Traditionally, one of the most informative and reliable methods for diagnosis of pathologies in organs and tissues are autopsy and histological examination. These methods allow us to estimate both the macroscopic and microscopic structural changes in organs and tissues of humans and animals, and the data obtained in the course of pathological studies, not rarely are of fundamental importance in the study of toxic effects of drugs in preclinical studies.
Sufficient qualification of our employees, including graduates of doctors pathologists, specialists in the field of veterinary morphology, as well as experienced technicians, histology, hard work, responsibility and love for his work allows us for many years to carry out pathological and histological examination at the highest level with the use of modern methods of macroscopic and microscopic diagnosis of pathological conditions and in experimental animals.
We are always glad to cooperate and expand scientific frontiers. We hope that together we can reach new stages in the development of Russian science and discover new unknown faces in the field of anatomy and histology of humans and animals.
The most important step in conducting a post mortem examination is to compare the macroscopic and the microscopic picture of pathologies. Particular attention is paid to the dynamics of the development of pathologies in animals in the experiment. We apply methods of pathologic diagnosis are common, and have yielded positive results in toxicological studies, and in assessing the impact of specific drugs and the study of reproductive and fetal toxicity.
Here are some aspects of our work.
The introduction of large volumes and high doses of drugs in the acute toxicity studies are often accompanied by dystrophic and necrotic lesions of liver tissues, which can be observed in postmortem histological examination of cadaveric material.
In the study of atherosclerosis applied macroscopic and microscopic staining of the aortic intima dye Oil Red (oil red), which allows good visualization of lesions atheromatosis.
In experiments to study the reproductive and fetal toxicity deviations in osteogistogeneze fetuses to determine who carried out staining of cartilage and bone skeletons Alizarin red.
To determine myocardial ischemic lesions we use complex macroscopic and microscopic methods coloring material and the coloring 2,3,5-triphenyltetrazolium chloride based on determining the overall activity of the tissue dehydrogenase.
Computed tomography (CT) of organs and tissues of the experimental animals is one of the most promising areas of instrumental diagnostics both in terms of a more complete picture of the disease, and from the standpoint of the humane treatment of animals. CT allows assessment of his lifetime lesions, study the dynamics of the disease and the effectiveness of treatment without the need evtanazirovaniya animals at all interested control points of the experiment. At the same time, the use of CT significantly expands the notion of localization of various lesions, the degree of their severity and pathogenesis of the disease, allows to obtain reliable data, correlated with the results of conventional morphological studies in real-time.
Histochemical methods of research for many years are used by researchers in the study of the normal structure of organs and diagnosis of pathological states. Histochemical staining techniques can detect tissue structures of interest, to identify and evaluate morphometrically organ damage such as tissue fibrosis, muscular dystrophy, and other pigments deposits. However, they do not exhibit species specificity, which is a great advantage when for different types of animal studies.
Immunohistochemistry studies of organs and tissues of animals used for experimental and diagnostic solutions tasks received in recent years widespread mainly due to the selectivity and high sensitivity of the used markers. These methods are relevant and most informative in studies on diabetes, neuropathy, pathologies of the cardiovascular system and endocrine organs.
In our work we use a wide range of immunohistochemical markers that are designed to not only the architecture of the tissue structure of the cells, but the mechanism of action study drugs.
The most important tasks of anatomical science and morphology as a whole is to define interspecies differences in the structure and syntopy organs in animals. These data are the basis for the development of surgical approaches and the implementation of a large number of experimental conditions such as ligation of arteries and veins, intravascular injection, and others. The information as relevant when selecting dosage forms that are often associated with the anatomical structure of organs in animals.
We aim to expand the existing knowledge of comparative anatomy and experimental animals receive the most striking anatomical specimens showing the structural features of the bodies of animals which can not be seen at autopsy.
The material is subjected to morphological studies, subject to archiving for 1 year in accordance with the Russian Ministry of Health Order from 06.06.2013 N354n “On the procedure for post-mortem autopsy.”
