- Email: [email protected]
Effect of Environmental Temperature on Immune Response of Broilers
2003 Poultry Science Association, Inc.
Effect of Environmental Temperature on Immune Response of Broilers E. Santin,1,2 A. Maiorka,2 W. J. C. Polveiro,3 A. C. Paulillo,3 A. C. Laurentiz, S. A. Borges, and A. V. Fischer da Silva4 Departamento de Patologia Veterina´ria Faculdade de Cieˆncias Agra´rias e Veterina´rias, Universidade Estadual Paulista, Jaboticabal, SP, Brazil
Primary Audience: Veterinarians, Flock Supervisors, Researchers
Several studies demonstrate that environmental temperature can influence the immune response of poultry. The objective of this research was to determine at which stage in the life of a bird this effect is greatest. In experiment 1, broiler breeder eggs were incubated at three different temperatures (36.8 ± 0.2, 37.8 ± 0.2, and 38.8 ± 0.2°C) from the 13th day of incubation to hatching. After hatching, birds were raised in thermoneutral temperature. In experiment 2, 144 1-d-old broiler chicks were distributed into three environmental chambers with different temperatures (18 ± 2, 24 ± 2, and 32 ± 2°C). In both experiments, the humoral immune responses to Newcastle disease virus (NDV) and infectious bursal disease (IBDV) were evaluated. NDV and IBDV antibody titers were not significantly different (P > 0.05) among treatments. Key words: broiler, temperature, humoral immunity 2003 J. Appl. Poult. Res. 12:247–250
DESCRIPTION OF PROBLEM Several studies have shown that the environmental temperature may influence the immune response of poultry [1, 2, 3]. The mechanism by which the environmental temperature may act as an immune suppressor is not fully understood. However, it is speculated that the increased activity of the adrenal gland due to stress increases the level of serum corticosteroids, which cause suppression of cell proliferation factor, or interleukin II [4]. The intensity and the duration of the stress induced by temperature may impair the immune system of poultry [1]; however, the effect of the environmental temperature depends on the degree of 1
habituation of the bird at the time of immunization. Arjona et al. [5] observed that environmental temperature stress early in life may induce resistance to weather variation in adult animals. The current study evaluated the effect of high or low environmental temperature before or after hatching on the humoral immune response of broilers vaccinated against Newcastle disease (NDV) and infectious bursal disease (IBDV).
MATERIAL AND METHODS Experiment 1 Fertilized eggs (Cobb commercial line) from broiler breeders immunized against NDV
To whom correspondence should be addressed: [email protected]. FAPESP fellowship. 3 CNPq fellowship. 4 Universidade Federal do Parana´ (UFPR). 2
Downloaded from http://japr.oxfordjournals.org/ by guest on April 13, 2015
SUMMARY
JAPR: Research Report
248
TABLE 1. Geometric mean titers against IBDVA and NDVB (expressed as log10) of broilers at 14, 35, and 42 d of age exposed to temperature changes during incubation, experiment 1 Geometric mean titers (log10) IBDV
NDV
Treatment
14 d
35 d
42 d
14 d
35 d
42 d
Control (37.8 ± 0.2°C) Hot (38.8 ± 0.2°C) Cold (36.8 ± 0.2°C)
2.81 2.79 2.68
3.51 3.62 3.55
3.66 3.81 3.84
1.58 1.54 1.54
2.10 2.07 2.33
2.03 1.84 1.84
Probability
0.791
0.656
0.206
0.956
0.262
0.528
A
Infectious bursal disease. Newcastle disease virus.
B
by centrifugation at 1,000 × g for 15 min and then inactivated at 56°C for 30 min and stored at −20°C until used. Hemagglutination inhibition reaction (HI) with live antigens, using the β-method [8] was used to measure NDV HI antibodies. An ELISA commercial kit [9] was used to measure IBDV antibodies. The antibody titers obtained in both serological tests were transformed in log10. A completely random experimental design, with three treatments (heat stress, cold stress, and control) and four repetitions, was used. Data were submitted to analysis of variance [10]. Experiment 2 One hundred forty-five, 1-d-old (Cobb commercial line) male broiler chicks from immunized breeders were used. Birds were distributed into three environmental chambers with different temperatures. Until 10 d of age, the temperature in the three chambers was similar (kept around 32 ± 2°C). From 10 d of age until the end of the trial, birds in chamber 1 were subjected to an environmental temperature of 18 ± 2°C (cold stress), birds in chamber 2 were subjected to an environmental temperature of 32 ± 2°C (heat stress), and birds in chamber 3 were subjected to an environmental temperature of 24 ± 2°C (control). Birds were fed a balanced feed based on corn and soybean meal [6], compatible with the different stages of their life cycles. In general, birds were subjected to conditions similar to the management practiced in the commercial broiler industry from 1 to 35 d of age. Vaccination, blood collection, serological analysis, and statistical analysis of data procedures were the same as those used in experiment 1.
