LABORATORY ANIMALS

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LABORATORY ANIMALS — the different types of animals which are specially divorced in the conditions of laboratories or nurseries for an experimental or work practice. L. use for diagnosis of diseases, modeling various fiziol, and patol, states, studying to lay down. - professional, drugs, volumetric and physical factors, production of biological drugs — diagnostic serums, vaccines, cultures of fabrics, etc.

Animals of various systematic groups treat laboratory: protozoa, worms, arthropods, erinaceouses, amphibians, birds, mammals. However most often L. subdivide into invertebrates and vertebrata.

Vertebrate laboratory animals

Use of vertebrate animals by the person with the cognitive purpose began, apparently, during development of cattle breeding. In the subsequent on animals began to study a structure and functions of various bodies of live organisms. In particular, observations of the Ancient Greek scientist Diogenes (5 century BC) who, opening corpses of animals, established different functions of auricles are known. Later the anatomy and physiology were studied on animals by Aristotle, K. Galen, U. Garvey, etc. Originally experiments were made on domestic animals. In 15 century white mice, rats and Guinea pigs became known. However the concept «laboratory animals» developed by the end of 19 century.

In total in medical - biol, researches use up to 250 animal species. One types constantly part in laboratories and nurseries for scientific research (white mice, white rats, Guinea pigs, rabbits, hamsters, cats, dogs, monkeys, mini-pigs, etc.). Others — periodically catch for an experiment (a vole, a sandwort, gophers, polecats, groundhogs, battleships, lemmings, amphibians, fishes, etc.). There is a group a lab. birds (hens, pigeons, canaries, quails, etc.). The part of medical experiments is made on page - x. animals (sheep, pigs, calfs, etc.). From total number of L. falls to the share of mice apprx. 70%, rats — 15%, Guinea pigs — 9%, birds — 3%, rabbits — 2% and other — 1%.

Interest of researchers in rodents is caused generally by the fact that many of them have the small sizes of a body, high fertility and the short period of life; for several months of life of a rodent it is possible to track processes which at the person proceed for years in an organism. Average life expectancy of white mice 1,5 — 2 years, rats 2 — 2,5 years, hamsters of 2 — 5 years, Guinea pigs of 6 — 8 years, rabbits of 4 — 9 years.

At cultivation of L. carry out control on genetic, ecological, morphological features, and also for health reasons.

Genetically L. are subdivided into nonlinear (heterozygous) and linear (homozygous). Nonlinear animals are bred on the basis of accidental crossings and owing to this fact they possess high degree of heterozygosity. Increase inbreeding (see) at this group L. no more than 1% for generation are allowed.

Linear animals (see) part on the basis of close inbreeding. Linear animals are subdivided into purely closed lines and mutant drains. The line is called purely inbredny, in a cut animals passed not less than 20 generations of crossing like «brother the X sister»; the kongenny (koizogenny) line — the closed line bearing a supplementary foreign gene. A mutant drain — posterity of animals at whom it is spontaneous or induced there were changes in inheritance of external or internal signs.

162 lines of rats, 16 lines of Guinea pigs, 66 lines of hamsters, 4 lines of sandworts and 7 lines of chickens are known apprx. 670 closed lines and sublines, kongenny lines and mutant drains of mice. In the USSR linear mice are bred since 1943. Each line has the features in gene pattern, reactivity on various antigenic and stressful factors. Linear animals are regularly controlled on homozygosity generally by method of transplantation of skin. The variety of linear animals is supported in the corresponding collections at scientific institutions. The largest collections of linear mice contain in the USSR, in department of genetics of Research laboratory of experimental and biological models of the USSR Academy of Medical Sciences, in the USA, in Jacksonian laboratory, and in some other countries.

On linear animals conduct researches in the field of microbiology, parasitology, oncology, immunology, genetics, physiology, morphology etc.

Ekol, control is directed to standardization of contents and feeding of L.

For feeding of L. use natural foodstuff or the bricketed concentrates on the developed norms (the order of the Ministry of Health of the USSR for No. 163 from 10/III 1966 g).

The bricketed forage and grain mix are set in bunker feeding troughs for several days. Other natural forages and water are given to animals daily.

