ÓÄÊ 579.62
DOI 10.33861/2071-8020-2023-3-32-38
Mukhammadiev Rish.S., Mukhammadiev Rin.S., Valiullin L.R., Baryshev M.G., Yarullin A.I., Dupleva L.Sh., Karimullina I.G., Timerbaeva R.R.
Summary. The results of studies on optimizing the parameters of co-cultivation of B. subtilis GA27 and B. subtilis RF-45 strains are presented in the article. The greatest accumulation of the biomass of the studied bacillary strains, corresponding to their maximum antagonistic action against pathogens of intestinal infections in young poultry, was provided on a soy medium with the addition of 0.5% wheat bran at a temperature of 35°C, the active acidity of the medium was 6.5, the rotation speed of the rocking chair was 180 rpm, the duration of the process of their joint deep cultivation is 16 hours. In this case, stationary cultures with a total titer of at least (6.12 ± 0.17) õ 106 CFU/ml should be used as seed material and applied in an amount of 10% on the volume of the medium for growth. The optimal ratio of the seed concentration of B. subtilis GA27 and B. subtilis RF-45 was 1:2. The cell suspension, culture liquid, and supernatant obtained after co-cultivation of bacilli were characterized by the ability to correct the intestinal microbiota of Wistar rats with gentamicin-induced dysbiosis. Compared with the cell suspension and the culture supernatant of the bacterial association, the culture liquid of the latter had the highest recovery rate of the intestinal microbiota of animals. Under the influence of the cultural liquid of the bacillary consortium, the rate of recovery of the total number of intestinal microorganisms relative to the similar indicator of the experimental groups of animals that received its cultural supernatant and cell suspension was 1.12 and 8.35 times higher, respectively, the number of lactic acid bacteria - 1.04 and 13.03 times, respectively. The cell suspension, culture liquid and supernatant of the association B. subtilis GA27 and B. subtilis RF-45 exceeded the indicated indicator of the control group of animals by 22-245 times in terms of the recovery rate of bacteria of the E. coli group, the most effective in this respect was the culture supernatant of the bacterial consortium.
Keywords: Bacillus subtilis, biocompatible strains, co-cultivation, optimization, antimicrobial potential, correction of intestinal microecological disorders, efficiency, probiotics, metaprobiotics, metabiotics, poultry farming.
References:
1. Mukhammadiev R.S., Mukhammadiev R.S., Mukhammadieva A.S. et al. Antagonisticheskaya aktivnost’ novyh shtammov mikroorganizmov v otnoshenii vozbuditelej kishechnyh infekcij molodnyaka sel’skohozyajstvennoj pticy [Antagonistic activity of new strains of microorganisms against intestinal infection pathogens of young farm poultry]. - In: Molodezhnye razrabotki i innovacii v reshenii prioritetnyh zadach APK [Youth developments and innovations in solving the priority tasks of the agro-industrial complex]. - Kazan’ (Russia), 2022. - pp. 348-350.
2. Chicherin I.Yu., Pogorel’skij I.P., Lundovskih I.A. et al. Autoprobiotikoterapiya [Autoprobiotic therapy]. - Journal Infectology. - 2013 (5(4)). - pp. 43-54. - DOI: 10.22625/2072-6732-2013-5-4-43-54.
3. Fisinin V.I., Il’ina L.A., Jyldyrym E.A. et al. Bakterial’noe soobshchestvo slepyh otrostkov kishechnika cyplyat-brojlerov na fone pitatel’nyh racionov razlichnoj struktury [Broiler chicken cecal microbiocenoses depending on mixed fodder]. - Mikrobiologiya. - 2016 (85(4)). - ðð. 472-480. - DOI: 10.7868/ S0026365616040054.
4. Valiullin L.R., Mukhammadiev Rish.S., Mukhammadiev Rin.S. et al. Bakterii -antagonisty vozbuditelej kishechnyh infekcij i producenty kompleksa cellyulaz kak osnova dlya sozdaniya dobavok, ob»edinyayushchih funkcii probiotika i kormovogo fermenta [Antagonistic bacteria of pathogens of intestinal infections and producers of a complex of cellulases as a basis for creating additives that combine the functions of a probiotic and a feed enzyme]. - Dostizheniya nauki i tekhniki APK. -2021 (35(9)). - ðð. 60-66. - DOI: 10.53859/02352451_2021_35_9_60.
