Human tissue from uninvolved liver of cancer patients was fractionated using differential centrifugation and characterized for 11betaHSD enzyme activity against corticosterone, dehydrocorticosterone, 7alpha- and 7beta-hydroxy-dehydroepiandrosterone, and 7-oxo-dehydroepiandrosterone. An enzyme activity was observed in nuclear protein fractions that utilized either NADP(+) or NAD(+), but not NADPH and NADH, as pyridine nucleotide cofactor with K(m) values of 12+/-2 and 390+/-2microM, compared to the K(m) for microsomal 11betaHSD1 of 43+/-8 and 264+/-24microM, respectively. The K(m) for corticosterone in the NADP(+)-dependent nuclear oxidation reaction was 102+/-16nM, compared to 4.3+/-0.8microM for 11betaHSD1. The K(cat) values for nuclear activity with NADP(+) was 1687nmol/min/mg/micromol, compared to 755nmol/min/mg/micromol for microsomal 11betaHSD1 activity. Inhibitors of 11betaHSD1 decreased both nuclear and microsomal enzyme activities, suggesting that the nuclear activity may be due to an enzyme similar to 11betaHSD Type 1 and 2.

In mammalian organs involved in sodium reabsorption, the 11-β hydroxysteroid dehydrogenases (11βHSDs) oxidize glucocorticoids (GC) from their 11-alcohol form to their 11-keto state and therefore prevent their binding to mineralocorticoid (MC) receptors (MR) and the development of a MC excess syndrome. In birds the information about 11βHSDs and GC metabolism in such organs is scarce. Herein, we report the expression and enzymatic activity of 11βHSDs in the kidney and colon of chickens. Both organs express 11βHSD2-like mRNA. With NAD+, microsomes from both tissues oxidized corticosterone (CS) into 11-dehydrocorticosterone (DHC) with Km of 200 and 20 nM and Vmax of 13 and 2 pmol/mg protein/min in the kidney and colon, respectively. Thiram, a specific 11βHSD2 inhibitor, suppressed this oxidation in kidney. The expression and action of the putative 11βHSD3 were also tested. The chicken colon, and to a greater extent the kidney, expressed 11βHSD3-like mRNA. Microsomal fractions from both tissues oxidized CS into DHC in the presence of NADP+ with Km of 150 and 4 nM and Vmax of 5 and 0.3 pmol/mg protein/min for the kidney and the colon, respectively. This oxidation was not affected when NADP+ conversion into NAD+ was inhibited by excess pyrophosphate or a phosphatase inhibitor cocktail. In microsomes of chicken’s duodenum, where 11βHSD1-like mRNA expression is high, NADP+-dependent oxidation of CS into DHC has a low-affinity Km of 1130 nM. This study documented the expression and activity of two enzymes that convert CS into DHC, one is 11βHSD2-like and the other is similar to the putative mammalian 11βHSD3.

The mammalian 11-β hydroxysteroid dehydrogenase type 1 (11βHSD1) reduces glucocorticoids (GC) at C11 from the 11-keto-GC nonactive form to the 11-hydroxy-GC active form, an action essential for survival. Whereas GC metabolism at C11 and the role of 11βHSD1 are studied extensively in mammals, information about these in birds is scattered. Herein, we report the GC bidirectional metabolism in chickens. In hens’ liver and duodenal mucosa, 11βHSD1-like mRNA expression was detected; and 11βHSD1-like immunoreactivity was found linked to membranes of hepatocytes and duodenal enterocytes. With either NADH or NADPH, the membranal fraction of liver and duodenal mucosa converted dehydrocorticosterone (A) into corticosterone (B) with Km (1.1–8.7μM) and Vmax (10–40pmol/mg protein/min) values similar to those reported for mammalian 11βHSD1. In the presence of NADP+ or NAD+, these membranal fractions oxidized B into A. With either NADPH or NADH, the cytosol of chicken liver and duodenal mucosa reduced A into B (Km of 1.1 – 2.3μM and Vmax of 260–960pmol/mg protein/min). These cytosolic fractions did not convert any amount of B into A when incubated with either NADP+ or NAD+. This may suggest that chicken liver and duodenal mucosa express 11βHSD1 that is a membrane-bound oxoreductase which uses both NADPH/NADP+ and NADH/NAD+ as cosubstrates. The substantial reduction of A into B (but no conversion of B into A) found in the cytosol is most likely executed by a unidirectional soluble reductase, different than 11βHSD1.

