Obuhovska O.V., Rudenko O.P., Candidate of Veterinary Science

Obuhovska O.V., Rudenko O.P., Candidate of Veterinary Science,

Matyusha L.V.,Junior Researcher,

PopovaO.M., veterinary doctor

National science centre “Institute of experimental and clinical veterinary medicine”

PROTEINFRACTIONS OF BLOOD SERUM INCHICKENS, VACCINATEDWITH INACTIVATED VACCINES AGANST RESPIRATORY MYCOPLASMOSIS OF POULTRY

Reviewer – Candidate of Veterinary Science Glebova K.V.

It is proved that the double intramuscular introduction of inactivated vaccines against respiratory mycoplasmosis of poultry will help to increase albumin and γ-globulin in the blood serum of chickens, indicating activation of the immune system.The use of a vaccine based on inactivated bacterinum increases to 47.0% albumin level and γ-globulin to 91.9% on the 21-st day after the second injection. Using by a similar scheme of subunit vaccine based on the unintegrated bacmass increases by 39.6% albumins loevel and γ-globulinsby 84.2%, respectively.

Keywords:respiratory mycoplasmosis of poultry, inactivated vaccines, albumin, γ-globulins.

Statement of the problem. Development of domestic vaccine preparations for the prevention of respiratory mycoplasmosis of poultry is actual direction of research. One of the most important indicators of the vaccine effect on the body of poultry is a change of the level of blood protein fractions during certain times after immunization. Significant increase in albumins and γ-globulins suggests activation of the immune system and confirms the effectiveness of the vaccine preparation. Therefore, the definition of these parameters, along with the level of protective antibodies and the definition of projective protection in the direct infection of immunized birds, is considered to be a necessarystage in the study of new vaccines [1, 2].

Analysis of the main research and publications which discuss the problem.The effectiveness and expediency of applicationof vaccination for the prevention of respiratory mycoplasmosis of poultryis undisputed [5, 6, 7, 9]. It is shown that economic costs on vaccination are minor compared to the losses that are caused by an outbreak of the disease [8]. Inactivated vaccines are widely used and are considered more safe and effective than the living ones [4, 10, 11, 12]. We have produced two series of inactivated vaccines against respiratory mycoplasmosis of poultry and conducted experiments to study the efficiency of their use in the experiments on chickens. It has been proved that they provide 100% protection of birds from the clinical manifestations of the disease and 95% of birds from the infection of strain-punch [3].

Aims and objectives. The aim was to determine the nature of the impacts of two inactivated vaccines against respiratory mycoplasmosison the body of birds and comparative analysis of the effectiveness of their application.

To achieve this, it was determined a goal – to study the dynamics of changes in albumin and γ-globulin in the blood serum of chickens after a double intramuscular administration of vaccines in comparison with the corresponding figures in unvaccinated birds.

Materials and methods. Experimental series of inactivated vaccines against respiratory mycoplasmosis were produced by two methods. In the first series as antigenic basis the inactivated bacterin of industrial strain of Mycoplasma gallisepticum S6 (VB)was used.In the second series as antigenic basis the disintegrated bacterial cell mass of production strain Mycoplasmagallisepticum S6 (VS) was used.To standardized inactivated antigen bases adjuvant at a rate of 30% antigenic basis (3x107 CFU) and 70% adjuvant (Mantanide ISA 70 VG) wereadded.The manufactured vaccines were tested for sterility (by ISO 4483) and harmless (by ISO 46.024).

Experiments were conducted on 3 groups of chickens. The first experimental group (n = 30) were immunized intramuscularly twice (at the age of 30 and 60 days) by inactivated vaccine against respiratory mycoplasmosison the basis of bacterin (VB). The second experimental group (n = 30) were immunized intramuscularly twice (at the age of 30 and 60 days) by inactivated subunit vaccine based on thedisintegrated bacmass of strain Mycoplasma gallisepticum S6 (VS). The control group (n = 30) were not subjected to immunization.

From birds of all groups blood samples were taken the day before vaccination and on the 7th and 14th day after the first administration of vaccines, and on the 7th, 14th and 21st day after the second injection. In the blood serum albumin level was determined by reaction with bromocresol green and γ-globulin by Turbodimetric method. All results were statistically calculated (program SPASS Statistics 17.0).

Studies. Analysis of the results of studying of the change of values of protein fractions of blood serum of hens of the research groups showed that the introduction of both vaccine preparations promoted a significant increase in albumin and γ-globulins. It should be noted that the values of these parameters and timing of the detected changes were somewhat different for the two experimental groups. These changes are shown in Fig. 1, 2.

As can be seen from Figure 1 the level of albumins in blood serum of chickens of both research groups gradually increased from the 7th day after the 1st vaccination. So, in group of VB we have found the increase of this figure by 14% compared to the control at the 14th day after the 1st injection. On the 7th day after the 2nd input it remained almost at the same level (21.97 g/l), but then gradually increased and reached its maximum value (28.33 g/l) on the 21st day after the second injection, which is 47% higher than the rate of control group.

In the group of VS similar changeswere observed. Albumins levels increased after the 1st vaccination (21.85 g/l on the 14th day). Then its slight increase to 22.29 g/l on day 7 after the 2nd injectionwas registered. Further improvement showed almost 33% compared to the control, the maximum level of albumins in this group was 26.97 g/l (21 st day), which was by 39.6% higher than in the control group, but 5% below than in group VB.

