The hen's egg: Prediction and reduction of downgrading in large producer groups

British Egg Marketing Board data on the incidence of downgrading in eggs received at packing stations during the second week in each month from April 1960 to March 1968 have been analysed in relation to:

1. the incidence of free‐range, deep‐litter and battery husbandry in flocks from which they came, as estimated by interpolation from annual census data;

2. season, as represented by fundamental and first‐harmonic sine and cosine terms;

3. age‐structure of the flocks, as indicated by the incidence of extra‐small eggs 10 months earlier;

4. time;

5. packing‐station throughput.

It is shown that 93–3 per cent of the variance of downgrading percentage is associated with variation in (i), (ii) and (iii). After these effects had been taken into account there was no significant long‐term time‐trend but there were small effects associated with certain years. There was no significant effect of (v). The regression equation was used to predict the incidence of downgrading in the second week of each month from April 1968 to January 1969; this it did accurately except in May and June 1968, when the downgrading percentage fluctuated to an unprecedented degree following a very cold and a very hot spell of weather.

It is suggested that large producer‐packer groups should systematically collect information on the incidence of downgrading and the percentages of birds of each strain and age‐group, of cages of each type, of nutritional regimes and of egg‐collection methods; and that these data should be analysed to yield (a) forecasts of downgrading incidence and (b) estimates of the effect of each strain, cage‐type, etc. on downgrading incidence, for use when deciding replacement policy.     

Risk Factors Associated with Salmonella Status of Broiler Flocks Delivered to Grow‐Out Farms

In a prospective field observational study in the southeastern USA, we sampled gastrointestinal (GI) tracts from chicks of 65 broiler flocks delivered to conventional grow‐out farms for rearing. The flocks were hatched at seven broiler hatcheries. The mean within‐flock prevalence of Salmonella‐positive samples was 6.5% and ranged from 0% to 86.7%. Of the 65 flocks studied, 25 (38.5%) had at least one Salmonella‐positive sample. Accounting for confounding variability among the hatcheries and broiler companies, we tested whether the probability of detecting Salmonella in GI tracts of the chicks delivered was associated with certain characteristics of parent breeder flocks; hatchery production volume; hatchery ventilation system; hatchery egg‐room conditions; egg incubation, candling, hatching, eggshell and bird separation, and bird‐processing procedures; management of hatchery‐to‐farm transportation; day of week of hatch; weather conditions during transportation; or season of the hatch. Two risk factor models were adopted. The first model indicated that a greater number of parent flocks, manual separation of eggshell and bird, and a greater amount of fluff and feces on tray liners used during hatchery‐to‐farm transportation at delivery were associated with increased probability of detecting Salmonella in chick GI tracts, whereas a greater number of birds in the delivery vehicle was associated with decreased probability. The second model indicated that broiler flocks hatched on Tuesdays versus either Mondays or Thursdays (with no hatches on Wednesdays, Fridays or week‐ends), increased average hatchability of the eggs from the parent flocks, and greater amounts of fluff and feces on the transport tray liners at delivery were all associated with increased probability of detecting Salmonella in chick GI tracts. The results of this study suggest potential management decisions to lessen Salmonella contamination of broilers supplied by commercial hatcheries and areas for further research.     

Feeding motivation on the incidence of floor eggs and extraneously calcified eggs laid by broiler breeder hens

