Biochemical activity of the microflora of a free-living bird

. Free-living birds have a wide range of habitats, can be present in the wild, near human homes, livestock and poultry complexes, farms. As a result of migration and migration of birds, the probability of spreading infectious diseases increases. In the body of birds, the pathogen can persist, multiply, be released into the external environment, and then enter the body of a healthy animal or bird, becoming the cause of new cases of the disease. Detection of pathogenic microflora in the body of free-living birds, determination of their biochemical activity is one of the aspects of studying pathogens in the parasitic phase of existence. The identification of biologically active bacteria allows us to predict the probability of free-living poultry participating in the epizootic process, and to plan appropriate preventive measures in poultry and animal husbandry. The purpose of the study is to study the species composition and biochemical activity of the microflora of the free-living birds of the Amur region. For research on the territory of the Amur region, impregnations from beaks and cloaks. The study identified such microorganisms as Enterococcus, Pseudomonas, Acinetobacter, Citrobacter, Actinobacillus, Escherichia, Micrococcus, Aspergillus, Enterobacter, Salmonella, Proteus, and Staphylococcus. Biochemical activity was detected in microorganisms of Enterococcus faecalis, Acinetobacter iwoffi, Actinobacillus species, Enterobacter aecum, Staph у lococcus к loossi, Staph у lococcus h у icus, Staph у lococcus aureus, Staph у lococcus epidermidis, and Staphylococcus Micrococcus candidus (1.2%) had weakly expressed enzymatic properties.


Introduction
The bird fauna of the Amur region is diverse and includes more than 320 species. About 60% of the avifauna are birds of the order sparrows. This group includes migratory, nomadic, and sedentary birds. Representatives of this Zoological group have a wide range of habitats, can be present in the wild, near human homes, livestock and poultry complexes, farms [1].
As a result of migration and migration of birds, the probability of spreading infectious diseases increases. In the body of birds, the pathogen can persist, multiply, be released into the external environment, and then enter the body of a healthy animal or bird, becoming the cause of new cases of the disease. Detection of microflora in the body of free-living birds, determination of its biochemical activity is one of the aspects of studying microorganisms in the parasitic phase of existence [2,3].
The works of many authors are devoted to the study of the body of free-living birds as a reservoir of infectious diseases [4][5][6][7][8][9][10]. Many publications describe modern methods of diagnostics of pathogenic and opportunistic microorganisms [11][12][13]. Studies of the species composition and properties of the microflora of free-living birds, taking into account the territorial nature, are not enough.
Relevance, scientific and practical significance determined the choice of the topic, goals and objectives of the study.
The scientific hypothesis is that the identification of biologically active bacteria will make it possible to predict the probability of free-living poultry participating in the epizootic process, and to plan appropriate preventive measures in poultry and animal husbandry.
In this regard, the purpose of our study was to study the species composition and biochemical activity of the microflora of the free-living bird of the Amur region.

Materials and methods
The object of the study was captured on the territory of the Amur region birds of the order of sparrows of different species in the number of 122 individuals. The material was selected from blue magpie (Cianopica ciana), common lentil (Caprodacus erithrinus), common hlycatcher (Scutigera coleoptrata), thick-billed warbler (Acrocephalus aedon), gray-headed bunting (Emberiza melanocephala), brown warbler (Phylloscopus fuscatus), siberian stingray (Lanius cristatus), nightingale common (Luscinia Luscinia), house sparrow (passer domesticus), magpie (pica pica), common tit (Parus Major); crows (Corvus corax); blue dove (Columba livia). The birds were caught in the spring and summer with the help of fishing nets.
The material was selected from the beaks and cloaks of birds by impregnating sterile cotton swabs with the contents of mucous membranes and further immersing them in test tubes with a sterile saline solution in the volume of 0.5 ml. The selected material was sown on nutrient media (meat-peptone agar, meat-peptone broth), placed in a thermostat and kept for 24 hours at a temperature of 37 °C.
Morphological properties of microorganisms by microscopy. Smears prepared from daily cultures were colored using the method of Gram, Romanovsky-Gimz, Peshkov, Kozlovsky, and ZIL-Nielsen.
Cultivation of bacteria was carried out on the following nutrient media: meat-peptone agar, meat-peptone broth, meat-peptone gelatin, Endo, Levina, Ploskireva, bismuth sulfite agar. To determine the cultural properties of microscopic fungi, Chapek and Saburo agar media were used. The growth pattern of colonies, color, edges, shape, profile, consistency, and structure were taken into account. The color change of colonies, the ability to hemolysis, and the growth pattern were observed in differential diagnostic media. In liquid nutrient media, the presence of sediment, its quantity, the presence of a film, ee thickness and consistency, and the degree of turbidity of the medium were noted [14,15].
The study of saccharolytic properties was determined by seeding microorganisms on GIS nutrient media with sucrose, lactose, glucose, maltose, mannitol, and dulcite. When evaluating the results, color changes and the presence of gas were taken into account [14,15].
Proteolytic properties were determined by determining the ability of microorganisms to liquefy gelatin. The degree of proteolysis was studied by determining the ability of microorganisms to release gases (hydrogen sulfide, ammonia and indole). The presence of gases was determined by using indicator papers impregnated with a 12% solution of oxalic acid, 10% solution of lead acetic acid. The results were taken into account after 18-24 hours [14,15]. To identify microbial cultures, chromogenic media were used, which allow determining the type of bacteria by detecting highly specific enzymes in the desired microorganisms [16].
The result of biochemical studies was compared with standard indicators of the determinant of bacteria Bergi.
The experimental material was processed using mathematical methods of variational statistics using the student's t-test.

