P-ISSN: 2304-3075; E-ISSN: 2305-4360 International Journal of Veterinary Science www.ijvets.com; [email protected]

Research Article

Influence of Probiotics "Stimix Zoostim" on the Microflora of Faeces, Hematological Indicators and Intensitivity of Growth of Calves of the Dairy Period

Fail Khaziakhmetov1, Ayrat Khabirov1, Maksim Rebezov2,3,4, Almaz Basharov1, Ilnur Ziangulov5 and Eleonora Okuskhanova6*

1Bashkir State Agrarian University, Ufa, Russia; 2Ural State Agrarian University, Yekaterinburg, Russia 3Russian Academy of Staffing of Agro-Industrial Complex, Moscow, Russia; 4KG Razumovsky Moscow State University of technologies and management (The First Cossack University), Moscow, Russia; 5Agrofirma Bayramgul, Uchaly, Russia; 6Shakarim State University of Semey, Semey, Kazakhstan *Corresponding author: [email protected]

Article History: Received: September 27, 2018 Revised: October 05, 2018 Accepted: October 12, 2018

ABSTRACT

Of great interest to calves are probiotics, which prevent gastrointestinal tract diseases, increase the activity of digestive enzymes, are able to produce a variety of antibiotic compounds that inhibit the growth of pathogens. Studies in the conditions of LLC "Agrofirm Bayramgul" of Uchalinsky district of the Republic of Bashkortostan found that the use of the probiotic "Stimix Zoostim" based on the microorganisms Escherichia coli, Lactobacillus acidophilus, Saccharomyces cerevisiae, Bacillus subtilis, Azotobaktervinelandii, Azotobakterchroococcum, positively influences the formation microbiocenosis of faeces, hematological blood indicators and growth rate of calves of the dairy period of the black-and-white Holstein breed of German breeding. The introduction of the probiotic "StimixZoostim" into the diets of the diary period in the amount of 10 ml per head at the age of 10-20 days, 15 ml - at the age of 21-90 days, contributes to the increase in the number of lactobacilli by 27.0%, bifidobacterium by 37.6%, with a decrease in the number of echerichia by 31.3%, compared with the first control group. The use of the probiotic "StimixZoostim" in rations of calves of the milk period, within the physiological norm, contributes to the increase in the number of red blood cells by 9.3%, hemoglobin - by 11.3%, γ - globulins - by 32.6%. These changes in the blood indicate a higher level of natural resistance and are characteristic of intensively growing calves. With 100% preservation of calves in the milk period in all groups, in the third and fourth experimental groups, the average daily growth in calves was higher by 8.9-10.2% with a decrease in feed costs per 1 kg of growth in live weight by 8.1-9.4% compared with the control group. However, in the fourth test group, the increase in the dose of the probiotic, equivalent to an increase in the growth rate of calves, did not occur. Thus, an effective dose of a probiotic is a dose of 10 ml per head per day at the age of 10-20 days, 15 ml at the age of 21-90 days. The results of the manufacturing inspection of the effective dose showed that the economic effect per head was 149.23 rubles.

Key words: Calves of diary period, Probiotic "StimixZoostim", Microflora of faeces, Hematological indicators, Average daily growth, Feed costs

INTRODUCTION young animals of farm animals and calves of the diary period (Biyashev et al.., 2016) attract special attention Since the 2000s, the antibiotic resistance of among the substances possessing an antimicrobial effect. microorganisms, according to the World Health The use of Optisaf and Prolam yeast probiotic additives Organization (WHO), has become a global problem. As on the basis of strains of lactobacilli, streptococcus of measures, the toughening of control over the use of lactic acid and bifidobacteria promoted the development antibiotics is discussed, as well as the development of the of normal intestinal microflora and the normal intestinal next generation of antibacterial drugs (Schinor et al., microflora affected the formation of the animal's immune 2007; Sharipova et al.., 2017). Probiotic preparations system and prevented the reproduction of pathogenic, based on beneficial microorganisms that influence the opportunistic bacteria (Cheremushkina et al., 2015; development of normal microflora of the intestines of Mikolaychik et al., 2016; Mikolaychik et al., 2017). In

Cite This Article as: Khaziakhmetov F, Khabirov A, Rebezov M, Basharov A, Ziangulov I and Okuskhanova E, 2018. Influence of probiotics "StimixZoostim" on the microflora of faeces, hematological indicators and intensitivity of growth of calves of the dairy period. Inter J Vet Sci, 7(4): 178-181. www.ijvets.com (©2018 IJVS. All rights reserved)

