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Effects of Feeding White Leghorn Hens Diets that Contain Deoxynivalenol (Vomitoxin)-Contaminated Wheat1,2
ENVIRONMENT AND HEALTH Effects of Feeding White Leghorn Hens Diets that Contain Deoxynivalenol (Vomitoxin)-Contaminated Wheat1'2 R. M. G. HAMILTON 3 , B. K. THOMPSON", H. L. TRENHOLM 3 , P. S. FISER 3 , and R. GREENHALGH 5 Animal Research Centre, Engineering and Statistical Research Institute, and Chemistry and Biology Research Institute, Research Branch, Agriculture Canada, Ottawa, Canada K1A 0C6 (Received for publication November 7, 1984)
1985 Poultry Science 64:1840-1852
INTRODUCTION R e c e n t survey results indicate t h a t cereal grains, particularly corn and w h e a t , m a y often b e c o n t a m i n a t e d w i t h 4-deoxynivalenol ( D O N ) (Cote et al, 1 9 8 4 ; Hagler et al, 1 9 8 4 ; Trenholm et al, 1 9 8 3 ; S c o t t et al, 1 9 8 1 ; Ueno,
'Contribution numbers: 1289 Animal Research Centre, 1-655 Engineering and Statistical Research Institute, and 1496 Chemistry and Biology Research Institute. 2 Preliminary report presented at the 70th Annual Meeting, Poultry Science Association, University of British Columbia, Vancouver, August 3—6, 1981 (Poultry Sci. 60:1666, 1981). 3 Animal Research Centre. 4 Engineering and Statistical Research Institute. 5 Chemistry and Biology Research Institute.
1 9 8 0 ; Vesonder et al, 1 9 7 8 ) , especially w h e n cool and w e t climatic conditions prevail ( S u t t o n , 1 9 8 2 ) . O n c e DON, a t r i c h o t h e c e n e m y c o t o x i n , is p r o d u c e d b y t h e mold Fusarium graminearium (Schwabe t e l e m o r p h Gibberella zeae [Schw.] Petch) (Neish et al, 1 9 8 3 ; Neish and Cohen, 1 9 8 1 ) , it will persist, b u t n o t necessarily at m a x i m u m levels (Miller et al, 1 9 8 3 ) . Efforts t o disguise DON w i t h feed additives have n o t b e e n productive (Mirocha et al, 1 9 7 7 ; Friend et al, 1 9 8 4 ) . DON has been r e p o r t e d t o cause inhibition of p r o t e i n and d e o x y r i b o nucleic acid ( D N A ) biosynthesis, inflammation of t h e skin, leukopenia, decreased feed i n t a k e and weight gains, and i m m u n o s u p p r e s s i o n (Newberne and Rogers, 1 9 8 1 ; Ueno, 1 9 8 0 ) . Emesis or vomiting is also associated w i t h con-
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ABSTRACT A short-term (10 weeks, Experiment 1) and a long-term experiment (24 weeks, Experiment 2) were done to determine effects of incorporating either white winter wheat, naturally contaminated with 1 mg deoxynivalenol (DON)/kg, or spring wheats, containing up to 6.5 mg DON/kg, into the diets of White Leghorn hens. Based on chemical analysis, the diets in Experiment 1 contained <.05 to .7 mg DON/kg, while those in Experiment 2 contained from .2 to 4.9 mg/kg. Incorporation of winter or spring wheat in the experimental diets had no effect (P>,05) on feed intake and efficiency, egg production and yield, the number of soft shell and cracked eggs observed in the laying house, body weight at the completion of the experimental period, fertility, hatchability of fertile eggs, and the proportion of malformed embryos and pips. In addition, presence of DON-contaminated wheat did not influence (P>.05) the organ weight to body weight ratio for a randomly selected sample of hens necropsied at the completion of each experiment. There was little evidence of lesions in the oral cavity, esophagus, proventriculus and gizzard, hemorrhaging in the viscera or skeletal muscles, or of changes in the appearance of spleen, heart, and kidney. However, the livers from DON hens were fatty in appearance. Furthermore, vomiting (emesis), diarrhea, or changes in behaviour were not apparent and mortality, normally very low, was not increased during either experiment. Inverse linear relationships were obtained in Experiment 1 between dietary DON concentrations and egg weight (P<.05), shell weight and thickness (P<.01), and percent shell (P<.05). Although egg and shell variables measured in Experiment 2 were not significantly influenced (P>.05) by DON treatment, trends towards lower values with higher dietary DON levels were evident. Egg specific gravity, nondestructive deformation, and quasistatic compression fracture strength of the egg's shell were not influenced (P>.05) by dietary DON levels. The results from these experiments indicate that laying hens can tolerate diets containing up to 5 mg DON/kg from white winter or spring wheat for extended periods of time without serious adverse effects on health and productivity. (Key words: mycotoxin, deoxynivalenol, hens, wheat, shell quality, performance)
DEOXYNIVALENOL AND LAYING HENS
MATERIALS AND METHODS
Experiment 1. White Leghorn pullets from two randombred control strains (7 and 10) and four 2-way strain crosses (1 X 4, 4 X 1, 4 X 8, and 8 x 4 ) were fed experimental diets for 70 days beginning when they were 192 days of age. The control strains have been previously described by Gowe and Fairfull (1980) and the strain crosses by Fairfull et al. (1983). The pullets were housed in individual wire cages (25.5 or 30.5 X 40.6 cm) in a light-tight room. During the experimental period, the length of the photoperiod was increased from an initial of 11 hr/day to 16 hr/day in increments of 30 min/ week. Feed and water were provided ad libitum. Composition of the control and experimental diets is given in Table 1. The experimental diets were formulated by replacing, on an equal weight basis, 50 or 100% of the white winter wheat in the control diet (CW) with white winter wheat containing 1 mg DON/kg of wheat; the DON-containing diets were designated CW/DON-W and DON-W, respectively. The supplies of white winter wheat were from the 1980 Ontario crop; details of their acquisition and storage were described by Friend et al. (1982). Samples of all mixes of the diets were analyzed for dry matter, Kjeldahl nitrogen, ether extract, ash, calcium, and total phosphorus by the methods of AOAC (1975). The method of Cohen and Lapointe (1982) was used to determine the DON content of the diets. Each diet was given to 102 hens distributed evenly, according to strain, among
three replicates; hence, each strain-diet group contained 17 hens in consecutive cages. The hens were weighed individually at the beginning and end of the experiment. Feed intake was measured over two 35-day periods. The numbers of normal shelled eggs collected from the egg trays and the numbers of broken, cracked, and soft shelled eggs observed in the egg trays and on the dropping boards were recorded daily. The birds were checked daily for evidence of vomiting and diarrhea. Mortality was recorded as it occurred and all hens that died were necropsied. One randomly selected hen from each diet-strain group was killed by carbon dioxide asphyxiation at the end of the experimental period, and the heart, spleen, proventriculus, one kidney, gizzard, and liver from each were removed and weighed. The oral cavity, esophagus, crop, proventriculus, and gizzard were examined for lesions. Egg shell and interior quality measurements were taken on all eggs laid for 5 consecutive days beginning when the hens were 243 days of age and had received the experimental diets for 52 days. The eggs were stored overnight in an egg cooler at 10 C. Upon removal from the cooler, the eggs were allowed to warm, over 3 hr, to room temperature. Subsequently, each egg was weighed in air, then submerged in tap water and reweighed to determine its specific gravity (SG) by Archimedes' principle. After each egg had been carefully dried with a paper towel, it was compressed at 20 mm/min using the egg shell tester described by Voisey and MacDonald (1978). Nondestructive deformation (DFM) was measured as the applied force increased from .98 to 10.79 newtons (N) and quasistatic compression fracture strength (CFS) was determined at the time of fracture. Next, the content of the eggs was broken out on a glass plate, the albumen height was measured by the electronic albumen meter described by Buckley et al. (1981), and the occurrence of blood spots was recorded. The shells were washed, dried, and weighed, including membranes, according to the procedure described by Hamilton (1978). Shell thickness was measured with a dial gauge comparator (see Voisey et al, 1969) on three pieces of shell taken near the point of fracture during the CFS measurement. To test the effects of dietary DON on egg hatchability, all eggs laid over a 7-day period, beginning when the hens were 251 days of age, were collected and incubated under standard conditions. The hens were artificially insemi-
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sumption of DON, especially with swine, hence, the origin of the common name vomitoxin. With the identification of DON in the white winter wheat crop grown in areas adjacent to Lake Huron and Lake Erie of southwestern Ontario (Trenholm et al., 1983), agricultural and health agencies became very concerned about the safety of animal and poultry feeds containing such DON-contaminated wheat (Trenholm et al., 1985). As there was little published data on the effects of feeding poultry diets that contained DON, the present experiments were done to determine the effects of incorporating white winter or spring wheat, naturally contaminated with DON, into diets for White Leghorn hens. Studies with Leghorn chicks, broiler chicks, and turkey poults have been reported separately by Hamilton et al. (1985).
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.63
.65
12.16 5 3.0
.66
11.14 48.3
10.5 3.51
2.6
89.1 12.4
.2 .02
95.0 195.0
710.0
HC2C
11.23 50.7
.64
3.31
2.5 8.4
88.8 15.5
1.3 1.0
477.5 177.5 150.0 195.0
HVRl
10.79 51.8
.63
11.3 3.63
2.8
89.1 15.5
3.7 2.0
322.0 355.0 128.0 195.0
HVR2
1 5
8 1
71 9 19
H
Exper
For Experiment 2, estimate of precision calculated from: SE = •y'2d , /2n, where d = the difference between duplica
Based on formulation No. 10 for laying hens published by Summers and Leeson.
.63
.61
12.2 3.34
2.9
12.4 3.09
2.4
2.6
12.9 3.24
2.2
89.6 15.4
.5 .02
170.0 195.0
635.0
HC1C
90.1 14.5
.7 .7
290.0
710.0
DON-W
91.2 14.2
12.4 3.04
90.7 13.5
0
.35 .35
290.0
290.0
<.05
355.0 355.0
710.0
CW/ DON-W
5
TME n = True metabolizable energy corrected to zero nitrogen balance.