The 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) activates glucocorticoids (GC) by reversibly converting 11-keto-GC to 11-hydroxy-GC, while 11βHSD2 and 11βHSD3 only catalyzes the reverse reaction. Recently, rat and human 11βHSDs were shown to interconvert 7α- and 7β-hydroxy-dehydroepiandrosterone (7α- or 7β-OH-DHEA) with 7-oxo-DHEA. We report that pig kidney microsomes (PKMc) and nuclei (PKN) oxidize 7α-OH-DHEA to 7-oxo-DHEA at higher rates with NAD+, than with NADP+. Corticosterone (CS), dehydrocoticosterone (DHC), 11α- and 11β-hydroxyprogesterone, and carbenoxolone completely inhibited these reactions, while 7-oxo-DHEA only inhibited the NAD+-dependent reaction. Conversely, CS oxidation was not inhibited by 7α-OH-DHEA or 7-oxo-DHEA. PKMc and PKN did not convert 7-oxo-DHEA to 7-OH-DHEA with either NADPH or NADH. Finally, PKN contained a high affinity, NADPH-dependent 11βHSD that reduces DHC to CS. The GC effects on interconversion of DHEA metabolites may have clinical significance, since DHEA and its 7-oxidized derivatives have been proposed for treatment of human autoimmune and inflammatory disorders.

1. This study compared the effect of bilateral electrolytic lesions of the basomedial hypothalamus (HL) in broiler and White Leghorn (WL) males.2. Hypothalamic lesions were placed in WL at 10 weeks of age (body weight 1.1 kg) and in broilers, either at 6 weeks (body weight 1.5 kg) or at 10 weeks of age (body weight 3.4 kg). They were fed ad libitum until autopsy at 16 and 17 weeks of age for broilers and WL, respectively.3. Hypothalamic lesions caused obesity (high percentage weight of abdominal adipose tissue) in both strains. Obese fowls with unimpaired reproductive systems were classified as OB and those with functional castration as FC (functionally castrated) or FCLC (functionally castrated with large comb).4. All post-HL syndromes-OB, FC and FCLC-were present in WL, whereas all obese broilers (which are immature at this age) were classified as OB.5. The percentage weight of abdominal adipose tissue in OB broilers was lower than in OB WL (3% vs 5%, respectively).6. Daily food intake of OB broilers was higher than control at 12 to 15 weeks of age, regardless of time of placement of HL, whereas daily food intake of OB WL was significantly higher than that of control WL only during the first 2 weeks following HL.7. Body weight of OB broilers at autopsy was 20% higher than control broilers, whereas body weight of OB WL was not significantly affected.8. An additional group of broilers was reared to sexual maturity under food restriction until 28 weeks of age. HL were placed at 10 weeks of age (body weight 1.7 kg). Autopsy was performed after a 4-week period of ad libitum feeding.9. There were OB as well as FC and FCLC among the HL, food-restricted broilers. Percentage weight of testes and spleen were reduced in OB fowls of both strains, but more so in OB WL.10. Hyperphagia and weight gain were not observed during the ad libitum feeding period of those obese broilers after HL, indicating that hyperphagia and weight gain are secondary to obesity.

