INFLUENCE OF LIGHT ON THE HEMATOLOGY, LYMPHOID ORGANS AND PRODUCTION OF THE BROILERS

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1 Sarhad J. Agric. Vol. 29, No.4, 2013 INFLUENCE OF LIGHT ON THE HEMATOLOGY, LYMPHOID ORGANS AND PRODUCTION OF THE BROILERS *YASSER SALEEM MUSTAFA 1 and MUHAMMAD AKRAM MUNEER 2 1. Provincial Diagnostic Laboratory, 16-Cooper Road, Lahore - Pakistan. 2. Project Director, NPCPAI, NARC, Islamabad - Pakistan. *Corresponding author: yasserbutt1@yahoo.com ABSTRACT Effect of light stress at various time intervals on broiler chicks was studied during January- February, 2010 by rearing a total of 400, one-day-old Hubbard X Hubbard experimental chicks at Department of Microbiology, University of Veterinary and Animal sciences, Lahore, Pakistan. It was observed that light stressed birds had showed non-significant (P >0.05) difference on blood cells population except lymphocytes percentage was found higher in groups D ( ) than group C. The effect on lymphoid organs studied and the mean thymus weight of chicks in group B ( ) was adversely effected as the chicks in this group had significantly (P<0.05) lower mean thymus weight than groups D ( ) and E ( ) indicating that increase in the period of off-light in chicks adversely effected the mean thymus weight and chicks reared on 24hr light had higher mean thymus weight. The bursa mean weight of group B ( ) was significantly (P<0.05) lower as compared to group D ( ). The spleen mean weight of group D ( ) was significantly (P <0.05) higher than group B ( ). The FCR values were significantly (P <0.05) different among groups in 6 th week and there was non-significant (P>0.05) difference among groups with treatment of Vitamin C, Vitamin E and glucose. Keywords: Light, Thymus, Spleen, Bursa of Fabricius, FCR, TLC, Heterophil, H/L ratio, Lymphocyte, Monocyte, Eosinophil, Basophil Citation: Mustafa. Y.S and M.A. Muneer. Influence of light on the hematology, lymphoid organs and production of the broilers. Sarhad. J. Agric. 29(4): INTRODUCTION Poultry, an important sub-sector of livestock, has emerged as a cheap source of protein for consumption by humans. Various types of poultry farmed in Pakistan are the commercial broiler, commercial layer, and the broiler or layer breeder chickens. Some farmers also keep the Grand-Parent flocks in various parts of the country. The indigenous locally bred non-descript poultry units each having birds (desi, fancy, golden, aseel, etc.) are reared as backyard poultry with other types of birds (pigeons, parrots, quails, peacock, pheasants, ducks, etc.) mainly in the rural and peri-urban areas throughout Pakistan. In Pakistan, the commercial broiler farming was initiated in the early 1960 s with the import of Shaver broiler chicks (Qureshi, 1993). The poultry industry is an important segment of livestock. This industry in Pakistan consists of over 25 thousand commercial farms, 200 chick hatcheries, and 120 Feed Mills, generating employment for about 1.5 million people. The current investment in Poultry industry is around Rs. 320 billion, and it is regarded as a vital source of eggs and meat for the common man. The present poultry production figures are estimated at million day old chicks, of which million are broiler chicks which provide metric tons of poultry meat per annum (Anonymous, ). With all the high potential of poultry development and its increasing consumption at the national level, efforts are required to keep its growth at a level that matches with the increasing human population of the country. The effect of light stress factor on poultry include various hormonal changes, reduction in feed and water intake, altered body metabolic behavior, poor growth and suppression of immune responses required for combating the infectious disease agents (Guyton and Hall, 1997). Upon exposure to the long term stress conditions, the exposed birds become fatigued and this condition of bird may lead to its starvation and being infected with various disease agents (Dohms, 1990). Provision of adequate light is required for controlling many physiological and behavioral body functions. Light allows the birds to acquaint themselves with the environment and synchronize various essential body functions