1. Commercial broiler breeder hens lay many eggs on the floor rather than in nest boxes provided. A study was conducted to determine whether feeding feed-restricted broiler breeder hens during the sitting phase of nesting results in a higher incidence of floor eggs and/or retained eggs. 2. Sixty broiler breeder females (Ross 308) were randomly assigned to 6 deep litter pens containing 10 nest-boxes. At 35 weeks of age and for 9 weeks, feed was distributed to all pens at lights-on every second day (fed normally, FN). On alternate days (feeding delay, FD), feed was distributed when 2-3 hens/pen were sitting in a nest box. Behaviour was sampled at 41 weeks of age, for 26 d. Eggs and egg location data were collected daily, and eggs were scored for extra-cuticular calcium. 3. Of 81 instances in which the hen was sitting firmly in a nest box at the time of feeding, on 80 instances the hen left the nest-box to feed, and on one instance the hen laid her egg then exited to the feeder. Of these 80 instances, on 58 occasions the hen returned to a nest-box to lay her egg; on 12 the hen returned to the nest-box but laid no egg; on 7 the hen did not return to the nest box and laid no egg; and on three the hen laid her egg on the floor. 4. Mean floor egg percentage was 13·3 ± 3·2% on FN and 13·3 ± 4·7% on FD days; these did not differ significantly. 5. The mean extra-cuticular calcium score over all pens was 0·9 ± 0·06 on FN days and 1·2 ± 0·06 on FD days; these differed significantly. 6. In conclusion, feeding broiler breeder hens during nesting results in a conflict between feeding and nesting motivation and higher numbers of extraneously calcified eggs, but does not result in a significant increase in floor eggs even though nesting hens will leave the nest box for food.     

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.     

Egg Shell Strength

Three experiments are reported which were designed to measure the relationship that exists between the specific gravity of an egg and deformation of its shell under load and also to determine the extent to which these two egg characteristics are related to shell breakage in the field.

A very close relationship was shown to exist between egg specific gravity and egg shell deformation.

In experiments 1 and 2, measurements of specific gravity and shell deformation were performed upon samples of eggs produced by groups of birds housed in wire‐floored pens and by individual birds kept in battery cages and were found, with one exception, to be significantly related to the incidence of cracking under these field conditions. The one non‐significant correlation was that between egg shell deformation and percentage cracks among eggs from the various pens of birds. However, too much emphasis should not be placed on this latter result because the incidence of floor‐laying in the different pens was not uniform and this increased chance variation in the occurrence of cracking.

In experiment 3 specific gravities and shell deformations were determined on 3000 sound eggs before they were subjected to treatments that simulated the hazards encountered during (a) laying in battery cages, (b) transport in the conventional travelling boxes. Both measurements were found to forecast fairly accurately the probability of a particular egg becoming cracked or remaining sound during the treatments.

The results confirm that egg specific gravity, as measured by the hydrometer method, is a reliable assessment of shell strength and suggest that the measurement of shell deformation is of comparable reliability. Other factors of practical significance that would influence the choice between the two assessments are discussed.     

Distribution and Diversity of Salmonella Strains in Shipments of Hatchling Poultry,United States, 2013

Multistate outbreaks of salmonellosis associated with live poultry contact have been occurring with increasing frequency. In 2013, multistate outbreaks of salmonellosis were traced back to exposure to live poultry, some of which were purchased at a national chain of farm stores (Farm store chain Y). This study was conducted at 36 stores of Farm store chain Y and was concurrent with the timing of exposure for the human outbreaks of salmonellosis in 2013. We used environmental swabs of arriving shipment boxes of hatchling poultry and shipment tracking information to examine the distribution, diversity and anti‐microbial resistance of non‐typhoidal Salmonella (NTS) across farm stores and hatcheries. Isolates recovered from shipment boxes underwent serotyping, anti‐microbial resistance (AMR) testing and pulsed‐field gel electrophoresis (PFGE). Postal service tracking codes from the shipment boxes were used to determine the hatchery of origin. The PFGE patterns were compared with the PFGE patterns of NTS causing outbreaks of salmonellosis in 2013. A total of 219 hatchling boxes from 36 stores in 13 states were swabbed between 15 March 2013 and 18 April 2013. NTS were recovered from 59 (27%) of 219 hatchling boxes. Recovery was not significantly associated with species of hatchlings, number of birds in the shipment box, or the presence of dead, injured or sick birds. Four of the 23 PFGE patterns and 23 of 50 isolates were indistinguishable from strains causing human outbreaks in 2013. For serotypes associated with human illnesses, PFGE patterns most frequently recovered from shipment boxes were also more frequent causes of human illness. Boxes positive for the same PFGE pattern most frequently originated from the same mail‐order hatchery. Only one of 59 isolates was resistant to anti‐microbials used to treat Salmonella infections in people. This study provides critical information to address recurrent human outbreaks of salmonellosis associated with mail‐order hatchling poultry.     

