Research Article

 

Immunomodulatory and Therapeutic Prospective of a Protein Supplement with Vitamins and Selenium (Multimune) against Chicken Infectious Anaemia in Broiler Chicks

 

Gopal Krishan1, Santosh Kumar Shukla2, Prakash Bhatt3, Rajesh Kumar4*, Ruchi Tiwari5, Yashpal Singh Malik6, Kuldeep Dhama7

1Department of Animal Husbandry, Himachal Pradesh, India; 2Department of Veterinary Medicine; 3Veterinary Clinics; 4Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, Udham Singh Nagar, Uttarakhand - 263 145, India; 5Department of Veterinary Microbiology, College of Veterinary Sciences, Uttar Pradesh Pandit Deen Dayal Upadhayay Pashu Chikitsa Vigyan Vishwa Vidyalaya Evam Go-Anusandhan Sansthan (DUVASU), Mathura, Uttar Pradesh – 281001, India; 6Division of Biological Standardization; 7Avian Diseases Section, Division of Pathology, Indian Veterinary Research Institute (IVRI), Izatnagar, Uttar Pradesh – 243 122, India.

Editor | Lakshmi Tulasi Sunkara (D.V.M, M.V.Sc, Ph.D), Research Assistant Professor, Animal Science Department, Oklahoma State University, Stillwater, OK-74078, USA.

Special Issue | 3(2015) “Poultry diseases and the strategies for their control and prevention”

Received | February 12, 2015; Revised | April 15, 2015; Accepted | April 16, 2015; Published | April 30, 2015

*Correspondence | Rajesh Kumar, Govind Ballabh Pant University of Agriculture and Technology, Uttarakhand, India; Email: [email protected]

Citation | Krishan G, Shukla SK, Bhatt P, Kumar R, Tiwari R, Malik YS, Dhama K (2015). Immunomodulatory and therapeutic prospective of a protein supplement with vitamins and selenium (Multimune) against chicken infectious anaemia in broiler chicks. Adv. Anim. Vet. Sci. 3(3s): 1-8.

DOI | http://dx.doi.org/10.14737/journal.aavs/2015/3.3s.1.8

ISSN (Online) | 2307-8316; ISSN (Print) | 2309-3331

Copyright © 2015 Krishan et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

 

INTRODUCTION

 

Chicken infectious anemia virus (CIAV), belongs to the genus Gyrovirus under the family Circoviridae (Todd et al., 2000). It causes acute anemic condition (chicken infectious anaemia, CIA) in young chicks below 4-5 weeks of age. CIA is a worldwide problem and acclaimed as the most economically important viral diseases of poultry (McNulty, 1991; McIlroy et al., 1992; Bulow and Schat, 1997; Farkas et al., 1998; Rosenberger and Cloud, 1998; Dhama et al., 2008; Schat, 2009; Bhatt et al., 2011; Wani et al., 2013). The birds exhibit symptoms of depression, severe anaemia, paleness, weakness, anorexia, ruffled feathers, poor weight gain, aplasia of the bone marrow, lymphoid atrophy, subcutaneous and muscular haemorrhages with increased case fatality (McNulty, 1991; Bulow and Schat, 1997; Hagood et al., 2000). The prominent sign of anaemia, prevails during the end of 2nd weeks post infection (PI) and manifest mainly the non-feathered areas that may extend to internal organs (Pope, 1991). There is marked damage of haematopoietic and lymphopoietic tissues viz. stem cells in bone marrow and precursor T-lymphocytes in thymus. The bursa, spleen and other lymphoid organs are also depleted of lymphoid cells though less severely (Goryo et al., 1989; Dhama, 2002). Repopulation of the bone marrow with proerythroblasts and promyelocytes, and recovery of haematopoietic activity (erythropoiesis) and lymphocyte repopulation occur during convalescent phase (Taniguchi et al., 1982; Liu et al., 1997). CIAV is a potent immunosuppressive agent for very young unprotected chicks thereby increasing their susceptibility to secondary infections (Van Den Berg, 1996; Adair, 2000; De Herdt et al., 2001). The CIAV infection makes birds prone to several other diseases like haemorrhagic syndrome, haemorrhagic anaemia syndrome, infectious/aplastic anaemia, anaemia-dermatitis syndrome, gangrenous dermatitis and blue wing diseases (Pope, 1991; Hagood et al., 2000; Toro et al., 2000). CIAV infections have been associated with a number of vaccination failures, vaccination reactions or aggravation of the residual pathogenicity of attenuated vaccine viruses viz. Newcastle disease virus (NDV), Marek’s disease virus (MDV), infectious laryngotracheitis virus (ILTV) and fowl pox virus (FPV) (Box et al., 1988; Otaki et al., 1988; Rosenberger and Cloud, 1989; De Boer et al., 1994; Zheng and Liu, 1996; Liu et al., 2001; Dhama, 2002). The notable characteristics such as vertical transmission, highly contagious, hardy and ubiquitous nature, and the potential for inducing marked immunosuppression have attracted the global scientific community towards the potential threat of CIAV infection in poultry industry (Todd, 2000; Dhama et al., 2008; Basaraddi et al., 2013). Alike other viral infections there is no specific treatment option for CIA and vaccination strategies of live-attenuated and inactivated vaccines are available with some limitations, also advanced vaccines like DNA vaccine and others have been developed but need to be applied yet (McNulty, 1991; Schat, 2003; Dhama et al., 2008; Sawant et al., 2015; Zhang et al., 2015). In such a case there is a need to evaluate other effective therapeutic regimens to subside the massive immunosuppression produced by the CIA virus. As an alternative approach birds can be supplemented with haematinics and immunostimulants to arrest anaemic stage and enhance the immune system (Bhatt et al., 2013; Latheef et al., 2013). With this aim, the present study was carried out to assess the immunomodulatory and therapeutic potential of a protein supplement containing essential vitamins and selenium (Multimune) in CIAV infected chicks.