178 Inter J Vet Sci, 2018, 7(4): 178-181. experiments on calves, the positive effect of various scientific and economic experiments, calves were selected probiotics on blood hematologic indicators was by pairs-analogues (by gender, date of birth, live weight) established, for example, an increase in phagocytic at the age of 7-10 days with 10 heads each (50% bull activity in leukocytes of peripheral blood with the calves + 50% calf). In each experiment, the conditions of addition of a multi-species multipurpose probiotic-MMP keeping and feeding were the same and corresponded to (Indartet al.., 2012), an increase in the total protein and the diet, developed according to the detailed standards IgG in groups with probiotic cultures of L. acidophilus (Kalashnikov et al.., 2003). The calves of the first control and (Al-Saiadyet al., 2010), erythrocytes and hemoglobin, group received the main diet (MD) without including reduction of heavy metals: zinc by 62.2%, cadmium by probiotic in it. The calves of the second test group in 63.7%, led by 56.7% compared to the control using a addition to the main diet were fed probiotic "Stimix probiotic preparation based on soy milk acidified by Zoostim" at a dose of 8 ml per head per day at the age of bifido-and propionic acid bacteria (Baeva et al., 2017). 10-20 days, 12 ml per head per day at the age of 21-90 Probiotic additives "Biodarin" and "Baikal EM-1" in days, the third test group - 10 ml per head per day at the rations of calves and young cattle lead to an increase in age of 10-20 days, 15 ml per head per day at the age of the digestibility and nutrient availability of nutrients in 21-90 days and the fourth test group - 12 ml per head per rations and an increase in the level of animal productivity day at the age of 10-20 days, 18 ml per head per day at the (Nurzhanova 2018; Malikova and Bagautdinova 2014). age of 21-90 days, periodicity - daily. During the The use of tested complex probiotic additives (milk feed manufacturing inspection the probiotic was tested as in additive contained lactobacilli and thermophilic the third test group - 10 ml per head at the age of 10-20 streptococci, Baikal EM-1, Vetosporin J) improves calf days, 15 ml at the age of 21-90 days, as an effective growth and milk production and can bring additional option. The probiotic was used with milk at the age of 10 economic benefits (Malikova and Bagautdinova 2014b; to 60 days and water at the age of 61 to 90 days 2 times a Malikova MG et al., 2012). day (50% of the daily norm). The duration of the As a probiotic for calves, those probiotics are of experiment was 83 days. The feeding of calves was interest that are capable of producing a variety of carried out taking into account the planned growth and the antibiotic compounds that inhibit the growth of pathogens, adopted calf feeding scheme, developed on the basis of for example the probiotic preparation "A2" (Chabayev et the actual nutritional value of the forages according to the al., 2013), for preventing calf and gastrointestinal diseases detailed standards. Up to 3 months of age (inclusively), - the complex use of the preparation Lactosept and during the experiment period, 290 liters of whole milk, 43 probiotics Zoostim - M (Karpin et al., 2015), increase the kg of hay, 95 kg of haystack, 33 kg of pre-feed mixed activity of digestive enzymes - the drug based on fodder and 89 kg of mixed feed KK-62 were consumed Lactobacillus and cottage cheese (Agarwal et al., 2002), for 1 head. The digestible protein consumption was 127 g contribute to the formation of antimicrobial proteins per 1 energy unit (EFU). produced by one strain of Escherichia coli to suppress Bacteriological study of faeces of calves was carried other E. coli strains (Tahamtan et al., 2012) and the out according to the methodological recommendations quality of feeds are the species-specific probiotic "Isolation and identification of bacteria of the Lactobacillus animalis SB310, Lactobacillus paracaseisub gastrointestinal tract of animals", approved by the sp, Paracasei SB137 and Bacillus coagulans in the ratio of Department of Veterinary Medicine of the Ministry of 30:35:35, respectively, probiotic "Prolam", "Bacell", etc. Agriculture of the Russian Federation No. 13-5-02 / 1043 (Agazzi et al., 2014; Khabirov and Khaziakhmetov, 2017). of May 11, 2004. Morphological composition and Despite the wide interest in the presented topic, the biochemical parameters of blood of experimental calves in following microorganisms included in the probiotic 3-month-old age were determined by conventional "StimixZoostim": Escherichia coli, Lactobacillus methods (Kondrakhin et al.., 2004). In carrying out the acidophilus, Sac-charomyces cerevisiae, Bacillus subtilis, studies the generally accepted methods for unifying Azotobaktervinelandii, Azotobakterchroococcum are research in the field of feeding farm animals using fairly new probiotic objects in cattle breeding. The detailed norms were followed. Biometric processing of influence of such a set of microorganisms on the state of the obtained material was carried out according to microbiocenosis, blood hematologic indicators and standard methods using Excel program of Microsoft growth rates of calves require extensive study. Office 2013 application. The aim of the study was to study the effect of the probiotic StimixZoostim on the microflora of faeces, hematological indicators and the growth rate of the calves RESULTS AND DISCUSSION of the dairy period. The microbiocenosis of faeces of calves at the age of MATERIALS AND METHODS three months is presented in Table 1.Studies have shown that the introduction of probiotic "StimixZooStim" into The research was carried out in the conditions of LLC the rations of calves in the third test group was 10 ml per "Agrofirma Bayramgul" in Uchalinsky district of the head at the age of 10-20 days, 15 ml at the age of 21-90 Republic of Bashkortostan, on calves of black-and-white days and the fourth test group - 12 ml per head day at the Holstein breed of German breeding during the period age of 10-20 days, 18 ml - at the age of 21-90 days, from October 1, 2017 to May 2, 2018. A total of 1 promoted an increase in the number of lacto- and scientific and 1 manufacturing inspection on calves of the bifodobacteria by 27.0-33.6 and 37.6-45.8%, respectively, dairy period were conducted to study and determine the reduction of the escherichia by 31.3 - 32.0%, compared effectiveness of the probiotic "StimixZoostim". For with the first control group (P˂0.05).