5. Feoktistova N.V., Mardanova A.M., Lutfullin M.T. et al. Biopreparaty mikrobnogo proiskhozhdeniya v pticevodstve [Microbial preparations in poultry farming]. - Uchenye Zapiski Kazanskogo Universiteta. Seriya Estestvennye Nauki. -2018 (160(3)). - ðð. 395-418.
6. Begunova A.V., Rozhkova I.V., Shirshova T.I. et al. Biosintez antimikrobnyh bakteriocinopodobnyh soedinenij shtammom Lactobacillus reuteri LR1: optimizaciya uslovij kul’tivirovaniya [Optimization of conditions for Lactobacillus reuteri LR1 strain cultivation to improve the biosynthesis of bacteriocin-like substances]. - Biotekhnologiya. - 2019 (35(5)). - ðð. 58-69. - DOI: 10.21519/ 0234-2758-2019-35-5-58-69.
7. Dobrynya Yu.M., Timchenko L.D., Bondareva N.I. et al. Vliyanie biologicheski aktivnoj substancii iz Medusomyces gisevii na pokazateli fagocitarnoj aktivnosti nejtrofilov krovi belyh krys [Influence of a biologically active substance from Medusomyces gisevii on the phagocytic activity of blood neutrophils of white rats]. -Agrarnyj vestnik Urala. - 2018 (1(168)). - ðð. 8-11.
8. Khadieva G.F., Lutfullin M.T., Nikolaeva A.A. et al. Vliyanie probiotikov Bacillus subtilis GM2 i GM5 na rost i usvoyaemost’ kormov u cyplyat-brojlerov [The effect of Bacillus subtilis GM2 and GM5 probiotics on the growth and fodder digestibility of broiler chickens]. - Uchenye Zapiski Kazanskogo Universiteta. Seriya Estestvennye Nauki. -2019 (161(3)). - ðð. 472-489. - DOI: 10.26907/2542-064X.2019.3.472-489.
9. Mukhammadieva A.S., Mukhammadiev R.S., Mukhammadiev R.S. et al. Vydelenie i izuchenie morfologicheskih i biohimicheskih svojstv novyh shtammov molochnokislyh bakterij, perspektivnyh dlya sozdaniya probioticheskih preparatov [Isolation and investigation of morphological and biochemical properties of new strains of lactic acid bacteria promising for creation probiotic preparations]. -Veterinarnyi vrach. - 2020 (3). - ðð. 39-46.
10. Gricinskaya V.L. Probiotiki: klassifikaciya, osnovnye harakteristiki, trebovaniya k probioticheskim shtammam i sfera ih primeneniya [Probiotics: classification, main characteristics, requirements for probiotic stains and their scope Nauchno-prakticheskij medicinskij zhurnal]. - Children’s Medicine of the North-West. - 2022 (10(3)). - ðð. 12-20.
11. Zorina A.S., Maksimova YU.G., Demakov V.A. Bioplenkoobrazovanie mono-i smeshannyh kul’tur shtammov Alcaligenes faecalis 2 i Rhodococcus ruber GT 1 [Biofilm formation by monocultures and mixed cultures of Alcaligenes faecalis 2 AND Rhodococcus ruber GT 1]. - Mikrobiologiya. - 2019 (88(2)). - ðð. 175-183. -DOI: 10.1134/S0026365619020149.
12. Ushakova N.A., Kotenkova E.V., Kozlova A.A. et al. Izuchenie mekhanizmov probioticheskogo dejstviya shtamma Bacillus subtilis 8130 [A study of the mechanisms of probiotic effect of Bacillus subtilis strain 8130]. - Appl. Biochem. Microbiol. - 2006 (42(3)). - ðð. 285-291. - DOI: 10.1134/S0003683806030057.
13. Neminushchaya L.A., Pavlenko I.V., Kazaku A.A. et al. Innovacionnye sinbiotiki dlya sel’skohozyajstvennyh zhivotnyh i pticy [Innovative synbiotics for farm animals and poultry]. - Veterinarnyj vrach. - 2023 (1). - ðð. 42-50.
