Morphological, Biological, and Biochemical Properties of Escherichia, Salmonella and Pasteurella Isolated from Environmental Objects

EurAsian Journal of BioSciences Eurasia J Biosci 14, 1567-1569 (2020)

Morphological, biological, and biochemical properties of Escherichia, Salmonella and Pasteurella isolated from environmental objects

D.G. Musiev 1*, G.Kh. Azaev 2, Sh.A. Gunashev 2, A.V. Volkova 3, R.O. Tsakhaeva 4, Iu.K. Zulfugarly 4 1 Ph.D. Veterinary, FSBEI HE “M.M.Dzhambulatov Dagestan State Agrarian University", Dagestan, RUSSIA 2 Candidate of Veterinary Sciences, FSBEI HE “M.M.Dzhambulatov Dagestan State Agrarian University", Dagestan, RUSSIA 3 Applicant, FSBEI HE “M.M.Dzhambulatov Dagestan State Agrarian University", Dagestan, RUSSIA 4 Graduate Student, FSBEI HE “M.M.Dzhambulatov Dagestan State Agrarian University", Dagestan, RUSSIA *Corresponding author: D.G. Musiev

Abstract The article describes studies on the isolation of bird salmonellosis, colibacteriosis and pasteurellosis pathogens from environmental objects, their morphological, pathogenic and biochemical properties. The cultures of Salmonella and Eshirichy grew well on the Endo, Levin and Ploskirev media, and pasteurellosis in the Hottinger medium. The data obtained show that in the poultry farm settings the pathogenicity of the isolated Escherichia strains was 53.3-80.0%, Salmonella - 73.3-93.3%, and Pasteurell - 66.6%. Biochemically, all strains were active.

Keywords: poultry, salmonellosis, colibacteriosis, pasteurellosis, external environment, pathogenesis, studies, morphology, pathogenic properties, biochemistry, strains

Musiev DG, Azaev GKh, Gunashev ShA, Volkova AV, Tsakhaeva RO, Zulfugarly IuK (2020) Morphological, biological, and biochemical properties of Escherichia, Salmonella and Pasteurella isolated from environmental objects. Eurasia J Biosci 14: 1567-1569.

INTRODUCTION of birds in Dagestan, salmonellosis was isolated in more than 13%, and pasteurellosis in more than 8% of cases. Industrial poultry farming, as one of the most cost- Colibacteriosis of birds often occurs in association with effective sectors of agriculture, has shown rapid salmonellosis and pasteurellosis. development in recent years. Old and new large broiler According to Panikar I.I. et al. (1993), an associative and egg poultry farms have been reestablished. course of colibacteriosis with salmonellosis in 21.0% However, the concentration of a large number of birds in was noted. A. Lebedev et al. (1973), described a mixed a limited area, for many reasons, causes a decrease in course of colibacteriosis and pasteurellosis. the body's resistance, which can lead to infection. This, The pathogens of infections that persist on infected in turn, leads to significant contamination of the external environmental objects, the virulence of which depends environment by pathogens of infectious diseases on the physical, chemical factors and biological secreted by a sick bird. The external environment, like a properties of the pathogen itself, have a significant sick bird, is a factor of the infection of a healthy bird. influence on the epizootic situation in infectious diseases Domestic and foreign scientists have shown in their of birds. Feed, water, care products and other studies that more than 60-70% of poultry mortality environmental objects play an important role in the occurs in bacterial infections, among which emergence and spread of bacterial and viral infections colibacteriosis, salmonellosis and pasteurellosis are of birds. In the external environment, under the influence most common. of various factors, bacteria can change their biological According to Andreev N.A. (2004), Borisenkova AN properties, which makes it difficult to identify such (2004), Bessorabov et al. (2008), Musieva D.G. (2008), cultures. Akpan and Udoh, 2017). the leading place among infectious pathologies of The study of the morphological, biological and bacterial etiology is colibacteriosis, which accounts for biochemical properties of pathogens that are most often more than 50% of infected birds. Among bacterial infections, researchers note a significant spread of Received: October 2019 salmonellosis, pasteurellosis. According to Azaev G.Kh. Accepted: May 2020 (2010), in the general pathology of infectious diseases Printed: June 2020