179 Inter J Vet Sci, 2018, 7(4): 178-181.

Table 1: Microbiocenosis of calves’ faeces at the age of 3 months, million CFU / g (X ± Sx, n = 3). Group Indicator 1 control 2 experimental 3 experimental 4 experimental Lactobacilli 5.48±0.36 6.48±0.38 6.96±0.32* 7.32±0.46* Bifidobacteria 6.12±0.38 7.12±0.42 8.42±0.44* 8.92±0.62* Escherichia 5.12±0.42 4.22±0.52 3.52±0.36* 3.48±0.38*

Table 2: Morphological composition and biochemical indicators of blood of experimental calves at the age of 3 months (X ± Sx, n = 3). Group Indicators 1 control 2 experimental 3 experimental 4 experimental White blood cells (WBC), 109/l 9.3±0.74 10.2±0.83 10.4±0.82 10.3±0.86 Red blood cells (RBC), 1012/l 7.5±0.16 7.8±0.18 8.2±0.19* 8.1±0.22 Hematoglobulin (Hb), g/l 102.6±2.52 108.6±3.82 114.2±3.12* 116.4±4.14* Total protein, g/l 62.4±3.12 63.1±3.04 65.8±3.18 65.9±3.28

albumins 41.8±2.26 40.3±1.96 38.8±1.82 38,6±1,64 α - globulin 14.8±1.14 11.4±0.83 11.1±0.73 11,2±0,93

β - globulin 14.6±1.82 12.1±0.86 11.9±0.78 11,1±0,89

fractions, fractions, % Protein Protein γ - globulin 28.8±2.22 36.2±2.12 38.2±2.26* 39,1±2,28* Glucose, mmol/L 3.2±0.18 3.3±0.24 3.4±0.22 3.4±0.21 AspAT, mmol/L (h*l) 0.66±0.06 0.67±0.08 0.68±0.12 0.68±0.16 АLТ, mmol/l (h*l) 0.38±0.18 0.40±0.12 0.42±0.16 0.45±0.12 Total calcium, mmol/l 2.6±0.14 2.7±0.21 2.8±0.22 2.9±0.24 Inorganic phosphorus, mmol / l 1.4±0.08 1.5±0.09 1.6±0.12 1.6±0.14

