Is it possible to increase the n-3 fatty acid content of eggs without affecting their technological and/or sensorial quality and the laying performance of hens?

1. The aim of this study was to increase the n-3 fatty acid (n-3 FA) content of eggs without affecting their sensorial and/or technological properties or the laying performance of hens. Laying hens from line 477 were divided into 5 groups corresponding to 5 different diets over the laying period: control diet (C) and diets containing extruded linseed with a high level of fibre (ELHF), extruded linseed with a low level of fibre (ELLF), microalgae, or a combination of 75% ELLF and 25% MA (ELLF+MA).

2. Dietary enrichment with n-3 FA had no effect on the laying performance, hen body weight or egg white viscosity. The egg yolks produced by hens fed the diet containing microalgae were redder than egg yolks from the other groups, suggesting the presence of red pigments in the microalgae preparation. However, the colour difference was low and not perceptible to the human eye. Moreover, colour measurement of egg yolks by sensorial analysis panellists using the Roche colour fan did not reveal a diet effect on this parameter.

3. Egg yolk lipid content was not affected by diet. The egg yolk of hens fed on diets containing linseed and/or microalgae had greater n-3 FA content (×2.5 to 2.9 compared to group C). Linseed mainly increased the linolenic acid content (×3.0 to 3.4 compared to group C) and the microalgae increased the LC n-3 FA content (×4.1 compared to group C). Dietary enrichment with n-3 FA had no effect on the sensorial quality of shell cooked eggs except for the “unusual flavour” criterion for which the score was higher for the MA group compared to the other groups and corresponded to a fishy flavour.

     

Egg collagen content is increased by a diet supplemented with wood charcoal powder containing wood vinegar liquid

1. The aims of the present study were to examine whether collagen exists in egg, particularly in egg yolk; to establish a Fourier transform-near infrared (FT-NIR) measurement method for collagen in egg and to assess the possibility of increasing the collagen content by feeding hens a diet containing wood charcoal powder containing wood vinegar liquid (WCV).

2. The collagen in eggs from 67-week-old hens fed on the dietary 0 and 9.9 g/kg WCV diets was investigated using a combination of histochemical, matrix-assisted laser desorption ionisation with time-of-flight mass spectrometry (MALDI-TOF MS), Fourier transform infrared (FT-IR) and FT-NIR approaches.

3. All approaches used to identify collagen in egg yolk yielded positive results.

4. The collagen in egg yolk measured using colorimetry, collagen in egg yolk, egg white and eggshell membrane using FT-NIR and collagen in egg yolk determined by treating the egg yolk with collagenase were abundant after feeding a dietary WCV (p<0.05).

5. These results suggest that egg yolk contains collagen, that the collagen in egg can be measured using FT-NIR, and that the collagen content of egg yolk can be increased by feeding dietary WCV diets.

     

Effect of starting body fat content and genotype of laying hens on the changes in their live weight,body fat content,egg production and egg composition during the first egg-laying period

1. A total 120 laying hens (60 TETRA BLANCA white egg layers and 60 TETRA SL brown egg layers) were selected from 250 TETRA BLANCA and 250 TETRA SL pullets based on their predicted body fat content by means of computed tomography (CT) at 16 weeks of age. Three groups of pullets were chosen for the investigation with the highest (n = 20), lowest (n = 20) and average (n = 20) body fat content.

2. Changes in the live weight, body fat content, egg production and egg composition of the chosen animals were recorded at 32, 52 and 72 weeks of age.

3. Based on the results, it was established that differences in starting body fat content of the hens remained the same during the experimental period. The differences between the two extreme groups were statistically significant at each age.

4. The starting body fat content of the hens affected the rate of egg production, i.e. hens with high starting body fat content produced 11–14 eggs fewer than the hens with a low or average body fat content but had no effect on the composition of the eggs.

5. Genotype affected almost all of the examined traits: TETRA BLANCA hens had lower live weight and higher body fat content during the experimental period and produced fewer eggs with lower albumen and higher yolk, dry matter and crude fat content than the TETRA SL hens.