Histological archive contains all the necessary information about the experiments to allow retrospective analysis micropreparations, paraffin blocks and organs of animals.
To archive animal organs we use HistoSafe system. The material is placed in a strong plastic bag containing 10% buffered formalin, after which the package is sealed and organs throughout the backup period is stored in a sealed package.
Histological slides and blocks are archived in separate shelves to store the identification numbers of the experiment.
Histological archive room is equipped with shelves, on which is placed the material to be stored.
- Muzhikyan AA, Makarova MN, Gushchin AA Features mortem examination group of experimental animals // International Bulletin of Veterinary Medicine. – 2014, №1. – P. 75-80.
- Muzhikyan AA, Makarova MN, Gushchin JA Features histological working of organs and tissues of laboratory animals // International Bulletin of Veterinary Medicine. – 2014, №2. – P. 103-108.
- Kashkin VA, Shekunova EV, Muzhikyan AA, Makarova MN, Makarov VG Comprehensive assessment of the degree of pathology in modeling the adjuvant-induced arthritis in rats // International Bulletin of Veterinary Medicine. – 2015, №1. – P. 92-103.
- Katelnikova AE, Kryshen KL, Muzhikyan AA, Makarova MN, Makarov VG acute inflammation model: carrageenan air pouch // International Bulletin of Veterinary Medicine. – 2015, №2. – P. 78-87.
- Muzhikyan AA, Makarova MN The use of computed tomography in the evaluation of the state of organs and tissues of laboratory animals // International Bulletin of Veterinary Medicine. – 2015, № 4. – P. 73-80.
- Makarova MN, Tesakova SV, Samusenko IA, Stolaschuk NV, Solovyov DE, Zhorina AS, Tikhonov VP, Makarov VG Experimental study of the effectiveness of biologically active food additives and drugs in a model of chronic prostatitis // Bulletin of St. Petersburg State Medical Academy. II Mechnikov. – 2007. № 2 (8). – P. 123-128.
- Tesakova SV, Makarova MN, Makarov VG, Samusenko IA, Gushchin JA The effectiveness of the use of drugs with a high content of phytosterols in the treatment of chronic prostatitis // Bulletin of the Research Center of Volgograd. -2009. Number 2. C. 35-39.
- Tesakova SV, Samusenko IA, Karachinskaya IV, Kryshen KL, Abrashova TV, Makarova MN, Makarov VG The experimental model of cervical lymphadenitis in rats to evaluate the anti-inflammatory efficacy // Preventive and clinical medicine. – 2011. – №1 (38), pp 57-63.
- Proshin SN, Makarov VG, Makarova MN, Kryshen KL, Kovshin AV, Samusenko IA Ulcerative nonsteroidal anti-inflammatory drugs and hepatotoxicity of nimesulide in an experiment on rats // Surveys on the wedge. Farmacol. and lectures. therapy. –2012. – Vol.10, №1. –P.28-34.
- Khodko SV, Makarova MN, Makarov VG, Samusenko IA, Shirunova MG The experimental model of acute rhinosinusitis in rats to assess the therapeutic efficacy of drugs // Preventive and clinical medicine. – 2013. – T. 46, number 1. – P. 57-62.
- Shedko VV, Gushchin JA, Muzhikyan AA, Makarova MN Retrospective analysis of histological archive // Development and registration of medicinal products. – 2016, № 3. – P. 162-165.
- Muzhikyan AA, Khod’ko SV, Gushchin Ya.A., Makarova MN, Makarov V.G. Histological changes in the lymph nodes of rats during the modeling of acute cervical lymphadenitis // International bulletin of veterinary Medicine. – 2017, No. 1.– P. 75-83.
- Gushchin YA, Muzhikyan AA, Selezneva AI, Makarova MN Complex morphological evaluation of atherosclerotic damage of the aorta of rabbits in the experiment // Atherosclerosis and dyslipidemia. – 2017. No. 1.– P. 50-59.