Rooms for L. shall be spacious, with 10 — 14-fold exchange of air in an hour and air humidity of 55 — 60%. More preferable and «dirty» speak rapidly the rooms of barrier type divided on «pure». Not less than 50% of space are reserved for utility rooms. In order to avoid mutual stressful influences and exchange of causative agents of infectious diseases contents in one room or a cell of animal different types is not allowed.

The designs and the sizes of cells are developed for each species of animals. Mice, rats, Guinea pigs and hamsters preferential contain in plastic cone-shaped trays with a mesh cover from stainless steel. Rabbits, dogs, monkeys, cats and birds contain in metal cells. The international standards of the sizes of cells, in particular for mice 30 X 20 X X 10 cm, for rats 42 X 32 X 15 cm, Guinea pigs 75 X 45 X 34 cm, cats 50 X 70 X 65 cm are offered. All cells are equipped with automatic drinking bowls and bunker feeding troughs, are placed on racks in 1 — 6 tiers, before use wash carefully and disinfected, and in the course of work at mice and rats are weekly replaced with pure. In one cell depending on a type of L. it is possible to support no more than 15 mice, 10 rats, 1 rabbit, 5 Guinea pigs and 5 hamsters.

Standardization of L. on mikrobiol. to the status caused their division into two groups: conventional animals (i.e. ordinary animals) and gnotobiotichesky animals (with the known microflora). Under the WHO recommendation (1973) conventional animals at cultivation shall be exposed mikrobiol, to control: mice, rats, hamsters, Guinea pigs and rabbits — on salmonellosis, tuberculosis, a pseudotuberculosis, listeriosis, leptospirosis, pasteurellosis, piyevmokokkoz, Bordetella bronchiseptica (except for rabbits), dermatomycoses, a lymphocytic choriomeningitis on all types of ectoparasites, helminths; in addition mice, rats and Guinea pigs — on Streptobacillus moniliformis, mice and rats — on mycoplasmas, mice — on Corynebacterium murium, streptococci of group A, a virus of smallpox, rabbits — on Treponema cuniculi.

Prevention of diseases of L. the dignity includes strict observance. - a gigabyte. governed, the maximum disinfecting of the environment (rooms, air, the equipment, forages, a laying, etc.), creation of conditions of keeping of the animals corresponding to their ecological features, holding quarantine actions during the movement of animals and also timely isolation of patients.

In prevention of intestinal infections much attention is paid to control of microflora of an intestinal path.

L. are subject to many infectious and invasive diseases: to salmonelloses, erizipeloid, listeriosis, pseudotuberculosis, pasteurellosis, tularemia, stafilokokkoza, smallpox, viral diarrhea, shigellosis, lymphocytic choriomeningitis, Tizzer's disease, mycoses, helminthoses, coccidioses, acarapidosis, etc. U L. the latent carriage of pathogenic elementary bacteriums and viruses occurs (especially at rats). Monkeys can be a source of infection of the person with viruses of herpes, hepatitis, smallpox, etc. In total more than 100 infections and invasions of L are described. Prevention of infection of the person from animals provides respect of personal hygiene by service personnel, and also systematic inspection of service personnel.

At cultivation of L. their treatment is not allowed. Elimination of the arisen disease is reached by endurance in a quarantine of again arriving L., timely establishment of the diagnosis and removal of patients. For this purpose in nurseries of L. there are control and diagnostic laboratories.

To gnotobiotichesky L. belong amicrobic, mono - and di - or polikontaminirovanny animals and animals without specific pathogenic factors or activators (SPF or SPV) which are widely used during the carrying out important experiments (see. Gnotobiologiya ).

In scientific institutions where researches on L are conducted., there have to be scientific and auxiliary divisions: vivarium (see) and experimental and biological clinic. Contain in a vivarium and partially separate animal species get divorced from their subsequent transfer for pilot study. The experimental and biological clinic only contains animals on whom researches are conducted. Vivariums and experimental and biological clinics are placed in the certain building (a complex of buildings). For the amphibious animals and fishes used in experiments the respective rooms are equipped.