 

MATERIALS AND METHODS

 

Virus

The cell culture (MDCC-MSB1 cells) adapted chicken infectious anaemia virus (CIAV), maintained in the Avian Disease Section, Division of Pathology, Indian Veterinary Research Institute (IVRI), Izatnagar, UP (India) was used to infect the chicks in the present study.

 

Immunomodulator (Multimune)

Multimune (Interface Pharmaceuticals, New Delhi, India), a protein supplement containing essential vitamins and selenium, was used as an immunomodulator in the study.

 

Experimental Chicks

Sixty day-old age broiler chicks were procured from Instructional poultry farm of the university. Chicks were reared in deep litter system under hygienic conditions and given normal basal ration and water ad libitum. The experiment on chicks was performed following the recommendations and approval of the Institute Animal Ethics Committee (IAEC) under the guidelines set forth by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA). Prior to experiment all birds were checked for presence of CIAV by using IDEXX FlockChek CAV test kit (IDEXX Laboratories, USA) and were found free of infection.

 

Experimental Design

Sixty day-old broiler chicks were distributed in three equal groups (Group I-III). Chicks of all three groups were vaccinated against Newcastle disease (ND) by live attenuated ‘F’ strain (NDV-F) and Georgia strain of infectious bursal disease (IBD), respectively as per manufacturer’s instructions. Group I chicks served as healthy control while chicks of group II and III were inoculated with 1 ml of CIAV infected cell culture supernatant (104.5 TCID50/0.1 ml) per bird intramuscularly. Group II chicks served as virus positive control and group III chicks were supplemented with Multimune at the dose of 6.0 gm / 60 birds in drinking water for 21 days.

 

Sampling for haematology and biochemical parameters was based on pooled blood samples (5 ml) collected from 3 birds of each group weekly up to 35th day of experiment (i.e. 7th, 14th, 21st, 28th and 35th day). One part of aseptically collected blood was separated in heparinised vials (10-20 IU heparin / ml) for haematological estimations and another part for serum analysis (biochemical and immunological studies). The mean live body weight was studied in experimental chicks of all the groups at regular intervals at weekly intervals up to 35th day of experiment after scarification of birds. At weekly intervals, three birds from each group were sacrificed to calculate ratio of lymphoid organs (thymus, bursa of Fabricius, spleen and liver) to body weight.

 

Haematological Parameters

Haemoglobin (Hb), packed cell volume (PCV) (Jain, 1986), total erythrocyte count (TEC), total leukocyte count (TLC) (Natt and Herrick, 1952) and differential leukocyte count (DLC) (Lucas and Jamroz, 1961) were estimated with pooled blood samples (5 ml from 3 birds) from each group at weekly intervals up to 35th day (i.e. 7th, 14th, 21st, 28th and 35th) of experiment.