2 Yasser Saleem Mustafa and Muhammad Akram Muneer. Influence of light on the hematology, lymphoid 580 including temperature and required metabolic steps that facilitate feeding, digestion, and conversion processes. In the poultry farming, various vitamin preparations for alleviating stress on birds are prescribed (Tibiletti, 1993). Vitamin E is fat soluble and is essential for maintaining the integrity and optimal functioning of the muscular, circulatory, nervous and immune systems. (Gershwin, et al.,1985). Vitamin C is also prescribed as an anti-stressor for the improvement of growth, feed efficiency, immune responses, and livability performance of birds. (Muneer, et al. 2001). These findings will further help the farmers to understand the importance of this factor in maintaining the overall health status of poultry. MATERIALS AND METHODS The effect of light on the performance of broiler chicks was evaluated by rearing a total of 400, one-day-old Hubbard X Hubbard experimental chicks obtained from M/S Sabir Poultry Breeders, at Department of Microbiology, University of Veterinary and Animal sciences, Lahore, Pakistan. All the experimental chicks were assigned randomly to five experimental groups i.e. A, B, C, D and E, each group consisting of 80 chicks. The chicks were separately housed and were offered the commercial broiler feed and water ad libitum. The chicks received vaccination against viruses like Infectious Bronchitis (IB), Newcastle Disease (ND); Infectious Bursal Disease (IBD) and Hydro-pericardium Syndrome (HPS). The experimental chicks in the group A were not provided any light and were reared in complete dark from day of their age; those in group B were provided light for 72 hrs and were reared in dark from 23 rd to 35 days; and chicks in the group C were provided light for 24 hrs and no light during the next 24 hrs (24 hrs light at alternate days); chicks in group D were reared under 24 hrs light throughout experiment and group E chicks reared on constant light for one week and only day light for (10 hrs- day) rest of the experimental period. The blood samples were randomly collected at 18 hours post exposure of chicks to various temperatures (in each experimental group) and the TLC and DLC values as per technique of Benjamin ( 1979) and the H/L ratio as per technique described by Gross and Siegel (1983) were recorded. The morphometery of lymphoid organs i.e. Spleen, Thymus and Bursa of fabricius of birds exposed to various stress conditions were recorded as per procedure suggested by Griffith et al. (1985). To observe the effect of various anti-stressors such as vitamin C, vitamin E, and the glucose in different light intervals given to the birds were administered these vitamins and the glucose after further sub-dividing each experimental group A, B, C and D in three sub-groups. The vitamins and glucose treatment were continued from days 36 th to 56 th and of the 03 subgroups in each experimental group (A1, A2, A3; B1, B2 B3; C1, C2, C3 and D1, D2, D3).The birds raised under different light intervals were also treated with vitamin C, Vitamin E, and glucose to determine whether these substances had any role in alleviating the stress on the chicks caused by light. These vitamins and glucose were administered via the drinking water as per recommended dosage level of VC 350mgL -, VE 250mgL - and glucose 1gmL - in drinking water. The control chicks in group E did not receive any of the vitamins or glucose. The data obtained through the experiment was analyzed using analysis of variance and the statistical differences among the various treatment means were determined using the Least Significant Difference (LSD) test at 05% probability level according to the method described by Steel and Torrie (1980). RESULTS AND DISCUSSION Lighting is a powerful exogenous factor in control of many physiological and behavioral processes. Light allows the bird to establish rhythmcity and synchronize many essential functions including body temperature, and various metabolic steps that facilitate the feeding and digestion. The use of continuous light is preferred because the growth rate is optimized through an increase in feed intake digestion and absorption of nutrients. Light intensity, color and the photoperiodic regime affect the physical activity of broiler chickens.