4 Based on chemical analysis, the DON content of the clean winter and spring wheat used to prepare diets CW and HC for diets HVR4, HVR2, and HVRl, 5.7 mg/kg; diet HVS, 6.5 mg/kg; and diet HVT 1.7 mg/kg; and the winter wheat mg/kg.
Basal diets for Experiments 1 and 2, respectively, contained (kg/1000 kg): barley, 189.0, 281.0; dehydrated alfa tallow, 34.0, 51.0; ground limestone, 262.0, 388.0; dicalcium phosphate, 40.0, 59.0; iodized salt, 9.0, 13.0; vitamin-m 4.0. Vitamin-mineral premix supplied per kg of diet: vitamin A, 3650 IU; vitamin D 3 , 725 IU; vitamin E, 3 IU; vitam thenate, 3.5 mg; niacin, 8.5 mg; folic acid, 375 Mg; biotin, 100 Mg; vitamin B i a , 5 ng; manganese, 59.0 mg; zinc, 37.5 mg
3
!
1
Composition (by analysis) DON (mg/kg) Analyzed Calculated" Dry matter (DM), % Crude protein, % Ether extract, % Ash, % Calcium, % Total phosphorus, % TME n , kj/g 5 Apparent DM digestibility, %
Ingredients (kg/1000 kg) Clean wheat DON wheat Soybean meal, 48% protein Basal3
CW
Experiment 1
TABLE 1. Composition of diets for laying hens that contained white winter wheat contaminated with deoxynivalenol (DON)1
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DEOXYNIVALENOL AND LAYING HENS
The hens were housed individually in cages in the same room as used for Experiment 1 and received 16 hr (0100 to 1700 hr) of light. There
were eight replicates of diets HC1C, HVR4, HVR2, HVR1, HVS, and HVT and four for diets HC2C and HDON-W. Each replicate contained seven hens housed as a group/diet, except for diet HC1C, which had six birds/ group. The hens were 361 days of age at the beginning of the experiment and were from three selected strains (2, 4, and 8) and eight crosses of these strains (14 and 4 1 , 18 and 81, 23 and 32, and 28 and 82). Each diet-treatment group contained approximately the same number of hens from each strain and cross. Feed and water were provided ad libitum, Feed intake was measured over 7-day intervals during the first 28 days, over 14-day intervals for the next 28 days, and at 28-day intervals to the end of the 168-day experimental period. Body weights were obtained at the beginning and at 42-day intervals thereafter during the experiment. Beginning 21 days after the start of the experiment and at 28-day intervals thereafter, the eggs laid on 3 consecutive days were sorted, according to dietary treatment group in each replicates, by weight into five groups: pee-wee (<42.49 g), small (42.50 to 49.69 g), medium (49.70 to 56.66 g), large (56.70 to 63.79 g), and extra large (>63.80 g) according to standard Canadian grades (Canada Gazette, 1974). Eggs laid over a 5-day period, starting when hens were 415, 471, and 511 days of age, were used to measure shell strength as described in Experiment 1, but albumen height was not determined. Two hatchability tests were done: eggs were saved from the hens between 419 and 425 days for the first and between 476 and 488 days of age for the second test. All hens were inseminated with pooled semen from Strain 4 roosters, twice before and once during the collection of the hatching eggs for each test. Necropsy examinations were done on one hen, randomly selected, from each replicate for diets HC1C, HC2C, HVR4, and HDON-W, and two hens/replicate for the remaining four diets. All other procedures were similar to those used in Experiment 1. Statistical Analyses. Analyses of variance were applied to means of the groups within replicate, rather than to individual observations. There was some concern that the imbalance of strains within groups in Experiment 2 might influence the analyses. However, the impact of strain differences was found to be minimal and hence strains were ignored in Experiment 2. In addition, previous results (Hamilton et al,
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nated (AI) with semen from male breeders from selected Strain 2 and control Strain 5, described by Gowe and Fairfull (1980), twice before and once during the collection of fertile eggs. Each hen was inseminated with semen from a different cock at each AI. The eggs set were candled after 8 days of incubation and after 18 days, as they were transferred from the "setter" to "hatchor" incubator. All of the eggs classified as infertile or dead, which were moved at candling after 8 and 18 days of incubation, were opened and visually examined, as well as all the unhatched eggs present at hatching. Counts were recorded according to strain and dietary treatment for the following: eggs set, infertile eggs and early dead germs (EDG) identified after 8 days of incubation, and late dead germs (LDG) and pips identified at hatching. Morphologically normal or malformed embryos were classified at 8 and 18 days of incubation and at hatching. Experiment 2. The eight diets used in this experiment were formulated by replacing the wheat and a portion of soybean meal in the control diet (HC1C) with white winter or spring wheat as indicated in Table 1. Diets HVR4, HVR2, and HVR1 were calculated to contain 4.0, 2.0, and 1.0 mg DON/kg of diet, respectively, from the same source of DON-containing (5.7 mg/kg) spring wheat, by replacing 0, 50, or 75% of this wheat with the "clean" wheat used for the control diet HC1C. Two other spring wheat samples that contained 6.5 and 1.7 mg DON/kg, respectively, were used in diets HVS and HVT. Diet HDON-W contained white winter wheat from the same lot as used for Experiment 1. Because the amount of both wheat and soybean meal varied in the experimental diets, diet HC2C was included as a control for diets HVR4, HVS, and HVT. All diets were formulated to be isocaloric and, except for diets HC2C, isonitrogenous. Samples of all diets were analyzed as described in Experiment 1, except the procedure of Scott et al. (1981) was used to measure the DON content of the Experiment 2 diets. The true metabolizable energy (TME) content of the diets was determined according to the method developed by Sibbald (1976) using adult cocks, except the excreta were collected over a 48-hr period and the TME values were corrected to zero nitrogen balance (TME n ) (Sibbald and Morse, 1983).
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1844
HAMILTON ET AL.
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wheat may also account for some of this difference (Trenholm et ah, 1985). The higher protein (N x 6.25) content of the DON-containing diets of Experiment 1 was largely due to the protein content of the DON wheats being higher than that of the clean wheat used in the control diet CW (Hamilton and Trenholm, 1984). Although the Experiment 2 diets were formulated to be isocaloric, the measured TME n content of these diets differed (P<.01) from the values for the DON-containing diets; the average was lower than the controls (11.34 vs. 11.65 kj/g). This is in contrast to Hamilton and Trenholm (1984) who observed that the TME n and true available amino acid content of the DON-containing wheats were higher than those of the clean wheat. However, the presence of DON wheat had no significant (P>.05) effect on the apparent dry matter digestibility of the diets in Experiment 2 (Table 1). Tables 2 and 3 show the effects on productive performance of laying hens that were given diets containing DON-contaminated white winter and spring wheats. Owing to an error in recording, it was not possible to calculate feed intake and efficiency for the mid-portion (57 to 111 days) of Experiment 2 (Table 3). Incorporation of white winter or spring wheat in the RESULTS AND DISCUSSION experimental diets providing up to 4.9 mg Trace levels of zearalenone (12 to 460 \1%I DQN/kg had no effect ''Pi*.05^ on feed intake kg) were found present in the lots of white and efficiency, egg production and yield, the winter and spring wheats in proportion to the number of soft shell and cracked eggs observed concentration of DON in the respective samples in the laying house, and final body weight at as reported by Hamilton and Trenholm (1984). the 10-week experimental period for ExperiThe DON content of the wheats are presented ment 1 and the 24-week period for Experiment in Table 1. Aflatoxin, ochratoxin, and T-2 2. In the latter experiment, the hens given the toxin were not detected in these wheats. diets that contained DON (HVR1, HVR2, As Allen et al. (1981) found that dietary HVR4, HVS, HVT, and HDON-W) tended zearalenone levels up to 800 mg/kg had little (P>.05) to consume less feed (2.4 and 4.4% for effect on egg production and weight, feed 1 to 56 and 112 to 168 days, respectively) and intake, body weight, fertility, hatchability of to produce more eggs (1.3%) and egg mass fertile eggs, organ weights, and egg interior and (2.1%), compared to those receiving the control shell quality, it was unlikely that the low levels diets (HC1C and HC2C). As a result, the DONof zeraralenone present in the experimental fed hens utilized their feed more efficiently (1.4 to 4.4%) for egg production than the birds diets influenced the results being reported. given the control diets. For Experiment 1, The measured DON content of the diets differed much more from the calculated values for however, the aforementioned variables were Experiment 2 than Experiment 1 (Table 1). As marginally improved for the hens receiving the Hamilton et al. (1985) noted, several reasons control diet CW than for those given the DONmay account for this lack of agreement be- containing diets. Mortality was very low in both tween the analyzed and calculated DON values experiments. The responses of the hens in the for the diets from Experiment 2, particularly present experiments (Tables 2 and 3) to diets diets HVR2, HVT, and HDON-W. However, that contained DON-contaminated wheat were recent results indicate that incomplete extract similar to the effects reported by Chi et ah of the DON from the naturally contaminated (1977) for Leghorn hens given diets that con1985) indicated that young Leghorn and broiler chicks, and turkey poults responded to diets containing DON in a similar manner. Those variables that involved repeated measurements at various stages of the experiments were analyzed using the split-plot approach described by Snedecor and Cochran (1967; Section 12.12). Trends with increasing levels of DON were examined using orthogonal polynomials as outlined by Snedecor and Cochran (1967) in their section 12.6. The egg grade data of Experiment 2 were expressed as percentages within group across the 5-day collection periods and then analyzed in the same manner as the group means. This approach was taken rather than analyzing the data as a contingency table because of the repeated eggs from each hen. There were several variables of interest, such as the egg grade data, which were reported as percentages w'here data transformations were considered. However, because the data analyzed were group means and the percentages for these variables tended to be in the mid-range, the distribution of these data were considered to approximate the normal distribution sufficiently for the purpose of this study.
