by John Haaland, PhD

Our Mission: To Control a Deadly Pathogen

MAP/PATH, LLC is an early-stage biotechnology company whose mission is to control and/or eliminate diseases caused by a single pathogen: Mycobacterium avium subspecies paratuberculosis (MAP.)

Intro to MAP 101

When I first met Dr. Tom Dow in February 2018, I had never heard of MAP.  I learned from him and his veterinarian associates that an incurable, infectious disease has been killing cattle, goats, sheep, camel, buffalo, deer, and elk for over 100 years,  affecting 3 billion animals per year globally. The pathogen responsible for this devastation is called Mycobacterium avium subspecies paratuberculosis (MAP) and the disease it causes in animals is called Johne’s disease. Farmers and veterinarians control Johne’s by herd management strategies and culling (killing) infected animals. For the North American dairy industry, the cost of Johne’s Disease is about $270 million per year. In the United States, hidden costs associated with Johne’s disease total $2 billion. Intensive research has been done over the past 35 years to cure this disease, without success. Antibiotics and vaccines were either not effective or were prohibited in food animals.

Since MAP can survive pasteurization, contaminated products are found in the human food supply. Milk, cheese, infant formula, livestock feed and calf milk replacers have tested positive for MAP. Beef cattle present an additional MAP source. MAP infections also originate from contaminated farm water that flows into municipal drinking water. It is a very resistant bacterium that can survive up to nine months in mud, a year in cow manure, and up to two years in water. Standard industrial water treatment, such as filtration systems and chlorination, may be ineffective to eliminate MAP in urban water supplies.

For decades, Dr. Dow has been researching human diseases associated with MAP. There is a general consensus among researchers that MAP is a zoonotic pathogen (a disease that can be transferred from animals to humans or vice versa). As a food safety system specialist, I am very familiar with biological, physical and chemical risks to our food supply, the various means to manage them, and the rules that apply to them. Control of MAP would have substantial global benefit to the agricultural industry and human health.

Why is it Hard to Kill MAP?

In evolutionary terms, MAP is unique in that it is slow, cunning and devious. MAP is a shape shifter that can alter its configuration between active and dormant forms. When stressed it can shift to its dormant state in a few hours. MAP cannot replicate outside of a host, so to survive, it hides inside eukaryotes (like amoeba) and immune system cells (like macrophages.) In its dormant state, it can survive in just about anything as it waits for animals, like cows or humans, to eat it. When conditions in the chosen host are favorable, it shifts to its active state and replicates, thus causing disease. These evolutionary mechanisms make MAP difficult to kill, and antibiotics are ineffective when MAP is protected inside a host cell. Mycobacteria are excellent mutators, which allows MAP to develop resistance to antibiotics, giving them an additional survival mechanism. Standard methods which are effective to reduce and eliminate many bacterial strains are ineffective when applied to mycobacteria.

What is a Bacteriophage?

Bacteriophage (phage) are viruses that target specific bacteria. They are among the most numerous bio-entities on earth and, unlike virulent viruses such as SARS-CoV-2, they are very important to our survival. However, the manner by which phages and their virulent counterparts detect and attack specific bacterial membranes is very similar.

Phages are highly specific to a target bacteria. They cannot infect eukaryotic cells of humans, animals and plants. An individual phage typically only infects a subgroup of strains within the same bacterial species, leaving the beneficial microbes intact. Once a phage has killed all of the targeted pathogen, it degrades into biomolecules leaving no residual DNA, which is why the FDA classifies them as Generally Recognized As Safe (GRAS).

Phages can be delivered by spray, liquid, pellet or injection and are inexpensive to produce. They work quickly and spread rapidly, usually requiring only one dose to eliminate the pathogen. Looking at what makes each phage attack a specific bacteria and the mechanisms that signal the release of the phage’s endolysin (which ruptures and kills the bacteria) is a key question for our company. This simple animation explains bacteriophages and their potential benefit to human health in more detail.

Can Phages Kill MAP?

Phage experts have historically agreed that phages cannot be used to target intracellular pathogens, such as MAP. (Remember from above that phages only target bacteria, not human or eukaryotic cells.)  Recent technological developments which utilize CRISPR have demonstrated that phages may be tailored to target intracellular pathogens. The trick is to get the phage into the infected macrophage using a commercially viable technique.

Our company began working on this challenge in 2018. The first step was to obtain the correct phage that could kill MAP. We then had to insert the phage into a benign carrier that a macrophage would “swallow.”  To test the viability of the phage, we added the phage to a liquid MAP culture, which showed significant reduction of the extracellular MAP at 4 days. (PCR identification of MAP via IS900 was conducted before and after the addition of our phage to the MAP cultures.)

The Future of Phages to Eliminate MAP

MAP/PATH, LLC plans to conduct further in vitro research to determine if our phage could be used to eliminate intra- and extracellular MAP in cow and human macrophages, as well as in food products. Our initial commercial focus is on infant milk replacers for goats and calves. By adding a phage solution to in process animal food products and using phage sprays for process equipment treatment, we expect MAP contamination will be controlled prior to consumption by the kid or calf. While the same approach could be taken for human baby formula, it will take longer to get FDA approval. This would be particularly valuable as applied to products fed to infants (both animals and humans) since high MAP exposure at a young age may be related to later immune dysfunction.

Today, more than 70% of calves consume milk replacers. As in human food products, studies have shown a significant presence of MAP in animal milk replacers – as many as 30% of samples were positive. If initial stages yield positive results, MAP/PATH, LLC plans to test our phages on a variety of food products and begin in vivo treatment in goats to eliminate Johne’s disease. Protocols have been established, but seed funding is necessary to continue our work, and we are actively seeking investors.

We have filed a patent for our discovery, and are working on modifications that will allow treatments of human diseases. We anticipate licensing our proprietary technology to farm food suppliers and pharmaceutical companies interested in adapting it to clinical trials for MAP related diseases.

For more information or to inquire about investment opportunities with MAP/PATH, LLC, please contact Dr. John Haaland via email at john(dot)haaland(at)mappathllc(dot)com and visit our website to view our patent application.

John Haaland, PhD is the Co-founder and CEO of MAP/PATH, LLC. He has over 40 years experience in aerospace, microelectronics, biotechnology and food businesses. As principal developer of manual controls for the NASA-funded Apollo space program while at Honeywell , his work led to the creation of Hazard Analysis and Critical Control Points (HACCP). As VP of Systems & Research at Health Service Systems, Inc. , he installed the first automated multiphasic health testing service in mainland United States. While a member of The Pillsbury Company’s senior management, he created the first corporate-wide product safety system, incorporating HACCP in diverse food segments including restaurants, food processing and agri-products.

Dr. C. Thomas Dow received his bachelor of science degree from the University of Wisconsin-Madison in microbiology, and attended medical school at the University of Wisconsin School of Medicine, Madison. Following medical school, he did an internship year at Bronson Methodist Hospital in Kalamazoo, Michigan, and completed his ophthalmology residency at the University of Wisconsin Hospital and Clinics. Dr. Dow is Board Certified in Ophthalmology, is licensed in Wisconsin, and is included in the Best Doctors in America database. He established Chippewa Valley Eye Clinic in 1978. He has served as a member of the surgical team, Free Rural Eye Clinic, in the Philippines several times. In addition to seeing patients, Dr. Dow has research interests in infectious causes of immune disease and in telomerase in oncology and regenerative medicine. His article, All Roads Lead From Crohn’s, was published on the Human Para website in 2017.