 

Biochemical Parameters

Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and uric acid (UA) were estimated with sera samples from 3 birds from each group at weekly intervals up to 35th day of experiment using reagent kits (IFCC Kinetic Method, Erba, Mannheim Ltd., Germany; Uricase PAP Method, Span Diagnostics Ltd., Surat, Gujarat, India).

 

Nd HI Titer

For determining the antibody titer against ND vaccination, Haemagglutination inhibition (HI) test was carried out on the sera samples obtained from 3 birds from each group on 7th, 14th, 21st, 28th and 35th day of experiment following method of Allan and Gough (1974). The mean HI titre (log2) for different experimental groups was calculated for comparison.

 

Mean Live Body Weight

To assess the immunomodulatory effect of ‘Multimune’ on growth performance of the CIAV infected chicks (group II and group III), the average live body weight of 5 chicks each from the three groups was recorded at weekly interval (day 0, 7th, 14th, 21st, 28th and 35th) of the study for comparison.

 

Organ: Body (O: B) Weight Ratio

To assess the protective effect of ‘Multimune’ on lymphoid organs of the virus infected birds, the Organ: body weight ratios of the thymus, bursa of Fabricius, spleen and liver were calculated from the 5 chicks each, from all three groups. Assessment was done on day 0, 7th, 14th, 21st, 28th and 35th of the study for comparison. This ratio was calculated by dividing respective organ weight with body weight (gm) and multiplying with 100.

 

Statistical Analysis

The statistical analysis was performed by analysis of variance ( two way ANOVA) (Snedecor and Cochran, 2004) by using SPSS (2007) version 16. All the values obtained in the study are represented as mean ± SE.

 

RESULTS AND DISCUSSION

 

Haematological Parameters

As shown in Table 1 a significant (P<0.05) decrease in the values of Hb, PCV and TEC was noticed in the chicks of CIAV infected group (group II) from 7th day PI where lowest values were seen at 21 day PI in comparison to the chicks of healthy group (group I). The normal value of Hb concentration of healthy group (116-121 g/L) was reduced to 52.67±0.29 in CIAV infected chicks on 21st day, and the improvement in Hb concentration was recorded in the immunomodulator treated (70.00±0.20 g/L) chicks (group III) on 21st day PI. The normal value of PCV was found to be 0.34-0.35 (v/v) in healthy chicks. CIAV infection was found to decrease normal PCV values on 7, 14, 21 and 28 DPI. Similar observations were recorded in the myeloid lineages with significant (P<0.05) decline in the TLC, percent lymphocyte count and percent eosinophil and basophil count (PE+BC) in group II birds from 7th day PI and lowest count was seen on 21 DPI. Immunomodulator treatment was found to significantly reduce the effect of CIAV induced reduction in PCV. Similar to the values of Hb and PCV, TEC also showed a significant reduction in CIAV challenged groups with maximum reduction on 21st day (1.52×1012/L), whereas the Multimune supplemented group showed significantly (P<0.05) improved values of TEC (2.09×1012/L) on the 21st day. The TLC value of healthy group was found to be 20-22×109/L, and CIAV infection was found to be reduced on days 7 (18.47±0.40), 14 (17.00±0.20), 21 (14.67±0.57), 28 (17.33±0.29) and 35 (17.97±0.12) PI, respectively. But immunomodulator group showed significantly (P<0.05) better values of TLC on day 7 (19.23±0.14), 14 (19.03±0.05), 21 (17.60±0.35), 28 (19.00±0.29) and 35 (19.43±0.27) PI, respectively. Similarly, the mean PLC and PE+BC values were significantly decreased while the PHC values showed an increasing trend in all the CIAV inoculated chicks. Our results are in agreement with earlier reports (Dhama, 2002; Schat, 2003; Bhatt et al., 2013). It is hypothesized that this decrease in the Hb, PCV, TEC and TLC levels in CIAV infected chicks might be due to destructive effect of CIAV on erythroid and myeloid tissues of bone marrow where it causes suppression of differentiation and proliferation of haemopoietic precursor cells, affecting erythropoiesis and myelopoiesis leading to anaemia and panleukopenia (Pope, 1991; Schat, 2003; Dhama et al., 2008; Bhatt et al., 2013).