Current research on dehydroepiandrosterone (DHEA) is limited due to lack of radiolabeled metabolites. We utilized pig liver microsomal (PLM) fractions to prepare [(3)H]-labeled 7 alpha-hydroxy-DHEA (7 alpha-OH-DHEA), 7 beta-hydroxy-DHEA (7 beta-OH-DHEA), and 7-oxo-DHEA substrates from 50 microM [1,2,6,7-(3)H]DHEA (specific radioactivity 60-80 mCi/mmol). The metabolites were separated by preparative thin-layer chromatography (TLC) using ethyl acetate:hexane:glacial acetic acid (18:8:3 v:v:v) as the mobile phase, extracted with ethyl acetate, and dried under a stream of nitrogen. Metabolites assayed by TLC and gas chromatography-mass spectrometry were observed to be pure. In the presence of an reduced nicotinamide adenine dinucleotide phosphate (NADPH)-regenerating system initiated with 1 mM NADPH alone, 1 mg/ml PLM produced 7 alpha-OH-DHEA with minor amounts of 7-oxo-DHEA (68 and 14 nmol/2h/2 ml, respectively; 82% conversion), while in the presence of 1mM NADPH and 1 mM oxidized nicotinamide adenine dinucleotide phosphate (NADP(+)), more 7-oxo-DHEA than 7 alpha-OH-DHEA (58 and 11 nmol/2 ml/120 min, respectively; 69% conversion) was formed. When longer reaction times were used with NADPH and NADP(+), a mixture of 7 alpha-OH-DHEA, 7 beta-OH-DHEA, and 7-oxo-DHEA was produced (19,14, and 35 nmol/180 min/2 ml, respectively; 62% conversion). Using pig liver microsomes, the radiolabeled metabolites of DHEA can be prepared in stable, pure form at 10mM concentrations and >0.5 mCi/mmol levels of radioactivity for biochemical studies.

The Mediterranean tortoise (Testudo graeca) is listed as “Vulnerable” on the IUCN Red List. Reproductive characteristics and means to increase offspring production were studied in T. graeca terrestris in a semi-natural environment. Courtship and mating occurred during early spring for about 4 weeks, followed by a laying season of approximately 2 months, with a second, shorter mating period in the fall. During the first mating, calcified eggs were already present in the uterus; we inferred that sperm from both mating seasons were stored in the oviduct for fertilization of eggs of the second laying cycle and of the following year. Average egg production was 3.8±0.3eggs/year. Most females laid all of their eggs in a single clutch, but 18% laid in a second clutch, 11–21 days later. X-ray radiography revealed calcified eggs in the uterus about 4 weeks before oviposition. All eggs in the uterus were calcified simultaneously and were laid in a single clutch; if a second clutch developed, those eggs were also calcified simultaneously. Based on endoscopic examinations, ovaries were active throughout the entire year. Plasma progesterone concentrations in females were very low and were detected only soon after oviposition (440±141pg/ml). Plasma estradiol concentrations in females varied from 4.1±1.5pg/ml to 70.2±29.4pg/ml, with no clear seasonal pattern. Maintaining tortoises at a low environmental temperature (9±1°C versus 28±1°C) reduced plasma estradiol concentrations. Giving 2mg/kg tamoxifen (TAM) increased plasma estradiol to 220±33pg/ml when treatment was given in September but not in late October, winter or spring. Treatment with TAM increased the number of eggs laid during the following laying season to 7.3±1.0eggs/year, laid in one to three clutches. In males, plasma testosterone concentrations had a seasonal pattern with the onset of a rise in July from 2 to >4ng/ml, a continued increase to a peak of 12.8±5.3ng/ml during November and a decline thereafter. Artificial incubation in sand at 29±1°C shortened the natural incubation time of 103±3.1 days to 83.5±1.3 days, increased hatching rate from 28 to 53%, and increased survival rate from 51 to 71% at 40 weeks of age. In summary, this study provides options for increasing reproductive performance, hatchability and offspring survival in captive Mediterranean tortoises, and may offer new tools for conservation of animals that are on the verge of extinction.