- Gushichin Y. , Myzhikyan A. , Shedko V. , Makarova M. , Makarov V. Comparative anatomy of the upper gastrointestinal tract of experimental animals and humans // International bulletin of veterinary Medicine. – 2017, No. 3.– P. 116-129. Сomparative anatomy of the upper gastrointestinal tract of experimental animals and humans [Full text is available in Russian].
- Zaikin K., Gayday E., Gayday D., Gushchin Ya., Muzhikyan A., Kryshen’ K., Makarova M. Spontaneous age-related pathology of the urinary system in rats. Laboratory Animals for Science. – 2018, №4. – P.1-15. https://doi.org/10.29296/2618723X-2018-04-03 SUMMARY. The article is devoted to spontaneous age-related pathologies of the urinary system of rats detected in animals of control group in a long-term carcinogenesis experiment. Interest in this group of pathologies is relevant during long-term experiments since the elderly rodents’ death is often associated with oncological diseases and pathologies of the kidneys. Knowing of pathologies with spontaneous manifestation in laboratory rats is necessary for correct interpretation of experimental results as well as for the differential diagnosis of «spontaneous» and «induced» pathologies. For the classification of pathologies INHAND nomenclature was used. Histological examination of the organs of urinary system (kidney and bladder) was carried out in 200 males and 200 female Wistar rats aged 24 months. Tumor pathology in the article is not considered. Spontaneous age-related pathology of the urinary system in rats is very common – kidney pathology was found in all 200 males studied and in 165 of 200 females studied. Pathological changes considered as manifestations of chronic progressive nephropathy (such as interstitial inflammatory cell infiltration, glomerulosclerosis, basophilia and atrophy of renal tubule epithelium, tubular dilatation, cylinders in the lumen of tubules, focal interstitial sclerosis of the renal cortex) are most common; all 200 males studied and 140 of 200 females studied were identified. Vesicular dystrophy (vacuolization) of renal tubules epithelium is only pathology of urinary system that is more common in females than in males. Vesicular dystrophy has a diverse pathogenesis its clinical and biological significance is ambiguous it can precede necrosis of epithelium, but it can also occur in normal conditions. Pathology of bladder is less common it was detected in 22 males and 1 female. Among the pathological changes were protein plugs in the lumen of bladder and focal inflammatory cell infiltration of bladder. In the experiments described pathologies should be considered as spontaneous associated with age-related changes in the body [Full text is available in Russian].
- Koptyaeva K., Muzhikyan A., Guschin Ya., Belyaeva E., Makarova M., Makarov V. Technique of dissection and extracting organs of laboratory animals. Message 2 (mouse). Laboratory Animals for Science. – 2018, №4. – P.1-24. https://doi.org/10.29296/2618723X-2018-04-05 SUMMARY. Post-mortem examination of animals (necropsy, autopsy) is a diagnostic procedure aimed at finding out the cause of death of an animal, as well as determining the spectrum of macroscopically visible changes in organs and tissues of cadaver. In scientific and research practice, one of the key stages of the experiment and at the same time a humane experimental point often is euthanasia of animals with subsequent pathoanatomical and histological examination of organs. From the point of view of the quality and reliability the obtained data, an important step is the standardization of the procedure to dissection and extracting organs of laboratory animals. A series of articles about the carrying out of necropsy of laboratory animals will highlight the main stages of autopsy. In this work, a technique to dissection and extracting organs of mice is described [Full text is available in Russian].
