steel parts. Since we are adding technology to our shell, it is natural that they would cost more. A calculation of the the raw cost of the materials that compose our shell comes to be about $100. With the addition of manufacturing costs (estimated to be $20 per shell), we can expect our shell to cost about $110. The raw materials can be broken down as follows. The steel tubing that we are using will cost approximately $12.75, which is the typical cost of the current plain milk shell. The cold shock would include a heat sink ($6.98) and thermoelectric cooler module ($35.00). The UV light sterilizing component would include the six UV lights ($5.39), a white LED ($1.04), and photocells ($7.91). The temperature sensor would consist of an adafruit thermistor ($4.00) and three springs ($1.42). The iodine dip will use sponges ($1.07) as well as steel cut from the steel tubing for arms. Additional materials will include another thermistor ($10.60), set screws ($3.78), a cooling fan ($1.17), machine screws ($.0.34), and Arduino Uno - R3 (one per every four shells) ($6.24), and copper wire ($0.73).
Although the price for our shell is higher, our feedback from farmers confirms that they would be willing to pay more for a shell since the cost of mastitis is so high. Shells are also a one time purchase, and are only replaced when damaged. Hence the shells are an investment for the farmers. Check out our discussion with farmers concerning cost here.
Does It Harm Me?
Whenever we perform any scientific protocol, we always have to think about the risks and impacts of the project, a task that is, indeed, not simple. One aspect of risks that we have to assess is the effects of the recombinant organisms and peptides that are used in the project on humans . Ideally, the E. coli cells that we used to take up the various genes to produce our bacteriocins would not have any impact on the hosts because they are kept within the laboratory setting. However, if the cells managed to leave the laboratory setting, then the biggest concern would be if another organism were to take up the DNA lost from these E. coli cells.
In addition, using bacteriocins is our approach to mastitis treatment in place of the traditional application of antibiotics. Milk that has antibiotic residues must be discarded, but it is safe to consume milk that has bacteriocins. One of the greatest applications of bacteriocins in today’s industry is food preservation in meat and fish products, fruits and vegetables, and beverages. Furthermore, bacteriocins produced by lactic acid bacteria (LAB) are generally recognized as safe (GRAS) and are effective against major Gram-positive pathogens in food-borne illnesses. One of the major concerns, however, is the maximum level of bacteriocins allowed in the milk. Nisin, a bacteriocin that we use in this project, is used worldwide in preservation of dairy products, sausages, and canned and packaged meat, but there is still no consensus on the maximum concentration at which it is still considered safe. More studies need to be conducted to further look into the maximum levels of Nisin, among other bacteriocins, that are considered to be safe .
Does It Harm the Cow?
In several experiments where bacteriocins have been used to treat mastitis, especially when they are injected into the mammary glands of the cows, the glands became irritated with the injection and often turning swollen and tender . In a particular study done by Wu et. al on the effect of nisin, a bacteriocin, on the treatment of mastitis, the cows’ mammary glands only became swollen after injection of a large dose of nisin.
The study also notes that it is common to have irritated mammary glands after an injection. Apart from that, the experiments have not shown to cause cows any other harm. Traces of bacteriocins have been found in the milk from cows that have been treated bacteriocins 24 hours and 48 hours after the treatment, but the residues were much lower than the limit set for using nisin to preserve milk. This supports that nisin was safe for subclinical mastitis therapy in lactating cows .
As Legendairy, we one day hope to take our project from proof of concept to a physical product, and knowing that this is a feasible option is important. To do so, we must examine the guidelines that the U.S. Public Health Service and Food and Drug Administration set. The FDA sets guidelines to regulate the approval of antimicrobial drugs and review equipment, in line with the Grade A Pasteurized Milk Ordinance . To approval bacteriocins as an antimicrobial drug, our next steps would be :
1. Demonstrating that bacteriocins are safe to the cow through an irritation study and analysis of milking during the treatment period
2. Determining the efficacy of the drug
3. Conducting clinical trials which set an appropriate dose
These tests involve treating a cow, and checking for both treatment and prevention of subclinical mastitis. Culture results test for the presence of pathogens such as Staphylococcus aureus and Streptococcus agalactiae, which are allowed to be generalized to other Staphylococcus and Streptococcus species. Since our bacteriocins each target different pathogens, some of which fall under the above species, and some of which don’t, each of the bacteriocins would have to undergo approval with their specific pathogen .
To manufacture the customizable shell, our next steps would be:
1. Testing by Registration, Evaluation, Authorization and Restriction of Chemicals (REACH) which ensures that the chemicals used to construct the liner are not harmful
2. Ensuring that the silicone of the liner falls in line with FDA CFR 177.2600, “Rubber articles intended for repeated use” 
3. Manufacturing the liner with a stainless steel metal of the American Iron and Steel Institute which is an FDA approved component 
Though we still have a ways to go until our products would be FDA approved, the process is certainly promising, and components of our design could easily be adjusted to match the FDA standards.
 Spectramast DC. Zoetis. Retrieved 4 Oct. 2016. Web. https://www.zoetisus.com/products/dairy/spectramast-dc.aspx
 US Milk Farm Price Received. YCharts. Retrieved 4 Oct., 2016. Web.
 Hoe, F.G., and P.L. Ruegg. 2006. Opinions and practices of Wisconsin dairy producers about biosecurity and animal well-being. J. Dairy Sci. 89:2297-2308.
 Oliveira, L., C. Hulland, and P.L. Ruegg. 2013. Characterization of clinical mastitis occurring in cows on 50 large dairy herds in Wisconsin. J. Dairy Sci. 96:7538-7549.
[ Sobrino-Lopez, A., O. Martin-Belloso. 2007. Use of nisin and other bacteriocins for preservation of dairy products. Retrieved from
 Taylor, J., A. Hirsch, and A. T. R. Mattick. 1949. The treatment of bovine streptococcal and staphylococcal mastitis with nisin. Vet. Rec. 61:197-198.
 Wu, J., Hu, S., & Cao, L. (2007, September). Therapeutic Effect of Nisin Z on Subclinical Mastitis in Lactating Cows. Antimicrobial Agents and Chemotherapy, 51(9). Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2043217/.
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 "CVM GFI #49 Target Animal Safety And Drug Effectiveness Studies for Anti-Microbial Bovine Mastitis Products (Lactating and Non-Lactating Cow Products)." U.S. Food and Drug Administration. U.S. Food and Drug Administration, 4 Apr. 1996. Web. 28 Sept. 2016.
 "CFR - Code of Federal Regulations Title 21." U.S. Food and Drug Administration. U.S. Food and Drug Administration, 1 Apr. 2016. Web. 28 Sept. 2016.