Results
In the course of bacteriological studies, only 516 samples were studied. The largest number of microorganisms was isolated from the cloacal cavities, which accounted for 54.5% of the total number of isolated microorganisms.
Microbial contamination of bird beaks with Enterococcus faecalis microorganisms was recorded to the greatest extent in common flycatchers and thick -billed warblers (20.7%), and to the smallest extent in siberian zhulan, common nightingale and blue magpie (6.9%).
Microorganisms of the species Pseudomonas aeroginosa, Enterococcus gallinarum and Pseudomonas species were more often detected in the thick-billed Warbler in 37.5%, 66.7% and 28.6% of cases, respectively. The least amount of Pseudomonas aeroginosa isolated from a brown leaf Warbler (12.5%); Enterococcus gallinarum -the flycatcher common (33.3%); Pseudomonas species -the common brown flycatchers and warblers (7.1 %).
Citrobacter freundi was recorded in 4 bird species: common lentil, thick-billed Warbler, grayheaded bunting and brown warbler in 25% of cases.
Esherichia coli and Micrococcus species are not isolated from the beak cavities. Cultures of Aspergillus fumigatus were isolated from the beak cavities of common flycatcher (50%) and thickbilled Warbler (50.0%).
Significant microbial contamination of cloacal cavities was observed in thick-billed warbler-29 cultures (23.9%), brown warbler -22 cultures (18.2%), common lentils and blue magpies -19 cultures for each species (15.70%). The lowest microbial contamination of cloaca was found in birds of the species gray-headed bunting -12 cultures (9.9%) and common flycatcher -4 cultures (3.3%).  The study of biochemical properties showed that the bacteria of the species Lactobacillus species (18.6%) fermented glucose to form acid, maltose, sucrose, lactose, did not break down dulcite, mannitol. Proteolytic properties were shown in the ability to release gas (hydrogen sulfide). Enterococcus faecalis (15.7%) cleaved glucose, maltose, sucrose and lactose. Escherichia coli (13.9%) changed glucose and lactose with the formation of acid and gas, curtailed milk, and isolated indole.
The enzymatic properties of Acinetobacter iwoffi (12.8%) were shown in the fermentation of glucose, maltose, sucrose, lactose; proteolytic properties -in the ability to release gas (ammonia).
The enzymatic properties of Pseudomonas aeruginosa (6.8%) were poorly expressed. Microorganisms did not change glucose, sucrose, maltose, or lactose.
The biochemical properties of Pseudomonas species (6.4%) were poorly expressed. The micro-organisms did not change glucose, lactose, maltose, or sucrose; they liquefied gelatin and converted nitrates into nitrites.
Citrobacter freundi (2.3%) had the ability to ferment glucose and lactose with the formation of acid and gas, to dilute gelatin and curdled milk, and to release hydrogen sulfide and ammonia Micrococcus candidus (1.2%) had weakly expressed enzymatic properties. The ability to ferment glucose with the formation of acid has been established.
Enterobacter agglomerans microorganisms (0.6%) fermented glucose, maltose, lactose and mannitol with the formation of acid and gas; indole, hydrogen sulfide were not isolated, and gelatin was slowly liquefied.
Salmonella enteritidis bacteria (6.3%) did not ferment sucrose or lactose. Fermented with the formation of acid and gas glucose, mannitol. They did not form indole, but isolated hydrogen sulfide.
All kinds of microorganisms of the genus Staphуlococcus fermented glucose, lactose, sucrose and maltose, except for the types Staphуlococcus кloosii and Staphуlococcus aureus -mannitol is changed. Hydrogen sulfide and ammonia were isolated.
The results of biochemical studies are presented in

Discussion
Birds break off sparrows have a large habitat range. This group includes families that are divided into migratory, nomadic and sedentary birds. As a result of migration and migration of birds, the probability of spreading infectious diseases increases. The species composition and its microflora of free-living birds of the Amur region are insufficiently studied. In this regard, the goal was set and the tasks of investigations were defined. When investigating 13 species of free-living birds poison sparrows isolated Enterococcus, Pseudomonas, Acinetobacter, Citrobacter, Actinobacillus, Escherichia, Microccocus, Aspergillus, Enterobacter, Salmonella, Proteus, Staphulococcus.
The bird that found the degree of contamination of the beak cavities was microorganisms of Enterococcus faecalis and Actinobacillus species. These types of microorganisms had stable properties of their own.
Cultures of Actinobacillus species (29.8%) were most often isolated from the cloacal cavities.
A large degree of contamination of cloacal cavities is represented by cultures of microorganisms of Actinobacillus species and Enterococcus faecalis. These types of microorganisms were isolated from the cavities of beaks and cloaks, which corresponds to the level of their circulation in the gastrointestinal tract of birds. When determining the correlation coefficient between microbial contamination of the oral cavity and the cloaca of wild birds, it was found that this coefficient is equal to + 0.97, which indicates a strong relationship. Biochemical