14. Muhammadieva A.S., Muhammadiev R.S., Usol’cev K.V. et al. Mikroflora kishechnika krys pri eksperimental’nom antibiotik-associirovannom disbioze i vozmozhnost’ ee korrekcii preparatom na osnove molochnokislyh i propionovokislyh mikroorganizmov [Rat intestinal microflora in experimental antibiotic-associated dysbiosis and the possibility of its correction with preparation based on lactic and propionic acid bacteria]. - Uchenye zapiski Kazanskoj gosudarstvennoj akademii veterinarnoj mediciny im. N.E. Baumana. - 2021 (246(2)). - ðð. 141-145.
15. Begunova A.V., Rozhkova I.V., Zvereva E.A. et al. Molochnokislye i propionovokislye bakterii: formirovanie soobshchestva dlya polucheniya funkcional’nyh produktov s bifidogennymi i gipotenzivnymi svojstvami [Lactic and propionic acid bacteria: the formation of a community for the production of functional products with bifidogenic and hypotensitive properties]. - Applied Biochemistry and Microbiology. - 2019 (55(6)). - ðð. -566-577. - DOI: 10.1134/S0555109919060047.
16. Khadieva G.F., Lutfullin M.T., Mochalova N.K. et al. Novye shtammy Bacillus subtilis kak perspektivnye probiotiki [New Bacillus subtilis strains as promising probiotics]. - Microbiology. - 2018 (87(4)). - ðð. 463-471. - DOI: 10.1134/S0026261718040112.
17. Valiullin L.R., Mukhammadiev Rish.S., Mukhammadiev Rin.S. et al. Novye shtammy Lactobacillus acidophilus kak perspektivnye probiotiki dlya pticevodstva [New Lactobacillus acidophilus strains as promising probiotics for poultry farming]. - Veterinariya Kubani. - 2022 (6). - ðð. 16-21. - DOI 10.33861/2071-8020-2022-6-16-21.
18. Mukhammadiev Rish.S, Valiullin L.R., Mukhammadiev Rin.S et al. Optimizaciya sostava pitatel’noj sredy probioticheskogo shtamma B. subtilis GA24 -producenta kormovyh fermentov [Optimization of nutrient medium composition of probiotic strain B. subtilis GA24 - producer of forage enzymes]. - Uchenye zapiski Kazanskoj gosudarstvennoj akademii veterinarnoj mediciny im. N.E. Baumana. -2022 (250(2)). - ðð. 155-159.
19. Latypov G.L., Muhammadieva A.S., Valiullin L.R. Parametry ostroj toksichnosti i kozhno-razdrazhayushchego potenciala kormovoj kompozicii s metaprobioticheskimi svojstvami [Parameters of acute toxicity and skin irritant effect of feed composition with metaprobiotic properties]. - Uchenye zapiski Kazanskoj gosudarstvennoj akademii veterinarnoj mediciny im. N.E. Baumana. -2023 (253(1)). - ðð. 174-179.
20. Feoktistova N.V., Mardanova A.M., Hadieva G.F et al. Probiotiki na osnove bakterij roda Bacillus v pticevodstve [Probiotics based on bacteria from the genus Bacillus in poultry breeding]. - Uchenye Zapiski Kazanskogo Universiteta. Seriya Estestvennye Nauki. - 2017 (159(1)). - ðð.85-107.
21. Siraeva Z.Yu., Zaharova N.G., Il’inskaya O.N. Tekhnologiya polucheniya i ocenka stabil’nosti pri hranenii zhidkoj preparativnoj formy biofungicida Bacizulin [Technology of production and storage stability evaluation of the liquid formulation of biofungicide Bacizulin]. - Uchenye Zapiski Kazanskogo Universiteta. Seriya Estestvennye Nauki. - 2010 (152(4.)). - ðð. 169-178.
22. Muhammadiev R.S., Muhammadiev R.S., Valiullin L.R. et al. A Sravnitel’naya ocenka in vitro effektivnosti metabioticheskoj i metaprobioticheskoj kompozicij, perspektivnyh dlya primeneniya v pticevodstve [Comparative in vitro evaluation of the efficiency of metabiotic and metaprobiotic compositions promising for use in poultry farming]. - In: Aktual’nye voprosy sovershenstvovaniya tekhnologii proizvodstva i pererabotki produkcii sel’skogo hozyajstva [Current issues of improving the technology of production and processing of agricultural products]. -Joshkar-Ola (Russia): Marijskij gosudarstvennyj universitet, 2023. - ðð. 813-816.