Table 3: The results of growing calves using different doses of probiotic "StimixZoostim" after 83 days of the experiment (X ± Sx, n = 10). Group Indicator Control 1 Experimental 2 Experimental 3 Experimental 4 Live weight at the beginning of the experiment, kg 34.7±0.73 35.0±0.96 34.6±0.62 34.8±0.56 Live weight at the end of the experiment, kg 95.1±1.55 97.9 ±1.75 100.4±1.87* 101.4±1.27* Absolute gain, kg 60.4±1.24 62.9±1.02 65.8±1.96* 66.6±1.98* Average daily weight gain, kg 728.0±14.92 758.0±12.56 793.0±23.66* 802.0±28.76* Tocontrol, % 100 104.1 108.9 110.2 EFU consumption per 1 kg of live weight gain 4.7 4.52 4.32 4.26 To control, % - 96.2 91.9 90.6 Preservation of livestock, % 100 100 100 100

Table 2 presents information on the morphological probiotic is a dose of 10 ml per head per day at the age of composition and biochemical indicators of blood of 10-20 days, 15 ml at the age of 21 -90 days. The results of experimental calves at the age of 3 months. the manufacturing inspection of the effective dose showed Morphological composition and biochemical that the economic effect per head was 149.23 rubles. indicators of blood of experimental calves at the age of 3 months were kept within the physiological norm specified Conclusions in the reference literature (Kondrakhin et al., 2004). According to the results of research it has been Within the limits of the physiological norm, the use of the established, that the probiotic "StimixZoostim" possesses probiotic "StimixZoostim" in calves' rations to the third high probiotic activity, having positive influence on a test group - a dose of a probiotic 10 ml per head at the age microbial landscape of faeces of calves. As a result of of 10-20 days, 15 ml - at the age of 21-90 days promoted feeding the probiotic, there was a significant increase in an increase in red blood cells by 9, 3%, hemoglobin - by the number of normofloras: lactobacilli and bifidobacteria, 11.3%, γ - globulins - by 32.6%, and in the fourth trial and a decrease in Escherichia. The use of the probiotic in group - 12 ml per head at 10-20 days, 18 ml - at the age of the amount of 10 ml per head per day at the age of 10-20 21-90 days contributed to an increase in hemoglobin by days, 15 ml - at the age of 21-90 days also positively 13.5%, γ - globulin by 35.8% compared to the first control influenced the growth rate of calves and feed consumption group (P˂0.05). Such changes in blood composition per 1 kg of live weight and the economic effect per head indicate a higher natural resistance of calves and are amounted to 149.23 rubles. characteristic of intensively growing animals (29). The results of growing calves are presented in Table 3. REFERENCES With 100% safety of the calves in all groups, in the third and fourth experimental groups, the average daily Agarwal N, Kamra DN, Chaudhary LC, Agarwal I, Sahoo calf gain was higher by 8.9-10.2%, while the feed costs A, Pathak NN, 2002. Microbial status and rumen per 1 kg of live weight decreased by 8.1-9.4% compared enzyme profile of crossbred calves fed on different to the first control group. However, in the fourth microbial feed additives. Letters in Applied experimental group, with an increase in the dose of the Microbiology: 34: 329-336. probiotic equivalent to an increase in the growth rate of Agazzi A, Tirloni E, Stella S, Maroccolo S, Ripamonti B, the calves did not occur, so the effective dose of the Bersani C, Caputo JM, Dell’Orto V, Rota N,