Design of laying nests in furnished cages: influence of nesting material, nest box position and seclusion

(1) Preferences for three nesting materials and nest box positions were investigated simultaneously in two trials using a furnished cage: one with 18 individual laying hens and one with 18 groups of 5 hens. Following a habituation period in pre-test cages, every hen or group of hens was tested for 2 d: once without and once with plastic flaps at the entrance of the nest boxes. (2) Hens preferred peat and artificial turf to coated wire mesh for egg laying. (3) One nest box position was clearly preferred to both other nest boxes. The hens' choice of nest box position was influenced by the pre-test cage in which they had been habituated. (4) The presence of plastic flaps at the entrance of the nest boxes had no influence on the proportion of eggs laid on the different nesting materials or on the proportion of floor eggs. (5) Individual and group testing resulted in the same overall results despite the presence of a distinct group effect.     

Variation in some shell membrane characteristics over different parts of the same shell

The variation in shell membrane thickness (μ), membrane weight (mg./cm.2), membrane nitrogen (μg. N/cm.2) and packing index (μg./cm.2/μ) from the broad pole, excluding the air space, to the narrow pole of the same egg has been studied. The pattern of variation for a given characteristic was the same in different eggs from the same bird. Two thickness patterns were found. In Group I thickness decreased from the B collar to the equator, increased again to the G collar and then decreased sharply at the H cap. In Group II the increase from the equator to the G collar was replaced by a continued, but less‐marked, decrease. There was a high, positive correlation between weight and nitrogen, but the relationship between weight and thickness varied within and between eggs. All eggs showed a very similar pattern of packing index variation, decreasing from the B to about the C collar and then increasing to the H cap, the pattern being the same whether measurements were made on wet or dry membranes.     

Egg trait variation in a large hawk‐cuckoo (Hierococcyx sparverioides) host population of Chinese babax (Babax lanceolatus)

Mutual interaction between brood parasites and their hosts is a well‐known model system for studying host–parasite coevolution. Both parties have acted reciprocally, resembling an evolutionary arms race, in which adaptations and counter‐adaptations have evolved as a result of host–parasite dynamics, such as the classical cuckoo–host system. Discrimination among parasite and cuckoo eggs and rejection of foreign eggs is regarded as an important anti‐parasitism strategy. The Chinese babax (Babax lanceolatus) is a large hawk‐cuckoo (Hierococcyx sparverioides) host distributed in southwest China. A previous study shows that the babax is an intermediate egg rejector, and most cuckoo eggs are accepted by the Chinese babax, although a small proportion of hosts reject cuckoo eggs. Interestingly, the large hawk‐cuckoo lays non‐mimetic eggs in contrast to the uniform blue eggs of babaxes. Because egg coloration is a critical cue used by host species in favor of the recognition of parasitic eggs by hosts, we used a spectrometer to quantify egg color variation to understand the differentiation in discrimination ability between the egg rejectors and acceptors. We found that the chroma of intra‐clutch variation of babax eggs was more consistent in egg rejectors than in acceptors. However, no statistical significance was found in inter‐clutch variation between these two types of hosts. Our results suggest that hosts lay eggs with a low level of intra‐clutch variation without the necessity of a high level of inter‐clutch variation simultaneously as predicted by the egg signature hypothesis. This study may further indicate that selection pressures from evolutionarily recent parasites can drive individual‐based differences in an anti‐parasitism strategy.     