23. Chicherin I.Yu., Pogorel’skij I.P., Lundovskih I.G. et al. Sravnitel’naya eksperimental’naya ocenka effektivnosti sovremennyh probiotikov, prebiotikov, sinbiotikov i metabiotikov pri korrekcii narushenij mikrobiocenoza kishechnika u zhivotnyh s antibiotiko-associirovannym disbiozom [Comparative experimental evaluation of the effectiveness of modern probiotics, prebiotics, synbiotics and metabiotics for correction of intestinal microbiocenosis in animals with antibiotic-associated dysbiosis]. - Eksp. Klin. Gastroenterol. - 2016 (7). - ðð. 106-120.
24. Stoyanova L.G., Ustyugova E.A., Netrusov A.I. Antimikrobnye metabolity molochnokislyh bakterij: raznoobrazie i svojstva (obzor) [Antibacterial metabolites of lactic acid bacteria: their diversity and properties]. - Prikl. Biokhim. Mikrobiol. -2012 (48(3)). - ðð. 259-275.
25. Pozolotina N.V., Darmov I.V., Marakulin I.V. et al. Tekhnologiya prigotovleniya zhidkoj formy veterinarnogo probioticheskogo preparata na osnove shtamma Lactobacillus paracasei [Technology of preparing the liquid form of the veterinary probiotic on the basis of the strain Lactobacillus paracasei]. - Fundamental’nye issledovaniya. 2015(2-10). - ðð. 2164-2169.
26. Mukhammadiev Rish.S., Mukhammadiev Rin.S., Valiullin L.R. et al. Fermentativnaya aktivnost’ ksilanaz i cellyulaz probioticheskih shtammov Bacillus subtilis [Enzymatic activity of xylanases and cellulases of probiotic strains Bacillus subtilis]. - Veterinarnyi vrach. - 2019 (3). - ðð. 19-23.
27-42. Vide supra.
Author affiliation:
Mukhammadiev Rinat S., Ph.D. in Biology, scientific researcher of the Laboratory of Applied and Experimental Microbiology of the All-Russian Scientific Research Institute of Phytopathology; scientific researcher of the Sector of Probiotic Preparations and Enzymes of the Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-987-4214127; e-mail: tanirtashir@mail.ru.
Valiullin Lenar R., Ph.D. in Biology, senior scientific researcher of the Department of Ecology of the All-Russian Scientific Research Institute of Phytopathology; Head of the Laboratory of Feeds and Feed Additives of the Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-950-9698469; e-mail: valiullin27@mail.ru.
Baryshev Mikhail G., D.Sc. in Biology, Professor, Professor of the Russian Academy of Sciences, Acting Director of the All-Russian Scientific Research Institute of Phytopathology; 5, Institutskaya st., Bolshie Vyazemy, Odintsovo district, Moscow region, 143050; phone: 8-495-5974228; e-mail: vniif@vniif.ru.
Yarullin Ainur I., Ph.D. in Biology, Senior Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-905-3174170; e-mail: abii@mail.ru.
Dupleva Liliya Sh., Ph.D. in Biology, Senior Research Fellow, Department of Bacteriology, Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-917-2782737; e-mail: dupleva.lilya@mail.ru.
Karimullina Ilsiyar G., Ph.D. in Biology, Senior Research Fellow, Department of Virological and Ultrastructural Research, Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-904-7699225; e-mail: 89047699225@mail.ru.
Timerbayeva Razaliya R., Ph.D. in Veterinary Medicine, Associate Professor of the Department of Epizootology and Parasitology, Kazan State Academy of Veterinary Medicine named after N.E. Bauman; 35, Sibirskiy Trakt st., Kazan, 420029; phone: 8-903-3401420; e-mail: 8-903-3401420@mail.ru.
Responsible for correspondence with the editorial board: Mukhammadiev Rishat S., Ph.D. in Biology, scientific researcher of the Laboratory of Applied and Experimental Microbiology of the All-Russian Scientific Research Institute of Phytopathology; scientific researcher of the Laboratory of Feeds and Feed Additives of the Federal Center for Toxicological, Radiation and Biological Safety; Nauchnyy gorodok-2 st., Kazan, 420075; phone: 8-939-3728789; e-mail: tashir9891@mail.ru.
http://vetkuban.com/en/num3_202306.html