DEOXYNIVALENOL AND LAYING HENS
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TABLE 2. Feed intake and efficiency, egg production and yield, laying house shell quality, mortality, and body weight of hens given diets that contained white winter wheat contaminated with deoxynivalenol (DON) in Experiment 1 Diets' CW/ DON-W
DON-W
SEM
122
121
125
.9
1.68 2.55 87.9 48.3 .9 .25 1
1.72 2.67 86.7 47.2 2.3 .45 1
1.70 2.64 88.4 47.8 1.1 .30 1
.020 .041 .76 .48 .25 .069 ...
1.57 1.64
1.58 1.64
1.59 1.64
.010 .011
1
CW = Control diet of clean white winter wheat (containing =.05 mg/kg DON; CW/DON-W and DON-W = winter wheat diets containing 1.0 mg/kg DON. tained purified T-2 toxin, which is anotherr member of the trichothecene class of mycotoxins. These researchers found that dietaryf T-2 toxin levels up to 4 mg/kg had no effectt (P>.05) on feed intake, egg production, andi feed efficiency, but at 8 mg, T-2 toxin/kg feedi intake and egg production were lower (P<.05)) than the values for the control hens. There weree differences among strains in Experiment 1 forr feed intake and efficiency (P<.01), number off soft shells (P<.05), body weights (P<.001), andi egg production and yield (P<.001). There weree no (P>.05) diet X strain interactions. Thee significant differences among strains for body/ weight, and egg production and yield agree withti the results of Fairfull et al. (1983). Examination of the egg size data fromi Experiment 2 indicated that the response to3 dietary DON of egg size may not have been ass consistent as indicated by the egg yield resultss (Table 3). While there was a tendency for thee majority of eggs to occur in the two largestt categories, extra large and large, the relativee proportions varied somewhat between thesee categories. The variation among diets wass significant (P<.05) only for the mediumi category. Age influenced (P<.05 to .001) thee distribution of eggs, but there were no age X< dietary treatment interactions. Measurements of egg shell quality were obtained three times during Experiment 2, butt diet X age interactions were significant only forr egg weight and percent shell. However, these:
interactions accounted for less than 6% of the total sum of squares. Hence, means over the three measurements are presented in Table 4 for the shell quality variables in Experiment 2. The corresponding means for Experiment 1 are also included. Dietary treatment influenced shell weight (P<.01) and thickness (P<.05), percent shell (P<.05), and DFM (P<.05) of eggs in Experiment 1. There were inverse linear relationships between dietary DON concentrations and egg weight (P<.05), shell weight (P<.01), shell thickness (P<.01), and percent shell (P<.05). Compared to the eggs from the hens receiving the control diet CW, those from the birds given diet DON-W containing .7 mg DON/mg weighed 1.8% less and their shells were lighter (3.6%), thinner (2.2%), and weaker (1.5% lower compression fracture strength). In contrast, the egg and shell variables were not significantly influenced (P>.05) by dietary treatment in Experiment 2 (Table 4). For these variables, the differences between the control diet HC1C and diet HDON-W, which contained DON-contaminated winter wheat from the same stock as used in Experiment 1, were the greatest (.2 to 6.9%, P>.05) of all pairs of dietary treatments, except egg weight for diets HVT and HC2C. These results indicate that the decreases observed in Experiment 1, as dietary DON levels increased, may have been due to wheat source, but further research would be required to confirm this hypothesis. There was little statistical evidence of a relationship be-
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Feed intake, g/hen/day Feed efficiency kg/12 eggs kg/kg egg Egg production, % hen day Egg yield, g/hen/day Soft shelled eggs, % Cracked eggs, % No. hens died Body weight, kg Initial Final
CW
1.82 1.67
1.92 1.77
265 26.13 52.70 21.17 0
2.57 2.85 61.8 37.8 1.9 1.1 1
2.65 3.26 61.0 38.8 5.4 1.2 3
466 47.63 42.39 9.73 .25
1.85 2.25
101 104
2.02 2.60
111 102
HC2C
513 52.44 42.00 5.56 0
1.79 1.72
2.60 3.17 58.8 37.6 5.3 1.2 1
1.93 2.50
103 93
HVRl
585 38.51 50.40 9.27 1.81
1.79 1.67
2.47 2.95 61.7 38.8 5.5 1.6 1
1.81 2.44
103 98
HVR2
Diets
591 41.52 48.30 9.98 .20
1.81 1.74
2.42 3.11 62.1 39.3 4.7 1.0 3
1.79 2.38
103 100
HVR4
582 42 50 6
1 1
2 2 63 40 3 1 1
1 2
103 99
HV
2 The standard error of the mean (SEM) for comparing the means of diets HC2C and HDON-W with those for the othe by 1.4.
1 Based on chemical analysis, the DON content of the clean winter and spring wheat used to prepare diets CW and HC1 for diets HVR4, HVR2, and HVRl, 5.7 mg/kg; diet HVS, 6.5 mg/kg; and diet HVT, 1.7 mg; and the winter wheat f mg/kg.
No. eggs graded Extra large, % Large, % Medium, % Small, %
Feed intake, g/hen/day 1-56 days 1 1 2 - 1 6 8 days Feed efficiency, kg/12 eggs 1-56 days 1 1 2 - 1 6 8 days Feed efficiency, kg/kg egg 1—56 days 1 1 2 - 1 6 8 days Egg production, % hen day Egg yield, g/hen/day Soft shelled eggs, % Cracked eggs, % No. hens died Body weight, kg Initial Final
HOC1
TABLE 3. Feed intake and efficiency, egg production and yield, laying house shell quality, mortality for hens given diets that contained spring or white winter wheat contaminated with deoxy in Experiment 2
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DEOXYNIVALENOL AND LAYING HENS
the conditions (temperature and humidity) were similar within the incubators. Thus, the actual cause(s) of the low hatchability for Hatch 2 is not known; however, all treatments were affected to about the same extent, except HDON-W group. Chi et al. (1977) found that dietary levels of up to 4 mg/kg T-2 toxin had little effect on fertility and hatchability of fertile eggs. The presence of DON-contaminated white winter or spring wheat in the experimental diets had little effect (P>.05) on the ratio of organ weight to body weight of the hens necropsied at the completion of Experiments 1 and 2 (Table 6). There was no apparent evidence of lesions in the oral cavity, esophagus, proventriculus, and gizzard or of hemorrhaging in the viscera or muscles of the hens. The spleen, heart, and kidney appeared normal, but the livers from DON-fed hens were fatty in appearance. Using the livers obtained at necropsy from the hens of Experiment 1, Farnworth et al. (1983) found that the total lipid and triglyceride content of the livers from the hens given the CW/DON-W and DON-W diets was higher (P<.05) than the values for those receiving the CW control diet; the relationship was quadratic. From daily observations of the hens during these two experiments, there were no apparent evidence of vomiting (emesis), diarrhea, or changes in the behaviour of the birds. Moran et al. (1982) found that dietary DON levels up to 49 mg/kg produced no lesions in the oral cavity and only minor erosion in the gizzard of male broiler chicks after the birds had received the experimental diets for 6 days. In addition, they found that alertness, coordination, and livability were not influenced by DON levels up to 210 mg/kg of diet nor, based on microscopic examination, did these dietary levels affect the livers and kidney. The results of Chi et al. (1977) indicate that levels of up to 4 mg/kg purified T-2 toxin had little (P>.05) effect on the organ-to-body weight ratios for heart, liver, gizzard, and spleen for White Leghorn hens. The results from these experiments with laying hens, fed DON-containing diets for up to 24 weeks, support the previous findings obtained by Huff et al. (1981), Moran et al. (1982), Hulan and Proudfoot (1982) with broiler chicks, and Hamilton et al. (1985) with young chickens and turkey poults; namely, low dietary levels of DON have little effect on feed intake and efficiency, production performance,
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tween dietary treatment and shell quality over diets HC1C, HVR1, HVR2, and HVR4, although the trends towards lower values with higher dietary DON levels were similar to those of Experiment 1. Data from Chi et al. (1977) indicate that hens given diets containing up to 4 mg T-2 toxin/kg produced eggs that were (P>.05) larger and had thicker shells than those from birds receiving the control diet (52.4 vs. 51.6 g, and 324 vs. 319 jum, respectively). Interior egg quality, expressed as Haugh units, was also higher (P>.05) for the T-2 toxin-fed hens and the controls (92.83 vs. 90.32). Results in Table 5 indicate that the presence of white winter (Experiment 1) or spring wheat (Experiment 2) had little (P>.05) effect on fertility, hatchability of fertile eggs, and on the proportion of malformed embryos and pips. The lower fertility for Hatch 1, diet HC1C in Experiment 2, was due to the fertility being inexplicably low for hens in two blocks which contributed 43 eggs to the total of 169 set. The average fertility for these blocks was 41 and 19%, respectively, compared to an average of 80.4% for the birds in the remaining 6 blocks for this treatment. Embryonic mortality was influenced by dietary treatment in Experiment 1 (P<.05) and in Hatch 2 of Experiment 2 (P<.01). There was little evidence from the morphological data that the mortality of the embryos was associated with any particular abnormality. In fact, visual examinations indicated that most of the dead embryos appeared normal. The. low hatchability of fertile eggs in Hatch 2 of Experiment 2 was due to high embryonic mortality, particularly to an increase in the proportion of LDG and pips. The shell quality data for the first and second measurement periods, which occurred 7 to 10 days before the eggs were collected for hatches 1 and 2, respectively, indicated that there were little differences among the treatments between periods for egg and shell weights, percent shell, shell thickness, specific gravity, deformation, and compression fracture strength. Therefore, the much lower hatchability of egg sets for Hatch 2 than Hatch 1 was likely not due to shell quality. In addition, while age of hen does influence egg hatchability (Tomhave, 1956, 1958; Martin and Wessels, 1965), the differences in the ages of the birds in the present study (419 vs. 476 days) would likely not account for the large decrease obtained in the hatchability between Hatches 1 and 2. Records maintained during these hatches indicate that
1847
56.2 5.22 372 9.30 1.080 81 27.4 7.54
SEM .40 .049 .092 .151 .0062 1.2 .45 .093
DON-W
55.2 5.03 364 9.12 1.079 82 27.0 7.67 64.3 5.41 357 8.41 1.080 77 15.3
HCIC 61.3 5.23 358 8.53 1.080 77 15.1
HC2C 64.2 5.45 357 8.49 1.081 79 15.2
HVR1 63.5 5.40 358 8.51 1.081 80 15.3
HVR2
Measurements taken on eggs laid for 5 consecutive days after t h e hens were 243 days of age and had received the diets for 52 days.