     

The correlation between height of thick albumen,yolk diameter,shape index and the position of the yolk after storage

From about 100 White Leghorn hens the following characteristics were determined on samples of four eggs per hen:

1. The shift of the yolk during a storage period of 7 d at a temperature of 20 °G

2. The height of the thick albumen directly after lay, and at the end of the storage period

3. The diameter of the yolk

4. The shape index of the egg.

The correlation, on a hen basis, between the shift of the yolk and the other above‐mentioned traits was only of very minor importance. This means that one of the arguments on the basis of which the height of the thick albumen was used as a criterion for the internal quality of eggs—the influence of the thick white on the central position of the yolk—has lost much of its significance.

In addition the same measurements were made on eggs from 12 entries at the Random Sample Testing Station at Putten. Here samples of 20 eggs per entry were taken. Contrary to expectation the combination of a large amount of thick albumen and a large shift of the yolk proved to be the rule rather than the exception.     

Comparison of moulting methods for layers: high-zinc diet versus fasting

1. The serum biochemical profiles, thyroid hormones, body weights and the production and quality of eggs subsequent to moulting, were compared in laying hens subjected to conventional forced moulting or forced moulting with a diet high in zinc.

2. A total of 200 Dekalb White laying hens in their second production cycle were studied. Blood sampling was conducted in a factorial experimental design (2 × 3) with two methods of moulting (fasting or zinc) and three sampling periods (pre-moult, moult and subsequent peak).

3. Total egg protein content, including globulins, was greater with the zinc diet, whereas egg weight and albumen percentage were greater after fasting. The zinc method resulted in an increased shell thickness and calcium percentage but lower percentage of phosphorus. During the moulting period, the hens in the zinc group had heavier mean body weights.

4. It was concluded that moulting with a high-zinc diet could replace fasting, without negative effects on body weight, biochemical variables or subsequent egg quality and production. The zinc method was also better for the birds’ welfare.

     

Effect of dietary supplementation of thymol,synbiotic and their combination on performance,egg quality and serum metabolic profile of Hy-Line Brown hens

1. An experiment was conducted to determine the effects of dietary supplementation of thymol, synbiotic (Biomin, IMBO) and their combination in laying hen diets on laying performance, egg quality and serum metabolic profile from 24 to 36 weeks of age.

2. Treatment groups were fed on a control diet, the control diet supplemented with thymol (250 mg/kg), the control diet supplemented with synbiotic (250 mg/kg) or the control diet supplemented with a combination of thymol (250 mg/kg) and synbiotic (250 mg/kg).

3. Supplementation of thymol and synbiotic, separately as well as combined, improved egg weight, egg production, egg mass and feed conversion ratio from 24 to 36 weeks of age. The eggs obtained from thymol, synbiotic or their combination treatments displayed higher values of shell thickness, Haugh unit and shell percentage compared to the control.

4. Serum cholesterol significantly decreased in the single or combined form of thymol and synbiotic supplementation treatments.

     

Changes in eggshell structure and predisposition of broilers to health problems: is there a common pathophysiology?

1. A study of broiler breeder eggs differing in eggshell matrix optical density was conducted to determine the association of eggshell structural quality and the risk of disease in broilers.

2. A total of 10 000 eggs from a broiler breeder flock were examined according to the pre-established criteria, and allocated to groups classified as having a high or low density shell matrix.

3. The eggs from respective groups were incubated and hatched in a commercial hatchery. Samples of unhatched eggs from each group were subjected to detailed examination to establish the cause of reproductive failure. First, quality chicks from each group were raised as separate flocks in a commercial broiler barn. Group performance, morbidity, and mortality were monitored throughout the growth period. All birds were processed in a commercial plant, and condemnation data were compiled.

4. There were significantly more unhatched eggs, and fewer quality chicks in the group classified as having a low density eggshell matrix, in comparison to the high-density group.

5. Embryo pathology accounted for a large proportion of the overall reproductive failure in both groups, with a large proportion of embryos showing anatomical anomalies. The eggs and embryos from the low-density eggshell matrix group were three times more likely to be infected.

6. Significantly higher production losses associated with mortality/morbidity, and condemnations of carcasses at processing were observed in broilers from the low-density eggshell group compared with the high-density eggshell group.

7. It is concluded that common metabolic/physiological changes in breeder hens associated with eggshell pathology may be also risk factors linked with predisposition of broiler chicks to some health problems.