/ Remarks:
1 – the introduction of vaccines;
2 – 7th day after the first injection;
3 – 14th day after the first injection;
4 – 7th day after the second infusion;
5 – 14th day after the second infusion;
6 – 21st day after the second administration

Fig. 1. Dynamics of changes in the level of albumins in blood serum of chickens of experimental and control groups

Fig. 2. Dynamics of changes in γ-globulins in the blood serum of hens of experimental and control groups

Of all the globulin fractions of blood serum in chickens most important for the assessment of the intensity of the immune response is γ-globulins, because in the birds this fraction consists of IgG, IgA and IgM. In the blood serum of chickens of research groups an increase in levels of these proteins after the 1st and the 2nd vaccines (Fig. 2)was observed.In group IB after the 1st vaccine on the 7th and the 14th day they were found in the amount of 7.84 g/l and 9.75 g/l, 20.6% and 50.0% higher than in control. After the 2nd input –in amounts 9.77 g/l on the 7th day and 12.57 g/l on the 21st day, which is almost twice higher than those in the control group.

In the group of VS increase in this protein fraction was less intense. On the 7th and 14th day after the 1st input – 7.24 g/l and 9.46 g/l. After the 2nd input of 9.64 g/l on the 7th day to 12.07 g/l on the 21st day, which is marginally (4%) lower than in group VB.

Maximum values of γ-globulins in the blood serum of hens of research groups exceeded similar to the control group at 91.9% (VB) and 84.3% (VS) respectively.

Conclusions:

1. Twice intramuscular introduction of inactivated vaccines against respiratory mycoplasmosis of poultry contributes to the level of albumin and γ-globulin in the blood serum of hens, indicating activation of the immune system. However, according to the analysis of biochemical parameters of blood serum of hens, the more appropriate is the use of vaccines based on the inactivated bacterinum.

2. The use of vaccines based on inactivated bacterin (VB) increases albumins level to47.0%and γ-globulins to 91.9% in the 21st day after the second injection.

3. The use of vaccines by the same scheme of subunit vaccine based on the disintegrated bacmass (VS) increases albumins level to 39.6% and γ-globulins to 84.2% on the 21st day after the second injection.

REFERENCES:

Белковый спектр крови цыплят-бройлеров при добавлении в рацион лизина сульфата/ С.Д.Чернявских, Ж.А.Боролаева, Н.А.Мусиенко, И.Н.Яковлєва// Научн. Ведомости Белгородск.госуд.ун-та. Серия: Естественные науки.–2012.–Т.19; №9.– С.156-158.
БолотниковИ.А. Практическая иммунология сельскохозяйственной птицы/ И.А.Болотников, Ю.В.Конопатов.–СПб.: Наука, 1993.- 204 с.
ОбуховськаО.В. Визначення рівня протективного захисту інактивованої вакцини проти респіраторного мікоплазмозу птиці в досліді на курчатах/ О.В. Обухівська// Вісник полтавської державної аграрної академії.-Полтава, 2013.– № 1.–С.103-106.
CrespoR. Facial cellulitis induced in chickens by Mycoplasma gallisepticum bacterin and its treatment/ R. Crespo, R. McMillan//Avian Dis.– 2008.– 52(4).–P.698-701.
Development and immunogenicity of recombinant GapA(+) Mycoplasma gallisepticumvaccine strain ts-11 expressing infectious bronchitis virus-S1 glycoprotein and chicken interleukin-6 / P.K. Shil, A. Kanci, G.F. Browning, P.F. Markham// Vaccine.–2011.–29(17).–P.3197-3205.
Development and immunogenicity of recombinant Mycoplasma gallisepticumvaccine strain ts-11 expressing chicken IFN-gamma/ Y.Muneta [et al.]// Vaccine.–2008.–26(43).–P.5449-5454.
Effects of an S6 strain of Mycoplasma gallisepticum challenge before beginning of lay on various egg characteristics in commercial layers/ T.A.Parker [et al.]//Avian Dis.–2002.–Vol.46, № 3.–P.593-597.
HalvorsonD.A. Biosecurity on a multiple-age egg production complex: a 15-year experience/ D.A. Halvorson //Avian Dis.– 2011.– 55(1).–P.139-142.

9.KlevenS.H. Control of avian mycoplasma infections in commercial poultry/ S.H.Kleven// Avian Dis.–2008.–52(3).–P.367-374.

10.OlanrewajuH.A. Effects of single and combined Mycoplasma gallisepticum vaccinations on blood electrolytes and acid-base balance in commercial egg-laying hens/ H.A.Olanrewaju, S.D. Collier, S.L. Branton//Poult. Sci.– 2011.–90(2).–P.358-363.

Protective immune response of Mycoplasma gallisepticumvaccines in poultry/ Ael-D. Hussein [et al.]// Egypt J. Immunol.– 2007. – 14(2).– P.93-99.

12.The efficacy of three commercial Mycoplasma gallisepticumvaccines in laying hens [text]/ N. Ferguson-Noel, K. Cookson, V.A. Labinis, S.H. Kleven// Avian Dis.– 2012.– 56(2).–P.272-275.