55.7 5.09 366 9.13 1.079 82 27.8 7.54
CW 3
white winter
Ex
or
6
5
4
Quasi-static compression fracture strength expressed in newtons (N).
Measured by Archimedes' principle.
T h e standard error of the mean (SEM) for comparing the means of diets HC2C and H D O N W with those for the other diets is obtained b y m
3 Based o n chemical analysis, t h e DON content of t h e clean winter and spring wheat used t o prepare diets CW and H C I C , respectively wa H V R 1 , 5.7 mg/kg; diet HVS, 6.5 mg/kg; and diet HVT, 1.7 mg; and t h e winter wheat for diets CW/DON-W, DON-W, and HDON-W, 1.0 mg/kg.
Mean of three measurements taken on eggs laid for 5 consecutive days beginning when the hens were 4 1 5 , 4 7 1 , and 511 days of age, respec experiment.
3
1
Egg weight, g Shell weight, g Shell thickness, fim Percent shell Specific gravity 5 Nondestructive deformation, (ira 6 Compression fracture strength, N Albumen height, m m
CW/ DON-W
Experirr lent 1'
TABLE 4. Shell and interior quality of eggs from hens given diets that contained contaminated with deoxynivalenol (DON)
ded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
92.8
Hatchability of fertile eggs, %
85.3
0
86.2
0
2.4
3.0
5.8
88.8
97.1
598
1.23
.35
.56
.69
1.19
.58
SEM
Hatch
60.6 23.9
.89 .39
65.8 33.1
0 5.4 0 1.3
1.9
11.1
27.8
8.3
1?.9 16.7 34.8
4.5
83.2 83.3 78.9 39.8
90 165
33.8
2.8
69.0 87.0 87.0 27.3
169 263
HC2C
6.7 24.5 5.0 36.0 3.5 12.0 1.1 .27 70.1 23.9
188 314 82.0 86.6 84.7 27.4
Hens were between 419 and 425 days of age when the eggs were collected for Hatch 1, and 476 and 488 days of age for Hatch 2.
The eggs were collected when the hens were between 244 and 250 days of age.
90.2
0
1.2
4.2
7.7
86.4
98.9
639
DON-W
Exper
12.1 16.8 6.5 36.4 1.1 8.3 .54 .31 64.1 32.2
195 338 79.1 84.2 80.3 38.5
HVR2
s
4
The standard error of the mean (SEM) for comparing the means of diets HC2C and HDON-W with those for the other diets is obtained by m Percent of fertile eggs. EDG = early dead germ. LDG = late dead germ.
Based on chemical analysis, the DON content of the clean winter and spring wheat used to prepare diets CW and HCIC, respectively w HVRl, 5.7 mg/kg; diet HVS, 6.5 mg/kg; and diet HVT, 1.7 mg; and the winter wheat for diets CW/DON-W, DON-W, and HDON-W, 1.0 mg/kg.
3
2
1
Overall hatchability of eggs set, %
Malformed embryos,s %
Pips, %
1.6
2.9
LDG,S %
5
2.7
EDG, S %
Embryonic mortality
97.1
605
Fertility of eggs set, %
No. eggs set
CW3
CW/ DON-W
Experiment l 1
TABLE 5. Fertility, batcbability, and embryonic mortality of eggs from bens given uncontaminated die or diets (CW/DON-W, DON-W, HVRl, HVR2, HVR4, HVS, HVT, and HDON-W) that contained white winter or spring wheat
ed from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
1850
HAMILTON ET AL.
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T h e a u t h o r s t h a n k Barbara B e t h u n e , Dina D ' E r m o , Lorraine R o b i n s o n , R. T. Poirier, and J. F . Shackleton for their capable technical assistance, K. E. Hartin for his veterinarian assistance, A. R. Morrison and staff for their careful daily care and observation of t h e birds, J. S. Racine and staff for their assistance w i t h t h e mixing of t h e diets, R. Ciok for assistance w i t h t h e statistical analyses of t h e data, and J. Carroll for her assistance with t h e d e o x y n i valenol analysis of t h e diets' from E x p e r i m e n t 2. Chemical analysis of t h e w h e a t samples and diets from E x p e r i m e n t 1 for deoxynivalenol was d o n e b y H. Campbell, L a b o r a t o r y Services Division, F o o d P r o d u c t i o n and Inspection Branch, Agriculture Canada. Analysis of t h e diets from b o t h e x p e r i m e n t s for p r o x i m a t e c o m p o n e n t s was d o n e b y M. Ihnat, Analytical Services, Chemistry and Biology Research Instit u t e , Research Branch, Agriculture Canada, Ottawa. REFERENCES
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Allen, N. K., C. J. Mirocha, S. Aakhus-Allen, J. J. Bitgood, G. Weaver, and F. Bates, 1981. Effect of dietary zearalenone on reproduction of chickens. Poultry Sci. 60:1165-1174. Association of Official Analytical Chemists, 1975. Official Methods of Analysis. 12th ed. Assoc. Off. Anal. Chem., Washington, DC. Buckley, D. J., G. St-Amour, and R. W. Fairfull, 1981. An improved electronic gauge for measuring egg albumen height. Poultry Sci. 60:777-780. Canada Gazette, 1974. Egg Regulations. Part II, Vol. 108, No. 8. Queen's Printer, Ottawa, Canada. Chi, M. S„ C. J. Mirocha, H. J. Kurtz, G. Weaver, F. Bates, and W. Shimoda, 1977. Effects of T-2 toxin on reproductive performance and health of laying hens. Poultry Sci. 56:628-637. Cohen, H., and M. Lapointe, 1982. Capillary gas-liquid chromatographic determination of vomitoxin in cereal grains. J. Assoc. Off. Anal. Chem. 65: 1429-1434. C6te, L. M., J. D. Reynolds, R. F. Vesonder, W. B. Buck, S. P. Swanson, R. T. Coffey, and D. C. Brown, 1984. Survey of vomitoxin-contaminated feed grains in midwestern United States, and associated health problems in swine. J. Am. Vet. Med. Assoc. 184:189-192. El-Banna, A. A., R.M.G. Hamilton, P. M. Scott, and H. L. Trenholm, 1983. Non-transmission of deoxynivalenol (vomitoxin) to eggs and meat in
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> E
u U
a n d livability of p o u l t r y . F u r t h e r m o r e , n o measurable quantities of DON were f o u n d in eggs from hens given diet H V R 4 (4.1 mg DON/kg) in E x p e r i m e n t 2 for a period of 2 4 w e e k s (El-Banna e t a / . , 1 9 8 3 ) .
DEOXYNIVALENOL AND LAYING HENS
Wyllie and L. G. Morehouse, ed. Marcel Dekker Inc., New York, NY. Moran, E. T„ Jr., B. Hunter, P. Ferket, L. G. Young, and L. G. McGirr, 1982. High tolerance of broilers to vomitoxin from corn infected with Fusarium graminearum. Poultry Sci. 61:1828— 1831. Neish, G. A., and H. Cohen, 1981. Vomitoxin and zearalenone production by Fusarium graminearum from winter wheat and barley in Ontario. Can. J. Plant Sci. 61:811-815. Neish, G. A., E. R. Farnworth, R. Greenhalgh, and J. C. Young, 1983. Observations on the occurrence of Fusarium species and their toxins in corn in eastern Ontario. Can. J. Plant Pathol. 5: 11-16. Newberne, P. M., and A. E. Rogers, 1981. Animal toxicity of major environmental mycotoxins. Pages 51—106, Vol. 1 in Mycotoxins and NNitroso Compounds Environmental Risks. R. C. Shank, ed. CRC Press, Boca Raton, FL. Scott, P. M., P.-Y. Lau, and S. R. Kanhere, 1981. Gas chromatography with electron capture and mass spectrometric detection of deoxynivalenol in wheat and other grains. J. Assoc. Off. Anal. Chem. 64:1364-1371. Sibbald, I. R., 1976. A bioassay for true metabolizable energy in feedingstuffs. Poultry Sci. 55:303 — 308. Sibbald, I. R., and P. M. Morse, 1983. Effects of the nitrogen correction and of feed intake on true metabolizable energy values. Poultry Sci. 62: 138-142. Snedecor, G. W., and W. G. Cochran, 1967. Statistical Methods. 6th ed. The Iowa State Univ. Press, Ames, IA. Summers, J. D., and S. Leeson, Poultry Nutrition Handbook. Ontario Ministry of Agric. and Food, Toronto, Agdex 450/50. Sutton, J. C , 1982. Epidemiology of wheat head blight and maize ear rot caused by Fusarium graminearum. Can. J. Plant Pathol. 4:195—209. Tomhave, A. E., 1956. Influence of age of New Hampshire female breeders upon hatchability of eggs. Poultry Sci. 35:236-237. Tomhave, A. E., 1958. Fertility and hatchability of eggs produced by New Hampshire breeders during their first 365 days of production. Poultry Sci. 37:27-29. Trenholm, H. L., W. P. Cochrane, H. Cohen, J. I. Elliot, E. R. Farnworth, D. W. Friend, R.M.G. Hamilton, J. F. Standish, and B. K. Thompson, 1983. Survey of vomitoxin contamination of 1980 Ontario white winter wheat crop: Results of survey and feeding trials. J. Assoc. Off. Anal. Chem. 6 6 : 9 2 - 9 7 . Trenholm, H. L., D. B. Prelusky, and R. M. Warner, 1985. Incomplete extraction as a source of error in the analyses of naturally contaminated grain products for deoxynivalenol (vomitoxin). J. Assoc. Off. Anal. Chem. 68:In press. Trenholm, H. L., B. K. Thompson, J. F. Standish, and W. L. Seaman. 1985. Mycotoxins in feeds and feedingstuffs. Pages 43—49 in Mycotoxins: A Canadian Perspective. P. M. Scott, H. L. Trenholm, and M. D. Sutton, ed. Natl. Res. Council of Canada, Ottawa, Ontario. Publ. No. 2284.