Downloaded from http://japr.oxfordjournals.org/ by guest on April 13, 2015
and IBDV were used. Eggs were incubated in three different automatic incubators at 37.8 ± 0.2°C until d 13. From d 14 of incubation until hatching, eggs in incubator 1 were subjected to an incubation temperature of 36.8 ± 0.2°C (cold stress), eggs in incubator 2 were subjected to an incubation temperature of 38.8 ± 0.2°C (heat stress), and eggs in incubator 3 remained at 37.8 ± 0.2°C (control). At hatching, chicks were sexed, and the males were housed in an environmentally controlled poultry house and according to the incubation temperature treatment (heat stress, cold stress or control), with four replicates of 12 birds each. Birds were fed a balanced feed based on corn and soybean meal [6], compatible with the different stages of their life cycles. In general, birds were subjected to similar conditions of management practiced in the commercial broiler industry from 1 to 35 d of age and were kept under thermoneutral temperature (33 ± 2°C up to 7 d of age and 24 ± 2°C until the end of the experiment). Chickens in all experimental groups were vaccinated by eye drop at 7 and 20 d of age with vaccines from the same laboratory and from the same batch, at the beginning of the shelf life. The lyophilized vaccines were prepared with NDV lentogenic strain LaSota (EID50 = 107.20/ 0.1 mL) and with IBDV strain S706 (EID50 = 103.50/0.1 mL). Lyophilized vaccines were diluted with distilled water at 30 mL/1,000 vaccine doses per 1,000 birds, corresponding to an eye drop vaccine dose of 0.03 mL [7]. Blood samples from eight birds per experimental group were collected at 14 and 35 d of age by ulnar vein puncture. Serum was separated
TEMPERATURE AND IMMUNE RESPONSE TABLE 2. Geometric mean titers against IBDVA and NDVB (expressed as log10) of broilers at 14 and 35 d of age exposed to temperature changes after hatching, experiment 2 Geometric mean titers (log10) IBDV
NDV
Treatment
14 d
35 d
14 d
35 d
Control (24 ± 0.2°C) Hot (32 ± 0.2°C) Cold (18 ± 0.2°C)
3.35 3.29 3.22
3.70 3.61 3.72
1.24 1.24 1.22
1.74 1.50 1.71
Probability
0.082
0.503
0.585
0.128
A
Infectious bursal disease. Newcastle disease virus.
B
Results of experiment 1 are shown in Table 1. Antibody titers against NDV and IBDV were not statistically different (P > 0.05) among treatments. In experiment 2 (Table 2), no significant differences (P > 0.05) were observed among treatments; however, IBDV antibody titers of 14-d-old birds were numerically higher (P = 0.08) than the titers of birds subjected to heat (32 ± 2°C) or cold (18 ± 2°C) stress. The incubation period is very important to the development of the immune system of poultry [11]. However, in the present experiment, changes in the incubation temperatures did not influence the humoral immune response. Although not fully elucidated, the mechanism by which changes in temperature affect the immune response is related to the increase of serum corticosteroids [4]. Therefore, it could be speculated that the humoral immune response of the broilers in these experiments was not affected because the environmental changes were not imposed when the birds were being immu-
nized. According to Henken et al. [1], environmental changes at immunization may be more important than temperature variation during the whole life of the bird. In experiment 2, there were no significant differences (P > 0.05) in serum antibody titers against NDV and IBDV after immunization. Similar results have been found in other studies [2, 12]. An important aspect of this trial is that immunization was carried out when the birds were 7 and 20 d old. Temperatures treatments in the environmental chambers started when the birds were 10 d old. Therefore, when these birds were vaccinated for the first time at 7 d of age, they had not been subjected to temperature stress. In this experiment, no differences were found among treatments (P > 0.05). Thaxton and Siegel [13], on the other hand, found that the influence of environmental temperature variations on the immune response depends on the degree of the adaptation of the animal at the time of immunization. Therefore, as the second vaccination occurred later in the life of the birds, it is possible that the birds were then more adapted to the environmental conditions and were less susceptible to immune-suppressive factors. Thus, no differences were found for antibody titers against NDV and IBDV in birds on any temperature regimens. Studies carried out by Siegel and Latimer [4] showed that the interference of high environmental temperatures on the humoral immune response of birds is related to the quantity and quality of the antigen used in vaccination. Beard and Mitchell [2] stated that immunization with live vaccines is more influenced by heat stress, as the host inherent factors (body temperature and changes in metabolism of immune system) may decrease the viral replication needed for the development of a proper immune response.
