EFFECT OF BETAINE WITH OR WITHOUT TWO ...plasma in White Leghorn hens during HS and that supplementation of 1000 ppm Vit C may further enhance in vitro lymphocyte proliferative responses

37

J.Agric.&Env.Sci.Dam.Univ.,Egypt Vol.10 (1) 2011

EFFECT OF BETAINE WITH OR WITHOUT TWO

ANTIOXIDANTS ON THE PERFORMANCE OF DUAL

PURPOSE BREEDING HENS EXPOSED TO HEAT

STRESS

YOUSSEF A ATTIA, AHMED A. ABDALLAH, MERVAT A. BRIEKAA,

ABD EL-HAMID E. ABD EL-HAMID AND BAHAA M. ABOU- SHEHEMA

Animal and Poultry Production Department, Faculty of Agriculture, Damanhour

University, Egypt

Animal Production Research Institute, ARC, Ministry of Agriculture and Land

Reclamation, Egypt

Corresponding author: Prof. Youssef A. Attia, Faculty of Agriculture Animal and Poultry

Production Department, Faculty of Agriculture, Damanhour University, Egypt

Tel:+2453318537- Fax: +2453316535

Email: [email protected]

ABSTRACT

The antioxidant capability of betaine (trimethylglycine,

Bet), ascorbic acid (Vit C) and vitamin E (α-Tocopherol, Vit

E) and their possible mixtures on improving productive,

reproductive, egg quality and physiological traits of dual

purpose hens exposed to heat stress (HS) was studied during

weeks 32-36 of age. A total of 288 hens and 36 cocks

Mandarah local developed strain were housed in

environmental controlled light proof house and randomly

distributed to 36 floor pens (2 x 1 x 2 m) furnished with

wheat straw. Pens were divided into nine treatment groups

and represented by 4 replicates (8 hens + 1 cock, each). The

first treatments was kept under optimum temperature 22-

24°C and relative humidity (RH) 55 – 65 % and used as

positive control (PC). Whereas the other eight treatments

were kept under HS condition (38o C; 65% RH) for three

successive days a week from 11.00 am until 15.00 pm. The

first group of the HS treatments was used as negative

control (NC). Both of PC and NC groups were fed the basal

diet without any antioxidant supplementations. The other

seven HS treatments were fed the basal diet supplemented

Tel:+2453318537-

37


with Bet (1000 mg/kg diet), Vit C (200 mg/kg diet), Vit E

(150 mg/kg diet as α- tocopherol acetate) and their possible

mixtures. Heat stress negatively affected productive

performance, exterior and interior egg quality traits, fertility

percentage and the relative weight of one day old chicks,

characteristics of red blood cells, blood pH and immune

indices. On the other hand, supplementation of either Vit C

or Bet improved productive performance of laying hens.

Betaine had a better effect than Vit C with respect to specific

gravity, Haugh unit score, hematological and immune

indices. However, Vit E was the most effective for the

heterophil, while, it had the least influence on the productive

traits. In conclusion, supplementation of diets for dual

purpose hens kept under HS condition (38°C and 65% RH)

with Vit C (200 mg/kg diet) or Bet (1000 mg/kg) improved

productive performance. Moreover, Bet supplementation

improved egg quality traits and immune indices, too

suggesting that Bet may be used alone as anti- heat stress for

dual purpose hens.

Key words: Heat stress, local developed strain, betaine, ascorbic acid, α-Tocopherol,

productive and reproductive performance.

INTRODUCTION

A major challenges facing the breeders and layers industry is the

HS, which negatively affects feed intake, laying performance,

reproductive performance and physiological traits of chickens (Attia

et al., 2006;2011 and Yoshida et al., 2011) and economic traits

(Mashaly et al., 2004; Daghir, 2008 and Attia et al., 2009). The

adverse effects of HS could be attributed to the complex inter-

response of low feed intake, endocrine system malfunction, acid-base

imbalance and poor physiological functions of organs and

mechanisms connected with the entire egg production process via

follicular recruitment and growth, ovulation, egg and shell formation,

eggs and yolk development, oviposition and oviposition interval

(Rozenboim et al., 2007; Oguntunji and Alabi, 2010). However, the

effect of HS depends on the environmental temperature, relative

37


humidity and severity of HS (Daghir, 2008; Yoshida et al., 2011 and

Tumová and Gous, 2012), and the strain of laying hens (Franco-

Jimenez et al., 2007). Several nutritional strategies have been suggested to relieve the

negative effects of HS, such as supplementation of Vit C, E as natural

antioxidant and Bet as a multi-nutritional agent (Daghir, 2008 and

Attia et al., 2009). However, their additive effects with Bet were not

addressed in the literature. Betaine is a tertiary amine formed by the

oxidation of choline (Kidd et al., 1997; Wang et al., 2004) and

implicated in methionine sparing, osmoprotective, fat distribution and

immune responses (Saunderson and Mackinlay, 1990; Petronini et

al., 1992; Kettunen et al., 2001; Türker et al., 2004; Remus et al.,

2004 and Graham 2002). However, Bet is not present in large

quantities in animal feedstuffs and dietary supplementation seems to

be important to improve productivity and resistance to stress (Wang et

al., 2004). Betaine has an energy sparing impact by reducing

maintenance requirement (Schrama and Gerrits, 2000), improves

growth, carcass yields and muscle protein (Virtanen and Rosi, 1995;

Esteve- Garcia and Mack, 2000; Türker et al., 2004 and Wang et

al., 2004). Furthermore, Miles et al. (1987) revealed that Bet could

substitute choline and methionine in broiler and laying hens diets

(Attia et al., 2005; Hassan et al., 2005). However, the use of Bet to

improve laying hens tolerance to HS deposit it's nutritional,

osmoprotective, and fat distribution and immune responses needs

further research.

Ascorbic acid, better known as Vit C, is water soluble, therefore,

had important task to protect the animals under stressors conditions

and had also an activity against oxidation (Lin et al., 2006 and Attia

et al., 2009). Vit C is necessary for various biosyntheses (collagen,

1,25-dihydroxy vitamin D and adrenaline) as well as for regulation of

diverse reactions (secretion of corticocosterones, regulation of body

temperature and activation of immune system). Adult poultry under

normal condition are able to synthesize adequate Vit C to meet their

requirements. However, supplemental dietary Vit C limits and

alleviates the metabolic sign of stress and improves performance,

immunological status and the behavior of birds. (Daghir, 2008 and

Attia et al., 2009). Laying hens have also shown responses to

supplemental Vit C at 200-400 mg/kg in terms of improving livability,

37


feed intake, egg production and egg quality (Lin et al., 2006; Daghir,

2008 and Çiftçi et al. 2005). Ahmed et al. (2008) found that hen-

housed egg production, feed conversion ratio (FCR), egg weight and

egg mass were improved (P<0.01) in hens receiving Vit C

supplementation (1000 and 1200 ppm/ L water).