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chickens fed deoxynivalenol-contaminated diets. J. Agric. Food Chem. 31:1381-1384. Fairfull, R. W., R. S. Gowe, and J.A.B. Emsley, 1983. Diallel cross of six long-term selected Leghorn strains with emphasis on heterosis and reciprocal effects. Br. Poult. Sci. 24:133-158. Farnworth, E. R., R.M.G. Hamilton, B. K. Thompson, and H. L. Trenholm, 1983. Liver lipid levels in White Leghorn hens fed diets that contained wheat contaminated by deoxynivalenol (vomitoxin). Poultry Sci. 62:832-836. Friend, D. W., H. L. Trenbolm, J. I. Elliot, B. K. Thompson, and K. E. Hartin, 1982. Effect of feeding vomitoxin-contaminated wheat to pigs. Can. J. Anim. Sci. 62:1211-1222. Friend, D. W., H. L. Trenholm, J. C. Young, B. K. Thompson, and K. E. Hartin, 1984. Effect of adding potential vomitoxin (deoxynivalenol) detoxicants or a F. graninearum inoculated corn supplement to wheat diets fed to pigs. Can. J. Anim. Sci. 6 4 : 7 3 3 - 7 4 1 . Gowe, R. S., and R. W. Fairfull, 1980. Performance of six long-term multi-trait selected Leghorn strains and .three control strains, and a strain cross evaluation of the selected strains. Pages 141—162 in Proc. 1980 South Pacific Poultry Sci. Convention, Auckland, NZ. Hagler, W. M., Jr., K. Tyczkowska, and P. B. Hamilton, 1984. Simultaneous occurrence of deoxynivalenol, zearalenone, and alfatoxin in 1982 scabby wheat from the Midwestern United States. Appl. Environ. Microbiol. 47:151—154. Hamilton, R.M.G., 1978. Observations on the changes in physical characteristics that influence egg shell quality in ten strains of White Leghorns. Poultry Sci. 57:1192-1197. Hamilton, R.M.G., and H. L. Trenholm, 1984. Observations on the chemical and nutritive content of white winter and spring wheats contaminated with deoxynivalenol (vomitoxin). Anim. Feed Sci. Technol. 9:293-300. Hamilton, R.M.G., H. L. Trenholm, B. K. Thompson, and R. Greenhalgh, 1985. The tolerance of White Leghorn and broiler chicks, and turkey poults to diets that contained deoxynivalenol (vomitoxin) contaminated wheat. Poultry Sci. 64:273—286. Huff, W. E., J. A. Doerr, P. B. Hamilton, and R. F. Vesonder, 1981. Acute toxicity of vomitoxin (deoxynivalenol) in broiler chickens. Poultry Sci. 60:1412-1414. Hulan, H. W„ and F. G. Proudfoot, 1982. Effects of feeding vomitoxin contaminated wheat on the performance of broiler chickens. Poultry Sci. 6 1 : 1653-1659. Martin, R. S., and J.P.H. Wessels, 1965. Seasonal fluctuation in hatchability of fowl eggs. S. Afr. J. Agric. Sci. 8:1069-1074. Miller, J. D., J. C. Young, and H. L. Trenholm, 1983. Fusarium toxins in field corn. I. Time course of fungal growth and production of deoxynivalenol and other mycotoxins. Can. J. Bot. 61:3080— 3087. Mirocha, C. J„ S. V. Pathre, and C. M. Christensen, 1977. Chemistry of Fusarium and Stachybotrys mycotoxins. Pages 365—420 in Mycotoxic Fungi, Mycotoxins, Mycotoxicosis. Vol. 1, Mycotoxic Fungi and Chemistry of Mycotoxins. T. D.
1851
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HAMILTON ET AL.
Ueno, Y., 1980. Trichothecene mycotoxins: Mycology, chemistry, and toxicology. Pages 301—353, Vol. 3 in Advances in Nutritional Research. H. H. Draper, ed. Plenum Press, New York, NY. Vesonder, R. F., A. Ciegler, R. F. Rogers, K. A. Burbridge, R. J. Bothast, and A. H. Jensen, 1978. Survey of 1977 crop year preharvest corn for vomitoxin. Appl. Environ. Microbiol. 36:885 — 888.
Voisey, P. W., J. R. Hunt, and P. E. James, 1969. A comparison of beta backscatter and quasi-static compression methods of measuring eggshell strength. Can. J. Anim. Sci. 49:157-168. Voisey, P. W., and D. C. MacDonald, 1978. Laboratory measurements of egg shell strength. 1. An instrument for measuring shell strength by quasi-static compression, puncture, and non-destructive deformation. Poultry Sci. 57:860-869.
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1985 Poultry Science 64:1840-1852
INTRODUCTION R e c e n t survey results indicate t h a t cereal grains, particularly corn and w h e a t , m a y often b e c o n t a m i n a t e d w i t h 4-deoxynivalenol ( D O N ) (Cote et al, 1 9 8 4 ; Hagler et al, 1 9 8 4 ; Trenholm et al, 1 9 8 3 ; S c o t t et al, 1 9 8 1 ; Ueno,
'Contribution numbers: 1289 Animal Research Centre, 1-655 Engineering and Statistical Research Institute, and 1496 Chemistry and Biology Research Institute. 2 Preliminary report presented at the 70th Annual Meeting, Poultry Science Association, University of British Columbia, Vancouver, August 3—6, 1981 (Poultry Sci. 60:1666, 1981). 3 Animal Research Centre. 4 Engineering and Statistical Research Institute. 5 Chemistry and Biology Research Institute.
1 9 8 0 ; Vesonder et al, 1 9 7 8 ) , especially w h e n cool and w e t climatic conditions prevail ( S u t t o n , 1 9 8 2 ) . O n c e DON, a t r i c h o t h e c e n e m y c o t o x i n , is p r o d u c e d b y t h e mold Fusarium graminearium (Schwabe t e l e m o r p h Gibberella zeae [Schw.] Petch) (Neish et al, 1 9 8 3 ; Neish and Cohen, 1 9 8 1 ) , it will persist, b u t n o t necessarily at m a x i m u m levels (Miller et al, 1 9 8 3 ) . Efforts t o disguise DON w i t h feed additives have n o t b e e n productive (Mirocha et al, 1 9 7 7 ; Friend et al, 1 9 8 4 ) . DON has been r e p o r t e d t o cause inhibition of p r o t e i n and d e o x y r i b o nucleic acid ( D N A ) biosynthesis, inflammation of t h e skin, leukopenia, decreased feed i n t a k e and weight gains, and i m m u n o s u p p r e s s i o n (Newberne and Rogers, 1 9 8 1 ; Ueno, 1 9 8 0 ) . Emesis or vomiting is also associated w i t h con-
1840
Downloaded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
ABSTRACT A short-term (10 weeks, Experiment 1) and a long-term experiment (24 weeks, Experiment 2) were done to determine effects of incorporating either white winter wheat, naturally contaminated with 1 mg deoxynivalenol (DON)/kg, or spring wheats, containing up to 6.5 mg DON/kg, into the diets of White Leghorn hens. Based on chemical analysis, the diets in Experiment 1 contained <.05 to .7 mg DON/kg, while those in Experiment 2 contained from .2 to 4.9 mg/kg. Incorporation of winter or spring wheat in the experimental diets had no effect (P>,05) on feed intake and efficiency, egg production and yield, the number of soft shell and cracked eggs observed in the laying house, body weight at the completion of the experimental period, fertility, hatchability of fertile eggs, and the proportion of malformed embryos and pips. In addition, presence of DON-contaminated wheat did not influence (P>.05) the organ weight to body weight ratio for a randomly selected sample of hens necropsied at the completion of each experiment. There was little evidence of lesions in the oral cavity, esophagus, proventriculus and gizzard, hemorrhaging in the viscera or skeletal muscles, or of changes in the appearance of spleen, heart, and kidney. However, the livers from DON hens were fatty in appearance. Furthermore, vomiting (emesis), diarrhea, or changes in behaviour were not apparent and mortality, normally very low, was not increased during either experiment. Inverse linear relationships were obtained in Experiment 1 between dietary DON concentrations and egg weight (P<.05), shell weight and thickness (P<.01), and percent shell (P<.05). Although egg and shell variables measured in Experiment 2 were not significantly influenced (P>.05) by DON treatment, trends towards lower values with higher dietary DON levels were evident. Egg specific gravity, nondestructive deformation, and quasistatic compression fracture strength of the egg's shell were not influenced (P>.05) by dietary DON levels. The results from these experiments indicate that laying hens can tolerate diets containing up to 5 mg DON/kg from white winter or spring wheat for extended periods of time without serious adverse effects on health and productivity. (Key words: mycotoxin, deoxynivalenol, hens, wheat, shell quality, performance)
DEOXYNIVALENOL AND LAYING HENS
MATERIALS AND METHODS
Experiment 1. White Leghorn pullets from two randombred control strains (7 and 10) and four 2-way strain crosses (1 X 4, 4 X 1, 4 X 8, and 8 x 4 ) were fed experimental diets for 70 days beginning when they were 192 days of age. The control strains have been previously described by Gowe and Fairfull (1980) and the strain crosses by Fairfull et al. (1983). The pullets were housed in individual wire cages (25.5 or 30.5 X 40.6 cm) in a light-tight room. During the experimental period, the length of the photoperiod was increased from an initial of 11 hr/day to 16 hr/day in increments of 30 min/ week. Feed and water were provided ad libitum. Composition of the control and experimental diets is given in Table 1. The experimental diets were formulated by replacing, on an equal weight basis, 50 or 100% of the white winter wheat in the control diet (CW) with white winter wheat containing 1 mg DON/kg of wheat; the DON-containing diets were designated CW/DON-W and DON-W, respectively. The supplies of white winter wheat were from the 1980 Ontario crop; details of their acquisition and storage were described by Friend et al. (1982). Samples of all mixes of the diets were analyzed for dry matter, Kjeldahl nitrogen, ether extract, ash, calcium, and total phosphorus by the methods of AOAC (1975). The method of Cohen and Lapointe (1982) was used to determine the DON content of the diets. Each diet was given to 102 hens distributed evenly, according to strain, among