The cytochrome P450-dependent formation and subsequent interconversion of dehydroepiandrosterone (DHEA) metabolites 7α-hydroxy-DHEA (7α-OH-DHEA), 7β-hydroxy-DHEA (7β-OH-DHEA), and 7-oxo-DHEA was observed in human, pig, and rat liver microsomal fractions. Rat liver mitochondria and nuclei also converted DHEA to 7α-OH-DHEA and 7-oxo-DHEA, but at a lower rate. With NADP+, and less so with NAD+, rat, pig, and human liver microsomes and rat liver mitochondria and nuclei converted 7α-OH-DHEA to 7-oxo-DHEA. This reaction was inhibited by corticosterone and the 11β-hydroxysteroid dehydrogenase (11βHSD) inhibitor carbenoxolone (CBX). The conversion of 7α-OH-DHEA to 7-oxo-DHEA by rat kidney occurred at higher rates with NAD+ than with NADP+ and was inhibited by corticosterone. With NADPH, 7-oxo-DHEA was converted to unidentified hydroxylated metabolites and low levels of 7α-OH-DHEA by rat liver microsomes. In contrast, pig liver microsomal fractions reduced 7-oxo-DHEA to nearly equal amounts of 7α- and 7β-OH-DHEA, while human fractions produced mainly 7β-OH-DHEA. Dehydrocorticosterone inhibited the reduction to both isomers by pig liver microsomes, but only to 7α-OH-DHEA by human microsomes; CBX inhibited both reactions. Rat kidney did not reduce 7-oxo-DHEA with either NADPH or NADH. These results demonstrate that DHEA is first converted in liver to 7α-OH-DHEA, which is subsequently oxidized to 7-oxo-DHEA in both liver and kidney. In liver, interconversion of 7-oxo-DHEA and 7-OH-DHEA isomers is largely catalyzed by 11βHSD1, while in kidney 11βHSD2 (NAD+-dependent) and 11βHSD3 (NADP+-dependent) likely catalyze the unidirectional oxidation of 7α-hydroxy-DHEA to 7-oxo-DHEA. Distinct species-specific routes of metabolism of DHEA and the interconversion of its metabolites obviate extrapolation of animal studies to humans.

The effects of glucocorticoids (GC) on embryonic mortality and posthatch BW were studied. Cortisol hemisuccinate or corticosterone in 0.1-mL vehicles were injected into the albumen of 7-d-old White Leghorn chicken embryos. Embryonic mortality rates and the age after injection at which death occurred were determined. When 0.02 to 20 microg cortisol per egg were injected in saline, total embryonic mortality rate increased in a doseresponse manner, with a median lethal dose (LD50) at 10 microg. Saline injection alone caused a similar mortality rate to that caused by injection of 2 microg cortisol (around 35%). However, whereas mortality among the cortisol-treated embryos was greatest on Days 16 to 18, most of the saline-treated embryos died around the time of injection. The lethal effect of corticosterone, which is endogenous GC in adult chickens, was compared to that of cortisol by injecting both in the same vehicle (a saline:ethanol mixture) and was found to be similar. However, when 2, 10, or 20 microg of corticosterone was injected in oil, mortality rates were lower than those caused by the matching doses of cortisol in saline, probably due to the lower diffusion rate of the steroid out of the oil carrier. Hatch weight was significantly lower in chicks treated with 10 and 20 microg cortisol, and BW of the latter was lower compared with control throughout the 3-mo observation. In conclusion, cortisol and corticosterone are equally active in causing embryonic mortality. Posthatch BW is affected only by GC doses that are equal to or greater than the LD50.

The effect of gonadal steroids (GS) on proliferation of lymphocytes and distribution of lymphocyte subpopulations in cell culture was examined. The involvement of protein kinase C (PKC) and calcium ionophore in the proliferative response was tested. Estradiol benzoate (EB) or testosterone propionate (TP) had no significant influence on proliferation of peripheral blood lymphocytes (PBL) when cells were not stimulated by mitogen. At high concentration (10-6 M), EB and dihydrotestosterone (DHT) decreased lymphocyte proliferative response to concanavalin A (ConA) and lipopolysaccharide (LPS) at 24 and 72 h of incubation. However, at physiological doses (10(-12) to 10(-16) M), EB significantly enhanced the proliferative response at 24 h of incubation, whereas DHT had no effect. The inhibitory effect of the high dose of EB or DHT on proliferation of T and B lymphocytes was independent of time of hormone presentation to the cells or age and gender of cell donor. In all cultures, pre-incubation of lymphocytes with 10(-6) M of EB or DHT significantly reduced their proliferative responses to ConA, phytohemagglutinin (PHA), and LPS. The percentage of CD3+ cells was significantly reduced by EB, whereas DHT had no such effect. In contrast to inhibition of proliferation in response to mitogens, 10(-6) M EB dramatically enhanced the proliferation of lymphocytes in response to the PKC activator, phorbol 12-myristate 13-acetate, and calcium ionophore, A23187. Results suggest that high doses of EB do not damage the viability or proliferation capability of lymphocytes and, therefore, suppress the proliferative response to mitogens in a different manner, perhaps by reducing gene transcription for receptors that recognize the mitogens, or suppressing some postreceptor events. The enhancement of proliferation in response to mitogens by low doses of EB may support this assumption, because the biphasic effects of steroids on gene transcription are well documented.

