Use of DIABAX feed additive and a biogenic stimulant in calves during their rehabilitation after gastrointestinal infections

INTRODUCTION

Livestock survivability and performance improvement is one of the important challenges facing the Russian Federation agro-industrial complex [1][2]. Calf rearing are challenging since the newborns’ bodies are poorly adapted to adverse environmental conditions due to the morphofunctional immaturity of immune system and gastrointestinal tract during their first days of life [3]. This, in turn, results in the various gastrointestinal diseases in calves and their deaths [4][5][6], as well as the disease recurrence [7][8].

Therewith, complex rehabilitation procedures including use of various biological stimulants enabling vital component deficiency filling and contributing to the metabolic normalization and full-calf body resistance enhancement are required to maintain the metabolic status and homeostasis of the calves during their recovery [9][10][11][12][13].

The mechanism of biogenic stimulant action includes changes in activity of some enzymes due to the biogenic stimulant attachment to the enzyme protein. Changes in the enzyme activity results in endocrine restructuring, an increase in tropic hormones secreted by pituitary gland that stimulate adrenal gland, thyroid and pancreas functions, etc. They have a favorable impact on trophic function of the nervous system, enhance thyroid tissue and adrenal gland functions, stimulate corticosteroid hormone secretion and pancreatic gland function, regulate gastrointestinal tract secretory and motor functions, gas and phosphorus metabolism, intermediary metabolism, reticuloendothelial system and regenerative processes, improve body state, appetite, assimilation processes that contribute to higher weight gains [14][15].

The study was aimed at testing of DIABAX feed additive and a biogenic stimulant for their effectiveness for correction of biochemical, hematological blood parameters in young cattle after gastrointestinal infections.

Test tasks:

1. To test used feed additive and biogenic stimulant for their effects on morphological and biochemical blood parameters in calves during their rehabilitation after gastrointestinal infections.
2. To test the feed additive and biogenic stimulant for their effects on disease rate and survival rate in calves, calf performance during their rehabilitation period.

MATERIALS AND METHODS

Bacteriological tests of calf feces were carried out for the purpose of diagnosis. Isolated microorganisms were tested for their sensitivity to antimicrobials with disk diffusion test [16].

A pilot batch of biogenic stimulant was prepared from raw materials (slaughter waste, category II offal) kept in a refrigerator at temperature of +2 up to +4 °С for 5–7 days, ground and then mixed with an extraction agent at the Department of the All-Russian Institute of Antler Reindeer Breeding, Altai Scientific Center for Agrobiotechnologies. The ultrasonic-assisted extraction was carried out; the final product was filtered, packed, and sterilized in an autoclave [17]. Administration of the biogenic stimulant enhances metabolism, body’s resistance and stimulates body’s functions.

DIABAX, a new feed additive, developed by the GK KONSTANTA (Saransk) [18] is a viscous fluid of light-brown color with slight odor, has a bactericidal, bacteriostatic effect on a wide range of microorganisms and pathogenic fungi owing to potassium iodide and electrostatically charged polyelectrolyte polydimethyldiallylammonium chloride contained in it. DIABAX feed additive is highly soluble in water, does not change medium pH level, remains active in acidic and alkaline media, as well as in protein and fatty media. The principle feature of the additive is inhibition of pathogenic microflora based on its physical, but not chemical effect, while its components has no adverse effect on normal gastrointestinal cells of animals. Polyelectrolyte has an electrostatic charge opposite to the charge of cells of pathogenic bacteria, fungi and other microorganisms. When Diabax feed additive interacts with the pathogen surface. This interaction leads to microorganism cell membrane is encapsulation that slows down and then completely inhibits microorganism breathing, nutrition and reproducibility.

For the trial aimed at testing of the restorative treatment for its effectiveness in calves during their rehabilitation after gastrointestinal infections, 3 groups of 10–30 day-old calves were formed according to the analogous pairs principle: two test (T-1, T-2) groups and one control (C) group. Tested products were administered to the calves according to the scheme presented in Table 1.

All procedures involving animals were carried out in accordance with ethical standards laid down by European Convention ETS No. 123.

The feed additive and biogenic stimulant were tested for their effectiveness based on the following: morphological tests of blood (total erythrocyte counts, total leukocyte counts, hemoglobin level) using conventional methods [19]; biochemical tests of sera: refractometric determination (IRF-22, Russia) of serum total protein, nephelometric determination of protein fractions [20]; ELISA determination of total immunoglobulin G using appropriate ELISA test-kit; determination of serum mineral content by unified method using Vital Diagnostics SPb kits (Russia) and Stat Fax® 1904+ biochemical photometer (Awareness Technology, Inc., USA); bacteriological tests of biological material samples in accordance with the Methodological Guidelines^{1, 2}.

Blood samples were collected before the trial and 10 days after the trial completion. Mean values was assessed for their reliability of using Student’s – Fisher’s test.

RESULTS AND DISCUSSION

The gastrointestinal disease incidence in calves kept on the farm periodically rises due to violation of zootechnical and veterinary rules for animal keeping, feeding and handling during mass calving. The etiological causes are: diseased animals, convalescent animals, dams – pathogenic microorganism strain carriers, infected environment. The following bacteria were detected in feces from diseased calves subjected to bacteriological tests: Salmonella Dublin, Mannheimia haemolytica, all isolated bacteria strains were pathogenic for white mice. Levofloxacin, marfloxin, enrofloxacin, norfloxacin, ofloxacin, polymyxin, kanamycin were found to be effective antimicrobials. At the final stage of the trial, no pathogenic microorganism strains were detected during the bacteriological tests of fecal samples from calves of test and control groups.