three replicates; hence, each strain-diet group contained 17 hens in consecutive cages. The hens were weighed individually at the beginning and end of the experiment. Feed intake was measured over two 35-day periods. The numbers of normal shelled eggs collected from the egg trays and the numbers of broken, cracked, and soft shelled eggs observed in the egg trays and on the dropping boards were recorded daily. The birds were checked daily for evidence of vomiting and diarrhea. Mortality was recorded as it occurred and all hens that died were necropsied. One randomly selected hen from each diet-strain group was killed by carbon dioxide asphyxiation at the end of the experimental period, and the heart, spleen, proventriculus, one kidney, gizzard, and liver from each were removed and weighed. The oral cavity, esophagus, crop, proventriculus, and gizzard were examined for lesions. Egg shell and interior quality measurements were taken on all eggs laid for 5 consecutive days beginning when the hens were 243 days of age and had received the experimental diets for 52 days. The eggs were stored overnight in an egg cooler at 10 C. Upon removal from the cooler, the eggs were allowed to warm, over 3 hr, to room temperature. Subsequently, each egg was weighed in air, then submerged in tap water and reweighed to determine its specific gravity (SG) by Archimedes' principle. After each egg had been carefully dried with a paper towel, it was compressed at 20 mm/min using the egg shell tester described by Voisey and MacDonald (1978). Nondestructive deformation (DFM) was measured as the applied force increased from .98 to 10.79 newtons (N) and quasistatic compression fracture strength (CFS) was determined at the time of fracture. Next, the content of the eggs was broken out on a glass plate, the albumen height was measured by the electronic albumen meter described by Buckley et al. (1981), and the occurrence of blood spots was recorded. The shells were washed, dried, and weighed, including membranes, according to the procedure described by Hamilton (1978). Shell thickness was measured with a dial gauge comparator (see Voisey et al, 1969) on three pieces of shell taken near the point of fracture during the CFS measurement. To test the effects of dietary DON on egg hatchability, all eggs laid over a 7-day period, beginning when the hens were 251 days of age, were collected and incubated under standard conditions. The hens were artificially insemi-
Downloaded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
sumption of DON, especially with swine, hence, the origin of the common name vomitoxin. With the identification of DON in the white winter wheat crop grown in areas adjacent to Lake Huron and Lake Erie of southwestern Ontario (Trenholm et al., 1983), agricultural and health agencies became very concerned about the safety of animal and poultry feeds containing such DON-contaminated wheat (Trenholm et al., 1985). As there was little published data on the effects of feeding poultry diets that contained DON, the present experiments were done to determine the effects of incorporating white winter or spring wheat, naturally contaminated with DON, into diets for White Leghorn hens. Studies with Leghorn chicks, broiler chicks, and turkey poults have been reported separately by Hamilton et al. (1985).
1841
.63
.65
12.16 5 3.0
.66
11.14 48.3
10.5 3.51
2.6
89.1 12.4
.2 .02
95.0 195.0
710.0
HC2C
11.23 50.7
.64
3.31
2.5 8.4
88.8 15.5
1.3 1.0
477.5 177.5 150.0 195.0
HVRl
10.79 51.8
.63
11.3 3.63
2.8
89.1 15.5
3.7 2.0
322.0 355.0 128.0 195.0
HVR2
1 5
8 1
71 9 19
H
Exper
For Experiment 2, estimate of precision calculated from: SE = •y'2d , /2n, where d = the difference between duplica
Based on formulation No. 10 for laying hens published by Summers and Leeson.
.63
.61
12.2 3.34
2.9
12.4 3.09
2.4
2.6
12.9 3.24
2.2
89.6 15.4
.5 .02
170.0 195.0
635.0
HC1C
90.1 14.5
.7 .7
290.0
710.0
DON-W
91.2 14.2
12.4 3.04
90.7 13.5
0
.35 .35
290.0
290.0
<.05
355.0 355.0
710.0
CW/ DON-W
5
TME n = True metabolizable energy corrected to zero nitrogen balance.
4 Based on chemical analysis, the DON content of the clean winter and spring wheat used to prepare diets CW and HC for diets HVR4, HVR2, and HVRl, 5.7 mg/kg; diet HVS, 6.5 mg/kg; and diet HVT 1.7 mg/kg; and the winter wheat mg/kg.
Basal diets for Experiments 1 and 2, respectively, contained (kg/1000 kg): barley, 189.0, 281.0; dehydrated alfa tallow, 34.0, 51.0; ground limestone, 262.0, 388.0; dicalcium phosphate, 40.0, 59.0; iodized salt, 9.0, 13.0; vitamin-m 4.0. Vitamin-mineral premix supplied per kg of diet: vitamin A, 3650 IU; vitamin D 3 , 725 IU; vitamin E, 3 IU; vitam thenate, 3.5 mg; niacin, 8.5 mg; folic acid, 375 Mg; biotin, 100 Mg; vitamin B i a , 5 ng; manganese, 59.0 mg; zinc, 37.5 mg
3
!
1
Composition (by analysis) DON (mg/kg) Analyzed Calculated" Dry matter (DM), % Crude protein, % Ether extract, % Ash, % Calcium, % Total phosphorus, % TME n , kj/g 5 Apparent DM digestibility, %
Ingredients (kg/1000 kg) Clean wheat DON wheat Soybean meal, 48% protein Basal3
CW
Experiment 1
TABLE 1. Composition of diets for laying hens that contained white winter wheat contaminated with deoxynivalenol (DON)1
ded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
DEOXYNIVALENOL AND LAYING HENS
The hens were housed individually in cages in the same room as used for Experiment 1 and received 16 hr (0100 to 1700 hr) of light. There
were eight replicates of diets HC1C, HVR4, HVR2, HVR1, HVS, and HVT and four for diets HC2C and HDON-W. Each replicate contained seven hens housed as a group/diet, except for diet HC1C, which had six birds/ group. The hens were 361 days of age at the beginning of the experiment and were from three selected strains (2, 4, and 8) and eight crosses of these strains (14 and 4 1 , 18 and 81, 23 and 32, and 28 and 82). Each diet-treatment group contained approximately the same number of hens from each strain and cross. Feed and water were provided ad libitum, Feed intake was measured over 7-day intervals during the first 28 days, over 14-day intervals for the next 28 days, and at 28-day intervals to the end of the 168-day experimental period. Body weights were obtained at the beginning and at 42-day intervals thereafter during the experiment. Beginning 21 days after the start of the experiment and at 28-day intervals thereafter, the eggs laid on 3 consecutive days were sorted, according to dietary treatment group in each replicates, by weight into five groups: pee-wee (<42.49 g), small (42.50 to 49.69 g), medium (49.70 to 56.66 g), large (56.70 to 63.79 g), and extra large (>63.80 g) according to standard Canadian grades (Canada Gazette, 1974). Eggs laid over a 5-day period, starting when hens were 415, 471, and 511 days of age, were used to measure shell strength as described in Experiment 1, but albumen height was not determined. Two hatchability tests were done: eggs were saved from the hens between 419 and 425 days for the first and between 476 and 488 days of age for the second test. All hens were inseminated with pooled semen from Strain 4 roosters, twice before and once during the collection of the hatching eggs for each test. Necropsy examinations were done on one hen, randomly selected, from each replicate for diets HC1C, HC2C, HVR4, and HDON-W, and two hens/replicate for the remaining four diets. All other procedures were similar to those used in Experiment 1. Statistical Analyses. Analyses of variance were applied to means of the groups within replicate, rather than to individual observations. There was some concern that the imbalance of strains within groups in Experiment 2 might influence the analyses. However, the impact of strain differences was found to be minimal and hence strains were ignored in Experiment 2. In addition, previous results (Hamilton et al,
Downloaded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
nated (AI) with semen from male breeders from selected Strain 2 and control Strain 5, described by Gowe and Fairfull (1980), twice before and once during the collection of fertile eggs. Each hen was inseminated with semen from a different cock at each AI. The eggs set were candled after 8 days of incubation and after 18 days, as they were transferred from the "setter" to "hatchor" incubator. All of the eggs classified as infertile or dead, which were moved at candling after 8 and 18 days of incubation, were opened and visually examined, as well as all the unhatched eggs present at hatching. Counts were recorded according to strain and dietary treatment for the following: eggs set, infertile eggs and early dead germs (EDG) identified after 8 days of incubation, and late dead germs (LDG) and pips identified at hatching. Morphologically normal or malformed embryos were classified at 8 and 18 days of incubation and at hatching. Experiment 2. The eight diets used in this experiment were formulated by replacing the wheat and a portion of soybean meal in the control diet (HC1C) with white winter or spring wheat as indicated in Table 1. Diets HVR4, HVR2, and HVR1 were calculated to contain 4.0, 2.0, and 1.0 mg DON/kg of diet, respectively, from the same source of DON-containing (5.7 mg/kg) spring wheat, by replacing 0, 50, or 75% of this wheat with the "clean" wheat used for the control diet HC1C. Two other spring wheat samples that contained 6.5 and 1.7 mg DON/kg, respectively, were used in diets HVS and HVT. Diet HDON-W contained white winter wheat from the same lot as used for Experiment 1. Because the amount of both wheat and soybean meal varied in the experimental diets, diet HC2C was included as a control for diets HVR4, HVS, and HVT. All diets were formulated to be isocaloric and, except for diets HC2C, isonitrogenous. Samples of all diets were analyzed as described in Experiment 1, except the procedure of Scott et al. (1981) was used to measure the DON content of the Experiment 2 diets. The true metabolizable energy (TME) content of the diets was determined according to the method developed by Sibbald (1976) using adult cocks, except the excreta were collected over a 48-hr period and the TME values were corrected to zero nitrogen balance (TME n ) (Sibbald and Morse, 1983).