Changes in the prevalence,genotypes and antimicrobial resistance phenotypes of non‐typhoidal Salmonella recovered from mail‐order hatchling poultry sold at US feed stores, 2013–2015

Every year, multiple outbreaks of salmonellosis in humans are linked to contact with mail‐order chicks and ducks. The objective of this study was to describe the temporal changes in the prevalence of serovars, genotypes and antimicrobial resistance (AMR) phenotypes of non‐typhoidal Salmonella (NTS) recovered from shipped boxes of mail‐order hatchling poultry in the United States during 2013 to 2015. In each year, a sample of feed stores belonging to a single national chain participated in the study. The store employees submitted swabs or hatchling pads from hatchling boxes and shipment tracking information of the arriving boxes to the investigators. NTS was cultured from the samples and isolates were sent to the National Veterinary Services Laboratories (Ames, IA) for serotyping, pulsed‐field gel electrophoresis (PFGE) and AMR phenotyping. The PFGE patterns of Salmonella serovars isolated from hatchling boxes were compared with those from human outbreaks of salmonellosis linked to live poultry contact. The box‐level prevalence of NTS was significantly higher in 2015 compared to 2014. Also, the population of Salmonella serovars recovered in 2015 was more diverse and substantially different from those recovered in the previous two years. Of PFGE patterns recovered from hatchling boxes, seven distinct patterns in 2015, three in 2014 and four in 2013 were indistinguishable from the PFGE patterns of human outbreaks‐associated strains in the respective years. Importantly, a significant positive correlation was found between the box‐level prevalence of PFGE patterns and the number of human illnesses associated with the same patterns. Also, the proportion of multidrug‐resistant isolates was higher in 2014 and 2015 compared to that in 2013. The results demonstrate that shipments of mail‐order hatchling poultry are frequently contaminated with Salmonella genotypes indistinguishable from human outbreaks‐associated strains each year, and control efforts at hatchery level are likely to have an important public health impact.     

Lying behavior in horses in relation to box size

To analyze if horses' lying behavior is influenced by the size of their lying area, 8 horses were kept, first, in a large box—(2.5 × height of the horse)²m²—and then in a small box—(1.5 × height of the horse)²m²—or vice versa. After a 5-day adaptation period, the lying behavior (frequency and duration of sternal and lateral recumbency, rotating behavior just before lying down, and rolling behavior just before getting up) was observed from video recordings on 3 consecutive nights. The results showed that the duration of sternal recumbency was significantly longer in the large boxes than in the small boxes (P = .002). Furthermore, box size exerted an influence on the frequency of the rolling behavior shown before getting up, but box size was not the only factor affecting this behavior. We conclude that observations of the resting behavior of horses are necessary to make recommendations for the minimal size of stalls for horses.     

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.     

Internal Egg Temperature in Response to Preincubation Warming in Broiler Breeder and Turkey Eggs

Various methods of preincubation warming have been used in commercial hatcheries to increase hatchability, decrease incubation time, and increase embryonic blastoderm development. A study was conducted to determine the effect of preincubation warming and egg size on the warming rate of eggs. Three egg groupings were evaluated: small broiler breeder eggs (52 to 57 g), large broiler breeder eggs (64 to 69 g), and turkey eggs (74 to 107 g). The eggs were subjected to 2 temperature treatments. The conventionally incubated eggs were moved directly from storage (17.5°C) to the incubator (adjusted to 37.5°C). The preincubation warmed eggs were moved from the storage room to the preincubation room (28°C) for 12 h and then to the incubator. Internal egg temperature was measured once per minute to calculate the rate of warming. The warming rate potential (k value) differed for egg type and temperature treatment. The values for small and large broiler breeder eggs and turkey eggs were 0.0506, 0.0488, and 0.0471 kJ/min per kJ of energy in the egg, respectively. This study characterizes the maximum rate of temperature change that occurs when eggs are transferred from a storage or preincubation area to the incubator, creating baseline values for future work with the modification of hatchery management protocols. These data can contribute to more informed decisions regarding hatchery temperature profile strategies and their effect on the developing embryo.     