Vitamin E is very important natural antioxidants and is an excellent

biological chain-breaking antioxidant that protects cells and tissues

from lipoperoxidative damage induced by free radicals (Bollengier-

Lee et al., 1998; 1999 and Metwally, 2003;2005). Poultry can not

synthesize Vit E, therefore requirements must be met from dietary

sources. Heat stress stimulates the release of corticosterone and

catecholamines and initiates lipid peroxidation in cell membranes. It is

suggested that vitamin E can reduce the negative effects of

corticosterone induced by stress (Metwally, 2003;2005). Vit E also

provides protection for those cells involved in immune responses

(lymphocytes, macrophages and plasma cells) against oxidative

damage and enhances proliferation and functions of these cells.

Therefore, Vit E supplementation in diets is necessary under HS

conditions. Bollengier-Lee et al. (1998 and 1999) concluded that a

dietary supplement of 250 mg Vit E/kg provided before, during and

after HS is optimum for alleviating, at least in part, the adverse effects

of HS in laying hens. In layers, Vit E supplementation at 125-

250mg/kg improves egg production, FCR and immune competence.

Heat stress depresses egg production due to lower plasma

concentrations of egg yolk precursors, vitellogenin and very low

density lipoprotein. Vit E improves egg production by facilitating the

release of vitellogenin from the liver and increasing the circulating

supply of this precursor for yolk formation by protecting the liver

from lipid peroxidation and cell membranes damage. Bollengier-Lee

et al. (1998) The additive, super additive or synergetic influence of different

antioxidants such synthetic source Vit E, C and/or Bet may enhance

laying hens' performance although it received little attention deposited

different root of action. In the literature, Vit C has been demonstrated

to enhance antioxidant activity of Vit E by reducing the tocopheroxyl

radicals back to their active form of Vit E (Jacob, 1995) or by sparing

available Vit E (Retsky and Frei, 1995). Yin et al. (1993) reported

that a mixture of α-tocopherol and ascorbate delayed myoglobin

http://www.tandfonline.com/action/doSearch?action=runSearch&type=advanced&result=true&prevSearch=%2Bauthorsfield%3A%28BOLLENGIER%5C-LEE%2C+S.%29




33


oxidation, whereas α-tocopherol or ascorbate alone did not delay

metmyoglobin formation. Schaefer et al. (1995) demonstrated that

oxidation of myoglobin is prone to retardation when α- tocopheroxyl

radical at the membrane-sarcoplasma interface is reduced by

ascorbate. Regarding antioxidant property, there is a positive

synergistic effect of Vits E and C on the immune response. Gonzalez-

Vega-Aguire et al. (1995) demonstrated that the combination of 200

ppm of Vit C plus 75 IU/kg of Vit E improved antibody levels of

broiler chickens against Brucella abortus and Newcastle modified live

and dead virus vaccine. In addition to antioxidation, Vit C was found

to enhance immune response by modifying corticosteroid synthesis in

adrenal glands (Pardue et al., 1985). Puthpongsiriporn et al. (2001)

observed that Vit E at 65 IU/kg diet may enhance production,

induction of in vitro lymphocyte proliferative responses to

concanavalin A (ConA) and salmonella typhimurium

lipopolysaccharide (LPS), and antioxidant properties of egg yolks and

plasma in White Leghorn hens during HS and that supplementation of

1000 ppm Vit C may further enhance in vitro lymphocyte proliferative

responses of laying hens during HS. Sahin et al. (2002) suggest that

dietary Vit C plus E at 250 mg/kg diet of each act synergistically as

Vit C protects Vit E from peroxidation. In addition, Çiftçi et al.

(2005) and Panda et al. (2008) reported that, Vit E (125 mg/kg diet)

plus C (200 mg/kg diet), have some protective actions against HS-

induced-deleterious effects and may increase egg production and

improve egg quality in laying hens during HS. Mohiti-Asli et al.

(2010) found that Vit E (α-Tocopheryl acetate) did not affect laying

hens performance exposed to high ambient temperature 33 ºC but

resulted in higher antibody titers and serum cholesterol concentration.

Also, dietary supplementation with 150 mg Vit C and/or 150 mg Vit E

increased production performance in heat stressed layer chickens

(Ajakaiye et al., 2011).

The aim of this study was to evaluate betaine as a natural product,

Vit E and C and their possible mixtures on productive and

reproductive performance, egg quality and heamatological traits of

laying hens exposed to heat stress.

37


MATERIALS AND METHODS

Birds and experimental design

A total of 288 hens and 36 cocks of Mandarah developed local

strain were divided into nine treatments groups and housed in 36 floor

pens (2× 1 x 2 m) furnished with wheat straw. Each treatment was

represented by 4 replicates (8 hens + 1 cock, each. Hens were housed

in environmental controlled light proof house. The first treatment was

kept under optimum temperature 22- 24° C and relative humidity

(RH) 55 -60 % and fed the basal diet only without supplementation of

additional Vit C, E or Bet and used as positive control (PC). Whereas,

the other eight treatments were kept under HS (38°C; 65% RH) for

three successive days a week from 11.00 Am until 15.00 Pm. The

first HS group was fed the basal diet only without supplementation of

additional Vit C, E or Bet and used as a negative control (NC).

Whereas the other treatments groups were fed the basal diet

supplemented with betaine (Bet, 1000 mg/kg; natural Betafin®

S6,

Danisco Animal Nutrition), Vit C (200 mg/kg diet, Ascorbic acid; a

heat stabilized product produced by Hoffmann-La Roche), Vit E (150

mg/kg diet as α- Tocopherol acetate produced by Hoffmann-La

Roche) and their possible mixtures.

The experimental was run during weeks 32-36 of age. Feed and

water were provided ad-libitum throughout the experimental period.

All birds were vaccinated every 45 days of age against Newcasle

Disease Virus (Lasota strain vaccine) during egg production period,

according to common veterinarian care practice. Birds were exposed

to photoperiod regimen 16 Light - 8 Dark cycle.

Data collection Laying rate (egg number/hen/d), egg weight (g), egg mass

(g/hen/day) and feed intake (g/hen/d) were measured daily for each

replicate. Feed conversion ratio was calculated as the amount of feed

required to produce 1 kg of eggs. Survival rate was calculated as

number of live hens at the end of the experiment divided by number of

hens at initiation of the experiment.

At weeks 36 of age, 9 eggs per treatment were collected for

determination of eggs and shell quality (Attia et al., 1994; 1995) and

dry matter in yolk and albumen according to AOAC (1995). Eggs

were collected for seven days, stored at room temperature, incubated

37


(37.6º C, 55% RH) and hatched (36.8º C, 65% RH) in an automatic

incubator. All hatched chicks were counted and fertility and

hatchability of total and fertile eggs were calculated according to

Attia et al. (1994; 1995). All unhatched eggs were broken out to

differentiate the infertile eggs from dead embryos. Pipped eggs were

counted and pipped percentage was calculated and hatched chicks

were weighted.

At 36 week of age, blood samples were withdrawn from the wing

vein from 6 hens per treatment for determination the number of red

blood cells (RBC's), white blood cell (WBC's) and different types of

leukocytes. Total RBC's were counted on an Ao bright line

hemocytometer using light microscope at 400x magnification. Blood

samples were diluted 200 times with physiological saline solution

before counting. Total WBCs were counted on a Ao bright line

hemocytometer using a light microscope at 100x magnification after

diluting the blood samples 20 times with a dilitry fluid (3ml acetic

acid glacial + 97 ml distilled water + some of Lushman stain)

according Hepler (1966).