1843
1844
HAMILTON ET AL.
Downloaded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
wheat may also account for some of this difference (Trenholm et ah, 1985). The higher protein (N x 6.25) content of the DON-containing diets of Experiment 1 was largely due to the protein content of the DON wheats being higher than that of the clean wheat used in the control diet CW (Hamilton and Trenholm, 1984). Although the Experiment 2 diets were formulated to be isocaloric, the measured TME n content of these diets differed (P<.01) from the values for the DON-containing diets; the average was lower than the controls (11.34 vs. 11.65 kj/g). This is in contrast to Hamilton and Trenholm (1984) who observed that the TME n and true available amino acid content of the DON-containing wheats were higher than those of the clean wheat. However, the presence of DON wheat had no significant (P>.05) effect on the apparent dry matter digestibility of the diets in Experiment 2 (Table 1). Tables 2 and 3 show the effects on productive performance of laying hens that were given diets containing DON-contaminated white winter and spring wheats. Owing to an error in recording, it was not possible to calculate feed intake and efficiency for the mid-portion (57 to 111 days) of Experiment 2 (Table 3). Incorporation of white winter or spring wheat in the RESULTS AND DISCUSSION experimental diets providing up to 4.9 mg Trace levels of zearalenone (12 to 460 \1%I DQN/kg had no effect ''Pi*.05^ on feed intake kg) were found present in the lots of white and efficiency, egg production and yield, the winter and spring wheats in proportion to the number of soft shell and cracked eggs observed concentration of DON in the respective samples in the laying house, and final body weight at as reported by Hamilton and Trenholm (1984). the 10-week experimental period for ExperiThe DON content of the wheats are presented ment 1 and the 24-week period for Experiment in Table 1. Aflatoxin, ochratoxin, and T-2 2. In the latter experiment, the hens given the toxin were not detected in these wheats. diets that contained DON (HVR1, HVR2, As Allen et al. (1981) found that dietary HVR4, HVS, HVT, and HDON-W) tended zearalenone levels up to 800 mg/kg had little (P>.05) to consume less feed (2.4 and 4.4% for effect on egg production and weight, feed 1 to 56 and 112 to 168 days, respectively) and intake, body weight, fertility, hatchability of to produce more eggs (1.3%) and egg mass fertile eggs, organ weights, and egg interior and (2.1%), compared to those receiving the control shell quality, it was unlikely that the low levels diets (HC1C and HC2C). As a result, the DONof zeraralenone present in the experimental fed hens utilized their feed more efficiently (1.4 to 4.4%) for egg production than the birds diets influenced the results being reported. given the control diets. For Experiment 1, The measured DON content of the diets differed much more from the calculated values for however, the aforementioned variables were Experiment 2 than Experiment 1 (Table 1). As marginally improved for the hens receiving the Hamilton et al. (1985) noted, several reasons control diet CW than for those given the DONmay account for this lack of agreement be- containing diets. Mortality was very low in both tween the analyzed and calculated DON values experiments. The responses of the hens in the for the diets from Experiment 2, particularly present experiments (Tables 2 and 3) to diets diets HVR2, HVT, and HDON-W. However, that contained DON-contaminated wheat were recent results indicate that incomplete extract similar to the effects reported by Chi et ah of the DON from the naturally contaminated (1977) for Leghorn hens given diets that con1985) indicated that young Leghorn and broiler chicks, and turkey poults responded to diets containing DON in a similar manner. Those variables that involved repeated measurements at various stages of the experiments were analyzed using the split-plot approach described by Snedecor and Cochran (1967; Section 12.12). Trends with increasing levels of DON were examined using orthogonal polynomials as outlined by Snedecor and Cochran (1967) in their section 12.6. The egg grade data of Experiment 2 were expressed as percentages within group across the 5-day collection periods and then analyzed in the same manner as the group means. This approach was taken rather than analyzing the data as a contingency table because of the repeated eggs from each hen. There were several variables of interest, such as the egg grade data, which were reported as percentages w'here data transformations were considered. However, because the data analyzed were group means and the percentages for these variables tended to be in the mid-range, the distribution of these data were considered to approximate the normal distribution sufficiently for the purpose of this study.
DEOXYNIVALENOL AND LAYING HENS
1845
TABLE 2. Feed intake and efficiency, egg production and yield, laying house shell quality, mortality, and body weight of hens given diets that contained white winter wheat contaminated with deoxynivalenol (DON) in Experiment 1 Diets' CW/ DON-W
DON-W
SEM
122
121
125
.9
1.68 2.55 87.9 48.3 .9 .25 1
1.72 2.67 86.7 47.2 2.3 .45 1
1.70 2.64 88.4 47.8 1.1 .30 1
.020 .041 .76 .48 .25 .069 ...
1.57 1.64
1.58 1.64
1.59 1.64
.010 .011
1
CW = Control diet of clean white winter wheat (containing =.05 mg/kg DON; CW/DON-W and DON-W = winter wheat diets containing 1.0 mg/kg DON. tained purified T-2 toxin, which is anotherr member of the trichothecene class of mycotoxins. These researchers found that dietaryf T-2 toxin levels up to 4 mg/kg had no effectt (P>.05) on feed intake, egg production, andi feed efficiency, but at 8 mg, T-2 toxin/kg feedi intake and egg production were lower (P<.05)) than the values for the control hens. There weree differences among strains in Experiment 1 forr feed intake and efficiency (P<.01), number off soft shells (P<.05), body weights (P<.001), andi egg production and yield (P<.001). There weree no (P>.05) diet X strain interactions. Thee significant differences among strains for body/ weight, and egg production and yield agree withti the results of Fairfull et al. (1983). Examination of the egg size data fromi Experiment 2 indicated that the response to3 dietary DON of egg size may not have been ass consistent as indicated by the egg yield resultss (Table 3). While there was a tendency for thee majority of eggs to occur in the two largestt categories, extra large and large, the relativee proportions varied somewhat between thesee categories. The variation among diets wass significant (P<.05) only for the mediumi category. Age influenced (P<.05 to .001) thee distribution of eggs, but there were no age X< dietary treatment interactions. Measurements of egg shell quality were obtained three times during Experiment 2, butt diet X age interactions were significant only forr egg weight and percent shell. However, these:
interactions accounted for less than 6% of the total sum of squares. Hence, means over the three measurements are presented in Table 4 for the shell quality variables in Experiment 2. The corresponding means for Experiment 1 are also included. Dietary treatment influenced shell weight (P<.01) and thickness (P<.05), percent shell (P<.05), and DFM (P<.05) of eggs in Experiment 1. There were inverse linear relationships between dietary DON concentrations and egg weight (P<.05), shell weight (P<.01), shell thickness (P<.01), and percent shell (P<.05). Compared to the eggs from the hens receiving the control diet CW, those from the birds given diet DON-W containing .7 mg DON/mg weighed 1.8% less and their shells were lighter (3.6%), thinner (2.2%), and weaker (1.5% lower compression fracture strength). In contrast, the egg and shell variables were not significantly influenced (P>.05) by dietary treatment in Experiment 2 (Table 4). For these variables, the differences between the control diet HC1C and diet HDON-W, which contained DON-contaminated winter wheat from the same stock as used in Experiment 1, were the greatest (.2 to 6.9%, P>.05) of all pairs of dietary treatments, except egg weight for diets HVT and HC2C. These results indicate that the decreases observed in Experiment 1, as dietary DON levels increased, may have been due to wheat source, but further research would be required to confirm this hypothesis. There was little statistical evidence of a relationship be-
Downloaded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
Feed intake, g/hen/day Feed efficiency kg/12 eggs kg/kg egg Egg production, % hen day Egg yield, g/hen/day Soft shelled eggs, % Cracked eggs, % No. hens died Body weight, kg Initial Final
CW
1.82 1.67
1.92 1.77
265 26.13 52.70 21.17 0
2.57 2.85 61.8 37.8 1.9 1.1 1
2.65 3.26 61.0 38.8 5.4 1.2 3
466 47.63 42.39 9.73 .25
1.85 2.25
101 104
2.02 2.60
111 102
HC2C
513 52.44 42.00 5.56 0
1.79 1.72
2.60 3.17 58.8 37.6 5.3 1.2 1
1.93 2.50
103 93
HVRl
585 38.51 50.40 9.27 1.81
1.79 1.67
2.47 2.95 61.7 38.8 5.5 1.6 1
1.81 2.44
103 98
HVR2
Diets
591 41.52 48.30 9.98 .20
1.81 1.74
2.42 3.11 62.1 39.3 4.7 1.0 3
1.79 2.38
103 100
HVR4
582 42 50 6
1 1
2 2 63 40 3 1 1
1 2
103 99
HV
2 The standard error of the mean (SEM) for comparing the means of diets HC2C and HDON-W with those for the othe by 1.4.
1 Based on chemical analysis, the DON content of the clean winter and spring wheat used to prepare diets CW and HC1 for diets HVR4, HVR2, and HVRl, 5.7 mg/kg; diet HVS, 6.5 mg/kg; and diet HVT, 1.7 mg; and the winter wheat f mg/kg.
