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336 EXTRA DIETARY CALCIUM studies showed that hatchability [lo] and embryo viability [ll] were si&icantly lower in fust-of-sequence eggs than in subsequent eggs within a laying sequence

Trang 1

EFFECTS OF OVIPOSITION TIME,

Instituto Superior de Agronomia, 1399 Lisboa Codex, Portugal

Phone: (01) 3638161

F M : (01) 3635031

Primarv Audience: Breeder Production Managers Researchers

Eggshell quality as measured by egg

specific gravity (SG) has frequently been

associated with hatchability of eggs from

broiler breeder hens [l, 2,3,4,5,6] Research

indicates that a SG of 1.080 is the dividing

point between good and poor hatchability

breeder hens fed in the afternoon exhibit a

sig&cant improvement in eggshell quality as

measured by SG and eggshell weight [8, 91

Reis et al [6] also observed that coarse lime-

stone supplied in the afternoon slightly in- creased the SG of broiler breeder eggs, but markedly improved embryo viability

The literature also indicates that SG of broiler breeder eggs increases as time of laying becomes later in the day [l, 21, although hatch- ability of eggs laid in the afternoon is not significantly greater than that of eggs laid

during the morning [l] However, further

Trang 2

336 EXTRA DIETARY CALCIUM

studies showed that hatchability [lo] and

embryo viability [ll] were si&icantly lower

in fust-of-sequence eggs than in subsequent

eggs within a laying sequence The fust egg of

a sequence is usually laid relatively early in

the day and the remainder laid later on each

following day; therefore, a high proportion of

eggs laid in the early morning of any given

day should be first-of-sequence eggs [12] In

addition, sequence length in feed-restricted

broiler breeder hens reaches a prime mean

length at 32 wk of age, then quickly shortens

with advancing hen age [13]; therefore, the

incidence of first-of-sequence eggs increases

with hen age

We conducted the present study to deter-

mine the effects of oviposition time, hen’s age,

and extra dietary calcium on egg weight (EW),

SG, EW loss during the first 18 days of in-

cubation, fertility, embryo viability, age of em-

bryonic death in fertile eggs, and hatchability

of eggs from feed-restricted broiler breeder

hens under commercial conditions Specific-

ally, we wished to ascertain whether the effect

of calcium supplementation varied with hen

age by affecting SG and with time of oviposi-

tion through interaction with time of calcium

absorption

SOURCE OF EGGS

The study was carried out over two pe-

riods, from November to December 1994 and

from March to April of the following year,

using eggs from one commercial flock of ap-

proximately 13,900 broiler breeder hens

(Cobb 500) at 46 and 65 wk of age Equal

numbers of birds were arbitrarily assigned to

each of two identical pens of one building at

effects of two treatments: no extra calcium

supplementation (NC) and extra calcium

supplementation (CS) Natural daylight was

supplemented with artificial light to give a

16 hr photoperiod (on 0790 hr, off 23:00 hr)

Daily low and high in-house air temperatures

ranged from 17 to 24°C and 16 to 24°C at ages

46 and 65 wk, respectively

All birds received a commercial broiler

breeder diet (16.50% CP, 2800 kcal ME,/kg,

formulated to meet or exceed National Re-

search Council [14] requirements, as pre-

viously described [6] Both sexes received the

same feed in separate sex feedings [15], as

diets used by the breeder hen appear to have

no detrimental effects on male performance

Hens received approximately 163 and 157 g

of feed per bird at 46 and 65 wk of age re- spectively, to maintain body weights recom-

mended by the primary breeder [16] The

hens had consumed their feed allowance by

approximately 11:OO hr each day A birds

consumed water ad libitum In the pen as- signed to CS treatment, coarse limestone

(4 g/bird daily) was scattered on the litter at

wood shavings (eucalyptus bark), so that white limestone particles were very visible; the birds rapidly consumed them The Limestone

supplement had particles ranging from 2 to

5 mm in diameter, with an average solubility of

described by Keshavarz et al [lq Extra di- etary calcium supplement was provided for

8 consecutive days at both 46 and 65 wk of flock age Calcium distribution began 3 days

before the date of first egg collection for

experimental purposes (Table 1)