Hemoglobin concentration was determined by the

cyanomethemoglobein method (Eilers, 1967). Wintrobe hematocrite

tubes were used for determination of the hematocrite value. Blood

samples were centrifuged for 20 minutes at 4000 (rpm), then

hematocrite values were measured by reading the packed cell volume

(PCV) on the graduated scale.

A thin blood film was prepared by using a small drop of blood

to determine different leukocytes count. The blood film was

completely dried before staining using Inshas stain. The film was

washed in distilled water and dried. Differential leukocyte counts were

examined in each blood film by using light microscope with 1000 x

magnification power. The number of any individual type of WBC's

was expressed as a percentage of the total (Lucas and Jamroz, 1961).

Determination of phagocytic activity and phagocytic index:

Phagocytic activity was determined according to Kawahara et

al. (1991).

Phagocytic activity (PA) = Percentage of phagocytic cells

containing yeast cells.

78


Statistical analysis Data were analyzed using the GLM procedure of SAS

® (SAS, 1996)

using one-way ANOVA. Before analysis, all percentages were

subjected to logarithmic transformation (log10 x + 1) to normalize

data distribution. Mean difference at P 0.05 was tested using student

Newman Values test.

RESULTS

Productive performance

The effect of HS and different supplements on egg production

traits are shown in Table (2). A significant negative effect of HS on

productive performance of laying hens was observed while

supplementation of Bet, Vit C or E and their combination significantly

improved the productive performance. However the level of positive

control was achieved only with egg weight and FCR. The results

showed that Bet mixtures and Vit C+E resulted in egg weight similar

to the PC treatment but these mixtures did not exceed the

supplementation of Bet or Vit C alone. In addition, Bet and Vit C

significantly improved FCR, thus difference from the PC was

diminished.

External egg quality traits

Data for external egg quality are shown in Table (3). Heat stress

and different supplementations had no significant effect on egg shape

index and absolute shell weight (g). However, there was a significant

impact of HS on specific gravity, shell thickness and SWUSA. The

results indicated that Bet, Vit C and E and their mixture improved

shell quality traits without significant differences between different

supplementations either alone or combined except for egg specific

gravity which was significantly lower of Vit C group than that of Bet,

Vit E, Bet+ Vit E and Bet +Vit C+E.

Internal egg quality traits

Data for internal egg quality are shown in Table (4). A negative

effect of HS was shown only in yolk dry matter (%), albumen dry

78


Table (1). Ingredient and chemical composition (g/kg) of the

experimental diet for laying hens. Ingredients g/kg

Yellow corn 663.30

Soybean meal 48% 242.0

Limestone 75.00

Dicalcium phosphate 13.20

Vit+Min Premix1 2.50

NaCl 2.50

DL-methionine 1.50

Total 1000

Calculated composition,%

ME, kcal/Kg 2775

CP, g/kg 172.6

Methionine, g/kg 4.4

Meth.+Cys. (TSAA, g/kg) 7.1

Lysine, g/kg 8.3

Calcium, g/kg 32.0

Av. Phos, g/kg 36.0

Determined Values (AOAC, 2000)

Dry matter, g/kg

Crude protein, g/kg 169.7

Crude fat, g/kg 24.5

Crude fibre, g/kg 39.6

Ash, g/kg 63.7

Nitorgen free extract, g/kg 1Vit+Min mixture provides per kilogram of diet: vitamin A, 12000 IU; vitamin E, 10 IU;

menadione, 3 mg; Vit. D3, 2200 ICU; riboflavin, 10 mg; Ca pantothenate, 10 mg; nicotinic

acid, 20 mg; choline chloride, 500 mg; vitamin B12, 10 g; vitamin B6, 1.5 mg; vitamin B1,

2.2 mg; folic acid, 1 mg; biotin, 50 g. Trace mineral (milligrams per kilogram of diet): Mn,

55; Zn, 50; Fe, 30; Cu, 10; Se, 0.10;

Anti oxidant, 3 mg.

78


77


77


77


77


73


matter (%), yolk index, yolk color and Haugh unit score but not in

yolk weight (%), albumin weight (%) and yolk albumin ratio.

However, Bet, Vit C and E resulted in significantly better egg quality

traits than the NC although Bet was more effective for Haugh unit

score. It should be noticed that different combinations between Bet,

Vit C and/or Vit E did not suppress that of Bet, Vit C or E alone.

Reproductive performance

Data for reproductive performance are presented in Table (5). A

negative effect of HS was observed only in fertility (%), but

supplementation with different additives diminished the adverse effect

of HS without significant differences between them. There was also a

significant effect of different supplements on relative weight of day-

old chicks but differences among various treatments were not

significant.

Blood heamatological traits

Data for heamatological traits are shown in Table (6). It was

found that HS adversely affect most of heamatological traits except

for monocyte, basophil, eosinophil and neutrophil and

supplementations had no significant effect on most of these traits

except for Bet that significantly decreased eosinophil compared to

only the PC and NC. On the other hand, Bet, Vit C or E similarly

relief the negative effect of HS on RBC's, Hgb, PCV, neutrophil, H/L

ratio, PA and PI. The two or three-way combination did not exceed

their respective single agent. The results indicated that Vit C had a

greater effect than Bet on WBC's but Bet had a greater effect on

lymphocyte than Vit C or E. On the other hand, Vit C without or with

Vit E resorted heterophil to the PC level while Bet and Vit C had the

strongest effect on blood pH but the pH value of the PC was not

achieved.

DISCUSSION

Worldwide, one of the major challenges facing the breeders/

layers industry is the higher ambient temperature in the summer

months that negatively affects feed intake, laying performance,

reproductive performance and physiological traits (Mashaly et al.,

2004; Attia et al., 2006; 2009;2011; Daghir, 2008 and Yoshida et

al., 2011). The response of laying / breeding hens to HS depends on

strain of hens (Franco-Jimenez et al., 2007), ambient temperature

77


and severity of HS chronic vs. acute stress (Daghir, 2008; Yoshida et

al., 2011; Tumová and Gous, 2012).

The negative effect of HS found herein on productive

performance, egg quality, reproductive performance and

hematological traits reflected the chronic effect of HS (High ambient

temperature, 38°C; 65% RH) for three successive days a week from

11.00 am until 15.00 pm weekly. In the literature the optimum

temperature for laying hens is ~ 25 ºC (Daghir, 2008; Tumová and

Gous, 2012). The latter authors reported, in accordance with the

present results, that egg shape index was significantly affected by

temperature (P ≤ 0.028), and specific gravity declined with increasing

temperature (P ≤ 0.013). The effect of HS could be attributed to the

complex inter-response of low feed intake, endocrine system

malfunction, acid-base imbalance and poor physiological functions of

organs and mechanisms connected with the entire egg production

process via follicular recruitment and growth, ovulation, formation of

egg and shell, development of eggs and yolk, oviposition and

oviposition interval (Rozenboim et al., 2007; Oguntunji and Alabi,

2010). In addition, Yoshida et al. (2011) found that feed intake in

hens exposed to 31 ºC significantly decreased compared with that at

23 and 27 ºC.