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EurAsian Journal of BioSciences 14: 1567-1569 (2020) Musiev et al. detected in the external environment will allow timely proteolytic activity of microbes was studied in a gelatin- planning of veterinary and sanitary measures and based medium. stopping the infection in the primary epizootic focus. The relatively long preservation of Salmonella, RESULTS Escherichia and Pasteurella in the external environment, To isolate E-Coli from environmental objects, 85 their potential ability to cause disease in birds make samples were taken: 25 washings from the walls, 20 these studies relevant from a scientific and practical samples from feeders, feed and floor. Samples after point of view. appropriate preparation were cultured on BEA, BEB and on differential diagnostic Endo and Ploskirev media. On MATERIALS AND METHODS solid nutrient media, mainly the formation of colonies Studies on the isolation of Escherichia, Salmonella, with smooth edges and a smooth shiny surface of 2-4 and Pasteurella were carried out at the Department of mm in size (S-shape) was noted. A small part of the Epizootology, DagSAU, Republican Veterinary colonies (0.5%) had rugged edges and a rough surface Laboratory. Material was taken from the objects of the (R-shape). Red or raspberry colonies with a metallic external environment of poultry farms of the sheen grew on Endo's medium, and pale pink with a Karabudakhkent and Khasavyurt districts. Samples for metallic sheen on Ploskirevo medium. the study were taken from feed, feeders, walls of the 22.3% of the 85 samples studied had esherichia poultry house, floor, water according to the “Guidelines isolated. O1, O2, O78, O26, O111 are identified by the for the accelerated indication of morganella, salmonella variant composition. Salmonella was isolated from 72 and enteropathogenic escherichia with adhesive samples taken from various environmental objects (22 antigens in pathological material, feed, environmental samples from the walls, 15 - from water, 25 - from the objects in the coagulation reaction approved by the floor, 10 - from the air). Department of Veterinary Medicine dated 11.10.1999, On BEA, salmonella formed round colonies with “Guidelines for the bacteriological diagnosis of smooth edges with a diameter of 1-2 mm, gray-white in colibacteriosis (Escherichiosis) of animals, approved by color with transparent edges. On the Endo medium, the Department of Veterinary Medicine on July 27, 2007. transparent pale pink colonies grew, on the Ploskirev In total, 85 samples, 72 samples and 32 samples were medium - small, colorless, slightly unclear colonies, on taken in various poultry farms to study for colibacillosis, Bismuth Sulfite Agar - black with metallic sheen salmonellosis, pasteurellosis, respectively. colonies, on the Levin medium - transparent, sometimes The samples after their appropriate preparation from with a purple tint colonies. Isolated salmonella was the environmental objects were cultured on Endo, identified as S. enteritidis and S. gallinarum. Ploskirev media, bismuth agar, broth and Höttinger agar. Pasteurella was isolated with 25 samples of material In the presence of characteristic colonies, serological taken from the walls of the houses (8 samples), from the identification was performed in RA on glass with floor (9 samples) and feed (8 samples). Cultures for monoreceptor absorption salmonella sera and with a set isolation of the pathogen were grown on a liquid and of typical agglutinating O-colimonovalent sera. solid Hottinger medium. The broth culture at the Identification of the isolated pasteurella cultures was beginning became turbid, and after 24-48 hours of carried out based on enzymatic properties. incubation, ir clarified and formed sediment at the Determination of the pathogenic properties of bottom of the tube. On a solid nutrient medium, bacteria was performed using agar cultures of transparent colonies with even edges developed, grown Salmonella and Escherichia. Suspended cultures of into agar. The resulting cultures were identified as P. each species of bacteria are infected. “Guidelines for the multocida. laboratory diagnosis of animal and bird pasteurellosis”, The pathogenic properties of the isolated cultures approved by the General Veterinary Administration of were determined by infection of white mice weighing 14- 08.20.1992 or three white mice weighing 14-15 g 16 g and aged 20-30 days, and chickens aged 18-21 intraperitoneally at a dose of 0.5 million microbial cells. days. The death of two or more mice within three days after A study of the pathogenicity of Escherichia cultures infection gives reason to argue about the pathogenicity isolated from environmental objects showed that the of this culture. To determine the pathogenicity of degree of pathogenicity of different serovariants was pasteurella, the daily broth culture is administered to different and ranged from 53.3% to 80.0%. white mice weighing 14-15 g intraperitoneally at a dose The pathogenicity of S. enteritidis and S. pullorum - of 0.3 cm³. Pathogenic cultures cause the death of mice gallinarum were 73.3% and 93.3%, respectively, and P. after 24-72 hours; slightly pathogenic cultures die within multocida - 66.6%. It should be noted that pathogenic 7 days. Saccharolytic properties were studied on the strains were detected in all studied samples, however, differential diagnostic environments of Giss. The the largest number of pathogenic samples in

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EurAsian Journal of BioSciences 14: 1567-1569 (2020) Musiev et al.

Escherichia, salmonella and pasteurellosis were CONCLUSION detected in the material taken from the floor. Thus, studies have shown that environmental objects Biochemical properties were studied in all studied in Dagestan poultry farms are contaminated by cultures of microorganisms. Pathogenic Escherichia pathogens of bacterial infections. From various ferment lactose, arabinose, decompose mannitol and environmental objects, S. enteritidis and S. sucrose to form acid and gas; some strains form pullorumgallinarum, E-Coli O1, O2, O78, O26, O111 hydrogen sulfide. Voges-Proskauer reaction was serovariants, and P. Multocida were isolated. Most negative, gelatin was not liquified. often, these bacteria were found in swabs from the floor All Salmonella strains were fermented to form acid and feeders. and gas glucose, fructose, maltose and mannitol. Salmonella, Escherichia and Pasteurella, isolated Salmonella ferment with the formation of acid and from environmental objects, grow well on ordinary gas, rhamnose, xylose, arabinose, maltose, dulcite, nutrient media (BEB, BEA) and differential diagnostic sorbitol, trehalose, mannitol, produce hydrogen sulfide Endo, Levin, Ploskirev, and Hottinger media. The and do not break down urea, fermented sucrose and degree of pathogenicity in mice and chickens of lactose, produced hydrogen sulfide. Escherichia was in the range of 53.-80.0%, Salmonella P. multocida ferments glucose, sucrose, mannitol, -73.3-93.3% and Pasteurellus - 66.6%. sorbitol, arabinose, dulcite with the formation of acid, without forming hydrogen sulfide and without diluting gelatin.

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