For satisfaction of constantly increasing demand for L. different types, lines and categories there was an independent industry of economy — laboratory livestock production with the relevant scientific and production bases. The corresponding training of personnel is organized. == Backboneless laboratory animals == In addition to vertebrate animals, in laboratories find application as well many invertebrates: protozoa, helminths, arthropods (insects, mites), etc. Purposes and methods of their use as L. are very various. Irreplaceable objects for various a lab. researches protozoa (Protozoa type) long since serve. Speed of their reproduction, the small sizes, comparative simplicity and convenience of contents in the conditions of laboratory do protozoa by the cheapest pilot models (see. Protozoa ).

Freeze-etch methods and long-term storage of some species of protozoa (trypanosomes, leyshmaniye, toksoplazm, etc.) in liquid nitrogen are developed. This method allows to create cryobanks of strains of protozoa that is convenient at their use as L.

Ability of many protozoa to breed is in the sexless way premises of receiving pure lines of the elementary organisms — clones which serve as an irreplaceable object for genetic, immunological and other researches.

At statement of experiments with protozoa it is necessary to consider not only their look, a strain or isolate, but it is frequent also belonging to a certain genetic line. Great value at a lab. contents has knowledge of a life cycle of development of the elementary and separate stages of this cycle (see. Life cycle ).

During the work with protozoa considerable impact is exerted by biotic and abiotic factors of the environment.

As L. from protozoa are widely used sarcodic (Sarcodoidea), and among them parasitic amoebas are Entamoeba histolytica (the activator of an amebiasis at the person) and E. invadens (the activator of an amebiasis at some reptiles), and also a free living amoeba (E. moshkovskii) representing morfol, the double of a dysenteric amoeba. E. histolytica and E. invadens in culture are convenient objects for tsitol, the analysis of effect of antiamoebic drugs. Infected E. histolytica infant rats, Guinea pigs, rabbits, hamsters, kittens and other L. are models for studying of questions of a pathogeny, immunity and chemotherapy of the diseases caused by parasitic amoebas.

Small free living amoebas of the limax group serve a lab. the model during the development of problems of parasitism, among them found the types capable to cause a serious illness of the person and animals (primary amoebic encephalomeningitis, etc.). Amoebas of types of Naegleria fowleri and Acanthamoeba culbertsoni and others are used in culture of cells for studying of their interaction with fabrics and cells of mammals. On cultures of these amoebas carry out searches of effective chemotherapeutic drugs and disinfectants, and on pilot models (the infected animals) study mechanisms of a pathogeny, immunity, etc.

Large amoebas (Amoeba proteus, Chaos, Pelomyxa, etc.) use at cytogenetic and other researches, in particular in the analysis of hereditary variability, emergence and a mutation rate. In mikrurgichesky experiences nucleocytoplasmic hybrids — heterokarionums on which study the phenomena of transplant incompatibility, epigenic variability etc. are received. On these objects make various observations on influence of the ionizing and ultraviolet radiation, a chemical mutagenesis.

Various researches are conducted on a lab. cultures of parasitic flagellar (Flagellata) — trichomonads, lyambliya, trypanosomes, a leushmania, etc. (see Flagellates). On these protozoa study features of mechanisms of antigenic variability, priming reactions of interaction with cells and their cytopathic effect. Flagellar Crithidia fasciculata and Strigomonas oncopelti close to promastigotiy forms of leyshmaniye from intestines of insects are widely used in biochemical, researches. These cultures serve as models during the studying of structure nucleinic to - t, various organellas zooflagellit, etc. Flagellates (trypanosomes, lyambliya, etc.) are widely used at a lab. modeling of relationship of a parasite and owner.

There are new data on cultivation of different groups of activators of the class Sporozoa (see. Sporozoa ), preparation with their help of vaccines, antigens for serological and allergic tests, on studying of relationship of an organism of a parasite and the owner at the cellular and molecular levels, mechanisms of effect of chemotherapeutic drugs, to creation of new effective drugs, studying of ecology of activators etc. Cultures of causative agents of malaria of the person are received that gives the chance of preparation of antimalarial vaccines. Pilot studies are conducted also on causative agents of malaria of rodents and especially malaria of birds. For cultivation of the last very convenient were embryos of birds (hens, etc.).