CONCLUSIONS AND APPLICATIONS 1. The findings of this study suggest that birds exposed to changes of 1 ± 0.02°C increments at hatching do not show interference in humoral immune response against NDV and IBDV vaccine. 2. After hatching, birds exposed to different environmental temperature (18 ± 2, 24 ± 2, and 32 ± 2°C) did not show difference in humoral immune response against NDV and IBDV vaccine.
Downloaded from http://japr.oxfordjournals.org/ by guest on April 13, 2015
RESULTS AND DISCUSSION
249
JAPR: Research Report
250
REFERENCES AND NOTES 1. Henken, A. M., A. M. J. Groote Schaarsberg, and M. G. B. Nieuwland. 1982. The effect of environmental temperature on immune response and metabolism of the young chicken. Poult. Sci. 62:51–58.
8. Cunnigham, C. H. 1971. Virologia Practica, ed. Acribia, Zaragoza.
2. Beard, C. W., and B. W. Mitchell. 1987. Influence of environmental temperatures on the serologic responses of broiler chickens to inactivated and viable Newcastle disease vaccines. Avian Dis. 31:321–326.
10. SAS. 1988. SAS User’s Guide: Statistics. SAS Inst. Inc., Cary, NC.
3. Donker, R. A., M. G. B. Nieuwland, and A. J. Van Der Zijpp. 1990. Heat-stress influences on antibody production in chicken lines selected for high and low immune responsiveness. Poult. Sci. 69:599–607. 4. Siegel, H. S., and J. W. Latimer. 1984. Interaction of high temperature and Salmonella pullorum antigen concentration on serum agglutinin and corticosteroid responses in White Rock chickens. Poult. Sci. 63:2483–2491.
11. Dibner, J. J., C. D. Knight, M. L. Kitchell, C. A. Atwell, A. C. Downs, and F. J. Ivey. 1998. Early feeding and development of the immune system in neonatal poultry. J. Appl. Poult. Res. 7:425–436. 12. Deyhiem, F., and R. G. Teeter. 1993. Dietary vitamin and/ or trace mineral premix effects on performance, humoral mediated immunity, and carcass composition of broilers during thermoneutral and high ambient temperature distress. J. Appl. Poult. Res. 2:347– 355. 13. Thaxton, P., and H. S. Siegel. 1970. Immunodepression in young chickens by high environmental temperature. Poult. Sci. 49:202–205.
6. National Research Council. 1994. Nutrient Requirements of Poultry. 9th ed. National Academic Press, Washington, DC.
Acknowledgments
7. Paulillo, A. C. 1987. Doenc¸a de Newcastle. IV Ensaio experimental de diferentes vias de vacinac¸a˜oCom a estirpe lentogeˆnica LaSota em frangos de corte. Ars Vet. 3:73–79.
The authors thank Fundac¸a˜o de Amparo a` Pesquisa do Estado de Sa˜o Paulo. Proc. n. 99/12952-7, and Conselho Nacional Pesquisa Proc. n. 300545/85-2 for their financial support.
Downloaded from http://japr.oxfordjournals.org/ by guest on April 13, 2015
5. Arjona, A. A., D. M. Denbow, and W. D. Weaver, Jr. 1990. Neonatally induced thermotolerance. Comp. Biochem. Physiol. 95A:393–399.