 

The levels of haematological parameters in comparison to group II (virus positive control) were significantly higher (P<0.05) in chicks of group III which were supplemented with Multimune for 21 days in drinking water. The reason for this could be that the protein and other ingredients of ‘Multimmune’ which possess immunostimulant properties and thus limited the progression of panleukopenia and immunosuppression during CIAV infection.

 

Biochemical Parameters

The biochemical analysis results of the chicks of all the three groups are summarized in Table 1. Assessment of activities of ALT, AST and ALP enzymes, and uric acid was significantly higher (P<0.05) in the CIAV inoculated birds (group II), which could be associated with the damage to the organs like liver, kidneys and muscles due to CIAV infection (McNulty, 1991; Dhama et al., 2008). AST, ALT and ALP are found in the liver and additionally ALP is also found in the kidney (Kaneko et al., 1997). CIAV infections are known to result in damage and focal necrosis of liver, kidney and spleen (McNulty, 1991; Dhama et al., 2008). The AST value of healthy group was found to be 172-175 IU/L, and CIAV infection was found to elevate it on 7th (228.10±5.13), 14th (291.83±3.77), 21st (322.00±4.44), 28th (296.50±2.84) and 35th (190.87±4.12) days post infection (DPI), respectively. But immunomodulator (Multimune) group showed significantly (P<0.05) improved values of AST on 7th (186.77±2.13), 14th (204.77±3.25), 21st (221.43±2.19), 28th (206.17±2.05) and 35th (172.83±1.92) DPI, respectively. The normal value of ALT (33-34 IU/L) in healthy chicks was increased to 71.06 IU/L in CIAV infected chicks, and the improvement was found in Multimune treated group (46.97 IU/L) on 21st DPI. Similar to above mentioned enzymes, ALP value was also found to be significantly (P<0.05) higher in group II (606.83 IU/L) as compared to the normal healthy group (489.40 IU/L) and immunomodulator supplemented group (534.67 IU/L) on the day of peak infection. The normal value of uric acid (UA) was found to be 5-6 mg/dl in healthy chicks. CIAV inoculation was found to increase the uric acid values on 7th (7.08 mg/dl), 14th (7.91 mg/dl), 21st (8.98 mg/dl) and 28th (8.37 mg/dl) DPI. Immunomodulator treatment was found to significantly reduce the effect of CIAV induced increase in the value of uric acid.

 

 

Table 1: Haematological and biochemical parameters of healthy, CIAV challenged and ‘Multimune’ treated groups of chicks (Mean±SE)