180 Inter J Vet Sci, 2018, 7(4): 178-181.

SavoiniG, 2014. Effects of species-specific probiotic lamb growth. Res J Pharmac Biolog Chem Sci, 9(3): addition to milk replacer on calf health and 866-870. performance during the first month of life. Annals of Kondrakhin IP, Arkhipov AV, Levchenko VI, 2004. Animal Science: 1(14):101–115. Methods of veterinary clinical laboratory diagnosis. Al-Saiady MY, 2010.Effect of probiotic bacteria on Moscow, KolosS. immunoglobulin g concentration and other blood Malikova MG, Akhmetova IN, Kuznetscova TN, Fisenko components of newborn calves. Journal of Animal NV, 2012. Efficiency of using new probiotic and Veterinary Advances; 9(3): 604-609. preparation “Vetosporin J” in calves diet. Feeding of Baeva ZT, Tedtova VV, Rekhviashvili EI, Tarchokov TT, agricultural animals and feed production, 11: 10-15. Shugusheva LK, Baragunova EA, 2017. Malikova MG, Bagautdinova AR, 2014. Effect of using Detoxification effect of soy milk-based probiotic on probiotics to nutrient digestion in the diet of calves. morphological and biochemical blood parameters in Feeding of agricultural animals and feed production; calves. Journal of Pharmaceutical Sciences and 7: 28-32. Research; 9(12): 2410-2413. Malikova MG and AR Bagautdinova, 2014b. Effect of Biyashev KB, Biyashev BK, Saribayeva DA, 2016. using “Baikal EM-1” probiotics in calves diet. Feeding Persistence of the Escherichia Coli 64g-probiotic of agricultural animals and feed production; 9: 20-24. strain in the intestine of calves. Biology and Merenkova VV and SV Moshkina, 2018. Physiological Medicine; 8(2): 2-3. approve of using “Bacell” probiotics in calves diet. Chabayev MR, Nekrasov R, Anissimova N, Gadjiyev AY, Proceeding of Int. Conf. “The ways of realization of Klementyeva Y, Grishenko V, 2013. Enrichment of Federal Science and Technology Program of agriculture calves’ diet with probiotic preparation “A2” and it development in 2017-2025”, Kurgan. P. 576-579. effect to productivity and metabolism. Dairy and Mikolaychik IN, LA Morozova, AG Koshchaev, ES meat cattle breeding: 4; 22-24. Stupina, 2016. Efficiency of intestinal Cheremushkina IV, Shakhov AG, Sashnina LY, microbiocenosis formation in calves by means of Chernitsky AE, Yerina TA, 2015. Antagonistic yeast probiotic supplements. Adv Agric Biolog Sci, activity of a probiotic Prolam in point of bacterial 2(6): 19-28. pathogens and its influence on an intestines Mikolaychik IN, LA Morozova, AG Koshchaev, Stupina microbiocenosis, the immune and clinical status of ES, 2017. Efficiency of the use of probiotic calfs. J Anim Vet Adv, 14(6): 182-191. supplements for the formation of digestive Indart M, Cerone S, Esteban EN, de Yaniz G, Inza AG, microbiocenosis in calves. Adv Agric Biolog Sci, Landi H, Mogni S, Igarza L, 2012. Multispecies 3(1): 35-43. multistrain probiotic effects on calves development Nurzhanova MA, 2018. Effect of probiotic additive and health. Open J Vet Med; 2: 225-229. Biodarin on nutrient intake and metabolism during Kalashnikov AP, Fisinina VI, Sheglova VV, Kleymenova feed consumption. Proceeding of Int. Conf. “The NI, 2003. The norms and feeding rations of livestock ways of realization of Federal Science and animals: manual. Moscow. Technology Program of agriculture development in Karpin SP, Kits YA, Lopteva MS, 2015. Combined use of 2017-2025”, Kurgan. pp: 605-608. Lactosept preparation and Zoostim-M probiotic for Schinor EC, MJ Silvador, H Ito, DA Dias, 2007. preventing colibacillosis in calves. Bulletin of All- Evaluation of the antimicrobial activity of crude Russian Research Institute of Sheep and Goat extracts and isolated constituents from Breeding: 8(2):169-174. Chrestascapigera. Braz J Microbiol, 38: 145-149. Khabirov AF, Khaziakhmetov FS, 2017. Effect of Sharipova A, Khaziev D, Kanareikina S, Kanareikin V, “Vitafort” and “Lactobifadol” probiotics to the Rebezov M, Okuskhanova E, Suychinov A, ducklings manure microbiocenosis. Proceedings of Yessimbekov Z, 2017. The effects of a probiotic International Conference “Current status, traditions dietary supplementation on the livability and weight and innovative technologies of agriculture”. Bashkir gain of broilers. Ann Res Rev Biol, 19(6): 1-5. State Agrarian University, 92-96. Tahamtan Y, Pourbakhsh A, Hyati M, Namvari MM, Khaziakhmetov F, Khabirov A, Avzalov R, Tsapalova G, Shirazi Z, Kargar M, Vesal Shirazi M, 2012. Rebezov M, Tagirov K, Giniyatullin S, Ishmuratov Detection of Colicin genes by PCR in escherichia coli K, Mishukovskaya G, Gafarova F, Yessimbekov Z, isolated from cattle in Shiraz-Iran. Archives of Razi 2018. Effect of probiotics on calves, weaned pigs and Institute: 1(67): 63-67.

181