A Long-Term Study of the Environment Within a Tunnel Incubator for Turkey Eggs

A 6-mo study of the temperatures within a tunnel incubator in a commercial turkey hatchery was conducted to determine the extent that the incubation environment varied with the type of egg incubated. The tunnel incubator moves air through the mass of eggs from the oldest to the youngest embryos, and air temperature was found to increase by approximately 1°F as it passed through the eggs. The temperature of the air entering the eggs was maintained at a constant temperature by the incubator, but the temperature of the air leaving the eggs showed considerable day-to-day variation. The main cause of temperature variation within the incubator was the setting of fresh eggs into and the transferring of d-25 eggs out of the incubator. However, the temperature was also found to vary with the predicted total embryo metabolic heat production within the incubator estimated from the age of the embryo, egg mass, and breeder flock fertility. The temperature of air within the egg mass was also measured and shown to correspond more closely to the temperature of the air where it exits the eggs rather than where it enters the eggs or the machine-operating temperature.     

Factors affecting egg production and layer bird mortality in private poultry farms in the subhumid zone of Nigeria

Egg production and layer bird mortality data obtained from five different private farms in Zaria within the subhumid zone of Nigeria, over a 6‐year period (1994–1999) were subjected to least squares analysis to determine the effects of age, season and year. It was demonstrated that age had a highly significant influence (P < 0.01) on egg production in that birds falling within the age group 30–39 weeks produced the highest number of eggs (3255 ± 109), while birds over 100 weeks of age produced the lowest number of eggs (1206 ± 412). Similarly, seasonal variation in egg production was also significant (P < 0.05); the highest egg production (2926 ± 90) was obtained during the early dry season, and the lowest (2423 ± 95) during the late wet season. Mortality was generally low (0.0–0.9%) and not significantly different from 20 to 49 weeks of age (P > 0.05). However, from 50 to 100 weeks of age, highly significant differences (P < 0.01) in mortality were observed, with the 90–100 weeks age group recording the highest mortality of 15.7 ± 1.3%. Furthermore, yearly variations in egg production and mortality were highly significant (P < 0.01) with 1999 producing the most eggs (4410 ± 102) and recording the lowest mortality (1.4%). It was concluded that due to the significant seasonal, age and yearly variations, data on egg production and mortality need to be adjusted for these effects to enable unbiased comparison between and within farms. Furthermore, farmers can make adequate feeding preparations commensurate with each season for optimal egg production.     

Relationship between egg‐shell colour and two measurements of shell strength in the Vasca breed

1. Egg‐shell colour, egg specific gravity, shell thickness and egg weight were determined for 2080 eggs from three varieties of the Vasca, a Spanish brown‐egg‐laying hen.

2. Intermediate, positive and significant (P < 0·01) correlation coefficients were found between egg‐shell colour and specific gravity (0·25), or shell thickness (0·21). There was a very high (0·84), significant (P < 0·01) correlation between the measures of shell strength.

3. Shell thickness was accurately predicted with both specific gravity and egg weight as independent variables (determination coefficient 0·74); eggshell colour did not explain any further variation.

4. All traits showed considerable departure from normality in the form of negative kurtosis (shell colour and specific gravity), positive kurtosis (shell thickness and egg weight), negative skewness (shell thickness) or positive skewness (egg weight).

5. Egg‐shell colour, specific gravity and shell thickness were less in July, in old birds and in eggs laid in the morning. Differences in egg quality traits between the three varieties were not significant.

     