EGG COLLECTION AND HANDLING

At 46 and 65 wk of age eggs were collected for 5 and 4 days, respectively (Table l), at 2 hr

intervals beginning at 08:OO hr and ending

at 18:OO hr Eggs laid before 08:OO hr were

removed from the nests and were not used

in this experiment Eggs that were cracked, very dirty, misshapen, or of extreme size were culled From each pen and collection

time, a random sample of 150 hatching eggs

was obtained and eggs were then individually marked Eggs from each day’s production

reached the hatchery at 12:45, 16:45, and

weighed to the nearest 0.1 g, and SG was de-

termined in graded sodium chloride solutions kept at room temperature (about 20°C) and

ranging from 1.060 to 1.090, with an increment

of 0.005 units between successive solutions

The concentration in which the egg barely floated beneath the surface indicated egg

SG Eggs that did not float in any of the so-

lutions were assigned a SG of 1.095, and those

that floated in all solutions were assigned a

SG of 1.060 After SG measurement, the eggs were rinsed in warm tap water and dried in a stream of warm air at approximately 20°C

Trang 3

NOVO et al

November 23

November 24

November 25

Field Report

337

TABLE 1 Egg collection and calcium supplementation for broiler breeders at two different periods (46 and

Eggs from each treatment and collection

time were then placed in setter trays

(150 eggshay)

INCUBATION AND HATCHING

Incubation was initiated at 2200 hr on the

day of egg collection in electronically con-

trolled single-stage incubators (Petersime

576) The ten experimental trays correspond-

ing to five different collection times and two

treatments were randomly placed in one

quadrant (front right) on the same trolley,

which was then filled with other eggs Quad-

rant and trolley positions were the same on all

9 collection days to reduce possible position

effects AU eggs were fumigated in the in-

cubator at incubation temperature on the

day of setting, using 20 g of potassium perman-

ganate and 30 mL formalin (400 g/L) per m3

incubator space with the ventilators closed for

20 min Eggs were automatically turned 90"

every hour and incubated at conventional

temperature and humidity conditions, as pre-

viously described by Roque and Soares [5]

The eggs were individually candled on the

18th day of incubation, using a hand candling

lamp Infertile eggs and eggs containing early

dead embryos were removed and broken out

for macroscopic examination The eggs with

apparently living embryos were individually

weighed to the nearest 0.1 g Weight loss was

calculated by difference and expressed in ab-

solute terms (g) and as a percentage of initial

fresh egg weight The eggs were quickly trans-

ferred to hatcher baskets, randomly distrib-

uted in the front part of the same trolley, and

placed in a hatcher (Petersime 192) operated

at conventional conditions, as outlined by Roque and Soares [5] Chicks and unhatched eggs were taken off at 21 days and 12 hr postincubation

Saleable hatched chicks and culls (un- marketable chicks) were recorded All eggs removed at candling, and unhatched eggs at the time of taking off the hatch, were broken out, examined macroscopically, and assigned

to one of the following categories: infertiles, early-dead germs (embryo deaths during 1 to

7 days of incubation), mid-dead germs (deaths during 8 to 18 days), late-dead germs (deaths after 18 days), pipped (beak penetrated eggshell but chick did not emerge), and rots (contaminated) From the data, percent hatchability (number of saleable chicks

hatched per all eggs set x 100) and percent

embryo viability (number of saleable chicks

hatched per number of fertile eggs set x 100)

were calculated A total of 7500 eggs were

used in the fust period (150 eggs per collection

time x 5 daily collection times x 5 days x 2 treatments), and 6000 eggs were used in the second period (4 days)

Data were statistically analyzed by analysis of variance [18,19,20]

The interaction between oviposition time and calcium supplementation was significant

only for embryo mortality up to 7 days and

contaminated eggs (Table 2); thus, it seems that the effect of oviposition time was basically the same for both treatments On the other

Trang 4

338

VARIABLE

JAPR

EXTRA DIETARY CALCIUM

SOURCES OF VARIATION Treatment Breeder Oviposition T X A O X T O X A

0 Age (A) Time (0)

TABLE 2 Significance of different sources of variation in the preliminary analysis

