The project aims to develop an innovative product: rapid immunochromatographic tests that will enable early and comprehensive diagnosis of parasitic hookworm infections in dogs. It aligns with the global trend of the new animal health strategy in the European Union, 'Prevention is better than cure.'

What hookworms are

Hookworms are nematodes that parasitize the digestive tract of carnivorous mammals, including dogs. They embed themselves in the walls of the small intestine or duodenum, leading to damage to the mucous membrane of the intestine. In Europe, two species of hookworms are of significant importance: Ancylostoma caninum and Uncinaria stenocephala. Analysis of the species composition of parasites in dogs in recent years in Poland has shown the widespread occurrence of nematodes from the Ancylostomatidae family, with a prevalence ranging from 2% to 75%. For example, after examining 500 samples from dogs in the Lublin region, 218 (43.6%) were assessed as negative, while 282 (56.4%) were positive (containing protozoan oocysts and nematode eggs). Studies have shown that, next to roundworms, hookworms are the most frequently diagnosed nematodes in dogs in Poland, with a prevalence ranging from 2% to 75%.

They also pose a zoonotic threat, despite being considered parasites with narrow specificity (monoxenous). There have been reported cases of invasions in humans. The individuals found in these invasions were mature representatives of both sexes, although no fertilized females were found. Infection of humans with invasive larvae of these parasites usually leads to the occurrence of cutaneous larva migrans (CLM) or eosinophilic enteritis. Less frequently, disorders such as eosinophilic pneumonia, localized muscle inflammation, hair follicles, or pathological ophthalmic conditions—ocular larva migrans (OLM) syndrome, and visceral larva migrans (VLM) may occur. Zoonotic hookworm species have been classified by the WHO as Neglected Tropical Diseases.

In parasitological diagnosis of hookworm invasions in canids, the flotation method is routinely used, followed by determining the species of the nematode based on morphometric characteristics of their eggs. This method does not allow for a certain differentiation of U. stenocephala and A. caninum species, so they are often described together without specifying the species affiliation. Additional disadvantages of the method include equipment requirements, the presence of qualified personnel, time of implementation, and cost. Despite often neglecting species determination in hookworm diagnostics, the medical and veterinary significance of A. caninum and U. stenocephala differs. These species exhibit different invasiveness due to their biology. Invasion by typical hematophages like A. caninum, especially with high intensity, may manifest as severe hemorrhagic enteritis and acute anemia with iron deficiencies. On the other hand, during U. stenocephala invasion, the intensity of acute symptoms is less pronounced.

Project Outcomes

• The creation of a rapid test allowing on-site diagnosis from a fecal sample, taking only a few minutes.
• A test designed to detect a specific and characteristic antigen or antibody of a particular hookworm.
• Not requiring specialized qualifications from the end user, thus making it possible for both veterinarians and breeders to perform the test.

The project aims to develop the world's first rapid strip test for the detection of pathogens associated with mastitis in bovine milk samples and, in accordance with the adopted R&D plan and pre-implementation work, to commercialize the results of the initiative in the economic activity of the applicant.

What mastitis is

Mastitis, inflammation of the udder in cows, is a defensive reaction of the organism to certain factors, mainly microorganisms, that have entered the mammary gland. The disease is caused by a range of infectious and environmental bacteria, as well as other microorganisms, with the main strategy for treatment being the use of antibiotics. It is estimated to affect approximately 20-50% of the dairy cow population annually and is a major cause of economic losses in breeding. Currently, as part of the EU's policy to reduce the use of antibiotics in animals, aiming to reduce the risk of antibiotic resistance, the condition for administering antibiotics is to identify the pathogens responsible for the existing infection. Meanwhile, commonly available methods for identifying the etiological factors of mastitis are time-consuming, such as classical microbiology, or costly, as is the case with PCR techniques.

In animal organisms, antibiotics undergo metabolism, often not being completely metabolized, and a portion of the dose is excreted unchanged into the environment. Antibiotics used in veterinary medicine, when excreted by animals, enter the soil with manure or slurry, and can then penetrate into groundwater, rivers, and lakes. There are also direct threats, such as the toxicity of drugs to humans and animals, as well as the risk of disrupting ecosystem balance. It has been proven that plants growing in soil containing antibiotics are able to absorb and accumulate them, posing a threat to consumers. Antibiotics have also been shown to affect the growth and development of plants.

Project Outcomes

• Reduction in the use of antibiotics, including broad-spectrum antibiotics. In situations where antibiotic use is unwarranted, unnecessary compounds causing antibiotic resistance will not be released into the environment.
• Acceleration of mastitis diagnosis from several days to a few minutes, thereby improving animal welfare.
• Possibility of using therapeutic compounds with a narrower spectrum of activity.
• This will prevent the excretion of unmetabolized antibiotic residues into the environment. Replacement of traditional laboratory methods and techniques with rapid diagnostic tests will reduce the material and energy consumption of services, leading to a reduction in electricity consumption.
• Its execution will not require any special training and can be performed by both veterinarians and breeders

The project aims to develop the world's first rapid strip test for the detection of Equine Herpesviruses type 1 and type 4 in swabs and tissues from horses and, in accordance with the adopted R&D plan, to commercialize the results of the initiative in the economic activity of the applicant.

What Equine Herpesviruses are

Equine Herpesviruses are commonly found among horses and are characterized by a high degree of host adaptation due to a shared evolutionary process. The main factor responsible for their success is the ability to establish latent infection. Horses, after the initial contact with the virus, remain infected for life, and subsequent reactivations of latent infection often occur without symptoms, making them challenging to detect even using real-time PCR, theoretically capable of detecting as few as 6 virus copies in a sample. This mechanism allows the disease to spread within a herd and between herds along with carrier horses. According to various data, around 30% of horses have had contact with EHV-1, and almost 100% of horses have had contact with EHV-4.

Reactivation of infection most commonly occurs due to a decrease in immunity caused by stress, such as the introduction of a new horse into the herd, transportation, training, illness, surgical procedures, or other infections. Out of the 9 types of herpesviruses occurring in horses (EHV), EHV-1 and EHV-4 have the greatest clinical significance, causing severe infections, potential neurological symptoms, and miscarriages in mares. EHV type 1 induces respiratory diseases, neurological symptoms, miscarriages (in EHV-1 infected herds, even 30-40% of pregnant mares may miscarry), and deaths among newborns. EHV type 4 causes upper respiratory tract infections in foals, can also lead to miscarriages and neurological symptoms. Therefore, rapid pathogen diagnosis is crucial for taking appropriate actions.

Currently used diagnostic methods (PCR, serological tests) require transporting the sample to a specialized laboratory.

Project Outcomes

• The innovative rapid immunochromatographic test developed as part of the project will allow for the diagnosis of Equine Herpesvirus type 1 and type 4 infections in horses in just a few minutes, without the need for specialized equipment, reagents, and can be conducted in the field without transporting samples to a laboratory.
• The test will be designed to detect the virus during symptomatic infection.
• Its execution will not require any special training and can be performed by both veterinarians and breeders.