     

Effect of excess dietary L-valine on laying hen performance,egg quality,serum free amino acids,immune function and antioxidant enzyme activity

1. The aim of this study was to evaluate the tolerance of laying hens for an excessive L-valine (L-val) supply on laying performance, egg quality, serum free amino acids, immune function and antioxidant enzyme activities of laying hens.

2. A total of 720 HyLine Brown hens were allocated to 5 dietary treatment groups, each of which included 6 replicates of 24 hens, from 40 to 47 weeks of age. Graded amounts of L-val were added to the basal diet to achieve concentrations of 0 (control), 1, 2, 3 and 4 g/kg, respectively, in the experimental diets.

3. Supplementing the diet with L-val did not affect egg production, egg mass, egg weight, feed conversion ratio (FCR) or egg quality. The average daily feed intake response to supplemental L-val was quadratic and was maximised at 2.0 g L-val/kg diet. No differences were observed for total protein, total amino acids, blood urea nitrogen (BUN), uric acid, lactate dehydrogenase (LDH), alkaline phosphatase (AKP), Ca and P concentrations among the treatments.

4. Serum albumin concentration increased significantly in response to supplemental L-val and was also maximised at 2.0 g/kg. In addition, serum glucose increased quadratically to peak at 2.0 g L-val/kg diet. Serum free valine increased as L-val concentration increased to 2.0 g/kg diet and then decreased linearly.

5. Supplementation of L-val did not affect the serum concentrations of total antioxidative capability (T-AOC), superoxide dismutase (SOD) and malondialdehyde (MDA). L-val supplementation did not affect the concentrations of immunoglobulins IgG, IgA, IgM and complements (C3 and C4). Serum concentration of triiodothyronine (T3) increased significantly at 2.0 g L-val/kg diet.

6. It is concluded that high concentrations of L-val are tolerated and can be successfully supplemented into diets without detrimental effects on laying performance or immune function of laying hens.

     

Dietary supplementation with sodium bicarbonate improves calcium absorption and eggshell quality of laying hens during peak production

1. The advantage of supplemental sodium bicarbonate (NaHCO₃) on eggshell quality in laying hens changes with age. Besides increasing calcium (Ca) secretion in the eggshell gland, it may improve Ca absorption in the intestine or kidney.

2. Hy-Line Brown layers (n = 384), 25 weeks of age, were allocated to two treatment groups in two experiments, each of which included 4 replicates of 24 hens. Hens were fed a basal diet (control) or the basal diet containing 3 g NaHCO₃ g/kg for 50 or 20 weeks in Experiment 1 or 2, respectively. A 24-h continuous lighting regimen was used to allow hens to consume the dietary supplements during the period of active eggshell formation.

3. In Experiment 1, particularly from 25 to 50 weeks of age, and in Experiment 2, NaHCO₃ supplementation favoured hen-d egg production at the expense of lower egg weight. The increased eggshell thickness should have nothing to do with the additional eggshell formation, because of the unchanged egg mass and daily eggshell calcification.

4. At 35 weeks of age in both experiments, NaHCO₃ supplementation increased duodenal expression of calbindin-d28k (CaBP-D28k) protein, contributing to higher Ca retention and balance.

5. From 50 to 75 weeks of age in Experiment 1, the hens had little response to NaHCO₃ supplementation and showed a negative trend on eggshell thickness and strength.

6. It is concluded that dietary supplementation with 3 g NaHCO₃ g/kg improves Ca absorption and eggshell quality of laying hens during the peak but not late production period, with the introduction of continuous lighting.

     

Some factors affecting the incidence of cracks in hens’ egg shells

Six blocks of data are analysed: British Egg Marketing Board (BEMB) data on second quality eggs‐in‐shell in packing station throughput; shell thickness and colour data on 1,440 eggs sampled in 4 packing stations; crack incidence data on 162,480 individually recorded eggs from a two‐strain experimental flock kept in battery cages; similar data on 34,611 eggs from another experimental flock; shell thickness, volume and shape data on 76 eggs from 19 pairs of hens, one of each pair with a high and one with a low shell‐crack record; and BEMB data on the prevalence of battery and deep‐litter husbandry. Conclusions reached are:

1. Over the last decade there has been a rise in the incidence of packing‐station egg downgrading in the United Kingdom; it is assumed to have been due largely to a rise in the incidence of cracked shells.

2. The rise in crack incidence was probably not due primarily to deterioration in shell strength associated with high production rates or white shell colour; such associations were looked for but not found.