9. Idexx Laboratories, BioBrasil, Sao˜ Paulo, SP, Brazil.
Effect of Environmental Temperature on Immune Response of Broilers E. Santin,1,2 A. Maiorka,2 W. J. C. Polveiro,3 A. C. Paulillo,3 A. C. Laurentiz, S. A. Borges, and A. V. Fischer da Silva4 Departamento de Patologia Veterina´ria Faculdade de Cieˆncias Agra´rias e Veterina´rias, Universidade Estadual Paulista, Jaboticabal, SP, Brazil
Primary Audience: Veterinarians, Flock Supervisors, Researchers
Several studies demonstrate that environmental temperature can influence the immune response of poultry. The objective of this research was to determine at which stage in the life of a bird this effect is greatest. In experiment 1, broiler breeder eggs were incubated at three different temperatures (36.8 ± 0.2, 37.8 ± 0.2, and 38.8 ± 0.2°C) from the 13th day of incubation to hatching. After hatching, birds were raised in thermoneutral temperature. In experiment 2, 144 1-d-old broiler chicks were distributed into three environmental chambers with different temperatures (18 ± 2, 24 ± 2, and 32 ± 2°C). In both experiments, the humoral immune responses to Newcastle disease virus (NDV) and infectious bursal disease (IBDV) were evaluated. NDV and IBDV antibody titers were not significantly different (P > 0.05) among treatments. Key words: broiler, temperature, humoral immunity 2003 J. Appl. Poult. Res. 12:247–250
DESCRIPTION OF PROBLEM Several studies have shown that the environmental temperature may influence the immune response of poultry [1, 2, 3]. The mechanism by which the environmental temperature may act as an immune suppressor is not fully understood. However, it is speculated that the increased activity of the adrenal gland due to stress increases the level of serum corticosteroids, which cause suppression of cell proliferation factor, or interleukin II [4]. The intensity and the duration of the stress induced by temperature may impair the immune system of poultry [1]; however, the effect of the environmental temperature depends on the degree of 1
habituation of the bird at the time of immunization. Arjona et al. [5] observed that environmental temperature stress early in life may induce resistance to weather variation in adult animals. The current study evaluated the effect of high or low environmental temperature before or after hatching on the humoral immune response of broilers vaccinated against Newcastle disease (NDV) and infectious bursal disease (IBDV).
MATERIAL AND METHODS Experiment 1 Fertilized eggs (Cobb commercial line) from broiler breeders immunized against NDV
To whom correspondence should be addressed: [email protected]. FAPESP fellowship. 3 CNPq fellowship. 4 Universidade Federal do Parana´ (UFPR). 2
Downloaded from http://japr.oxfordjournals.org/ by guest on April 13, 2015
SUMMARY
JAPR: Research Report
248
TABLE 1. Geometric mean titers against IBDVA and NDVB (expressed as log10) of broilers at 14, 35, and 42 d of age exposed to temperature changes during incubation, experiment 1 Geometric mean titers (log10) IBDV
NDV
Treatment
14 d
35 d
42 d
14 d
35 d
42 d
Control (37.8 ± 0.2°C) Hot (38.8 ± 0.2°C) Cold (36.8 ± 0.2°C)
2.81 2.79 2.68
3.51 3.62 3.55
3.66 3.81 3.84
1.58 1.54 1.54
2.10 2.07 2.33
2.03 1.84 1.84
Probability
0.791
0.656
0.206
0.956
0.262
0.528
A
Infectious bursal disease. Newcastle disease virus.
B
by centrifugation at 1,000 × g for 15 min and then inactivated at 56°C for 30 min and stored at −20°C until used. Hemagglutination inhibition reaction (HI) with live antigens, using the β-method [8] was used to measure NDV HI antibodies. An ELISA commercial kit [9] was used to measure IBDV antibodies. The antibody titers obtained in both serological tests were transformed in log10. A completely random experimental design, with three treatments (heat stress, cold stress, and control) and four repetitions, was used. Data were submitted to analysis of variance [10]. Experiment 2 One hundred forty-five, 1-d-old (Cobb commercial line) male broiler chicks from immunized breeders were used. Birds were distributed into three environmental chambers with different temperatures. Until 10 d of age, the temperature in the three chambers was similar (kept around 32 ± 2°C). From 10 d of age until the end of the trial, birds in chamber 1 were subjected to an environmental temperature of 18 ± 2°C (cold stress), birds in chamber 2 were subjected to an environmental temperature of 32 ± 2°C (heat stress), and birds in chamber 3 were subjected to an environmental temperature of 24 ± 2°C (control). Birds were fed a balanced feed based on corn and soybean meal [6], compatible with the different stages of their life cycles. In general, birds were subjected to conditions similar to the management practiced in the commercial broiler industry from 1 to 35 d of age. Vaccination, blood collection, serological analysis, and statistical analysis of data procedures were the same as those used in experiment 1.