K L. began to carry also different types infusorians (see). Free living and parasitic ciliary (Ciliata), including Balantidium coli — the activator of a balanthidiasis, are used at biochemical, physiological, cytologic, ecological researches, during the studying of action of a penetrating radiation and other physical. and chemical factors, and also at permission of other problems obshchebiol. and medical values.

Infusorians represent classical objects and for cytogenetic researches, including the genetic analysis during the studying of some problems of variability and heredity. Infusorians serve as convenient objects at toxicological researches, and also during the studying biol, effect of action of ultraviolet rays, a penetrating radiation and other factors. At the same time changes of speed and the nature of the movement, a pulsation of sokratitelny vacuoles, the nuclear device, disturbance of rates of division etc. are considered. In recent years some types of infusorians found broad application in experiments on molecular biology, in particular in genetic engineering. Wednesdays, various on structure — from the simplest herbs and leaves in the form of infusions to difficult synthetic with in advance defined chemical structure are developed for the maintenance of infusorians of in vitro.

On various helminths — helminths (see) — study many questions of effect of chemotherapeutic drugs, some questions connected with biology of helminths, etc. Observations are made on the helminths parasitizing at the person and animals. The helminths extracted from an organism of the owner wash Ringer's solution which is warmed up to t ° 38 ° and place in the special vessels allowing to register their movement, food, changes in a condition of a cuticle etc. Short-term overseeing in vitro helminths is made, using Ringer's solutions, Ringer — Locke, Tirode, Heddon — Fleyga, Krotova.

In many laboratories of the world as L. use various arthropods, such as a fruit fly (Drosophila melanogaster), a bee moth (Galleria melonella), a bug (Rhodnius prolixus), etc. Standard lab. widely use cultures of blood-sicking insects and mites in various parazitol, laboratories.

Necessary condition of use in an experiment of arthropods is check of parent natural population (the ancestor a lab. culture) on purity of the line — lack of natural infectiousness causative agents of diseases as blood-sicking arthropods have the defining value as carriers and keepers of causative agents of many transmissible infections (rickettsioses, arboviral infections, leyshmanioz, filariases, malyariya, etc.). Definition of extent of participation of any type of arthropods in transfer of causative agents of infections or its true role in epidemiology and an epizootology requires conducting pilot studies with blood-sicking arthropods and causative agents of diseases.

Blood-sicking arthropods collect from animals, and also in rooms for page - x. animals, in the nature — from vegetation, from holes, caves, from nests of rodents and birds by means of developed parazitol, methods. In a lab. rearing cages of blood-sicking arthropods contain in specially mounted flasks, test tubes, cages.

Argasidae and ixodic (Ixodidae) mites use for long preservation of activators of spirokhetoz, rickettsioses, arboviral infections, etc.

Mites, mosquitoes, mosquitoes, flies and other arthropods use in experiments on testing of efficiency of insecticides, acaricides and repellents, and also for development biol, methods of controlling with carriers of causative agents of diseases of the person and animals and wreckers of agriculture.

For experimental studying as carriers of causative agents of natural and focal diseases of the person (encephalitis, hemorrhagic fevers, rickettsioses, etc.), and also during the testing of efficiency of effect of acaricides and development of specific methods biol, fight use ixodic mites (sorts Ixodes, Haemaphysalis, Hyalomma, Rhipicephalus, Dermacentor). Ixodic mites are easily cultivated in a lab. conditions. For creation a lab. cultures of ixodic mites collect with page - x. the animals (who already got drunk blood) or from vegetation in natural stations (hungry). Full ticks are placed in specially mounted humidified test tubes for a yaytsekladka. Hungry ticks are fed on L. under caps made of cloth which paste on a back to an animal-prokormitelyu (pigs, rabbits, mice, hamsters, and also sheep and cattle). At the correct leaving mites of one line are cultivated in laboratory for years.