The nutritional strategies may be easy and applicable way to

improve birds tolerance to HS and supplementation of Bet, Vit C

and/or E have been suggested (Metwally, 2003; Daghir, 2008 and

Attia, 2009). It is interesting to report that Bet had similar effect to

Vit C on the productive traits. Besides, Bet was the most effective

agents for egg quality traits (specific gravity and albumen weight

percentage), immune response (lymphocyte and PI), acid base balance

(blood pH) and reproductive performance (relative weight of day old

chicks). Similarly, Attia et al. (2009) reported that 0.15 g Vit C and 1

g of Bet/ kg diet was equally potent for partial relief (P<0.05) of the

negative effect of HS on productive, digestibility, physiological and

humoral immune competence to sheep red blood cell (SRBC's). The

enhancing effect of Bet could be attributed to multi-nutritional effects

such as methionine sparing, osmoprotective, fat distribution and

immune responses (Saunderson and Mackinlay, 1990; Petronini et

al., 1992; Kettunen et al., 2001; Türker et al., 2004; Remus et al.,

2004 and Wang, 2004), water balance or osmoregulatory property

http://ps.fass.org/search?author1=E.+T%C5%AFmov%C3%A1&sortspec=date&submit=Submit

http://ps.fass.org/search?author1=R.+M.+Gous&sortspec=date&submit=Submit

77


(Kettunen et al. 2001 and Graham 2002), and energy sparing role by

reducing maintenance requirement (Schrama and Gerrits, 2000).

Thereby, Bet has shown to improved productive traits and product

quality (Virtanen and Rosi, 1995; Esteve- Garcia and Mack, 2000;

Türker et al., 2004 and Wang et al., 2004). Furthermore, Miles et al.

(1987) revealed that Bet had a sparing effect for choline and

methionine in chicken's diets (Attia et al., 2005 and Hassan et al.,

2005). The present results indicated that Bet at 1000 mg/kg diet had

the capability to improve productive and reproductive, egg quality

traits and immune response similar to Vit C and better than Vit E.

For productive traits, the present results indicated that Vit C

was the most effective supplementation as similar FCR to the positive

control was obtained. On the other hand, Bet and Vit E

supplementation and any combination between Vit C, E and/or Bet

were also effective but did not surpassed the effect of Vit C on the

productive traits. Vitamin C is necessary for various biosyntheses

(collagen, 1, 25-dihydroxy vitamin D and adrenaline) as well as for

regulation of diverse reactions such as secretion of corticocosterones,

regulation of body temperature and activation of immune system (Lin

et al., 2006 and Attia et al., 2009). In addition, Vit C enhances the

antioxidant activity of Vit E by reducing the tocopheroxy radicals

back to their active form of Vit E. Thus, Vit C requirement is higher

during stress and supplemental Vit C limits and alleviates the

metabolic sign of stress and improves performance, immunological

status and the behavior of birds. In the literature, laying hens have also

shown responses to supplemental vitamin C at 200-400mg/kg in terms

of improvement in livability, feed intake, egg production and egg

quality (Jacob, 1995; Lin et al., 2006 and Daghir, 2008). In addition,

Ahmed et al. (2008) found that egg production, FCR, egg weight and

egg mass were improved (P<0.01) in hens exposed to 34.5°C (28-

42°C) when supplemented with Vit C at either 1000 or 1200 ppm/ L

water as compared to the control birds. Shell thickness was also higher

(P<0.01) in birds receiving 1000 ppm Vit C supplementation

compared to those of other groups. The birds received Vit C

supplementation had significantly (P<0.01) increased plasma ascorbic

acid while decreasing blood glucose and serum cholesterol levels as

compared to the birds of control group. The enhancing effect of Vit C

was attributed in addition to its antioxidation, Vit C was found to

78


enhance immune response by modifying corticosteroid synthesis in

adrenal glands (Pardue and Thaxton, 1985).

The Vit E was less effective than Bet and Vit C for productive

traits but had similar effect to Bet and Vit C for egg quality,

reproductive traits and heamatological traits. In addition, Vit E

induced the greatest effect on heterophil and blood pH than the other

agents but had the least effective on lymphocyte. The present results

are in agreement with those reported by Puthpongsiriporn et al.

(2001) who indicated that Vit E supplementation at 65 IU/kg diet may

enhance production traits, induction of in vitro lymphocyte

proliferation by ConA and LPS, and antioxidant properties of egg

yolks and plasma of heat stressed White Leghorn hens. In addition,

Kirunda et al. (2001) and Bollengier-Lee et al. (1998) showed that

Vit E addition reduced the negative impact of high ambient

temperature on laying hen performance and blood plasma

constituents. Also, Puthpongsiriporn et al. (2001) and Metwally

(2003) revealed highly significant increase in the eggshell thickness in

hot climates due to addition of Vit E at 310 mg/kg diet. Moreover,

Leeson et al. (1998) reported that high levels of Vit E (100 mg/kg) in

the diet of hens fed flaxseed improved the overall egg acceptability.

In the literature, the positive effect of Vit E on HS hens was

attributed to its antioxidants and biological chain-breaking antioxidant

that protects cells and tissues from lipoperoxidative damage induced

by free radicals (Çiftçi, et al., 2005 and Daghir, 2008). In addition to

corticosterone lowering effect of Vit E (Tengerdy,1989), maintaining

the supply of egg precursors lipoprotein in plasma for yolk formation

(Bollengier-Lee et al., 1998 and Yoshida et al., 2011) and decreasing

free radicals in biological system (Morrissey et al., 1994;1998). Also,

Bollengier-Lee (1999) reported that Vit E supplementation at 125-250

mg/kg diet to laying hens improves egg production, feed efficiency,

immune competence, facilitating the release of vitellogenin from the

liver and increasing the circulating supply of this precursor for yolk

formation by protecting the liver from lipid peroxidation and damage

to cell membranes (Bollengier-Lee, 1999), and also provides

protection for those cells involved in immune responses (lymphocytes,

macrophages and plasma cells) against oxidative damage and

enhances proliferation and functions of these cells (Mohiti-Asli et

al.,2010). On the other hand, Metwally (2003; 2005) and Mohiti-





78


Asli et al. (2010) found that Vit E (α-tocopherol acetate) did not affect

laying hens performance exposed to high ambient temperature 33 ºC

but resulted in higher antibody titers and serum cholesterol

concentration. Poultry requirements for Vit E must be met from

dietary sources. Heat stress stimulates the release of corticosterone

and catecholamines and initiates lipid peroxidation in cell membranes

and Vit E may decrease the negative effects of corticosterone induced

by stress (Daghir, 2008).

The combined effect of using different antioxidants such

synthetic source of Vit E and C with Bet as a mutli-nutritional and

osmoregulatory agent in the laying hens diets under HS has received

little attention deposited different root of action. In general, several

positive effects was noticed due to mixtures of Bet and/or Vit C over

Vit E showed improving laying rate, egg mass and FCR than those of

Vit E alone. In addition, supplementation of Bet over Vit C improved

egg specific gravity, also the combination of three agents improved

egg shell thickness to the level of the PC, thus the shell thickness was

significantly better than the NC. Similarly, Yin et al. (1993) reported

that a mixture of α-tocopherol and ascorbate delayed myoglobin

oxidation, whereas α-tocopherol or ascorbate alone did not delay

metmyoglobin formation. In addition, Schaefer et al. (1995)

demonstrated that oxidation of myoglobin is prone to retardation when

α- tocopheroxyl radical at the membrane-sarcoplasma interface was

reduced by ascorbate. Moreover, Vit C has been demonstrated to

enhance antioxidant activity of Vit E by reducing the tocopheroxyl

radicals back to their active form of Vit E (Jacob, 1995) or by sparing

available Vit E (Retsky and Frei, 1995). The combination of both

vitamins showed small effect but of little additive effect on

performance of laying/breeding hens exposed to chronic HS herein,

indicating that Vit C or Bet may be adequate. Similarly,

supplementation with 150 mg Vit C and/or 150 mg Vit E increased

production performance in heat stressed layer chickens (Ajakaiye et

al., 2011).