Downloaded from http://japr.oxfordjournals.org/ by guest on April 13, 2015
and IBDV were used. Eggs were incubated in three different automatic incubators at 37.8 ± 0.2°C until d 13. From d 14 of incubation until hatching, eggs in incubator 1 were subjected to an incubation temperature of 36.8 ± 0.2°C (cold stress), eggs in incubator 2 were subjected to an incubation temperature of 38.8 ± 0.2°C (heat stress), and eggs in incubator 3 remained at 37.8 ± 0.2°C (control). At hatching, chicks were sexed, and the males were housed in an environmentally controlled poultry house and according to the incubation temperature treatment (heat stress, cold stress or control), with four replicates of 12 birds each. Birds were fed a balanced feed based on corn and soybean meal [6], compatible with the different stages of their life cycles. In general, birds were subjected to similar conditions of management practiced in the commercial broiler industry from 1 to 35 d of age and were kept under thermoneutral temperature (33 ± 2°C up to 7 d of age and 24 ± 2°C until the end of the experiment). Chickens in all experimental groups were vaccinated by eye drop at 7 and 20 d of age with vaccines from the same laboratory and from the same batch, at the beginning of the shelf life. The lyophilized vaccines were prepared with NDV lentogenic strain LaSota (EID50 = 107.20/ 0.1 mL) and with IBDV strain S706 (EID50 = 103.50/0.1 mL). Lyophilized vaccines were diluted with distilled water at 30 mL/1,000 vaccine doses per 1,000 birds, corresponding to an eye drop vaccine dose of 0.03 mL [7]. Blood samples from eight birds per experimental group were collected at 14 and 35 d of age by ulnar vein puncture. Serum was separated
TEMPERATURE AND IMMUNE RESPONSE TABLE 2. Geometric mean titers against IBDVA and NDVB (expressed as log10) of broilers at 14 and 35 d of age exposed to temperature changes after hatching, experiment 2 Geometric mean titers (log10) IBDV
NDV
Treatment
14 d
35 d
14 d
35 d
Control (24 ± 0.2°C) Hot (32 ± 0.2°C) Cold (18 ± 0.2°C)
3.35 3.29 3.22
3.70 3.61 3.72
1.24 1.24 1.22
1.74 1.50 1.71
Probability
0.082
0.503
0.585
0.128
A
Infectious bursal disease. Newcastle disease virus.
B
Results of experiment 1 are shown in Table 1. Antibody titers against NDV and IBDV were not statistically different (P > 0.05) among treatments. In experiment 2 (Table 2), no significant differences (P > 0.05) were observed among treatments; however, IBDV antibody titers of 14-d-old birds were numerically higher (P = 0.08) than the titers of birds subjected to heat (32 ± 2°C) or cold (18 ± 2°C) stress. The incubation period is very important to the development of the immune system of poultry [11]. However, in the present experiment, changes in the incubation temperatures did not influence the humoral immune response. Although not fully elucidated, the mechanism by which changes in temperature affect the immune response is related to the increase of serum corticosteroids [4]. Therefore, it could be speculated that the humoral immune response of the broilers in these experiments was not affected because the environmental changes were not imposed when the birds were being immu-
nized. According to Henken et al. [1], environmental changes at immunization may be more important than temperature variation during the whole life of the bird. In experiment 2, there were no significant differences (P > 0.05) in serum antibody titers against NDV and IBDV after immunization. Similar results have been found in other studies [2, 12]. An important aspect of this trial is that immunization was carried out when the birds were 7 and 20 d old. Temperatures treatments in the environmental chambers started when the birds were 10 d old. Therefore, when these birds were vaccinated for the first time at 7 d of age, they had not been subjected to temperature stress. In this experiment, no differences were found among treatments (P > 0.05). Thaxton and Siegel [13], on the other hand, found that the influence of environmental temperature variations on the immune response depends on the degree of the adaptation of the animal at the time of immunization. Therefore, as the second vaccination occurred later in the life of the birds, it is possible that the birds were then more adapted to the environmental conditions and were less susceptible to immune-suppressive factors. Thus, no differences were found for antibody titers against NDV and IBDV in birds on any temperature regimens. Studies carried out by Siegel and Latimer [4] showed that the interference of high environmental temperatures on the humoral immune response of birds is related to the quantity and quality of the antigen used in vaccination. Beard and Mitchell [2] stated that immunization with live vaccines is more influenced by heat stress, as the host inherent factors (body temperature and changes in metabolism of immune system) may decrease the viral replication needed for the development of a proper immune response.
