Dwarf pigs are the preferred test system for studying chronic toxicity, immunogenicity, pharmacokinetics, pathophysiological conditions, and pathology induction and are widely used worldwide as a model for preclinical studies. However, the use of large animals in preclinical studies has several limitations, primarily the personality of the personnel working with the animals, the cost of each animal in the experiment, the cost of their maintenance and manipulation. Therefore, a very important aspect of the use of large animals in research is to find ways to reduce the number of experimental animals used, without compromising the quality of research and its scientific value. Given the facts established, our study aimed to test additional vital methods for the diagnostic evaluation of the condition of dwarf pigs during the intermediate points of the experiment. The study was conducted on 12 dwarf pigs of both sexes, aged from 36 to 60 months and weighing from 40 to 71 kg. Minimally invasive examination methods such as abdominal ultrasound, fine needle soft tissue biopsy, collection of bile, gastric juice, and urine were tested. Using ultrasound, clinical examination, hematological, biochemical blood analysis, and coagulation system assessment, the rate of animal rehabilitation after interventions were assessed. The risks of the procedures on the integrity and validity of the preclinical study were evaluated. It was determined that: a) ultrasound, as an intermediate point of the investigation, is an easy and informative manipulation, through which, it is possible to assess the anatomical and structural changes in organs and tissues; b) fine-needle biopsy is a convenient and diagnostically significant method to assess the effects of the test drug on organs (liver, spleen, lungs, prostate); c) percutaneous puncture of the gallbladder is a manipulation requiring certain skills, but allowing to obtain a sufficient amount of biomaterial; d) obtaining gastric juice in anesthetized pygmy pigs, as well as cystocentesis was not difficult.
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