3 Sarhad J. Agric. Vol. 29, No.4, Effect of Light on Various Cell Populations The hematological values of chicks in groups A, B, C, D and E are presented in Table-I. The relative values of TLC 10 3 /mm, the heterophil, lymphocyte, monocyte, basophil, and eosinophil percentages at 18 hrs post light restriction, in group A were recorded as 17.04±0.48, 55.1±2.38, 40.4±2.51, 4±0.30, 0.1±0.10 and 0.4±0.22, respectively. At 18 hrs post light restriction, the relative values of TLC 10 3 /mm, the heterophil, lymphocyte, monocyte, basophil and eosinophil percentages in the control chicks (group D) were 17.66±0.75, 29.8±0.84, 61.4 ±1.16, 7.5±0.45, 0.4±0.16, and 0.9±0.41, respectively. At 18 hrs post light restriction, the relative values of H/L ratio in groups A, B, C, D and E were recorded as 1.36±1.10, 1.41±1.14, 1.5±1.02, 0.48±1.24 and 0.54±1.52, respectively. This study indicated that the heterophil percentages of chicks in group A was significantly (P 0.05) higher (55.1±2.38) than that of the chicks in group D ( 29.8±0.84) which were provided 24 hrs/day light). Similar, heterophil percentages observations in relation to groups B (56.2±3.51) and C (57.1±1.48) compared to group D (29.8±0.84) chicks were recorded. There were non-significant (P 0.05) differences amongst the overall TLC values of that were reared under 24hrs/day light or those which were provided light for various time intervals during this experiment. The lymphocyte percentages of the group D chicks (61.4±1.16) were significantly (P 0.05) higher those of group A (40.4±2.51), group B (39.6±1.44) or group C (37.6±1.47). The lymphocyte percentages in chicks in groups A, B and C (groups deprived of light for various time intervals) did not indicate any significant (P 0.05) differences. From the above findings, it can be concluded that there were non-significant (P 0.05) differences in mean TLC values amongst the various treatment groups. There were non-significant (P 0.05) in the heterophil percentages in groups A (55.1±2.38), B (56.2±3.51) and C (57.1 ±1.48). However, the heterophil percentages of chicks in groups D (29.8±0.84) and E (32.8±0.11) were significantly (P 0.05) lower than those of groups A (55.1±2.38), B (56.2±3.51) and C (57.1±1.48). The lymphocyte percentages of chicks in experimental gr oups B (39.6±1.47) and C (37.6±1.47) were significantly (P 0.05) lower from those of groups D (61.4±1.16) and E (60.6±0.16). There were non-significant (P 0.05) differences in monocyte percentages in chicks of groups A (4±0.30) and C (4.8±0.25). Group D (7.5±0.45) showed significantly higher (P 0.05) monocyte percentages than other treatment groups. However, the differences in monocyte percentages of chicks in groups D (7.5± 0.45), B (6.4±0.27) and E (6±0.20) and those in groups A (4±0.30) and C (4.8 ±0.25) was significant (P 0.05). There were also non-significant (P 0.05) differences in the basophil percentages amongst the chickens of groups A (0.1±0.10), B (0.2±0.13), C (0.0±0.00), D (0.4±0.16) and E (0.4±0.10). Moreover, the eosinophilic percent ages of chickens in groups A (0.4±0.22) and C (0.4±0.22) were similar and significantly (P 0.05) lower than those in groups B (0.6±0.34), D (0.9±0.41) and E (1±0.30). The H/L ratio was significantly (P 0.05) higher in group C (1.5±1.02) than groups A (1.36±1.10) and B (1.41±1.14) indicated the effect of light stress on chicks. Abbas et al. (2008) reported that intermittent light treatment caused significant elevation of total white blood cells (WBC) population compared to control broiler chicken. Zulkifli et al. (1998) reported higher H/L ratio in broilers reared under continuous availability of light. Effect of Light on the Weight of Lymphoid Organs During this study the effect of light on the development of various lymphoid organs (thymus, bursa of Fabricius and spleen) was also determined (Table 2). Thymus It was concluded that the mean thymus weight showed significant (P 0.05) differences among groups. The chicks in groups D (4.64±0.74) and E (4.34±0.25) gained significantly (P 0.05) higher mean thymus weight than groups A (2.56±0.44), B (1.90±0.53) and C (2.14±0.30) while group B (1.90±0.53) showed significantly (P 0.05) lower mean thymus weight than other groups. Bursa of Fabricius