Egg specific gravity

Egg weight at set, g

Fertility, %

Hatchability, %

NS

NS

* * * .** * t

* * ** I * * *.*

* * *

NS

* * *

EmbIyo viability, % NS

* P < .os; * *P < 01; ***P < 001

hand, the interaction between oviposition time

and age of the hen was signifcant not only

for early mortality and contaminated eggs but

also for egg weight loss, embryo viability,

and hatchability Interaction of hen age and

calcium treatment significantly affected all

traits except intermediate and late embryonic

mortality and contaminated eggs, which sug-

gests that the effect of calcium supplementa-

tion differs according to flock age

INFLUENCE OF OVIPOSITION TIME

The effects of oviposition time, expressed

as the linear regression coefficients of the vari-

ables analyzed on time of oviposition, are sum-

marized in Table 3, either within treatment or

age, or pooled, depending on interaction sig-

nificance These results clearly demonstrate

that regardless of hen age, EW decreases

(P < Ol) gradually as time of laying becomes

later in the day These data substantiate previ-

ous findings that eggs laid in the morning are

heavier than eggs laid during the later periods

of the day [12, 21, 22, 23, 241 Research in-

dicates that this variation in EW with time of

laying is related to the sequence of eggs in the

ovulatory cycle, but not directly related to the

time of oviposition [12] However, under usual

photoperiodic conditions, it is likely that early

morning ovipositions would include most of the first-of-sequence eggs [12], which are heavier than subsequent eggs [lo, 21,251 This probably occurs because an ovarian follicle destined to become the first egg in a clutch

resides as the largest follicle (F1) on the ovary

for about 16 hr longer than mature FI follicles

of subsequent eggs in the same sequence [26]

Such a long period of follicular growth is presumably associated with a greater yolk deposition, as it occurs in the ovary [27

Egg specific gravity increased signifi- cantly (P< 001) during the day at both ages, with a slightly higher increase in older hens (linear regression coefficients on time were

O.OOO4 and O.OOO5 at 46 and 65 wk of age, re- spectively) These findings are in general agreement with previous studies involving laying hens [22, 28, 29, 30, 311 and broiler breeder hens [l, 21, which showed that eggs laid in the afternoon have an average SG greater than eggs laid in the morning The reason for this variation is not clear This in- crease in SG may be due to a more adequate calcium carbonate supply directly from the feed administrated in the morning Eggshell calcification of eggs laid in the afternoon oc- curs in part when the hen has access to feed, whereas in eggs laid early in the morning the

Trang 5

Field Report

NOVO et al

Embryo viability, 70 46 wk

65 wk

339

0.9247 0.0023 +0.0013 NS

* * *

0.8344 -0.0075f0.0018

aDo = Intercept

%,+sb = Linear regression coefficient f linear regression coefficient standard error

D ~ NO ~extra calcium supplementation =

= With extra calcium supplementation

'Pi.05; **P<.Ol; ***P<.OOl

calcium carbonate of the shell is formed at

night when the hen is not eating This hypoth-

esis is also supported by the work of Farmer et

fed at 16:OO hr had significantly better eggshell

quality (SG) than buds fed at 0530 hr

Flock age affected the relationship of ovi-

position time to egg weight loss during the first

18 days of incubation For hens 46 wk of age,

absolute and relative egg weight loss de-

creased significantly with later laying time

For hens at 65 wk of age no significant changes

for different laying times were observed when

weight loss was expressed in absolute terms;

however, weight loss as a percentage of fresh egg weight increased significantly for eggs laid later in the day The results obtained at 65 wk

of age are consistent with the findings of

Robinson et al [lo], who used eggs from broiler breeders 45 to 53 wk of age and ob-

served that eggs laid later in a sequence lost significantly more weight during the 7-day incubation period than did first-of-sequence eggs This relationship of weight loss to ovi- position time may not be attributable to differ- ences in shell quality, since SG increased through the day (Table 3), and previous studies showed that eggs with the highest SG

Trang 6

340 EXTRA DIETARY CALCIUM

had the lowest weight loss [2,5] Present data

suggest that besides SG, other egg compo-

nents may also play an important role in water

loss during incubation The work of Meur and

Baumann [32] indicates viscous quality of

albumen during the initial stages of incubation

as one possibility

At both ages, oviposition time did not

sigruficantly influence fertility, intermediate

or late mortality, or percentage of pips In

their work with broiler breeders housed in

individual laying cages, Robinson et al [lo]

and Fasenko et al [ 111 had similar fmdings and

reported that fertility was not significantly

different between first-of-sequence and sub-

sequent eggs

At the younger age, oviposition time did

not significantly influence early embryonic

mortality, embryo viability, percentage of

contaminated eggs, or hatchability These

results are in general agreement with the

work of McDaniel et al [2], who suggested

that in eggs with SG higher than 1.080 there

appeared to be little relationship between

SG and hatchability In the present study,

younger hens produced eggs with a higher

average SG (1.084) (Table 4) In older hens,

on the other hand, there was a si&icant trend for hatchability and embryo viability

to decrease as oviposition occurred later in the day, mostly because of increased early embryonic mortality and mortality due to infections These results were unexpected, because previous studies [2, 3, 4, 5] have re- ported that thick-shelled eggs had better hatchability than thin-shelled eggs due to their greater fertility and lower embryonic mortal- ity, and that the SG of eggs from older hens increased with oviposition time Concerning the relationship between oviposition time and

embryonic mortality, Robinson et al [lo] re-

ported no significant difference for embryo viability between first (89.70%) and subse- quent (92.75%) eggs in a sequence, and