Parameters	Groups	Days of observation
		7th day	14th day	21st day	28th day	35th day
Hb (g/L)	I	116.67±0.18A	118.00±0.12A	120.67±0.07A	121.33±0.07A	118.67±0.18A
	II	100.33±0.30Ca	86.33±0.12Cb	52.67±0.29Cc	88.33±0.18Cb	102.67±0.29Ba
	III	103.67±0.24Ba	94.67±0.47Bb	70.00±0.20Bc	103.33±0.48Ba	104.67±0.30Ba
PCV (L/L)	I	0.34±0.01A	0.34±0.01A	0.35±0.01A	0.35±0.01A	0.34±0.01A
	II	0.28±0.00Ca	0.26±0.00Cb	0.18±0.01Cc	0.25±0.00Cb	0.29±0.01Ba
	III	0.31±0.00Ba	0.29±0.01Ba	0.26±0.00Bb	0.29±0.00Ba	0.31±0.01Ba
TEC (x1012/L)	I	3.29±0.06A	3.37±0.02A	3.39±0.01A	3.40±0.01A	3.39±0.12A
	II	2.97±0.12Ba	2.43±0.06Bb	1.52±0.07Cc	2.60±0.03Cb	2.99±0.10Ba
	III	3.00±.012Ba	2.49±0.13Bb	2.09±0.09Bc	3.08±0.12Ba	3.18±0.12Ba
TLC (x109/L)	I	20.60±0.42A	21.20±0.70A	21.53±0.37A	21.50±0.32A	21.50±0.32A
	II	18.47±0.40Ba	17.00±0.20Cb	14.67±0.57Cc	17.33±0.29Cab	17.97±0.12Bab
	III	19.23±0.14Ba	19.03±0.35Ba	17.60±0.35Bb	19.00±0.29Ba	19.43±0.27Ba
PLC (%)	I	60.67±0.58A	59.00±0.58A	59.34±0.67A	59.00±0.58A	59.00±0.58A
	II	52.67±0.58Ba	49.67±0.88Cb	48.33±0.33Cb	49.67±0.88Cb	50.00±0.58Cb
	III	53.00±0.58Ba	53.67±1.20Ba	53.67±1.20Ba	54.67±0.88Ba	55.67±2.18Ba
PHC (%)	I	33.00±0.58B	34.67±0.88B	32.67±0.34B	33.00±1.00B	32.67±0.88B
	II	40.00±0.58Ac	43.33±0.33Ab	47.33±0.33Aa	44.33±0.88Ab	43.67±0.67Ab
	III	39.00±0.58Aa	39.00±0.58Ba	39.67±1.20Ba	38.67±0.67Ba	38.00±1.00Ba
PMC (%)	I	3.67±0.34	4.00±0.58	4.67±0.33A	4.33±0.34A	4.67±0.58A
	II	4.33±0.34	4.33±0.88	2.67±0.33B	3.67±0.33B	3.67±0.58A
	III	4.67±0.34	4.67±1.20	4.00±1.20AB	5.00±0.00A	4.67±0.58A
P (B+E) (%)	I	2.67±0.33	2.33±0.33	3.67±0.33A	3.67±0.33A	3.67±0.33
	II	3.00±0.58	2.67±0.33	1.67±0.33B	2.33±0.33B	2.67±0.33
	III	3.33±0.33	3.33±0.33	3.33±0.33A	3.33±0.33AB	3.33±0.33
AST (IU/L)	I	172.50±3.00C	174.80±2.30C	174.33±1.96C	174.33±1.42C	175.30±1.85B
	II	228.10±5.13Ac	291.83±3.77Ab	322.00±4.44Aa	296.50±2.84Ab	190.87±4.12Ad
	III	186.77±2.13Bc	204.77±3.25Bb	221.43±2.19Ba	206.17±2.05Bb	172.83±1.92Bd
ALT (IU/L)	I	33.35±0.43C	33.51±0.34C	33.58±0.31C	33.62±0.32C	33.50±0.33B
	II	47.64±0.63Ac	60.03±0.66Ab	71.06±1.08Aa	59.47±0.32Ab	36.23±0.62Ad
	III	38.07±0.42Bc	42.89±0.31Bb	46.97±0.88Ba	43.56±0.53Bb	33.63±0.27Bd
ALP (IU/L)	I	485.33±4.15C	487.00±3.33C	489.40±2.46C	492.23±1.52C	491.07±2.69B
	II	527.33±4.87Ac	571.83±3.65Ab	606.83±3.17Aa	561.50±2.77Ab	502.33±1.30Ad
	III	503.73±2.15Bc	515.00±1.32Bb	534.67±1.32Ba	523.67±3.17Bb	490.17±3.21Bd
UA (mg/dl)	I	5.89±0.13C	5.92±0.06C	5.94±0.08C	5.93±0.03C	6.01±0.04B
	II	7.08±0.06Ad	7.91±0.09Ac	8.98±0.06Aa	8.37±0.06Ab	6.29±0.05Ae
	III	6.23±0.04Bd	6.74±0.06Bb	7.18±0.03Ba	6.52±0.03Bc	6.09±0.03Be

 

The values (Mean±SE) having atleast one common superscript (Capital letters in columns and small letters in rows) does not differ significantly (P<0.05) for a parameter.

 

 

Table 2: ND HI titre (log2) in healthy, CIAV challenged and ‘Multimune’ treated groups of chicks (Mean±SE)

Groups	Days of observation
	7th day	14th day	21st day	28th day	35th day
I	1.67±0.33Ad	9.67±0.33Aa	8.67±0.33Aab	7.67±0.33Abc	6.67±0.33Ac
II	1.00±0.58Ad	6.33±0.33Ca	5.00±0.58Bab	4.67±0.33Bbc	3.33±0.33Bc
III	1.33±0.33Ac	8.00±0.58Ba	7.67±0.33Aa	7.00±0.00Aab	6.00±0.58Ab

 

The values (Mean±SE) having atleast one common superscript (Capital letters in columns and small letters in rows) does not differ significantly (P<0.05)