4 Yasser Saleem Mustafa and Muhammad Akram Muneer. Influence of light on the hematology, lymphoid 582 It was concluded that the mean weight of burse of fabricius showed significant (P 0.05) differences among groups. The chicks in groups D (1.59±0.17) and E (1.32±0.10) gained significantly (P 0.05) higher mean bursa weight than groups A (0.90±0.13) and C (0.85±0.18) while group B (0.43±0.05) had significantly (P 0.05) lower mean bursa weight. Spleen It was concluded that the mean spleen weight showed significant (P 0.05) differences among groups. The chicks in group D (1.88±0.15) gained significantly (P 0.05) higher mean spleen weight than groups A (1.38±0.09), B (1.18±0.08) and C (1.22±0.10). Effect of Light on the Feed Efficiency of Chicks The weekly FCR values of chicks in various treatment groups are presented in Table 3. In the first week of their age, birds in all experimental groups had same FCRs (1.14±0.08). The mean FCR values at week 6 of age of chicks in the experimental groups A, B, C, D and E were recorded as 2.10±0.10, 1.98±0.08, 2.29±0.83, 2.20±0.59 and 2.15±0.65, respectively. At this age, the differences in the FCR values of chicks in various groups were significant (P 0.05). The use of vitamin C, vitamin E and glucose did not indicate any improvement in FCR values of the treated groups (Table-III). The FCR values of chicks, on day 56 th, in the vitamin C treated groups A, B, C, and D were recorded as 2.40±0.52, 2.20±0.12, 2.58±0.52 and 2.27±1.03, respectively. In the 7 th week the effect of VC showed significant (P 0.05) differences among groups and group D (2.32±0.73) has significantly (P 0.05) higher FCR than group B (2.15±0.11). The effect of VE showed signifi cant (P 0.05) differences among groups A (2.24±0.04), B (2.10±0.05), C (2.50±0.12) and D (2.40±0.21). The treatment of glucose showed no differences between groups C (2.53±0.15) and D (2.50±0.07) but differences in groups A (2.30±0.44) and B (2.25±0.49). In the 8 th week the effect of VC showed non-significant (P > 0.05) differences in groups B (2.20±0.12) and D (2.27±1.03) but differences from group C (2.58±0.52). The effect of VE showed significant (P 0.05) differences in groups A (2.44±0.16), B (2.08±0. 17), C (2.64±0.17) and D (2.30±0.29). The effect of glucose showed non-significant (P 0.05) differences in groups A (2.50±0.19) and B (2.49±0.55) but significant (P 0.05) differences from group C (2.74±0.28).The differences in the FCR values of chicks in g roup B and group C were significant (P 0.05). Lewis and Morris (1998) reported that FCR is not significantly affected by light source. Buyse et al. (1996) reported no inferior feed conversion in intermittent light in broilers. Al- Gharib et al. (2008) reported that affect of intermittent light increased FCR in broilers. Abbas, et al. (2008) reported that both intermittent and non-intermittent restricted light improved feed conversion significantly compared to continuous light. Classen et al. (1991) reported increased FCR in increased light provided to broiler. CONCLUSIONS AND RECOMMENDATIONS From the findings of this research study, it was concluded that the stress factor light has not adverse effect on the overall hematology of the bird except the heterophil/lymphocytes ratios. Moreover, the stressed birds have lesser functional lymphoid organs compared to non-stressed chickens. The anti-stressor including Vitamin C, Vitamin E and Glucose has positive effect on stressed birds body weight and FCR. Based on finding of this study following recommendation can be made for the poultry farmers. 1. Every effort be made to avoid stress of light on the growing chicks. 2. Chicks must be vaccinated as per schedule to avoid immuno-suppression during stress periods caused by light. 3. Recommended light should be provided to the broiler chicks in view of increased cost of production in Pakistan. 4. The determination of cell population, FCR, weight of lymphoid organs is good indicators of stress condition which may affect the broiler performance.