- Koptyaeva K., Guschin Ya., Belyaeva E., Makarova M., Makarov V. Technique of dissection and extracting organs of laboratory animals. Message 3 − hamster. Laboratory Animals for Science. – 2019, №1. – P. 1-19. https://doi.org/10.29296/2618723X-2019-01-02 SYMMARY. The third message from the series of articles on the methodology of necropsy and extraction organs of laboratory animals, describing in detail and illustrating the technique of sequential and complete autopsy and extraction of organs of laboratory hamsters. Among laboratory animals, hamsters occupy a special place due to the presence of large cheek pouches in their digestive tract. A large area of the mucous membrane lining the cheek pouches from the inside, allows to visually demonstrate and explore the local irritant effect of orally dispersible dosage forms (lozenges), which can easily be placed in the cavity of the cheek pouches. This article demonstrates the procedure for laboratory hamster necropsy. It describes the process of preparing an animal for an necropsy, conducting a primary examination of a corpse after euthanasia for the presence of external changes and injuries, fixing the corpse on the dissecting table, opening and examining the internal cavities of the body, as well as studying the surface lymph nodes. Methods for the extraction of organs of the oral cavity (including the extraction of the tongue, pharynx and cheek pouches along with the adjacent salivary glands), all organs of the thoracic, abdominal, and pelvic cavities are described and illustrated. The topographic location of some organs of the thoracic, abdominal and pelvic cavities is illustrated. Described two methods to extract the brain, along with olfactory bulbs, the method of separating the preparations of the chest bone for histological examination of the bone marrow and maxilla for histological examination of the nasal cavities. The procedure for preparing the spinal cord for fixation in formalin without removing it from the spinal canal is described. Described the extraction of the eyes together with the lacrimal gland and eyelids, the extraction of the muscles and the peripheral nerves adjacent to them. The methods of extraction and the sequence of actions performed, described in this article, minimize the damage to extracted organs, prevent their contamination in order to prevent the occurrence of certain artifacts detected during subsequent histological examination. Conclusions are presented at the end of the article [Full text is available in Russian].
- Vavilova V., Gushin Ya. Model of The Global Cerebral Ishemia in Mongolian Gerbil. Laboratory Animals for Science. – 2019, 2. https://doi.org/10.29296/2618723X-2019-02-03 ABSTRACT. The aim of this study was to develop a global cerebral ischemia simulation protocol (GCI, global cerebral ischemia) with subsequent reperfusion, which was carried out on Mongolian gerbil. The choice of a gerbil as a biological model is due to the anatomical feature of the blood supply to the brain. Gerbils, unlike other laboratory animals and humans, do not have a posterior communicating artery (do not have the full circle of Willis). Mortality, neurological status, pathological and histological studies were evaluated as criteria for the pathology degree estimation. Visualization of the ischemic region after the induction of pathology was carried out by staining brain sections with 2, 3, 5 triphenyl tetrazolium chloride (TTC). The method of the neurological status scoring which takes into account the presence or absence of pathological changes is described. The model is confirmed by pathological and histological examination. The severity of damage prevails on the 2nd and to a lesser extent on the 3rd day after the pathology induction. Histological examination of the gerbil brain identified neurons with signs of dystrophy (polymorphic, hyperchromic cells with fuzzy contours – “red” neurons) and reduced in size – “shadow cells”, as well as fields with mild gliosis in the cerebral cortex and cerebellar cortex. Thus, the results of the study showed that the 20-minute bilateral occlusion resulted in animal survival of about 60%, most animals demonstrated pronounced neurological deficit. Staining with TTC has shown that it is advisable to evaluate volume of the ischemic lesion in the first 24 hours after the induction of pathology. In addition, results showed that the neurological deficit was most pronounced during first 4 days after occlusion. To estimate long-lasting pathological changes it is advisable to conduct a histological examination of brain tissue samples. [Full text is available in Russian].