There is a positive synergistic effect of Vit E and C on only

WBC’s and neutrophil, and the same was observed when Bet and Vit

C was combined. Similarly Gonzalez-Vega-Aguire et al. (1995)

demonstrated that the combination of 200 ppm Vit C and 75 IU/kg Vit

E improved antibody levels of broiler chickens against Brucella

78


abortus and Newcastle modified live and dead virus vaccine. Also,

Sahin et al. (2002) suggested that Vit E and C at 250 mg of each,

offering a good management practice in laying hens reared at high

temperatures and Vit C and E act synergistically as Vit C protects Vit

E from peroxidation. Along the same line, Çiftçi et al. (2005)

reported that 125 mg Vit E plus 200 mg Vit C/kg of diet have some

protective actions against HS-induced-deleterious effects, thus egg

production and egg quality in laying hens were improved during HS.

In addition, Panda et al. (2008) concluded that Vit E (125 IU/kg) and

Vit C (200 mg/kg) could independently alleviate the effects of HS on

production performance and immunological variables of layers.

However, combination of both Vit C+E at the above-mentioned

concentrations is beneficial in eliciting higher antioxidant status in

laying hens exposed to tropical summer conditions. On the other hand,

the contradiction between the abovementioned results and the present

one could be attributed to strain of layers, severity of HS and dose and

form of additive used as anti-stress agents and their root of actions.

In conclusion, supplementation of the diet for local developed

strain kept under HS condition (38o C and 65% RH) with Vit C (200

mg/kg diet) and Bet (1000 mg/kg) improved productive performance

and Bet improved also egg quality traits and immune indices,

suggested that Bet may be used alone as anti- heat stress for local

developed chickens strain.

REFERENCES

Ahmed, W., Ahmad, S., Ahsan-ul-haq. and Kamran, Z. (2008). Response of laying hens to vitamin C supplementation through

drinking water under sub-tropical conditions. Avian Bio.Res,

Vol. 1., 59-63

Ajakaiye, J. J., Cuesta-Mazorra, M. and Garcia-Diaz, J. R

(2011).Vitamins C and E can Alleviate Adverse Effects of

Heat Stress on Live Weight and Some Egg Quality Profiles of

Layer Hens.Pak Vet J, 31(1): 45-49.

Association of Official Analytical Chemists, AOAC. (1995). Official methods of analysis. 16

th edn. published by the

A.O.A.C, Washington, D.C., USA.

http://www.ncbi.nlm.nih.gov/pubmed?term=Panda%20AK%5BAuthor%5D&cauthor=true&cauthor_uid=18836906






http://www.ingentaconnect.com/content/stl/abr;jsessionid=l4dee6wf84y8.victoria

77


Attia, Y. A., Burke, W. H. and Yamani, K. A. (1994). Response of

broiler breeder hens to forced molting by hormonal and dietary

manipulations. Poultry Sci. 73:245-258.

Attia, Y. A., Burke, W.H., Yamani, K. A. and Jensen, L. S. (1995).

Energy allotments and performance of broiler breeders. 2-

Females. Poultry Sci.74: 261-270.

Attia, Y. A., Böhmer Barbara, M. and Roth-Maier, Dora, A.

(2006). Responses of broiler chicks raised under constant

relatively high ambient temperature to enzymes, amino acid

supplementations, or diet density. Archiv Für Geflügelkunde

70: 80-91.

Attia, Y. A., Hassan, R. A. and Qota, E. M.A (2009). Recovery

from adverse effects of heat stress on slow-growing chicks in

the tropics 1: Effect of ascorbic acid and different levels of

betaine. Tropical Animal health and Production, 41:807-818.

Attia, Y. A., Hassan, R. A., Shehatta, M. H. and Abd El-Hady, S.

B. (2005). Growth, carcass quality and blood serum

constituents of slow growth chicks as affected by betaine

additions to diets containing 2. Different levels of methionine,

International Journal of Poultry Science, 11, 856–865.

Attia, Y. A, Hassan, R. A., Tag El-Din, A. E. and Abou- Shehema,

B. M. (2011). Effect of ascorbic acid or increasing

metabolizable energy level with or without supplementation of

some essential amino acids on productive and physiological

traits of slow-growing chicks exposed to chronic heat stress.

Journal of Animal Physiology and Animal Nutrition, 95:744-

755.

Bollengier-Lee, S., Mitchell, M. A., Utomo, D.B., Williams, P.E.

(1998). Influence of high dietary vitamin E supplementation

on egg production and plasma characteristics in hens subjected

to heat stress. Br. Poult. Sci. 39: 106-112.

Bollengier-Lee, S., Williams P.E.V. and Whitehead, C. C. (1999). Optimal dietary concentration of vitamin E for alleviating the

effect of heat stress on egg production in laying hens. Br.

Poult. Sci. 40: 102-107.

Çiftci, M., Nihat -Ertas, O. and Guler, T.(2005). Effects of vitamin

E and vitamin C dietary supplementation on egg production

77


and egg quality of laying hens exposed to a chronic heat

stress.Vet.,156: 107-111.

Daghir, N.J. (2008). Poultry Production in Hot Climates, 2nd Edition,

Published by CAB International, Wallinford, Oxfordshire, UK,

pp.387.

Eilers, R. J., (1967). Notification of final adoption of an international

method and standard solution for hemoglo-binometry

specifications for preparation of a standard solution. American

Journal of Clinical Pathology, 47: 212–214

Esteve-Garcia, E. and Mack, S. (2000). The effect of DL-

methionine and betaine on growth performance and carcass

characteristics in broilers, Animal Feed Science and Tech-

nology, 87, 85–93. doi:10.1016/S0377-8401(00)00174-7

Franco-Jimenez, D. J., Scheideler, S. E. , Kittok, R. J. , Brown-

Brandl, T. M., Robeson, L. R., Taira, H., and Beck, M. M.

(2007). Differential effects of heat stress in three strains of

laying hens. J. Appl. Poult. Res. 16:628–634.

Gonzalez-Vega-Aguirre, D., Conteras, B.P.A., Klein, R.,

Bohmwald, H.(1995) .Effect of vitamin C and E

supplementation in the diet of broiler chicks on performance

and immune response.Veterinaria., 1995, 26 : 333-340.

Graham, H. (2002). Betaine-Combating heat stress in poultry, Afma

Matrix, December, 15, 16–17.

Hassan, R.A., Attia, Y. A. and El-Ganzory, E. H. (2005).Growth,

carcass quality, and blood serum constituents of slow growth

chicks as affected by betaine additions to diets containing 1.