Convenient lab. model are argasovy mites (sorts Ornithodoros, Alveonasus, Argas). They are used for experimental studying of the relations of mites with activators (spirochetes, viruses, rickettsiae), and also for long (long-term) preservation of causative agents of diseases in an active state. Argasovy ticks at cultivation are fed or on L., or blood of animals through the membrane prepared from leather of a mouse or a chicken. The way of feeding of argasovy mites on a chicken embryo by their replanting in an air chamber of egg is developed. Mites of Alveonasus lahorensis, Ornithodoros papillipes, etc. are cultivated in laboratories already many decades.

As L. use also gamazovy mites (Gamasoidea). Among them mites of Ornithonyssus bacoti (a rat tick), Dermanyssus gallinae (a chicken tick), Allodermanyssus sanguineus (a mouse tick) are especially convenient for contents in the conditions of laboratory. Gamazovy ticks are used for modeling inf. process at rickettsioses, a tick-borne encephalitis, a tularemia, hemorrhagic fevers. The laboratory arranges the so-called plant — an artificial nest, in a cut place ticks and L. (mice, hens, etc.) for their feeding. As required ticks are selected from the plant and in the course of the experiment and observation contained in the special humidified cameras.

For experimental works in various laboratories part blood-sicking mosquitoes (Culicidae) of different childbirth (Aedes, Anopheles, Culex). In some cases it is convenient to use mosquitoes of the sort Culex pipiens molestus who are easy for parting in the conditions of laboratory; the impregnated females in favorable conditions do not fall into a diapause and can lay eggs without preliminary food blood. The larvae which left eggs develop in the water rich with organic matters.

From mosquitoes of the sort Aedes it is the easiest to part mosquitoes of a type of Aedes aegypti which are carriers of viruses of yellow fever and other diseases of the person and also a plasmodium of birds, etc. They can be supported in rather small cages; females of mosquitoes are fed with blood of rabbits or other animals. The eggs laid by females Aedes it is possible to store a long time in a dry state; for removal of larvae they are placed in a vessel with water. As a forage for larvae serves rice powder, powder from water fleas, egg yolk, etc. Water in a vessel with larvae shall be pure and not become soiled a forage. Vessels in which dolls were formed place in gauze cages for breeding of mosquitoes.

For the most various pilot studies, in particular for studying of transfer of causative agents of plague, rickettsioses and other bacterial diseases of people and animals, studying of action of various insecticides, repellents, etc., use the cultures of fleas (Aphaniptera) removed in the conditions of laboratory. In the conditions of laboratory fleas of rats — Xenopsylla cheopis are the most convenient for cultivation, Ceratophyllus fasciatus, etc. B laboratories cultivate them in the special plants — glass jars in which they place animals-prokormiteley; as L. apply also louses — carriers of pathogenic spirochetes and rickettsiae.

For studying of relationship of a parasite and the owner at leyshmanioza and mosquito fever use mosquitoes of Phlebotomus papatasii, Lutzomyia longipalpis and Sergentomyia arpaklensis. A number of effective ways of cultivation of mosquitoes is offered. The main condition of successful cultivation of these insects is selection of nutritious substrate for larvae and creation of mosquitoes of temperature and humidity, optimum for development. For mass cultivation of mosquitoes in stationary conditions use the various cages lodges offered P. A. Petrishcheva (1954) and other authors. A. I. Lisova (1952) offered an original method of feeding and infection of mosquitoes. For this purpose use the skins of a tail removed from svezheubity white mice which fill with blood or the infecting material (e.g., washout of culture of leyshmaniye or other microorganisms). Such skins place in test tubes where place the mosquitoes willingly feeding on contents of a skin.