3. It was probably due primarily to an increase in “ environmental insults “ to packing‐station eggs accompanying the spread of deep‐litter and battery husbandry; a close association was found between their incidence and that of downgrading.

Implications for cage designers, breeders, egg producers and research workers are discussed.     

Assessment of caecal parameters in layer hens fed on diets containing wheat distillers dried grains with solubles

1. There is much interest in quantifying the nutritional value of UK wheat distillers dried grains with solubles (W-DDGS) for livestock species. A study was designed to evaluate caecal parameters (pH, short chain fatty acids (SCFAs) and bacterial diversity) in layer hens fed on balanced diets containing graded concentrations of W-DDGS.

2. A total of 32 layer hens (Bovans Brown strain at 27 weeks of age) were randomly allocated to one of 4 dietary treatments containing W-DDGS at 0, 60, 120 or 180 g/kg. Each treatment was fed to 8 replicate individually housed layer hens over a 5-d acclimatisation period, followed by a 4-week trial. Individual feed intakes were monitored and all eggs were collected daily for weeks 2, 3 and 4 of the trial, weighed and an assessment of eggshell “dirtiness” made. All hens were culled on d 29 and caecal pH and SCFAs measured. Polymerase chain reaction denaturing gradient gel electrophoresis of the bacterial 16 S rDNA gene was used to assess total bacterial diversity of luminal caecal content from hens fed the 0 and 180 g W-DDGS/kg diets. Unweighted pair group method with arithmetic mean (UPGMA) dendrograms were generated from DGGE banding patterns.

3. Increasing W-DDGS dietary concentrations resulted in a more acidic caecal environment. Caecal SCFAs were unaffected by diet aside from a quadratic effect for molar proportions of isobutyric acid. Diversity profiles of the bacterial 16S rRNA gene from luminal caecal contents were unaffected by W-DDGS inclusion.

4. The results of the current study suggest that W-DDGS can be successfully formulated into nutritionally balanced layer diets (supplemented with xylanase and phytase) at up to 180 g/kg with no detrimental effects to the caecal environment.

     

Dietary but not in ovo feeding of Silybum marianum extract resulted in an improvement in performance,immunity and carcass characteristics and decreased the adverse effects of high temperatures in broilers

1. A total of 360 fertile eggs from a broiler breeder strain (Ross 308) were used for in ovo feeding (IOF) of 0, 100 and 200 mg/kg Silybum marianum water extract at 17.5 d of incubation.

2. After hatching, 240 chicks were transferred to the experimental cages. The diets consisted of two types; one of them without extract and the other one containing 100 mg/kg of Silybum marianum extract. Then chicks were exposed to elevated temperatures 4°C above optimum from 7 to 28 d of age for 4 h per d and after 28 d they were kept at optimum temperature. The chicks were divided into 6 treatments with 4 replicates as a completely randomised 2 × 3 factorial design.

3. There was no effect of IOF of Silybum marianum extract on hatchability, body weight (BW) of hatched chicks or hatched chick BW/initial egg weight ratio. Chickens fed on the diet containing the extract had the highest feed intake, daily weight gains (DWGs), final BW and lowest feed conversion ratio.

4. At 28 d, the weights of heart, spleen and bursa of birds in the treatment groups given extract were higher, but abdominal fat weights were lower. At 42 d in those fed extract in the diet, the weights of spleen and bursa were higher and abdominal fat weights were lower than the other groups.

5. No differences were observed between groups in body temperature before application of the higher temperature but at 21 d and 28 d the body temperature of chicks given the Silybum marianum extract was lower than the other treatments during the elevated temperatures.

6. In birds fed on the extract in the diet, HDL and cholesterol concentrations were lowest at 28 d, whereas blood glucose levels were higher in the other treatments. At 42 d the cholesterol concentration was significantly lower in birds given extract in their diet.

7. These results indicated that dietary feeding of Silybum marianum extract resulted in an improvement in performance, immunity and carcass characteristics and decreased the adverse effects of the higher temperatures, specifically in reducing blood fat reduction, but IOF of the extract did not have an effect.

     

Estimation of genetic parameters related to eggshell strength using random regression models

1. This study examined the changes in eggshell strength and the genetic parameters related to this trait throughout a hen’s laying life using random regression.