Estimation of the Rate of Egg Contamination from Salmonella‐Infected Chickens

Salmonella enterica serovar Enteritidis (S. Enteritidis) is one of the most prevalent causes for human gastroenteritis and is by far the predominant Salmonella serovar among human cases, followed by Salmonella Typhimurium. Contaminated eggs produced by infected laying hens are thought to be the main source of human infection with S. Enteritidis throughout the world. Although previous studies have looked at the proportion of infected eggs from infected flocks, there is still uncertainty over the rate at which infected birds produce contaminated eggs. The aim of this study was to estimate the rate at which infected birds produce contaminated egg shells and egg contents. Data were collected from two studies, consisting of 15 and 20 flocks, respectively. Faecal and environmental sampling and testing of ovaries/caeca from laying hens were carried out in parallel with (i) for the first study, testing 300 individual eggs, contents and shells together and (ii) for the second study, testing 4000 eggs in pools of six, with shells and contents tested separately. Bayesian methods were used to estimate the within‐flock prevalence of infection from the faecal and hen post‐mortem data, and this was related to the proportion of positive eggs. Results indicated a linear relationship between the rate of contamination of egg contents and the prevalence of infected chickens, but a nonlinear (quadratic) relationship between infection prevalence and the rate of egg shell contamination, with egg shell contamination occurring at a much higher rate than that of egg contents. There was also a significant difference in the rate of egg contamination between serovars, with S. Enteritidis causing a higher rate of contamination of egg contents and a lower rate of contamination of egg shells compared to non‐S. Enteritidis serovars. These results will be useful for risk assessments of human exposure to Salmonella‐contaminated eggs.     

Individual variation in prelaying behaviour and the incidence of floor eggs

1. Floor eggs are a problem in non‐cage systems for laying hens, as they require secondary egg collecting. Failure to lay in a well‐defined nest site may also be a welfare problem for the hens, but only if their nesting motivation has been thwarted.

2. We investigated the relationships between a hen's prelaying behaviour and its tendency to lay on the floor by recording the behaviour of 20 hens housed individually in wire cages with single littered nest boxes.

3. Most floor eggs (80%) were laid by the same 6 hens. These 6 “floor‐layers” performed more nest seeking behaviour, less nest‐building behaviour and less sitting prior to oviposition than the 14 hens that consistently laid in nest boxes.

4. The incidence of floor eggs declined with age. Both nest and floor laying hens performed less nest seeking behaviour with age. Floor layers, however, increased their performance of nesting behaviour, whilst nest layers performed less nesting behaviour with age.

5. Floor laying hens behaved as if they found the nest box less attractive than nest‐laying hens; perhaps because they had lower nesting motivation, or perhaps because their nesting motivation was as high, but they less readily perceived the nest box as an appropriate nest site.     

Microbiological survey of the incubated eggs of chickens and waterfowl

Coliform organisms were the principal contaminants of rotten eggs from commercial duck hatcheries and incubated eggs of 45 species of waterfowl. Micrococci were the main contaminants of “incubator clears” obtained from a chicken hatchery. Pseudomonas, Acinetobacter and Bacillus spp were minor contaminants of the eggs of waterfowl. All the coliforms recovered from waterfowl eggs formed a large, well‐defined capsule when grown on nutrient agar containing glucose or lactose.     

Influence of quantity of utter on nest box selection and nesting behaviour of domestic hens

1. Twenty two hens (Gallus gallus domesticus) were given a free choice, for egg laying, between 4 nest boxes containing different quantities of wood shavings (litter): 3180 cm³ (large), 1060 cm³ (medium), 350 cm³ (small) and none (zero).

2. There was considerable individual variation in the quantity of litter selected for egg laying, but 17 hens laid 50% or more of their eggs in a single litter quantity. Hens selected the large quantity in preference to the small and zero. More searching, nest selection and nesting behaviours tended to be directed to the boxes with the largest quantities and also to the quantities that the hens finally selected for egg laying.

3. In a second experiment, the hens from experiment 1 which showed a preference for litter, and a further 22 naïve hens were given a choice of three nest boxes containing no litter and one containing one of 6 quantities. Over 30 trials there were 5 replicates using each of the three quantities used in experiment 1 plus 115 cm³, 40 cm³ and 10 cm³.

4. There was a wide range in the proportion of trials in which hens selected litter; for experienced hens the range was 0.43 to 0.97, but only two naïve hens were within this range, all others being between 0 and 0.43.

5. The pattern of choosing litter quantities was comparable but at different levels of preference in experienced and naive hens and a contour map was derived enabling predictions to be made of the probability of a particular quantity being chosen.

6. The variability of individual responses suggest that no single type of nest box system can cater for the requirements of all hens.