Fasenko et al [ l l ] found that viability was sig-

nificantly reduced in first-of-sequence eggs compared to subsequent eggs The data from the present research suggest that oviposition time (egg sequence position) affects viability

of fertile eggs in older flocks through higher early embryonic mortality, regardless of shell thickness

TABLE 4 Breeder age x treatment interaction effect on egg specific gravity, egg weight, survival, hatchability,

and embryo viability

B~~ = NO extra calcium supplementation

‘CS = With extra calcium supplementation

Means for the same combination effect and row with different suuescriuts differ at P < .OS

a x

Trang 7

NOVO et al

Field Report

341

As Table 4 shows, average EW increased

with hen age (64.96 vs 67.34 g at 46 and

65 wk of age, respectively) Higher EW is

usually associated with an increase in yolk

deposition [33], as the largest preovulatory

follicle is heavier in older hens [34]

Average specific gravity of eggs was

greater for the eggs from younger hens (1.0840

vs 1.0799) Other studies with broiler breeder

hens [2,4,6] and with layer strain hens [35,36]

have also associated reductions in SG with

increased bird age Shell quality (SG and shell

thickness) declines as the hen ages chiefly be-

cause EW increases with no proportionate in-

crease in shell deposition [36,37l Dysfunction

of the shell gland may also be a contributing

factor [34]

Average weight loss during the first

18 days of incubation was slightly higher in

absolute terms in eggs from older hens (7.17

vs 6.99 gin treatment NC), but relative weight

loss decreased slightly with flock age (10.78

and 10.67% at 46 and 65 wk of age, respec-

tively), probably because of the associated

increase in egg weight These data support the

suggestion of Reis et al [6] that the same in-

cubator humidity might be recommended for

eggs from broiler breeder hens from 46 to

64 wk of age

Fertility, embryo viability, and hatchabil-

ity declined significantly ( P < Ol) with hen

age, phenomena that are well documented in

the literature [6, 11, 38, 39, 401 The present

study’s very high level of early embryonic mor-

tality for eggs from hens 65 wk of age was

unexpected, since lower levels have been re-

ported at similar broiler breeder ages [6, 391

However, embryonic viability per 100 eggs set

(fertility x viability) reported in the present

investigation (91.98 and 72.11% at 46 and

65 wk of age, respectively) is consistent with

findings on eggs from flocks of similar ages

[6, 391 The discrepancy in the relative im-

portance of early embryonic mortality and

infertility may result from difficulties in dis-

tinguishing between infertile blastodiscs and

very early dead blastoderms by macroscopic

examination

EFFECTS O F CALCIUM SUPPLEMEN-

TATION

As shown in Table 4, the effects of extra

calcium fed in the afternoon varied mark-

edly with flock age: there was a significant

treatment x age interaction for most traits

analyzed At 46 wk of age, none of the de- pendent variables were significantly (P < Ol) affected by calcium supplementation, with the exception of SG, which increased slightly

In contrast, at 65 wk of age, calcium supplementation significantly affected all response variables but SG and relative egg weight loss It could be anticipated that cal- cium supplementation would have a higher effect on SG of eggs from the same birds at

an older age, but that was not the case, as was also found by Reis et al [6] Presumably thin- shelled eggs from aged birds are associated with a disorder of vitamin D metabolism, pos- sibly associated with reduced synthesis of

1,25-dihydroxycholecalciferol by the kidney [34,41] and also related to a decreasing ability

to absorb calcium from the intestine and to mobilize skeletal calcium [42] In agreement with Reis et al [6], our results suggest that extra dietary calcium does not increase SG but still improves hatchability This improvement

is probably attributable to both increased fer-

tility and decreased early embryonic mortality

Early embryonic mortality and pipped

eggs were significantly (P c .01) reduced when older hens received extra calcium Therefore,

a significant improvement in embryo viability (5.88%) also occurred in eggs from broiler breeders receiving supplemental calcium at

65 wk of age Results from this experiment supported the previous finding [6] that coarse limestone fed in the afternoon caused an im- provement in embryo viability; however, the improvement observed in this experiment was due to lower early embryonic mortality and pipped eggs In contrast, Reis et al [6] attrib- uted the higher viability to a reduction in con- taminated eggs (the suspected contaminant was Pseudomonas spp)