Different levels of choline, International Journal of Poultry

Science, 4, 840–850.

Hepler, O.E. (1966). Manual of Clinical Laboratory Method.

Thomas: Springfield, Illinois.

Kawahara, E., Ueda, T.and Nomura S. (1991). In vitro phagocytic

activity of white-spotted shark cells after injection with

Aermonas salmonicida extra cellular products. Gyobyo

Kenkyu, Japan, 26: 213 - 214.

Kettunen, H., Tiihonen, K., Peuranen, S., Saarinen, M.T. and

Remus, J. C. (2001). Dietary betaine accumulates in the liver

and intestinal tissue and stabilizes the intestinal epithelial

structure in healthy and coccidia-infected broiler chicks,

77


Comparative Biochemistry and Physiology, A 130,759–769.

doi:10.1016/S1095-6433(01)00410-X

Kidd, M. T., Ferket, P. R. and Garlich, J. D. (1997). Nutritional

and osmoregulatory functions of betaine. Poultr. Sci. 53:125-

139.

Kirunda, D. F. K., Scheideler, S.E. and Mckee, Sr. (2001). The

efficacy of vitamin E (Dl-Alpha-tocophery acetate)

supplementation in hen diets to alleviate egg quality

deterioration associated with high temperature exposure. Poult.

Sci., 80: 1378-1383.

Lin, H., Jiao, H. C., Buyse, J. and Decuypere, E. (2006). Strategies

for preventing heat stress in poultry, World’s Poultry Science

Journal, 62, 71–85.doi:10.1079/WPS200585

Lucas, A. M., Jamroz, C. (1961). Atlas of Avian Hematology.

Washington, United States Department of Agriculture,

Agriculture Monograph25,271p.

Mashaly, M. M., Hendricks, G. L., Kalama, M. A. , Gehad, A. E.,

Abbas, A. O., and Patterson P. H. (2004). Effect of heat

stress on production parameters and immune responses of

commercial laying hens. Poult. Sci. 83:889–894.

Metwally, M. A. (2003). Effect of vitamin E on the performance of

Dandarawi hens exposed to heat stress. Egypt. Poultry Sci. J.

23: 115-127.

Metwally, M. A. (2005). Efficacy of different dietary levels of

antioxidants vitamin C, E and their combination on the

performance of heat-stressed laying hens. Egypt Poult. Sci. J.,

25:613-636.

Miles, R. D., Ruiz, N.and Harms, R.H. (1987). Dietary conditions

necessary for methyl donor response in broiler and laying hens

fed a corn - soybean meal diet.Poult.Sci.,66:29(abst).

Mohiti-Asli, M., Shariatmadari, F. and Lotfollahian, H. (2010). The influence of dietary vitamin E and selenium on egg

production parameters, serum and yolk cholesterol and

antibody response of laying hen exposed to high

environmental temperature. Arch. Geflügelk., 74: 43–50.

National Research Council, NRC, (1994). Nutrient requirements of

poultry. 9th rev.

edn. National academy press, Washington, DC., USA.

77


Oguntunji, O.M. and Alabi, A.O. (2010). Influence of high

environmental temperature on egg production and shell

quality: a review World's Poult.

Sci. J., 66:739- 74

Panda, A. K., Ramarao, S. V., Raju, M. V. L. N. and Chatterjee,

R. N. (2008). Effect of dietary supplementation with vitamins

E and C on production performance, immune responses and

antioxidant status of White Leghorn layers under tropical

summer conditions. Brit Poul Sci.,49 :592-599.

Pardue, S. I., Thaxton, J. P. and Brake, J. T. (1985). Influence of

supplemental ascorbic acid on broiler performance following

exposure to high temperature, Poultry Science, 64,1334–1338

Petronini, P.G., Deanglis, E.M., Borghetti, P. and Borghetti,

A.F.(1992). Modulationby betaine of cellular response

to osmotic stress. J. Biochem, 282:69-73.

Puthpongsiriporn, U., Scheideler, S.E., Sell, J.L. and Beck, M.M.

(2001). Effect of vitamin E and C supplementation on

performance, in vitro lymphocyte proliferation and

antioxidant status of laying hens during heat stress. Poult. Sci.,

80: 1190-1200.

Remus, J. C., Pierson, E. E. M. and Hruby, M. (2004). Theevaluation of betaine and enzymes in coccidian challenged

broilers. XXII Poultry Congress, Istanbul, Turkey 8-13 June,

2004.

Retsky, K. L. and Frei, B. (1995). Vitamin C prevents metal

iondependent initiation and propagation of lipid peroxidation

in human low-density lipoprotein. Biochem. Biophys.

Acta.1257:279–287.

Rozenboim, I., Tako, E., Gal-Garber, O., Proudman, J. A. and

Uni, Z. (2007). The effect of heat stress on ovarian function of

laying hens. Poult. Sci., 86:1760–1765.

SAS Institute (1996). SAS User's Guide. Version 6.12. SAS, Institute

Cary, NC.

Sahin K.; Sahin N. and Yaralioglu, S. (2002) . Effects of vitamin C

and vitamin E on lipid peroxidation, blood serum metabolites,

and mineral concentrations of laying hens reared at high

ambient temperature. Biol Trace Elem Res.,85(1):35-45.

http://www.researchgate.net/researcher/60017408_OGUNTUNJI

http://www.researchgate.net/researcher/66265370_OM_AO_AND_ALABI

http://www.researchgate.net/researcher/39963576_A_K_Panda

http://www.researchgate.net/researcher/14971441_S_V_Ramarao

http://www.researchgate.net/researcher/39121143_M_V_L_N_Raju

http://www.researchgate.net/researcher/14971442_R_N_Chatterjee

http://www.researchgate.net/journal/1466-1799_British_Poultry_Science

http://www.ncbi.nlm.nih.gov/pubmed?term=Sahin%20K%5BAuthor%5D&cauthor=true&cauthor_uid=11881797

http://www.ncbi.nlm.nih.gov/pubmed?term=Sahin%20N%5BAuthor%5D&cauthor=true&cauthor_uid=11881797

http://www.ncbi.nlm.nih.gov/pubmed?term=Yaralioglu%20S%5BAuthor%5D&cauthor=true&cauthor_uid=11881797

http://www.ncbi.nlm.nih.gov/pubmed/11881797

73


Saunderson, C. L. and Mackinlay, J. (1990). Changes in body

weight, composition and hepatic enzyme activities in response

to dietary methionine, betaine, and choline levels in growing

chicks. Br. Poult.Sci.,31:339-349.

Schaefer, D. M., Liu, Q., Faustman, C. and Yin, M. C. (1995). Supranutritional administration of vitamin E and C improves

oxidative stability of beef. J. Nutr. 125(Suppl. 6):S1792–

S1798.

Schrama, J. W. and Gerrits, W. J. J. (2000). Effect of dietary

betaine supplementation on energy partitioning in pigs.

Internal report, wagening en Agricultura. University, the

Netherlands 24P.

Tengerdy, R. P. (1989). Vitamin E, immune response and disease

resistance. Ann. N.Y. Acad. Sci., 57: 335-344.

Tumová E.and Gous R. M. (2012).Interaction of hen production

type, age, and temperature on laying pattern and egg quality.