- Koptyaeva K., Guschin Ya., Belyaeva E., Makarova M., Makarov V. Technique of Dissection and Extracting Organs of Laboratory Animals. Message 4: Guinea Pig, Gerbil, Degu. Laboratory Animals for Science. – 2019, 2. https://doi.org/10.29296/2618723X-2019-02-05ABSTRACT. The fourth message from the series of articles on the methodology of necropsy and extraction organs of laboratory animals, describing in detail and illustrating the technique of sequential and complete autopsy and extraction of organs of laboratory guinea pigs, gerbils and degu. Guinea pigs are often chosen to study the allergenic properties of drugs and to determine the pharmacological activity, for example, in experiments that model hypercholesterolemia, airway hyperresponsiveness, etc. The high susceptibility of gerbils makes it possible to be a model for various infectious diseases. Also on them examine the effect of drugs on the level of steroid hormones and cholesterol. Degus that predisposed to the development of diabetes mellitus are a good model for studying this disease. In addition to this, on them conduct studies of cataracts, atherosclerosis, and Alzheimer’s disease. This article demonstrates the procedure for necropsy of laboratory guinea pigs, gerbils, and degu. It describes the process of preparing an animal for an autopsy, conducting a primary examination of a corpse after euthanasia for the presence of external changes and injuries, fixing the corpse on the dissecting table, necropsy and examining the internal cavities of the body, as well as examining the superficial lymph nodes. Possible methods for the extraction of organs of the oral cavity (including extraction of the tongue, pharynx), all organs of the thoracic, abdominal and pelvic cavities are described and illustrated. The topographic location of some organs of the thoracic, abdominal and pelvic cavities is illustrated. Prescribed two ways to extract the brain, the method of separation of the chest bone for histological examination of the bone marrow and upper jaw for histological examination of the nasal passages. The procedure for preparing the spinal cord to formalin fixation without removing it from the spinal canal is described. Eye extraction with the lacrimal gland and eyelids, extraction of muscle and adjacent peripheral nerves is described. The methods of extraction and the sequence of actions performed, described in this article, minimize the damage of the organs being removed and prevent their contamination in order to prevent the occurrence of certain artifacts detected by subsequent histological examination. Conclusions are presented at the end of the article.[Full text is available in Russian].
- Guschin Ya., Kovaleva M. Comparative Morphology of Human and Laboratory Animals. Laboratory Animals for Science. – 2019, 2. https://doi.org/10.29296/2618723X-2019-02-06 ABSTRACT. Knowledge the characteristics of the morphological structure, the main structural and functional components of human skin and laboratory animals is the foundation of rational planning of pharmacological and toxicological preclinical studies of skin dosage forms in aspect. The choice of laboratory animals must be justified for accurate translation of the results of pre-clinical studies into clinical studies (early phase). A brief report shows the comparative morphology of human skin and certain types of laboratory animals: mouse, rat, guinea pig, hamster, rabbit, ferret, minipig. Structural similarities and differences of skin layers (epidermis, dermis, gmpoderm) are described. A comparative characteristic of the thickness of the epidermis and dermis in different species, carried out on the analysis of literature data and results obtained in their own research. A comparative analysis was conducted both between humans and laboratory animals, and between species.[Full text is available in Russian].
- Koptyaeva K., Ustenko J.U., Belyaeva E.,Guschin Ya.A., Makarova M., Makarov V. Technique of dissection and extracting organs of laboratory animals. Message 5 – rabbit, ferret // Laboratory Animals for Science. – 2019, 3. https://doi.org/10.29296/2618723X-2019-03-05 ABSTRACT. The fifth message from a series of articles on the methodology of necropsy and extraction of organs of laboratory animals, describing in detail and illustrating the technique of sequential and complete necropsy and extraction of organs of laboratory rabbits and ferrets, is presented. Rabbits are suitable for long-term ongoing research, and are the only laboratory rodents from which you can get recombinant pharmaceutical proteins and explore the oral or intragastrically forms without destroying the integrity of the tablets. In addition, they are used to develop and study new surgical methods, physiological tests and studies of various types of toxicity for drug testing. Various viral and bacterial diseases were studied on ferrets, as well as pathology of the organs of hearing, vision, the vomiting reflex due to the similarity of many anatomical, metabolic and physiological characteristics to humans. This article describes the method of necropsy from the procedure of external examination of a corpse, sequential dissection and examination of the thoracic and abdominal cavities, and before the extraction of organs, the selection of various bone preparations. Possible methods for the extraction of organs of the oral cavity (including extraction of the tongue, pharynx), all organs of the thoracic, abdominal and pelvic cavities are described and illustrated. Describes the method of extraction of the brain, the method of selection of the chest bone for histological examination of the bone marrow and upper jaw for histological examination of the nasal passages. A procedure for preparing a spinal cord site for fixation in formalin without removing it from the spinal canal is described. Demonstrated methods for extracting the eyes, together with the lacrimal gland and eyelids by a single organocomplex, extracting the muscles and adjacent peripheral nerves. The methods of extraction and the sequence of actions performed, described in this article, minimize the damage to the organs being removed and prevent their contamination in order to prevent the occurrence of certain artifacts detected by subsequent histological examination [Full text is available in Russian].