5 Sarhad J. Agric. Vol. 29, No.4, Table 1. Post light stress hematology of groups g at 18 hour Hematological counts Group A Group B Group C Group D Group E Mean ± S.E Mean ± S.E Mean ± S.E Mean ± S.E Mean ± S.E TLC10 3 /mm 17.04±0.48 a 17.05±0.47 a 16.4±0.58 a 17.66±0.75 a 17.66±0.20 a Heterophils (%) 55.1±2.38 a 56.2±3.51 a 57.1±1.48 a 29.8±0.84 b 32.8±0.11 b Lymphocytes (%) 40.4±2.51 c 39.6±1.44 b 37.6±1.47 b 61.4±1.16 a 60.6±0.16 a Monocytes (%) 4.00±0.30 c 6.4±0.27 a 4.8±0.25 c 7.5±0.45 b 6±0.20 a Basophils (%) 0.1±0.10 a 0.2±0.13 a 0±0.00 a 0.4±0.16 a 0.4±0.10 a Eosinophils (%) 0.4±0.22 a 0.6±0.34 a 0.4±0.22 b 0.9±0.41 a 1.00±0.30 c H/L ratio 1.36±1.10 c 1.41±1.14 b 1.5±1.02 a 0.48±1.24 e 0.54±1.52 d Groups g =Group A, chicks provided no light throughout experimental period; Goup B, chicks provided light for 72 hrs during days 1-3 of their age and no light thereafter; Group C, chicks provided light for 24 hrs and no light during the following 24 hrs, alternatively, throughout experimental period; Group D, chicks raised on 24 hrs constant light throughout experimental period. Group E, chicks reared on constant light for one week and only day light for (10 hrs/day) rest of their life. a, b, c Any two means carrying the same superscript are not significantly different from each other at 5% probability level using LSD test. Table 2. Morphometery of lymphoid organs of light stressed and unstressed chickens Groups g Body weight Thymus Weight Bursa Weight Spleen Weight Mean ± S.E Mean ± S.E Mean ± S.E Mean ± S.E A ±10.01 b 2.56±0.44 b 0.90±0.13 c 1.38±0.09 b B ±9.89 a 1.90±0.53 d 0.43±0.05 d 1.18±0.08 c C ±10.15 b 2.14±0.30 c 0.85±0.18 c 1.22±0.10 c D ±10.02 a 4.64±0.74 a 1.59±0.17 a 1.88±0.15 a E ±14.36 a 4.34±0.25 a 1.32±0.10 b 1.38±0.10 b Groups g =Group A, chicks provided no light throughout experimental period; Goup B, chicks provided light for 72 hrs during days 1-3 of their age and no light thereafter; Group C, chicks provided light for 24 hrs and no light during the following 24 hrs, alternatively, throughout experimental period; Group D, chicks raised on 24 hrs constant light throughout experimental period. Group E, chicks reared on constant light for one week and only day light for (10 hrs/day) rest of their life. a, b, c, d Any two means carrying the same superscript are not significantly different from each other at 5% probability level using LSD test. Table 3. Weekly body weight/fcr (Mean ± S.E) of birds of light stress and control groups g Groups Age in Weeks g Vitamin C Vitamin E Glucose Vitamin C Vitamin E Glucose weight(fcr) weight(fcr) weight(fcr) weight(fcr) weight(fcr) weight(fcr) weight(fcr) A ±10.01 (2.10±0.10) b ±10.21 (2.20±0.07) b 1560±10.15 (2.24±0.04) b 1560±12.21 (2.30±0.44) a 1650±9.79 (2.40±0.52) b 1580±11.21 (2.44±0.16) c 1595±12.01 (2.50±0.19) b B ±9.89 (1.98±0.08) a ±11.02 (2.15±0.11) a 1535±12.21 (2.10±0.05) a 1540±12.34 (2.25±0.49) a 1675±10.45 (2.20±0.12) a 1575±10.45 (2.08±0.17) a 1625±10.45 (2.49±0.55) b C ±10.15 (2.29±0.83) c ±10.45 (2.20±0.35) b 1535±12.12 (2.50±0.12) c 1530±11.45 (2.53±0.15) b 1650±10.21 (2.58±0.52) c 1595±12.32 (2.64±0.54) d 1642±9.69 (2.74±0.28) c D ±10.02 (2.20±0.59) c 1615±10.21 (2.32±0.73) c 1558±12.41 (2.40±0.21) d 1540±12.10 (2.50±0.07) b 1700±11.09 (2.27±1.03) a 1605±11.02 (2.30±0.29) b 1665±10.08 (2.4±0.10) a ± ±15.21(2.22±0.7) 1710±15.89(2.20±0.18) E * (2.15±0.65) b Groups g =Group A, chicks provided no light throughout experimental period; Goup B, chicks provided light for 72 hrs during days 1-3 of their age; Group C, chicks provided light for 24 hrs and no light during the following 24 hrs, alternatively, throughout experimental period; Group D. chicks raised on 24 hrs constant light throughout experimental period. Group E, chicks reared on constant light for one week and only day light for (10 hrs/day) rest of their life. * Chicks note given treatment with Vitamin. C, Vitamin. E or glucose a, b, c, d Any two means carrying the same superscript are not significantly different from each other at 5% probability level using LSD test. REFERENCES Abbas, A. O., A. K. A. El-Dein, A. A. Desoky and M. A. A. Galal The effects of photoperiod programs on broiler chicken performance and immune response. Intl. J. Poult. Sci. 7 (7): Al Gharib, H. B., A. A. Desoky, M.A. El-Menawey, A. O. Abbas, G.L. Hendricks and M.M. Mashaly The role of photoperiod and melatonin on alleviation of the negative impact of heat stress on broilers. Intl. J. Poult. Sci. 7 (8):

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