Calcium supplementation was associated with a slight increase of fresh egg weight in older hens, in agreement with the results

obtained by Bootwalla et al [8] with broiler

breeders, but in contrast to those of Reis et al

[6] with broiler breeders, and Muir et al [43]

and Watkins et al [44] with laying hens In

older hens, extra dietary calcium was also associated with a slight increase of absolute weight loss during incubation, but not with weight loss as a percentage of egg weight at set (Table 4)

Trang 8

342

JAPR

EXTRA DIETARY CALCIUM

At 65 wk of age, the feeding of coarse

limestone scattered on the litter in the

afternoon was associated with a significant

(P < 01) improvement in fertility (93.08 vs

96.37%) This improvement could be due to a

stimulus to heavy males to be sexually active

Spreading of coarse limestone increased the

birds’ activity, which may have led to more females being mated The increased fertility and reduction in embryonic mortality associ- ated with calcium supplementation resulted

in an 8.2% improvement in hatchability for eggs from older hens, whereas no improve- ment was observed in eggs from younger birds

1 Egg weight decreased, specific gravity increased, and fertility was unaffected as time of

laying became later during the day

2 In contrast to younger (46 wk) birds, at the older age (65 wk) hatchability and embryo

viability declined during in eggs laid later in the day

3 Fertility, embryo viability, and hatchability declined with hen age In older hens, early

embryonic mortality was a greater source of overall loss in hatchability than was infertility

4 The most significant effects of extra dietary calcium supplementation were observed at

65 wk of age: egg weight, fertility, embryo viability, and hatchability were improved

1 McDaniel, G.R and D.A Roland, 1977 The influ-

ence of time of oviposition and eggshell quality on hatch-

ability of e from broiler breeder hens Poultry Sci

56:1354 ( A b 7

2 McDaniel, G.R, D.A Roland, Sr., and M.A Cole-

man, 1979 The effect of e shell qualityon hatchability

and embryonic mortality &try Sci 5810-13

3 McDaniel, G.R, J Brake, and M.K Ekman, 1981

Factors affecting broiler breeder performance 4 The

interrelationship of some reproductive traits Poultry Sci

60:1792-1797

4 Bennett, C.D., 1992 The influenceof shell thickness

on hatchability in commercial broiler breeder flocks

J Appl Poultry Res 1:61-65

quali and broiler breeder age on hatchability #%:$

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6 Reis, LH., P Feio, LT Gama, and M.C %ares,

1995 Extra dietary calcium su lement and broiler

breeders J Appl Poultry Res 4:$?&282

7 North, M.O and D.B Bell, 1990 Commercial

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Reinhold, New York, NY

8 Bootwalla, S.M., R H Harms, and H.R Wilson,

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breeders Poultry Sci 61:1421 (Abs)

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5 Roque, L and M.C Soares, 1994 Effects of e

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Poultry Sci 62465471

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BJ Williams, 1991 The influence of egg sequence posi-

tion on fertility, embryo viability, and embryo weight in

broiler breeders Poultry Sci 70760-765

11 Fasenko, G.M., RT Hardin, F.E Robinson, and

J.L Wilson, 1992 Relationship of hen age and egg se-

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12 Choi, J.H., R.D Miles, AS Arafa, and R.H

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60824-828

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The effect of solubility and particle size of calcium sources

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18 Data were analyzed by analysis of covariance with the General Linear Models procedure (PROC GLM) of SAS 1191 Hatchability, embryo viability and sources of mortality =re coded as 0 or 1 and analyzed accordingly,

as indicated by Hawey [20] In a preliminary analysis the linear model included the effects of calcium supplemen- tation and breeder age as discrete independent variables

and time of oviposition as a linear covariate, with 3 two-

way interactions between these factors If an interaction was not statistically significant (P =- .OS) for a given re- sponse variable, it was dropped and the reduced model

run in order to obtain least-squares means and estimates

of regression coefficients on time of oviposition The latter were obtained either as pooled coefficients or

within age or treatment, according to the significance of the interactions

Trang 9

NOVO et al

Field Report

343

Results for total eggs were based on data from 13,500

eggs; results for fertile eggs, on 13,145 eggs

19 SAS Institute, 1985 SAS Users Guide: Statistics

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comparative value of five calcium sources for laying hens

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44 Watkins, RM., B.C Dihvorth, and EJ Day, 1977

Effect of calcium supplement particle size and source

on the performance of laying chickens Poultry Sci

561641-1647

ACKNOWLEDGEMENT

The authors express their appreciation to Valouro Group, Portugal for providingboth facilities and financial support for this study

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