- Gushchin Y.A. Additional histological methods of stainig in preclinical studies. Laboratory Animals for Science. – 2019, 4. https://doi.org/10.29296/2618723X-2019-04-07 ABSTRACT. In clinical practice and in pharmacological studies, microscopic analysis is an integral part of the study of normal structure of tissues and pathologically altered organs. To prepare the material for histological examination, visualization is carried out by staining the tissue with dyes. The most common routine staining is hematoxylin and eosin, which is not enough to reveal the full picture of the process. Therefore, it is necessary to use a number of additional stains. Some of them can be used as routine instead of classical hematoxylin and eosin, for example, van Gieson stains or Mallory trichrome. Most stains are more specific, and they are used to identify specific structures or chemical compounds in cells and tissues. This allows you to obtain a significant amount of information, which facilitates an understanding the course of normal and pathological processes. Methods for the detection of mucopolysaccharides with Alcian blue are used in the study of the gastrointestinal tract and respiratory system. PAS staining is necessary in the diagnosis of accumulation diseases, some oncological processes and fungal infections. Fat-soluble dyes, primarily Sudan III and scharlach red are used in the study of dystrophic diseases and atherosclerosis. The Oil Red O dye is applicable for macroscopic assessment of the area of atherosclerotic lesions of the aorta. Congo red is the most popular dye for the amyloid detection. Specialized dyes for the diagnosis of myocardial damage were considered, such as (HBFP) (Haematoxylin-Basic Fuchsin-picric acid) and tetrasolium salts to visualize the area of damage to the heart muscle and brain. In this review, we examined several histological dyes, some features of their application, as well as mechanisms of the dye-substrate interaction in tissues. These staining methods can be used during histological work, and consider the possibility of their use, which will help to identify, as well as comprehensively study the pathological processes modeled in preclinical studies [Full text is available in Russian].
- Belyaeva E.V., Ustenko J.U., Guschin Ya.A. Technique of dissection and extracting organs of laboratory animals. Message 6 – minipigs. Laboratory Animals for Science. – 2019, 4. https://doi.org/10.29296/2618723X-2019-04-08 ABSTRACT. The sixth report from a series of articles on the methodology of necropsy and organ extraction o laboratory animals is presented. The report describes and illustrates in detail the methodology for sequential and complete necropsy and organ extraction of miniature pigs. Currently, miniature pigs are widely used in biomedical research as laboratory animals, since their anatomical and physiological characteristics are most similar to human ones. Miniature pigs are considered among the basic models for studying stroke, diabetes, organ transplantation, surgical treatment methods, and also used for testing of pharmaceutical products and biomechanical devices. Also, these animals are essential in studies of the structure and physiology of skin, cardiovascular, digestive and urinary systems. The size and weight of minipigs are perfect for modeling of many pathologies. Also they provide a sufficient amount of biomaterial, e.g. nternal organs, blood or other body fluids, for further laboratory researches. This article describes the step-by-step procedure of animal necropsy from preparation, external examination, sampling of mammary glands and skin, sequential opening and examination of the thoracic and abdominal cavities, and to the extraction of separate systems and organs. Possible methods of extracting organs of the oral cavity, thoracic, abdominal and pelvic cavities are described and illustrated, and the fastest, most effective and safe method of extraction of the brain from the cranial cavity is presented. The methods of eye extraction along with the optic nerve and the extraction of muscles and adjacent peripheral nerves are demonstrated. All methods of organ extraction, as well as the sequence of actions described in this article minimize damage of harvested organs and prevent their contamination in order to avoid the formation of some artifacts, which might be found during subsequent histological examination and could lead to conflicting results [Full text is available in Russian].