2. The data were collected from a crossbred population between 2011 and 2014, where the eggshell strength was determined repeatedly for 2260 hens.

3. Using random regression models (RRMs), several Legendre polynomials were employed to estimate the fixed, direct genetic and permanent environment effects. The residual effects were treated as independently distributed with heterogeneous variance for each test week.

4. The direct genetic variance was included with second-order Legendre polynomials and the permanent environment with third-order Legendre polynomials.

5. The heritability of eggshell strength ranged from 0.26 to 0.43, the repeatability ranged between 0.47 and 0.69, and the estimated genetic correlations between test weeks was high at > 0.67.

6. The first eigenvalue of the genetic covariance matrix accounted for about 97% of the sum of all the eigenvalues.

7. The flexibility and statistical power of RRM suggest that this model could be an effective method to improve eggshell quality and to reduce losses due to cracked eggs in a breeding plan.

     

Increasing persistency in lay and stabilising egg quality in longer laying cycles. What are the challenges?

1. In the past 50 years, selection starting initially at the breed level and then using quantitative genetics coupled with a sophisticated breeding pyramid, has resulted in a very productive hybrid for a variety of traits associated with egg production.

2. One major trait currently being developed further is persistency of lay and the concept of the “long life” layer. Persistency in lay however cannot be achieved without due consideration of how to sustain egg quality and the health and welfare of the birds in longer laying cycles. These multiple goals require knowledge and consideration of the bird’s physiology, nutritional requirements, which vary depending on age and management system, reproductive status and choice of the selection criteria applied.

3. The recent advent of molecular genetics offers considerable hope that these multiple elements can be balanced for the good of all in the industry including the hens.

4. The “long life” layer, which will be capable of producing 500 eggs in a laying cycle of 100 weeks, is therefore on the horizon, bringing with it the benefits of a more efficient utilisation of diminishing resources, including land, water, raw materials for feed as well as a reduction in waste, and an overall reduced carbon footprint.

     

The hen's egg: Shell cracking at oviposition in battery cages and its inheritance

It is known that there is much variation between individual hens and between strains of hen in (a) the incidence of egg‐shell cracking and (b) the height through which the egg is dropped at oviposition. Pilot experiments are described from which it is concluded that:

1. most cracks that occur in a battery cage are produced when the egg drops on to the floor at oviposition;

2. they are located predominantly in the two polar thirds of the shell;

3. their incidence is determined mainly by the effective mass of the cage floor, the mean drop height and the mean attitude of the eggs at emergence from the cloaca (broad or narrowpole first) ;it is affected only slightly by the mean shell thickness and the overall mean shell curvature (egg size);

4. if a hen is kept on a floor with a high effective mass, an indirect estimate of her mean drop height can be obtained quickly, easily and with high precision from observations made on her eggs after they have been laid; the observations required are the incidence of eggs with poorly formed shells, the type, location and incidence of cracks in eggs with well‐formed shells and the mean weight of the eggs;

5. the incidence of cracking on a heavy cage floor can be used to predict the incidence on a normal floor;

6. the incidence of cracking is heritable and responds rapidly to downward selection.

It is suggested that breeders should test the technique described, with a view to adopting it as a method of selecting for low crack incidence.     

Dietary conjugated linoleic acid affects blood parameters,liver morphology and expression of selected hepatic genes in laying hens

1. The objective of this research were to investigate the effect of a conjugated linoleic acid (CLA)-enriched diet on Isa Brown laying hen health status and to provide a comprehensive analysis of changes in blood parameters, liver morphology and selected hepatic gene expression.

2. Hens were allocated to the control and experimental group (diet enriched with 0.75% CLA) for a total period of 4 m. At the end of the experiment half of the hens from each group were slaughtered for analyses. The remaining hens were transferred to an organic farm for the next 5 m and fed on the diet without CLA supplementation.