Prolonged stress inhibits the hypothalamus-pituitary-gonadal (HPG) axis and reduces plasma testosterone (T). However, enhanced secretion of luteinizing hormone (LH) and T has been documented during the initial stages of acute stress in mammals. This study assayed the effect of short-term stress on plasma T and corticosterone (B) in juvenile, pubertal, and adult White Leghorn cockerels. Stress was induced by brief physical restraint of caged juvenile (7 weeks), pubertal (17 weeks), and adult (40 weeks) cockerels, as well as 40-week-old adults reared together in a room lined with wood shavings (group reared). Blood was sampled immediately before restraint (0 time), at the end of a 10-min restraint period, and at 30, 60, and 180 min after 0 time. Restraint resulted in an initial increase in plasma T in all groups, along with a rise in B. Whereas B generally reached its peak level at the end of the restraining period, T peaked 20 min later. The maximum increase of T and B relative to prestress levels (T and B ratios) was similar in all groups, with median T ratio reaching 1.25-1. 5-about half that of the B ratio. Thus, the extent of T and B response to short-term stress was not influenced by basal levels of T, which were highest in adults, and basal levels of B, which were higher in caged adults than in group-reared adults. Injection of ACTH did not induce a greater increase in plasma T than in sham-injected controls. Further, the elevation of T in response to stress was extinguished in castrated adults, indicating that T is secreted from the testes rather than the adrenals in response to stress. When the same regime of blood sampling was applied to adults not subjected to restraint, the T ratio rose by up to 11 times. It can therefore be stipulated that T response depends on the type of stress applied, a factor that should be considered when investigating androgen levels in plasma.

Inherited overfeeding and fattiness reduce laying performance in broiler breeder pullets. Although feed restriction is used to compensate for overeating and weight gain, this management practice leads to increases in BW variation, labor cost, and bird stress. Dietary supplementation of anorectic agents, such as fenfluramine, may be an alternative. Anak female prebreeder hens (19 wk of age; n = 10 per group) were treated as follows: daily oral administration of 5, 10, 20, or 40 mg DL-fenfluramine/kg BW or saline with food provided for ad libitum intake or administration of saline and feed restriction. Daily feed intake (FI), laying rate, egg composition, and BW were measured. At 40 wk of age, adipose tissue and ovary weights were measured. Fenfluramine depressed (P < 0.05) BW and FI in a dose-dependent manner, but was less effective in reducing BW than feed restriction. Suppression of FI occurred in two phases: a dynamic phase, coinciding with the rapid growth phase, during which FI declined progressively and a static phase during which FI reached a plateau at a significantly low level until the end of the experimental period. Egg production peaked first in saline-treated hens fed for ad libitum intake, but soon after started to decline. In all fenfluramine-treated and feed-restricted hens, egg production peaked 3 to 4 wk later and remained high until the end of the experiment. There were no differences in egg and egg component weights among the experimental groups. Abdominal adipose tissue weight was reduced by fenfluramine in a dose-related manner, and its weight in the group treated with the highest dose was similar to that of feed-restricted hens. In these two groups, ovarian weight was significantly higher than in the saline-treated hens fed for ad libitum intake, and a small, nonsignificant increase in ovary size was observed in groups treated with the two median doses of fenfluramine. The effect of fenfluramine on egg production was similar to that of feed restriction, but it was not dose-dependent and, thus, not directly related to its leaning effect. In broiler breeder hens, oral fenfluramine may be used for chemical feed restriction and diminution of fattiness without reducing egg production relative to manually feed-restricted hens.