No. eggs graded Extra large, % Large, % Medium, % Small, %
Feed intake, g/hen/day 1-56 days 1 1 2 - 1 6 8 days Feed efficiency, kg/12 eggs 1-56 days 1 1 2 - 1 6 8 days Feed efficiency, kg/kg egg 1—56 days 1 1 2 - 1 6 8 days Egg production, % hen day Egg yield, g/hen/day Soft shelled eggs, % Cracked eggs, % No. hens died Body weight, kg Initial Final
HOC1
TABLE 3. Feed intake and efficiency, egg production and yield, laying house shell quality, mortality for hens given diets that contained spring or white winter wheat contaminated with deoxy in Experiment 2
oaded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
DEOXYNIVALENOL AND LAYING HENS
the conditions (temperature and humidity) were similar within the incubators. Thus, the actual cause(s) of the low hatchability for Hatch 2 is not known; however, all treatments were affected to about the same extent, except HDON-W group. Chi et al. (1977) found that dietary levels of up to 4 mg/kg T-2 toxin had little effect on fertility and hatchability of fertile eggs. The presence of DON-contaminated white winter or spring wheat in the experimental diets had little effect (P>.05) on the ratio of organ weight to body weight of the hens necropsied at the completion of Experiments 1 and 2 (Table 6). There was no apparent evidence of lesions in the oral cavity, esophagus, proventriculus, and gizzard or of hemorrhaging in the viscera or muscles of the hens. The spleen, heart, and kidney appeared normal, but the livers from DON-fed hens were fatty in appearance. Using the livers obtained at necropsy from the hens of Experiment 1, Farnworth et al. (1983) found that the total lipid and triglyceride content of the livers from the hens given the CW/DON-W and DON-W diets was higher (P<.05) than the values for those receiving the CW control diet; the relationship was quadratic. From daily observations of the hens during these two experiments, there were no apparent evidence of vomiting (emesis), diarrhea, or changes in the behaviour of the birds. Moran et al. (1982) found that dietary DON levels up to 49 mg/kg produced no lesions in the oral cavity and only minor erosion in the gizzard of male broiler chicks after the birds had received the experimental diets for 6 days. In addition, they found that alertness, coordination, and livability were not influenced by DON levels up to 210 mg/kg of diet nor, based on microscopic examination, did these dietary levels affect the livers and kidney. The results of Chi et al. (1977) indicate that levels of up to 4 mg/kg purified T-2 toxin had little (P>.05) effect on the organ-to-body weight ratios for heart, liver, gizzard, and spleen for White Leghorn hens. The results from these experiments with laying hens, fed DON-containing diets for up to 24 weeks, support the previous findings obtained by Huff et al. (1981), Moran et al. (1982), Hulan and Proudfoot (1982) with broiler chicks, and Hamilton et al. (1985) with young chickens and turkey poults; namely, low dietary levels of DON have little effect on feed intake and efficiency, production performance,
Downloaded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
tween dietary treatment and shell quality over diets HC1C, HVR1, HVR2, and HVR4, although the trends towards lower values with higher dietary DON levels were similar to those of Experiment 1. Data from Chi et al. (1977) indicate that hens given diets containing up to 4 mg T-2 toxin/kg produced eggs that were (P>.05) larger and had thicker shells than those from birds receiving the control diet (52.4 vs. 51.6 g, and 324 vs. 319 jum, respectively). Interior egg quality, expressed as Haugh units, was also higher (P>.05) for the T-2 toxin-fed hens and the controls (92.83 vs. 90.32). Results in Table 5 indicate that the presence of white winter (Experiment 1) or spring wheat (Experiment 2) had little (P>.05) effect on fertility, hatchability of fertile eggs, and on the proportion of malformed embryos and pips. The lower fertility for Hatch 1, diet HC1C in Experiment 2, was due to the fertility being inexplicably low for hens in two blocks which contributed 43 eggs to the total of 169 set. The average fertility for these blocks was 41 and 19%, respectively, compared to an average of 80.4% for the birds in the remaining 6 blocks for this treatment. Embryonic mortality was influenced by dietary treatment in Experiment 1 (P<.05) and in Hatch 2 of Experiment 2 (P<.01). There was little evidence from the morphological data that the mortality of the embryos was associated with any particular abnormality. In fact, visual examinations indicated that most of the dead embryos appeared normal. The. low hatchability of fertile eggs in Hatch 2 of Experiment 2 was due to high embryonic mortality, particularly to an increase in the proportion of LDG and pips. The shell quality data for the first and second measurement periods, which occurred 7 to 10 days before the eggs were collected for hatches 1 and 2, respectively, indicated that there were little differences among the treatments between periods for egg and shell weights, percent shell, shell thickness, specific gravity, deformation, and compression fracture strength. Therefore, the much lower hatchability of egg sets for Hatch 2 than Hatch 1 was likely not due to shell quality. In addition, while age of hen does influence egg hatchability (Tomhave, 1956, 1958; Martin and Wessels, 1965), the differences in the ages of the birds in the present study (419 vs. 476 days) would likely not account for the large decrease obtained in the hatchability between Hatches 1 and 2. Records maintained during these hatches indicate that
1847
56.2 5.22 372 9.30 1.080 81 27.4 7.54
SEM .40 .049 .092 .151 .0062 1.2 .45 .093
DON-W
55.2 5.03 364 9.12 1.079 82 27.0 7.67 64.3 5.41 357 8.41 1.080 77 15.3
HCIC 61.3 5.23 358 8.53 1.080 77 15.1
HC2C 64.2 5.45 357 8.49 1.081 79 15.2
HVR1 63.5 5.40 358 8.51 1.081 80 15.3
HVR2
Measurements taken on eggs laid for 5 consecutive days after t h e hens were 243 days of age and had received the diets for 52 days.
55.7 5.09 366 9.13 1.079 82 27.8 7.54
CW 3
white winter
Ex
or
6
5
4
Quasi-static compression fracture strength expressed in newtons (N).
Measured by Archimedes' principle.
T h e standard error of the mean (SEM) for comparing the means of diets HC2C and H D O N W with those for the other diets is obtained b y m
3 Based o n chemical analysis, t h e DON content of t h e clean winter and spring wheat used t o prepare diets CW and H C I C , respectively wa H V R 1 , 5.7 mg/kg; diet HVS, 6.5 mg/kg; and diet HVT, 1.7 mg; and t h e winter wheat for diets CW/DON-W, DON-W, and HDON-W, 1.0 mg/kg.
Mean of three measurements taken on eggs laid for 5 consecutive days beginning when the hens were 4 1 5 , 4 7 1 , and 511 days of age, respec experiment.
3
1
Egg weight, g Shell weight, g Shell thickness, fim Percent shell Specific gravity 5 Nondestructive deformation, (ira 6 Compression fracture strength, N Albumen height, m m
CW/ DON-W
Experirr lent 1'
TABLE 4. Shell and interior quality of eggs from hens given diets that contained contaminated with deoxynivalenol (DON)
ded from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
92.8
Hatchability of fertile eggs, %
85.3
0
86.2
0
2.4
3.0
5.8
88.8
97.1
598
1.23
.35
.56
.69
1.19
.58
SEM
Hatch
60.6 23.9
.89 .39
65.8 33.1
0 5.4 0 1.3
1.9
11.1
27.8
8.3
1?.9 16.7 34.8
4.5
83.2 83.3 78.9 39.8
90 165
33.8
2.8
69.0 87.0 87.0 27.3
169 263
HC2C
6.7 24.5 5.0 36.0 3.5 12.0 1.1 .27 70.1 23.9
188 314 82.0 86.6 84.7 27.4
Hens were between 419 and 425 days of age when the eggs were collected for Hatch 1, and 476 and 488 days of age for Hatch 2.
The eggs were collected when the hens were between 244 and 250 days of age.
90.2
0
1.2
4.2
7.7
86.4
98.9
639
DON-W
Exper
12.1 16.8 6.5 36.4 1.1 8.3 .54 .31 64.1 32.2
195 338 79.1 84.2 80.3 38.5
HVR2
s
4
The standard error of the mean (SEM) for comparing the means of diets HC2C and HDON-W with those for the other diets is obtained by m Percent of fertile eggs. EDG = early dead germ. LDG = late dead germ.
Based on chemical analysis, the DON content of the clean winter and spring wheat used to prepare diets CW and HCIC, respectively w HVRl, 5.7 mg/kg; diet HVS, 6.5 mg/kg; and diet HVT, 1.7 mg; and the winter wheat for diets CW/DON-W, DON-W, and HDON-W, 1.0 mg/kg.
3
2
1
Overall hatchability of eggs set, %
Malformed embryos,s %
Pips, %
1.6
2.9
LDG,S %
5
2.7
EDG, S %
Embryonic mortality
97.1
605
Fertility of eggs set, %
No. eggs set
CW3
CW/ DON-W
Experiment l 1
TABLE 5. Fertility, batcbability, and embryonic mortality of eggs from bens given uncontaminated die or diets (CW/DON-W, DON-W, HVRl, HVR2, HVR4, HVS, HVT, and HDON-W) that contained white winter or spring wheat
ed from http://ps.oxfordjournals.org/ at New York University on June 1, 2015
1850
HAMILTON ET AL.
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T h e a u t h o r s t h a n k Barbara B e t h u n e , Dina D ' E r m o , Lorraine R o b i n s o n , R. T. Poirier, and J. F . Shackleton for their capable technical assistance, K. E. Hartin for his veterinarian assistance, A. R. Morrison and staff for their careful daily care and observation of t h e birds, J. S. Racine and staff for their assistance w i t h t h e mixing of t h e diets, R. Ciok for assistance w i t h t h e statistical analyses of t h e data, and J. Carroll for her assistance with t h e d e o x y n i valenol analysis of t h e diets' from E x p e r i m e n t 2. Chemical analysis of t h e w h e a t samples and diets from E x p e r i m e n t 1 for deoxynivalenol was d o n e b y H. Campbell, L a b o r a t o r y Services Division, F o o d P r o d u c t i o n and Inspection Branch, Agriculture Canada. Analysis of t h e diets from b o t h e x p e r i m e n t s for p r o x i m a t e c o m p o n e n t s was d o n e b y M. Ihnat, Analytical Services, Chemistry and Biology Research Instit u t e , Research Branch, Agriculture Canada, Ottawa. REFERENCES
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a n d livability of p o u l t r y . F u r t h e r m o r e , n o measurable quantities of DON were f o u n d in eggs from hens given diet H V R 4 (4.1 mg DON/kg) in E x p e r i m e n t 2 for a period of 2 4 w e e k s (El-Banna e t a / . , 1 9 8 3 ) .
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