Poult Sci., 91:1269-1275.

Türker, M., Alp, M. and Kocabacli, N. (2004). Performance of

broiler chicks fed on reduced methionine diets supplemented

with betaine, XXII Poultry Congress, Istanbul,Turkey8–

13June,2004.

Virtanen, E. and Rosi, L. (1995). Effects of betaine on methionine

requirement of broilers under various environmental

condition. Proc. of the Austral. Poult.

Sci.Symp.,Sydney,pp:88-92 .

Wang, Y. Z., Xu, Z. R. and Feng, G. (2004). The effect of betaine

and DL- methionine on growth performance and carcass

characteristics in meat ducks,

Animal Feed Scienc and Technology, 116, 151–159.

Yin, M. C., Faustman, C. Riesen, J. W. and Williams, S. N. (1993). Alpha- tocopherol and ascorbate delay oxymyoglobin and

phospholipid oxidation in vitro.J.FoodSci.58:1273–1276.

Yoshida, N., Fujita, M., Nakahara, M., Kuwahara, T., Kawakami,

S. I. and Bungo, T. (2011). Effect of high environmental

temperature on egg production, serum lipoproteins and follicle

steroid hormones in laying hens. J. Poult. Sci., 48: 207-211

http://www.ncbi.nlm.nih.gov/pubmed?term=Tumov%C3%A1%20E%5BAuthor%5D&cauthor=true&cauthor_uid=22499888

http://www.ncbi.nlm.nih.gov/pubmed?term=Gous%20RM%5BAuthor%5D&cauthor=true&cauthor_uid=22499888

http://www.ncbi.nlm.nih.gov/pubmed/22499888

77


الملخص العربى

و بدون اثنين من مضادات االكسدة على اداء أاستخدام البيتايين مع تأثير الحرارى لإلجهادالغرض المعرض ثنائيالدجاج

*يوسف عطية ، احمد عبد العزيز عبد هللا* ، مرفت بريقع*، عبد الحميد السيد ، بهاء ابو شحيمة

مصز -جامعة دمنهىر –عة كلية الزرا –قسم االنتاج الحيىانى والدجنى

مصز –مزكز البحىث الزراعية –معهد بحىث االنتاج الحيىانى –* قسم تغذية الدواجن

تم دراسة قدرة كال من البيتايين وفيتامين ج وفيتامين هـ والخلطات الممكنة بينها على تحسين اجات ثنائية الغرض صفات االداء االنتاجى والتناسلى وجودة البيض والصفات الفسيولوجية للدج

222تم تسكين عدد 0اسبوع من العمر 36الى 32المعرضة لالجهاد الحرارى خالل الفترة من ديك من ساللة المندرة ثنائية الغرض فى عنبر محكم من ناحية الظروف البئية 36دجاجة ،

شوش وزعت الع .م ( وفرشت بفرشة من التبن x1 x2 2عشة ارضية ) 36ووزعت عشوائيا الى دجاجات 2عشوش كل منها يحتوى على 4معامالت تجربية بحيث تحتوى كل معاملة على 9على .وديك

% رطوبة نسبية 60 - 55م ، 24o - 22المجموعة االولى تم تربيتها تحت درجة حرارة بينما 0واستخدمت ككنترول ايجابى وغذيت على عليقة اساسية بدون اضافة االضافات التجربية

نية مجاميع االخرى تم تربيتها تحت ظروف االجهاد الحرارى حيث عرضت لدرجة حرارة الثما32o ، 0% رطوبة نسبية وذلك لمدة ثالث ايام متتالية من كل اسبوع خالل فترة التجربة65م

المجموعة االولى من المجاميع المعرضة لالجهاد الحرارى غذيت على العليقة االساسية بدون التجربية واستخدمت ككنترول سلبى بينما باقى المجاميع تم تغذيتها على العليقة اضافة االضافات

200كجم علف او فيتامين ج بمعدل /ملليجرام 1000االساسية مضافا اليها البيتايين بمعدل 0كجم علف او الخلطات الممكنة منها/ملليجرام 150كجم علف او فيتامين هـ بمعدل /ملليجرام

رارى الى اثار سلبية على االداء االنتاجى وصفات جودة البيض الخارجية والداخلية أدى االجهاد الحوالخصوبة ووزن الكتكوت عند عمر يوم وخصائص كرات الدم الحمراء ودرجة حموضة الدم

فى المقابل أدى اضافة كالمن فيتامين ج اوالبيتايين منفردا الى تحسين االداء 0والمقايس المناعيةجاجات البياضة وعلى اية حال فان البيتايين لة قدرة اكبر فى تحسين الكثافة النوعية االنتاجى للد

من ناحية اخرى فان فيتامين 0للبيضة ووحدات هيو وصفات الدم الهيماتولوجية والمقايس المناعية 0نتاجىهـ كان لة تأثير كبير على نسبة الهيتيروفيل بينما أعطى اقل النتائج بالنسبة لصفات االداء اال

-الخالصة :كجم علف( لعلف /ملليجرام 1000كجم علف( او البيتايين ) /ملليجرام 200اضافة فيتامين ج )

يؤدى الى 0% رطوبة نسبية (65م ، 32oالدجاجات ثنائية الغرض المعرضة لالجهاد الحرارى )لذلك 0س المناعيةتحسن ادائها االنتاجى كما ان البيتايين يحسن كذلك صفات جودة البيض والمقاي

فانة يمكن استخدام البيتاين منفردا كمضاد لالجهاد الحرارى للدجاجات ثنائية الغرض.

77


Table 2: Productive performance of Mandara laying hens as affected by heat stress and addition of Betaine with or

without Vitamin: C and Vitamin: E supplementation (mean± SEM).

abc Means within a row not sharing a common a superscripts differ significantly (P<0.05); ND, not done.

SEM= Standard error of means, P value= Probability level.

P

Value

SEM

Heat stress treatments

Control

( + )

Criteria + Bet

+ Vit C

+ Vit E

+ Vit C

+ Vit E

+ Bet

+ Vit E

+ Bet

+ Vit C

+Vit E

+Vit C

+Bet

( - )

0.0001 0.747 126cd

130b

127cd

129bc

128cd

125 d

127cd

121e

133a

Feed intake, g/h/d

0.0001 1.35 63.0de

66.3b

64.9bc

63.5cd

61.8e

66.4b

64.3cd

57.5f

70.0a

Laying rate,%

0.0026 0.433 46.9abc

47.1abc

46.2bc

46.8abc

46.2bc

47.8abc

48.1ab

45.5c

48.5a

Egg weight, g

0.0001 0.363 29.5de

31.2bc

30.0cde

29.7de

28.6e

31.7b

30.9bcd

26.4f

33.9a

Egg mass, g/h/d

0.0001 0.052 4.25cd

4.18cd

4.23cd

4.34bc

4.46ab

3.93e

4.10de

4.61a

3.92e

FCR, g feed/g egg

ND ND (32/0) (32/0) (32/0) (32/0) (32/1) (32/0) (32/0) (32/2) (32/0) Survival rate

888


Table 3: External egg quality traits of Mandara laying hens as affected by heat stress and addition of Betaine with or

P

Value

SEM


Control

( + )

Criteria + Bet

+ Vit C

+ Vit E

+ Vit C

+ Vit E

+ Bet

+ Vit E

+ Bet

+ Vit C

+Vit E

+Vit C

+Bet

( - )

0.131 1.83 81.4 76.4 77.6 73.5 77.3 75.5 75.5 75.1 77.9 Egg shape index, %

888



abc Means within a row not sharing a common a superscripts differ significantly (P<0.05).