1. The aim of this study was to determine the effect of different light sources and light schedules on the growth and quality of commercial broilers.2. In each experiment 810 broiler chicks were divided into 3 groups, 3 replicates per group. All were reared at 20 lux. Body weight and food consumption were recorded weekly3. Experiment 1. Birds were reared under 3 light sources: incandescent light bulb, warm-white fluorescent light tube or warm-white mini-fluorescent light bulb. Experiment 2. Birds were reared on 3 light schedules. 23 h light and 1 h dark (23L:1D) throughout; an increasing light schedule with initial 23L:1D then 8L:16D increasing daylight gradually to 16L:8D or an intermittently increasing daylight schedule (16:8P) where light and dark periods were shorter but portioned to achieve the same total hours per day up to 16L:8D.4. Broilers reared under mini-fluorescent light bulb were heavier than those under fluorescent tubes or incandescent bulbs by 49 d.5. Until 42 d of age, photoperiod had no effect an growth. However, at 49 d broilers reared under 16:8P and 16L:8D regimens were heavier than those or 23L:1D.6. At 42 d, female broilers on 23L:1D, were heavier than those on 16L:8D and 16:8P.7. Mortality was higher in groups on 23L:1D than on 16L:8D on 16:8P.8. At 49 d incidence of leg condemnation was higher in the 16:8P group. However, skin damage was lower in this group than in those on 23L:1D and 16L:8D.

Artificial illumination, including light quality, is crucial in modem broiler management. In the present study, a new, highly efficient, monochromatic light system has been developed for broilers. One hundred and eighty male broiler chicks (Anak) were divided into four light treatment groups (n = 45) in three replicates each. All birds were housed in a single room previously divided by wooden bars into 12 sealed cells of 1 m2. Feed and water were provided for ad libitum consumption. Light intensity was 0.1 W/m2 at the height of birds' heads and was scheduled for 23 h of light and 1 h of dark during the entire experimental period. Light treatments were: control white (mini-incandescent light bulbs), blue (480 nm), green (560 nm), and red (660 nm). Body weight was recorded periodically, feed consumption was measured daily, and feed efficiency was calculated. Blood samples were taken at 1, 9, and 32 d of age and plasma testosterone was determined. Two necropsies were conducted, at 23 and 35 d of age, and selected glands and organs were weighed. In the group reared under green light, a significant enhancement in weight gain was observed as early as 3 d of age; this gain was maintained during the entire experimental period. Broilers reared under blue light had a later onset of growth enhancement and were significantly heavier than those reared under white and red light at 20 d of age. Plasma testosterone levels were significantly higher in birds reared under blue light. Breast muscle weights were significantly higher in the birds reared under green light at 23 and 35 d of age. These results suggest that green and blue light stimulate growth.

Embryonic and posthatch long-term exposure to the odor of 2-methoxy-3-isobutyl-pyrazine (2M3IP) was examined for its potential physiological consequences as reflected in changes in BW and organ weights in domestic chicks (Gallus gallus domesticus). Experiments were run from Day 1 of incubation to the age of 3 wk with a total of 360 fertile chicken eggs. The experimental design consisted of four treatment groups: PP chicks were exposed to 2M3IP during both incubation and posthatch rearing; PC chicks were exposed to 2M3IP during incubation only; CP chicks were exposed to 2M3IP during rearing period only; CC control chicks were not exposed to 2M3IP. Chicks were weighed immediately after hatch and at 3 wk of age, when they were necropsied. Various organs (thyroid, adrenal, testes, comb, liver, spleen, abdominal fat, and the bursa of Fabricius) were removed and weighed. Body weights of both sexes in the PP group were reduced. This reduction was significant in males relative to both CP and CC groups and in females only relative to the CP group. Effects of 2M3IP exposure on the examined organs were as follows: in males, adrenal gland weight significantly increased in the PP group vs all other groups. No weight differences were found between the other inspected organs among the four treatments. In females, comb weight significantly decreased compared with the rest of the groups when 2M3IP exposure occurred during incubation (PC). Further investigation is needed to study the mechanisms that underlie the differential effects of pyrazine odor on male and female chicks.

1. Mule ducks were produced by naturally mating Muscovy drakes and Khaki Campbell ducks. 2. Semen was collected from 6-month-old mule drakes via an artificial vagina. The fluid was clear without any spermatozoa or spermatids. 3. Testes from 27-week-old mule drakes were smaller in size than those of Khaki Campbell drakes but heavier than Muscovy males of the same age. Histological sections of these testes revealed that spermatogenesis was not complete. 4. Testosterone concentration in the mule drakes was higher than in Muscovy males but similar to Khaki Campbell drakes. 5. Mule drakes have strong sexual drives as a result of high concentrations of testosterone, but, because spermatogenesis is incomplete, their semen had no sperm.