3. The CLA-enriched diet resulted in significant changes in blood and serum parameters; specifically, haematocrit (HCT), mean corpuscular volume (MCV) and white blood cells (WBC) count were decreased compared to the control. The total cholesterol (TC) was not significantly affected while the triacylglycerol’s (TG) concentration was elevated. The activity of alanine aminotransferase (ALT) was significantly increased in the CLA-supplemented group, while aspartate aminotransferase (AST) showed an increasing tendency. Liver biopsies showed pathological changes classified as non-alcoholic fatty liver disease (NAFLD). Additionally, the expression of hepatic genes involved in fatty acids synthesis (ME1, ACLY, ACC, FASN, SCD1), oxidation (CPT1α, PPARA), detoxification processes (Cytochrome P450, CYP, Flavin-containing monooxygenase, FMO3), oxidative stress (NOX4, XbP1) and inflammation (IL6, TNFα) were elevated. Cessation of CLA supplementation for 5 m of organic farming resulted in normalisation of blood and hepatic parameters to the levels observed in control hens.

4. The results of this study indicate that dietary CLA triggers an integrated stress response in laying hens and activates mechanisms involved in liver detoxification.

     

Assessment of the effectiveness of head only and back-of-the-head electrical stunning of chickens

1. The study assesses the effectiveness of reversible head-only and back-of-the-head electrical stunning of chickens using 130–950 mA per bird at 50 Hz AC.

2. Three trials were conducted to compare both stunning systems: (a) behavioural assessment of return of consciousness, (b) insensibility to thermal pain, and (c) assessment of return of brain activity with visually evoked potentials (VEPs).

3. Assessment of behaviour suggested that the period of unconsciousness following head-only electrical stunning was shorter in hens compared to broilers.

4. Stunning across the back-of-the-head delayed the time to return of brainstem function compared to stunning with standard head-only electrodes. Additionally, back-of-the-head stunning produced a more prolonged period of electroanalgesia compared to head-only.

5. Based on examination of return of brain function with VEPs in hens, back-of-the-head stunning produced a shorter-lasting stun than standard head-only. However, even for standard head-only, the stun was notably shorter than previously reported. In some birds, brain function had returned within 9 s after the end of stunning.

6. The results suggest that some birds may recover consciousness prior to or during the neck cut. Based on these findings, back-of-the-head stunning and standard head-only stunning of hens should not be recommended without further development.

     

Effects of fermentatively recovered fish waste lipids on the growth and composition of broiler meat

1. The aim of the present study was to evaluate the effects of feeding fermentatively recovered fish oil (FFO) from fish processing waste (FPW), on the performance and carcass composition of broilers. A total of 60 one-d-old VenCobb^® broiler chicks randomly assigned to 5 treatment groups were studied.

2. The birds were randomly assigned to treatment groups and fed with a normal commercial diet (control, T1), a diet with 2% groundnut oil (positive control, T2), a diet with 1% FFO (T3), a diet with 1.5% FFO (T4) and a diet with 2% FFO (T5). Performance and growth parameters (feed intake and body weight) and fatty acid composition of serum, liver and meat were determined.

3. The performance characteristics of broiler meat did not differ among treatments. Feeding FFO reduced total cholesterol concentration in serum, meat and liver of the FFO-fed groups (T3 to T5) as compared to both the controls (T1 and T2), but there was no significant difference in triglyceride concentration between treatments. Increased concentrations of EPA and DHA in serum, liver and meat of FFO-fed groups, as compared to both controls, were observed as the FFO concentration increased.

4. The study clearly demonstrates the value of oil recovered from FPW in addition to addressing the environmental issues related to disposal of such biological waste.

     

The Hen's egg: Factors affecting the shearing strength of shell material

Experiments are described from which it is concluded that

1. the force required to shear egg shell material can be measured with high precision by pressing a narrow, flat‐ended plunger on to the shell;

2. shearing force falls and the precision of its measurement rises as the radius of the plunger is reduced;

3. shearing force is related linearly to shell thickness ;

4. the regression line of shearing force on shell thickness does not pass through the origin but has a substantial positive intercept on the thickness axis, at about a third of the mean thickness ;

5. there are differences between hens and between strains of hen in the magnitude of the intercept, but not in the slope of the regression;

6. shearing force is affected also by the glossiness of the shell;

7. shearing force was not significantly affected by stripped membrane thickness, number of mammillae per unit area of shell, shell organic content (standardised for shell thickness), shell curvature, shell colour, mottling score, translucent patch score, speckling score, age of the egg or age of the hen that laid it;

8. measured with a plunger of radius 165 μm, the mean shearing strength of incremental shell was 57 kg wt/mm² (3–7 ton/in²).

The findings suggest that the shearing strength of egg shells might be increased by direct selection and by selection for shell glossiness.