1*SWUSA = shell weight per unit of surface area; SEM= Standard error of means, P value= Probability level.

0.0001 0.0001 1.092a

1.088ab

1.092a

1.087ab

1.091a

1.085b

1.093a

1.078c

1.091a Specific gravity,

g/cm3

0.164 0.147 5.38 5.13 5.18 5.34 5.40 5.23 5.58 4.93 5.41 Shell weight, g

0.001 0.127 11.0a

11.1a

11.2a

10.8a

11.0a

10.9a

11.1a

10.4b

10.7ab

Shell weight, %

0.062 0.001 38.0a

36.9ab

36.9ab

36.1ab

36.0ab

37.4ab

36.8ab

34.0b

38.4a Shell thickness,

mm

0.015 1.20 88.5a

87.8a

89.0a

87.5a

88.9a

87.3a

90.0a

83.0b

87.3a

SWUSA1, mg/cm

2

888


Table 4: Internal egg quality traits of Mandara laying hens as affected by heat stress and addition of Betaine with or


P

Value

SEM


Control

( + )

Criteria + Bet

+ Vit C

+ Vit E

+ Vit C

+ Vit E

+ Bet

+ Vit E

+ Bet

+ Vit C

+Vit E

+Vit C

+Bet

( - )

887




0.004 0.279 30.7ab

31.7a

31.6a

31.1ab

30.9ab

30.9ab

30.9ab

30.0b

31.1ab

Yolk weight, %

0.0001 0.316 58.4b

57.2b

57.1b

58.3b

58.1b

58.2b

58.0b

59.6a

58.2b

Albumin weight, %

0.0007 0.755 52.6ab

55.5a

55.4a

53.5ab

53.3ab

53.0ab

53.4ab

50.4b

53.4ab

Yolk albumin ratio

0.002 0.337 15.9ab

15.9ab

16.8a

16.8a

16.1ab

16.7a

16.4a

14.9b

16.4a

Yolk dry matter, %

0.092 0.217 7.58ab

7.69ab

7.78ab

7.56ab

7.67ab

7.74ab

8.14a

7.09b

8.07a Albumin dry matter,

%

0.0001 1.30 48.1a

46.2a

47.5a

48.8a

48.4a

47.5a

43.5a

36.3b

48.5a

Yolk index, %

0.0005 0.292 5.67a

5.67a

5.36a

6.11a

5.78a

5.56a

6.00a

4.44b

6.56a

Yolk color

0.0001 4.41 65.8b 65.0

b 67.0

b 57.4

b 74.2

b 62.7

b 89.7

a 43.6

c 86.1

a Haugh unit score

887


Table 5: Hatchability traits of Mandara laying hens as affected by heat stress and addition of Betaine with or


P

Value

SEM


Control

( + )

Criteria + Bet

+ Vit C

+ Vit E

+ Vit C

+ Vit E

+ Bet

+ Vit E

+ Bet

+ Vit C

+Vit E

+Vit C

+Bet

( - )

0.0001 1.24 97.0a

95.9a

95.6a

95.9a

96.9a

95.2a

96.5a

80.2b

96.6a

Fertility, %

0.0659 1.95 83.8 82.5 84.6 82.5 83.0 81.1 83.4 89.1 83.4 Hatchability of

total eggs, %

887




0.001 1.57 86.4 86.0 88.5 86.0 85.7 85.2 86.4 90.0 86.3 Hatchability of

fertile eggs, %

0.3670 1.12 4.77 6.49 6.14 5.12 5.81 6.48 6.13 3.40 4.44 Dead, %

0.1140 1.64 10.88 10.56 8.52 11.60 10.60 11.58 9.91 5.43 11.57 Piped, %

0.1043 0.371 35.2 35.2 35.1 34.3 35.2 34.8 35.0 34.1 35.6 Chick weight, g

887


Table 6: Blood heamatological parameter of Mandara laying hens as affected by heat stress and addition of Betaine with or


P Heat stress treatments Criteria

883



RBC=Red blood cells, Hgb=Haemoglobin, PCV=Packed-cell volume, WBCs =White blood cell, H/L ratio= Heterophil / lymphocyte ratio, PA=

Phagocytic activity, PI= Phagocytic index; SEM= Standard error of means, P value= Probability level.

Value

SEM

+ Bet

+ Vit C

+ Vit E

+ Vit C

+ Vit E

+ Bet

+ Vit E

+ Bet

+ Vit C

+Vit E

+Vit C

+Bet

( - )

Control

( + )

0.0001 0.067 1.47a

1.58a

1.30a

1.38a

1.40a

1.58a

1.55a

1.07b

1.60a

RBCs, x106/mm

3

0.0007 0.390 9.67ab

10.33a

10.67a

11.00a

9.50ab

9.50ab

9.50ab

8.67b

11.17a

Hgb, g/dl

0.0007 0.805 31.5a

31.7a 32.7

a 32.3

a 29.7

ab 29.8

ab 29.7

ab 27. 3

b 32. 7

a PCV, %

0.0001 0.470 22.2bc

24.5a

21.3c

24.5a

22.2bc

23.7ab

21.5c

20.5c

24.7a

WBCs, x103/mm

3

0.0001 0.540 44.00ab

41.70cd

45.30a

41.80cd

41.50cd

42.64bc

44.80a

40.20d

45.00a

Lymphocyte, %

0.192 0.432 7.67 6.67 7.50 7.83 6.67 7.83 7.33 6.33 7.33 Monocyte, %

0.076 0.190 0.360 0.800 0.370 0.800 0.500 0.500 0.833 0.667 0.333 Basophil, %

0.0030 0.508 7.67ab

8.33ab

7.83ab

8.67ab

9.17ab

8.83ab

7.00b

9.67a

9.83a

Eosinophil, %

0.007 1.18 16.3ab

20.3a

14.7b

16.1b

19.7ab

15.5ab

15.2ab

18.2ab

16.2ab

Neutrophil, %

0.0001 0.473 24.0abc

22.8bcd

24.3ab

24.8a

22.5cd

24.7a

24.8a

25.0a

21.3d

Heterophil, %

0.0001 1.19 57.2b

58.8b

57.2b

58.5b

58.3b

57.5b

58.0b

64.5a

50.1c

H/L ratio

0.0001 0.017 7.76b

7.72b

7.72b

7.71b 7.72

b 7.65

c 7.62

c 7.84

a 7.56

d Blood pH

0.0006 0.593 17.50abc

16.80bc

19.50a

17.70abc

18.70ab

18.70ab

18.80ab

15.70c

19.70a

PA, %

0.0001 0.053 1.33b

1.35b

1.52ab

1.40b

1.48ab

1.50ab

1.65a

1.13c

1.65a

PI, %

Documents

EFFECT OF BETAINE WITH OR WITHOUT TWO ...plasma in White Leghorn hens during HS and that supplementation of 1000 ppm Vit C may further enhance in vitro lymphocyte proliferative responses