The ontogeny of the tonic immobility (TI) response in domestic fowl chicks was studied during the first week of life. The TI response of naive White-Leghorn Gallus domesticus male chicks (N=5–9), was tested at the age of 1, 2, 3, 5 and 7 days. TI was induced dorsally and its duration, the number of induction trials and the latency of peeping were recorded. The TI response was strongly affected by age. It was poorly developed during the first 3 days of life, when the median TI duration in control chicks was 10 s and the mean number of induction trials 2.3±0.3. After the third day of life, TI duration increased by up to 15× and susceptibility by about two. Peeping latencies were very short throughout the first week and in many cases, peeping started long before the termination of TI. Immediately following recovery from TI, chicks were put in an open field and the latencies of walking and jumping and the number of steps, jumps and peeps were observed. No changes in either locomotion or vocalization in an open field were found between the third and fifth day. Furthermore, there was no correlation between any of the parameters of the TI and OF tests. The effect of habituation, which is known to attenuate the TI response, was studied by repeatedly subjecting chicks to TI and OF tests, once on each day of the experiment. Habituation prevented the increase in TI duration and susceptibility after the third day of life, but did not affect the OF response. The effect of aversive treatment, which was expected to increase TI, was examined by placing chicks in 5-cm deep tap water for 5 min, prior to testing. Treatment significantly attenuated TI on Day 1 and increased overall locomotion and peeping in open field.

Groups of 9 or 10 cows were assigned to one of three treatments 1) machine-milking three times daily, 2) machine-milking six times daily, and 3) suckling three times daily in addition to machine-milking three times daily. Treatments were conducted during the first 6 wk postpartum. During wk 5, digestibility of the diet was estimated by the indigestible neutral detergent fiber method. During wk 6, milk yield and dry matter intake (DMI) were recorded daily, and plasma concentrations of glucose, nonesterified fatty acids, urea, protein, growth hormone, insulin, insulin-like growth factor I, oxytocin, and prolactin were determined. Milk yields were 38.5, 46.8, and 52.7 kg/d, and DMI were 18.1, 21.2, and 17.2, for cows on treatments 1, 2, and 3, respectively. Plasma glucose concentrations decreased, and plasma nonesterified fatty acid concentrations increased, for cows on treatments 2 and 3 compared with cows on treatment 1. Digestibility of dry matter was 57.5, 60.5, and 60.6%; of organic matter was 62.6, 64.6, and 66.8%; and of crude protein was 59.3, 62.7, and 64.6% for cows on treatments 1, 2, and 3, respectively. Concentrations of all assayed hormones, except insulin, increased moderately for cows on treatment 2 compared with cows on treatment 1 and increased dramatically for cows on treatment 3. Insulin concentrations followed the opposite trend. The DMI were positively related to milk yields and negatively related to oxytocin concentrations. Digestibility was negatively related to plasma glucose concentrations in a nonlinear pattern. The possible involvement of hormones in improvement of digestibility is discussed.

Forty Holstein heifer calves were assigned to two treatments. Control calves (n = 20) were fed milk replacer in open buckets, and calves that were allowed to suckle (n = 20) were paired and suckled the same dam three times daily. Treatments were conducted during the first 6 wk following birth; thereafter, all calves received the same management, and weaning was at 60 d of age. During treatment, calves that were allowed to suckle had significantly higher average daily gains than did control calves. However, at 12 wk of age, calves that were allowed to suckle had significantly lower body weights (BW) than did control calves. Age at conception was significantly lower, and BW at conception and conception rate tended to be higher, for calves that were allowed to suckle. Calving age was significantly earlier for heifers that had been allowed to suckle as calves, and BW at calving also tended to be higher. Height at the withers after calving was also significantly higher for those heifers. Milk production during first lactation tended to be higher for the heifers that had been allowed to suckle as calves. Our results indicated that heifer calves that suckled milk during the first 42 d of age had higher average daily gains, higher height at the withers, an earlier age at calving, and a tendency for greater milk production than did calves fed milk replacer.