 

 

Table 3: Mean live body weight (g) of healthy, CIAV challenged and ‘Multimune’ treated groups of chicks (Mean±SE)

Groups	Days of observation
	0 day	7th day	14th day	21st day	28th day	35th day
I	39.80±0.51Ae	84.40±4.48Ae	153.20±6.51Ad	256.32±9.77Ac	366.75±9.01Ab	730.00±19.64Aa
II	39.70±0.51Ae	66.85±2.13Ce	121.20±4.97Cd	194.38±8.75Cc	252.50±5.32Cb	443.00±18.83Ca
III	39.50±0.75Ae	72.90±3.68Be	133.20±4.68Bd	223.65±8.86Bc	324.50±7.32Bb	578.00±17.56Ba

 

The values (Mean±SE) having atleast one common superscript (Capital letters in columns and small letters in rows) does not differ significantly (P<0.05)

 

 

Table 4: Organ: body weight ratios of all lymphoid organs in healthy, CIAV challenged and ‘Multimune’ treated groups of chicks (Mean±SE)

Organs	Groups	Days of observation
		7th day	14th day	21st day	28th day	35th day
Thymus	I	0.21±0.01A	0.21±0.01A	0.20±0.00A	0.20±0.00A	0.19±0.01A
	II	0.13±0.01Ba	0.10±0.00Bb	0.09±0.00Cb	0.09±0.01Cb	0.11±0.01Cb
	III	0.14±0.01Bab	0.12±0.01Bc	0.13±0.00Bbc	0.13±0.01Bbc	0.15±0.02Ba
Bursa	I	0.18±0.01A	0.17±0.01A	0.17±0.01A	0.18±0.00A	0.18±0.00A
	II	0.12±0.00Bb	0.10±0.00Bc	0.09±0.01Bc	0.10±0.01Bc	0.14±0.01Ba
	III	0.14±0.00Bb	0.13±0.01Bb	0.14±0.01Ab	0.16±0.00Aa	0.17±0.00Aa
Spleen	I	0.18±0.00A	0.18±0.01A	0.18±0.01A	0.17±0.00A	0.19±0.00A
	II	0.12±0.00Ba	0.11±0.01Ba	0.11±0.01Ba	0.12±0.01Ca	0.12±0.01Ba
	III	0.13±0.00Bb	0.13±0.00Bb	0.13±0.01Bb	0.13±0.00BCb	0.14±0.00Ba
Liver	I	3.66±0.03B	3.64±0.07B	3.63±0.07C	3.68±0.06B	3.69±0.06B
	II	3.92±0.03Ac	4.13±0.04Ab	5.02±0.10Aa	4.28±0.03Ab	3.90±0.04Ac
	III	3.68±0.02Bc	3.88±0.03Bb	4.04±0.03Ba	3.99±0.01Ba	3.60±0.03Bc

 

The values (Mean±SE) having at least one common superscript (Capital letters in columns and small letters in rows) does not differ significantly (P<0.05) for a parameter

 

ND Antibody Titre (HI Test)

The humoral immune response (HIR) to ND vaccination in chicks of the three groups of birds was measured by Haemagglutination inhibition (HI) test at weekly intervals, i.e. day 7, 14, 21, 28 and 35. The mean HI antibody titers in chicks of all three groups obtained at different days post vaccination, expressed as HI titre (log2) are presented in Table 2. A significant decrease (P<0.05) in the mean HI ND antibody titre was seen on 14th, 21st, 28th and 35th days post vaccination in chicks of group II challenged with CIAV, while statistically no significant difference was observed in the mean HI antibody titres in all chicks on 7th day of the experiment. The group III chicks supplemented with ‘Multimune’ showed significantly higher values of HI titres as compared to group II CIAV challenged birds only which might be due to immunomodulatory potential of protein and immunoglobulins present in the immunomodulator used. This effect could be due to suppression of the population of both helper (CD4+) and cytotoxic (CD8+) T-Lymphocytes in the thymus due to CIAV infection, as suggested by Hu et al. (1993) and Adair (2000). Herbal and protein supplementation has been reported to have ameliorative effects against immunosuppression induced by CIAV (Bhatt et al., 2013; Latheef et al., 2013).

