It's been a busy few years for Tridelta. Thanks to an extensive range of assays for Acute Phase Proteins (APP), Tridelta is focusing on three main market sectors: academic and industrial research, mastitis and animal welfare. It has already co-founded another company - TriMed Research.

Academic and Industrial Research: The assays have been used by academic research institutions around the world for a variety of studies, ranging from environmental influence on polar bears in the arctic, to the health of buffaloes in Sri Lanka. In addition, major pharmaceutical companies, as part of drug development and toxicity studies, are using the assays routinely.

Mastitis: Through a collaboration with the University of Nebraska, the company has discovered, and patented, a new APP: MAA. The new protein has been shown to be a highly sensitive marker of bovine mastitis. The company has developed a range of assays that will help farmers detect individual cows with mastitis and monitor the overall health of the herd.

Animal Welfare: recent publications have shown the measurement of APP's to be a major factor in improving animal welfare and food safety. The company is working with a number of academic, governmental and retail bodies to develop systems that will incorporate the benefits of these findings into routine use.

Everything started when the company was researching further applications of the Serum Amyloid A (SAA) protein. A unique APP, it was taken from a cow's udder, when previously it had been thought to be only produced in the liver.

ACUTE PHASE PROTEINS Acute Phase Proteins are produced by all mammals in varying degrees in response to infection, trauma, stress or disease. Depending on the kind of event they are responding to, the proteins are set of by the body's cytokine systems, rising to clearly measurabl levels. The level and pattern of increase varies according to species and to disease. It also reflects both response to infection but also response to a foreign substance such as a drug. For example, the day may be not too distant when an APP kit from Tridelta could be applied to cattle suspected of suffering from FMD (foot and mouth disease), to clarify which animals are infected or which have been vaccinated. The APP's which show a pronounced and rapid rise in most species, have particular beneficial applications for food safety and animal welfare. The most sensitive and accurate appear to be: * Haptoglobin (Hp) * Serum Amyloid A (SAA) * Carbon Reactive Protein (CRP)

Table 1: Benefits Of APP At Slaughter: Differentiates between healthy and non-healthy animals. APP are markers for active disease, responding especially to bacterial infection. APP can identify individual animals with disease for thorough post-mortem examination or for screening on the farm prior to transport to the abattoir. Potentially harmful animals can then be excluded from the human food chain. APP testing can now be automated utilising simple, easy to operate instruments to allow for high throughput and rapid testing of animals either at the farm or at the point-of-slaughter. In Production: Screening can identify animals with infectious diseases for treatment, allowing therapeutic substances to be used appropriately. With antibiotics banned as growth promoters, APP will identify animals for therapeutic treatment and herds where hygiene should be improved. A raised mean APP in a herd identifies sub-clinical infection or poor hygiene. APP testing provides an objective test of hygiene for Quality Assurance schemes. APP testing complements organic farming as a means of disease detection prior to therapeutic treatment (which is allowed under organic rules for confirmed cases). Disease Specific Markers: Specific and sensitive test for Bovine Mastitis, new marker APP (MAA) discovered and measureable. Potential role in milk quality testing (compared with current Somatic Cell Count method). Testing can be automated during milking process. Further disease specific markers may be discovered as experience grows and research progreses.

THE MARKET FOR MASTITIS

The SAA protein may prove to be a US$7 billion per year market in Europe and the USA. Mastitis is a condition where udder inflammation in cows is characterised by sudden onset, redness, swelling, hardness, pain, grossly abnormal milk and a reduced milk yield.
Tridelta has been working with the University of Nebraska to develop a commercial mastitis test and this will be on the market by the end of the year. Using an assay that can pick up minute traces of SAA protein, it can give results in just ten minutes.

TRIMED RESEARCH

With technology owned by the University of Nebraska and licensed to Tridelta, TriMed Research Inc. was set up as a joint venture. It was started in October 2000, and has two leading researchers in Dr. T. McDonald and Dr. D. Mack, from the university. Having discovered and patented the naturally occurring peptide called MAA, the company's primary focus is necrtotising enterocolitis (NEC - see box), which is a severe, life threatening condition which affects children's intestinal tract. The condition particularly affects premature and low-birth-weight infants, especially if there has been injury to the intestinal wall. That allows bacteria to stick to and colonise the wall where there is an excess of protein. The resulting localised intestinal necrosis allows further bacterial growth, and the subsequent vicious cycle can end in shock, sepsis and death. The benefit of MAA is that it significantly reduces bacterial and viral gastro-intestinal infections because, when given prophylactically, it reduces the ability of organisms to bind to the gut wall. The incidence of NEC has risen in recent years and the market is estimated at over $100 million per year. Further applications could include "travellers tummy" and infant and adult diarrhoea.

Necrotising Enterocolitis (NEC) NEC is a serious worldwide complication that occurs in premature and very low birth weight infants. In the USA alone there are over 440,000 premature infants born each year and NEC can be expected to develop in 8% of them. Additionally, as a result of current reproductive practices, the number of infants that are born with with very low birth weight is rapidly increasing. These infants are of particular concern because NEC may develop in 30% of them. Many forms of therapy targeted at many of the complications of that occur in these infants have shown continual improvements. However, the same cannot be said for NEC where therapy has remained the same for the last decade: discontinuation of enteral feeds and the administration of broad-spectrum antibiotics. These therapies are used in the hopes of preventing bacterial growth, adhesion and colonisation and the development of bowel necrosis and perforation. The repeated cessation of enteral feeds, especially in cases of recurring episodes of NEC, creates a major medical issue since growth of the infant is directly integrated with its overall development. Furthermore, medical therapy fails in about 20% of the patients, necessitating surgical resections of the affected bowel. Unfortunately, despite surgical management, about 25% of this group of infants die. For those that survive the surgical management, the massive bowel resections lead to significant problems including malnutrition, complications of intravenous catheters (infections and blood clots) and liver damage from the solutions used to provide nutrition. The overall consequences of these therapies include a high incidence of infections, high hospital costs and potential long-term neurodevelopmental disability as a result of poor nutrition during this most vulnerable period of growth and development.

THE FUTURE BECKONS So what does the future hold for this multi-faceted company? Further work continues to extend the range of Acute Phase Protein tests - Tridelta is now working with a major UK food retailer to develop further APP's for food safety and animal welfare. The intention is to provide quantitative data on which to base meat inspection. In addition, the SAA protein can not only be applied to cattle, but its protective human therapeutic applications are also being further explored, particularly through TriMed.

Table 2 Estimated Relative Risks of Neonatal and Postnatal Mortality for IUGR Infants Birthweight (g) Neonatal Post-neonatal 1500-1999 18.0 5.0 2000-2499 4.0 2.0 2500-2999 1.0 1.0 3000-3499 0.4 0.5 3500+ 0.3 0.5

Reproduced with kind permission of Helix Magazine