Tests of the blood samples collected before the trial aimed at testing DIABAX feed additive and biogenic stimulant for their effectiveness for correction of biochemical, hematological blood parameters in calves during their rehabilitation after gastrointestinal infections showed decrease in phosphorous level by 19.1%, calcium level by 9.2%, magnesium level by 46.3% as compared with the physiological norm and simultaneous increase in zinc level by 3.5% as compared to the physiological norm (Table 2).

Calves of both test groups demonstrated normalization of serum phosphorus level, a significant increase in serum calcium level by 19.0% in the T-1 group and by 23.8% in the T-2 group (p < 0.05) as compared to that ones in sera from control group calves, in serum magnesium level – by 57.1% in T-1 group and by 61.2% in T-2 group (p < 0.05), serum potassium level – by 2.2% in T-1 group and by 4.3% in T-2 group 10 days after completion of tested feed additive and biogenic stimulant administration. There was a positive dynamics of zinc level decrease by 0.8% in sera from control group calves, by 5.5% in sera from T-1 test group calves, by 6.3% in sera from T-2 test group calves as compared to initial values during the rehabilitation period.

Biochemical tests of calf sera collected before the trial showed slight decrease in albumin protein fraction by 4.7%, and α-globulin protein fraction by 23.3%. Decrease in albumin-globulin ratio by 31.3% is indicative of imbalance between the protein fractions, 14.0% decrease in the amount of immunoglobulins G mainly responsible for humoral immunity is indicative of body protective function suppression in animals used for the trial (Table 3).

At the final stage of the trial, albumin and α-globulin protein fractions in the calf sera were found to normalize to the physiological norm with a significant increase (p ≤ 0.05) in albumin levels in T-1 and T-2 groups and α-globulin fraction in T-2 group. In calves of control group, serum β-globulin level increased by 34.2% as compared to the initial values and significantly differed (p ≤ 0.05) from the test group values. Albumin-globulin ratio in animals of control group and test groups increased by 10.5% and 29.8–35.1% during the period of rehabilitation of the calves after gastrointestinal infections.

Immunoglobulin G amount reached its physiological norm in sera from test group calves. Immunoglobulin G amount increased by 72.1% in calves of control group and by 2.3–2.7 times in calves of test groups as compared to the initial values.

Analysis of hematological blood parameters in calves at the beginning and at the end of the trial showed no significant differences between the control and test groups. There was a positive trend towards an increase in the tested blood parameters within the physiological norm at the final stage of rehabilitation of calves in test groups as compared to the initial values and blood parameters of calves in control group (Table 4).

In test groups all calves survived (100% survival rate) during rehabilitation period when the tested feed additive and biogenic stimulant were administered to the calves and then during 60-day clinical observation period (Table 5). Disease rate was 66,7% in control group calves that was higher by 14.4 and 40.0% than that one in T-1 and T-2 groups, respectively.

Increase in average daily weight gains by 33.5% in T-1 group and by 27.9% in T-2 group as compared to daily weight gains in control group was recorded during the first control weighing of the calves used for the trail (30 days after the trial start). During the second control weighing of the calves preformed 60 days after the trial start, 55.5–67.7% increase was recorded. Average daily weigh gain in T-1 group calves and in T-2 group calves was by 10.6% and 4.3% higher than that one in control group calves during the whole calf raising period (Table 6).

The test results show that administration of the biogenic stimulant of animal origin by injections and DIABAX feed additive (in T-2 group) with milk contribute to 100% survival rate in calves, significant increase in calcium and magnesium levels in the animal sera by 14.5–23.8% and 61.2–79.5%, respectively, as compared to the initial levels and levels in control group, an increase in albumin and α-globulin protein fractions by 10.1 and 43.2% (p ≤ 0.05), respectively, as compared to that ones in control group, increase in albumin/globulin ratio by 17.5%, color index – by 1.1% as compared to that ones in control group, the total immunoglobulin G amount was 2.7 times higher than the initial value. Daily administration of DIABAX feed additive to calves of T-1 group for 15 days reduced recurrent disease rate in the calves by 14.4%, resulted in significant increase in serum calcium and magnesium levels by 10.1 and 75.0% (p ≤ 0.05), respectively, as well as in increase in immunoglobulin G level by 2.3 times, erythrocyte level – by 3.8%, hemoglobin level – by 8.0%, leukocyte level – by 21.8%, albumin/globulin ratio – by 35.1% as compared to the initial values.

CONCLUSIONS

1. Daily administration of DIABAX feed additive to calves for 15 days contributed to significant increase in serum calcium, magnesium, albumin, β-globulin levels (p ≤ 0.05) in T-1 group calves, decrease in number of diseased animals by 14.4% and 100% survivability in T-2 group calves as compared to control group calves, increase in average daily weigh gain by 55.5% as compared to that ones in control group calves during 60-day observation period.
2. Combination of the biogenic stimulant injections to calves on day 1, 5, 10 of the trial and adding DIABAX feed additive to the calf diet for 15 consecutive days contributed to a significant increase in calcium, magnesium, albumin levels, α-, β-globulin protein fractions (p ≤ 0.05), decrease in disease rates by 40.0 and 29.9%, as well as an increase in average daily weight gain by 67.7 and 7.9% as compared to that ones in control and T-1 groups, respectively, during 60-day observation period with 100.0% survival rate in T-2 test group calves.

Contribution: Shanshin N. V. – collection of biological materials and preparation of pilot biogenic stimulant batch, collection of blood samples from the animals, morphological and biochemical tests, analysis of the obtained data, systematization and statistical processing, preparation of the paper for publication.

Вклад автора: Шаньшин Н. В. – лично автором произведен отбор биоматериала и изготовлена опытная партия биогенного препарата, осуществлен забор крови у животных, проведены морфо-биохимические исследования, анализ, систематизация и статистическая обработка полученных данных, подготовка статьи к публикации.