For the purpose of development of scientific bases of cultivation and the reasonable choice on a specific research of animal species in the USSR, England, the USA, France, Germany, Japan and other countries scientific centers in comparative biology of L are organized. In the USSR such center is the Research laboratory of experimental and biological models of the USSR Academy of Medical Sciences. Coordination of work is in the field carried out by the International committee on laboratory animal (Personal computers L A), about the Crimea more than 40 countries, including the USSR cooperate. Scientific conferences on different questions of biology of L are annually held. and biol, modeling. On these questions more than 30 periodicals are abroad published. The international and regional centers are organized: The ACORUS V OZM international help center for providing with animals with spontaneous development of tumors (The Netherlands, Amsterdam, Institute of cancer), the International help center FAO/WHO for mycoplasmas of animals (Denmark, Aarhus, medical f-t un-that). Regional help center for viruses of monkeys (USA, State of Texas, department of microbiology and inf. diseases). The help centers are available at IKLA: on histocompatability of mice (Party of Russian Taxpayers), rats (Germany and the USA), Guinea pigs (USA), dogs (Germany), on viruses of rodents (ChSSR, England, Germany, Japan), on hairless mice (Denmark), on causative agents of malaria of birds (Canada), etc.

In publications of results of the researches received on L., under the WHO recommendation, it is required to specify their type, the line, age, sex, a source of acquisition, a condition of keeping and feeding.

From additional materials

Battleships (addition to article of the same name published in the 12th volume) — the mammal Dasypodidae Bonaparte families, 1838 from group nepolnozuby — Edentata.

In family of battleships of 9 childbirth (21 look). Battleships (a synonym armadillos) — the most ancient of the living mammals, are widespread in South and Central America, in the south of the USA. They lead a nocturnalism, live in holes. The name «battleships» is connected with existence on a dorsal body surface of the armor consisting of the separate bone plates covered with a corneous layer (the so-called skin skeleton which is not found at other mammals). Length of a trunk of battleships of different types fluctuates from 12 to 100 cm, weight to 55 kg.

Fig. 1. Devyatipoyasny battleship (Dasypus novemcinctus Linnaeus).

Battleships are used in medicine and biology as laboratory animals, is especially frequent — a devyatipoyasny battleship — Dasy-pus novemcinctus Linnaeus, 1758 (fig. 1). Length of a body of an adult nine-zone battleship of 40 — 55 cm, weight is 3 —-7 kg; the armor consists of the chest and pelvic guards divided by 9 mobile belts. The lowered body temperature (32 — 35 °), the big duration of a delay of implantation of a blastocyst — up to 4,5 months (the general duration of pregnancy apprx. 9 months), reproduction of four monozygotic cubs, ability to transfer a long absence of exogenous oxygen, reduced reactions of cellular immunity at the expressed humoral immune responses belongs to features of biology of nine-zone battleships; life expectancy — up to 15 years.

Fig. 2. Extremity of a nine-zone battleship: the leproma on an inner surface of a hip which developed in 20 months after infection of an animal with material from the patient with lepromatous type of a leprosy.

In the nature nine-zone battleships are owners of activators of a number of zoonoz — a leushmaniosis (see), Chagas diseases (see. Shagasa disease ), salmonelloses, nek-ry mikobakterioz, etc. In an experiment on nine-zone battleships it was possible to model many bacterial, viral and parasitic infections, lepromatous type of a leprosy (see) — fig. 2, and also sypny, returnable and mouse typhus, African sleeping sickness, schistosomatosis, trichinosis, foot-and-mouth disease, mycoses, etc.

At battleships of the sort Dasypus from one oospore several embryos (a true polyembriony) develop that does them by unique natural model for studying of mechanisms of appearance of twins, and also many questions of heredity and variability. Monozygotic twins of battleships are an object for researches in the field of transplantation, and also for immunological, toksikol, and teratol. researches. The pharmacokinetics of pharmaceuticals in an organism of battleships is very close to that at the person. E.g., it is established that the thalidomide causes uglinesses of a fruit of battleships that was not observed at others a lab. animals.

Battleships easily adapt to bondage. It is the best of all to support them in small (2 — 4 sq.m) open-air cages with the doghouse for a nest and a box for sand. As covering material usually use scraps of paper or a moss. In the nature they eat generally insects, worms, vegetable food makes less than 10% of a diet. In their diet include mincemeat, eggs, milk, vegetables, fruit in vivariums. Battleships are not aggressive therefore leaving and experimental work with them come easy. In bondage nine-zone battleships do not breed (nek-ry other types, napr, setaceous battleships, breed).




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V. A. Dushkin; D.N. Zasukhin, L. M. Gordeeva; A. A. Yushchenko.

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