CONCLUSIONS AND APPLICATIONS 1. The findings of this study suggest that birds exposed to changes of 1 ± 0.02°C increments at hatching do not show interference in humoral immune response against NDV and IBDV vaccine. 2. After hatching, birds exposed to different environmental temperature (18 ± 2, 24 ± 2, and 32 ± 2°C) did not show difference in humoral immune response against NDV and IBDV vaccine.
Downloaded from http://japr.oxfordjournals.org/ by guest on April 13, 2015
RESULTS AND DISCUSSION
249
JAPR: Research Report
250
REFERENCES AND NOTES 1. Henken, A. M., A. M. J. Groote Schaarsberg, and M. G. B. Nieuwland. 1982. The effect of environmental temperature on immune response and metabolism of the young chicken. Poult. Sci. 62:51–58.
8. Cunnigham, C. H. 1971. Virologia Practica, ed. Acribia, Zaragoza.
2. Beard, C. W., and B. W. Mitchell. 1987. Influence of environmental temperatures on the serologic responses of broiler chickens to inactivated and viable Newcastle disease vaccines. Avian Dis. 31:321–326.
10. SAS. 1988. SAS User’s Guide: Statistics. SAS Inst. Inc., Cary, NC.
3. Donker, R. A., M. G. B. Nieuwland, and A. J. Van Der Zijpp. 1990. Heat-stress influences on antibody production in chicken lines selected for high and low immune responsiveness. Poult. Sci. 69:599–607. 4. Siegel, H. S., and J. W. Latimer. 1984. Interaction of high temperature and Salmonella pullorum antigen concentration on serum agglutinin and corticosteroid responses in White Rock chickens. Poult. Sci. 63:2483–2491.
11. Dibner, J. J., C. D. Knight, M. L. Kitchell, C. A. Atwell, A. C. Downs, and F. J. Ivey. 1998. Early feeding and development of the immune system in neonatal poultry. J. Appl. Poult. Res. 7:425–436. 12. Deyhiem, F., and R. G. Teeter. 1993. Dietary vitamin and/ or trace mineral premix effects on performance, humoral mediated immunity, and carcass composition of broilers during thermoneutral and high ambient temperature distress. J. Appl. Poult. Res. 2:347– 355. 13. Thaxton, P., and H. S. Siegel. 1970. Immunodepression in young chickens by high environmental temperature. Poult. Sci. 49:202–205.
6. National Research Council. 1994. Nutrient Requirements of Poultry. 9th ed. National Academic Press, Washington, DC.
Acknowledgments
7. Paulillo, A. C. 1987. Doenc¸a de Newcastle. IV Ensaio experimental de diferentes vias de vacinac¸a˜oCom a estirpe lentogeˆnica LaSota em frangos de corte. Ars Vet. 3:73–79.
The authors thank Fundac¸a˜o de Amparo a` Pesquisa do Estado de Sa˜o Paulo. Proc. n. 99/12952-7, and Conselho Nacional Pesquisa Proc. n. 300545/85-2 for their financial support.
Downloaded from http://japr.oxfordjournals.org/ by guest on April 13, 2015
5. Arjona, A. A., D. M. Denbow, and W. D. Weaver, Jr. 1990. Neonatally induced thermotolerance. Comp. Biochem. Physiol. 95A:393–399.
9. Idexx Laboratories, BioBrasil, Sao˜ Paulo, SP, Brazil.