 

Mean Live Body Weight

The observations of mean body weights recorded in all three groups of chicks are presented in Table 3. The healthy group I chicks showed highest weight gain in comparison to CIAV infected groups II and III. The findings show that chicks of CIAV affected group II revealed minimum weight gain, while the immunomodulator supplemented group III showed significantly (P<0.05) higher weight gains from group II chicks on all the experimental days, which may be due to growth stimulation with the immunomodulator used. The mean live body weights of all the groups on 21st day were in the range of 194.38±8.75 and 256.32±9.77 g and on 35th day were in the range of 443.00±18.83 and 730.00±19.64 g. The healthy group showed maximum weight gain as compared to CIAV infected group, whereas group III showed significantly (P<0.05) higher weight gain than group II. The mean live body weight of the chicks of group II challenged with CIAV showed a significant (P<0.05) decline as compared to the control group I which is in accordance with the findings of Dhama (2002).

 

Organ: Body (O: B) Weight Ratios

The mean percent ratios of the lymphoid organs (thymus, bursa, spleen and liver) determined in chicks of all 3 groups are summarized in Table 4. The group II chicks revealed a significant (P<0.05) decline in their mean percent ratios of thymus, bursa and spleen to body weight on 7th, 14th, 21st, 28th and 35th day PI, with the maximum decline on day 21. The CIAV challenged group showed a significant (P<0.05) elevation in their mean percent ratio of liver to body weight as compared to control group I on all the days intervals studied. These observations are in accordance with the experimental CIAV infection studies as documented by Dhama (2002) in day old chicks. The ‘Multimune’ supplemented group III of chicks showed minimal alterations in mean percent ratios of lymphoid organs as compared to group II chicks, and had significantly (P<0.05) improved ratios for lymphoid organs. These findings of the organ: body weight ratios in the present investigation are in accordance that CIAV causes atrophy of the thymus, bursa and spleen, and increases the size of liver (Goryo et al., 1987; Dhama, 2002; Sommer and Cardona, 2003). Dhama (2002) and Vachhani (2005) also reported similar findings in accordance with the present observations. The ‘Multimune’ supplemented group III of chicks exhibited minimal alterations in mean percent ratios of lymphoid organs as compared to CIAV inoculated group II chicks which can be attributed to the presence of proteins in the immunomodulatory agent used.

 

Immunomodulatory, growth performance enhancing and protective effects of the components of ‘Multimune’ viz. proteins like lactalbumins, betaglobulins, serum albumin, immunoglobulins and lactoferrins, vitamins (A, D3, E, C, B) and selenium are well known ingredients which enhance the immunity and act as immunomodulator in livestock and poultry (Sahin et al., 2001; Konjufca et al., 2004; Wintergerst et al., 2007; Baker, 2009; Peric et al., 2009; Saad et al., 2009; Dhama et al., 2014a, 2014b). Therefore, ‘Multimune’ was selected to explore its immunomodulatory and ameliorative potential in CIA affected birds. CIAV infection suppresses the immunity and reduces the size of lymphoid organs in young birds with stunted growth performance, and in this situation the results of the present trial indicate that ‘Multimune’ could serve as a useful immunomodulatory and treatment formulation to ameliorate CIAV infection in broiler chicks.

 

CONCLUSION

 

Overall, our results revealed the immunomodulatory effects of “Multimune” during immunosuppressive condition (CIA) reducing the pathogenicity of CIAV and up surging the depressed immune response. Furthermore, there is need to assess the impact of “Multimune” supplementation on growth performance of the virus affected birds. Adoption of such alternative approach along with use of available effective vaccines is the most attractive way of minimizing economic losses being suffered by poultry producers due to CIAV infection. In view of the preliminary findings, we propose that supplementation of immunomodulators could be adopted to minimize the pathological effects and immunosuppressive potential of CIAV and other similar diseases in young chicks.

 

ACKNOWLEDGEMENTS

 

Authors acknowledge the Director, Experiment Station, Dean, College of Veterinary and Animal Sciences, and Dean, Post Graduate Studies, G. B. Pant University of Agriculture and Technology, Pantnagar, Udham Singh Nagar, Uttarakhand and Director, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, for providing basic services to complete the present work. This research work is partially funded by a research grant of Directorate of Experiment Station, G. B. Pant University of Agriculture and Technology, Pantnagar.

 

CONFLICT OF INTEREST

 

Authors declare no conflict of interest.

 

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