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Pregnancy following transfer of ooplasm from cryopreserved-thawed donor oocytes into recipient oocytes.

OBJECTIVE: To determine if frozen-thawed donor oocytes could be used to provide cytoplasm for transfer into patients' oocytes to improve subsequent embryonic development. DESIGN: Prospective evaluation of the procedure in consenting IVF patients. SETTING: Assisted reproductive technology program. PATIENT(S): The study was open to consenting IVF patients (of any age) with a history of poor embryo quality or those couples in which the wife's age was > or = 40 years. INTERVENTION(S): Transfer of donor egg cytoplasm from frozen-thawed oocytes into the oocytes of infertile recipients. MAIN OUTCOME MEASURE(S): Donor oocyte survival following cryopreservation, fertilization following cytoplasmic transfer into recipient oocytes, embryo quality, and pregnancy outcome. RESULT(S): Oocytes collected from four donors were cryopreserved and 61% (28/46) survived the thaw procedure. Cytoplasmic transfer was performed on the eggs of four patients, with fertilization occurring in 70.3% (26/37). Twin pregnancy was established in one patient (35 years of age) with a history of poor embryo quality. CONCLUSION(S): Cryopreserved donor oocytes may provide a source of cytoplasm for transfer into recipient oocytes, eliminating the need for cycle synchronization between donor and infertile patient.

Adult↗

Effect of turning and age of egg on hatchability in the pheasant, chukar, and Japanese quail.

Eggs from pheasants, Chukar partridge and Japanese quail were stored 1 to 28 days in a cold box where temperature and relative humidity were maintained at approximately 16 degrees C. and 70 percent, respectively. Half the eggs from each species was turned once daily from a 45 degree angle from the verticle to the opposite angle. Turning eggs prior to incubation had no appreciable effect on hatchability of eggs stored up to 28 days. Length of storage for eggs from the pheasant and quail was critical, with significant declines in fertility in eggs stored beyond 14 days. Storage of Chukar eggs up to 28 days had little effect on subsequent hatchability, and their holding quality was comparable to that for commercial strains of chickens and turkeys.

Animals↗

Dietary interaction of 1,4-diaminobutane (putrescine) and calcium on eggshell quality and performance in laying hens.

An experiment was conducted to evaluate the potential dietary interaction between 1,4-diaminobutane (putrescine) and calcium on eggshell quality and overall laying performance. One hundred ninety-two 30-wk-old White Leghorn hens were fed a corn-and soybean-meal-based diet supplemented with 0.00, 0.05, 0.10, or 0.15% putrescine and 2.5, 3.0, 3.5, or 4.0% calcium in a factorial design (12 birds per diet) for 4 wk. The percentage of egg production increased linearly (P < 0.05) with increasing levels of dietary calcium. Significant interactions (P < 0.05) were observed between dietary putrescine and calcium for eggshell thickness, eggshell deformation, percentage of eggshell, calcium intake, total calcium retention, total eggshell calcium, and percentage of eggshell calcium. Interactions were due to quadratic effects of putrescine or calcium on these parameters. Eggshell thickness and percent eggshell increased when hens were fed 3.5% calcium in combination with 0.1% putrescine; however, calcium intake and calcium retention were significantly lower (P < 0.05). Eggshell quality improved with increasing dietary levels of calcium due to increased calcium retention and calcium balance. Increasing levels of dietary putrescine did not have a negative effect on eggshell quality; however, calcium intake was lower at higher-supplemented levels of putrescine. It was observed that dietary calcium in excess of requirements resulted in increased egg production and eggshell quality. Eggshell quality improved when hens were fed 3.5% calcium diet in combination with 0.10% putrescine. It was concluded that small supplements of dietary putrescine may improve eggshell quality, depending on dietary calcium concentration.

Animal Feed↗

Food, energy and macronutrient intake of postmenopausal women from a menopause program.

OBJECTIVE: This study aimed to analyse the food, energy and macronutrient intake of a group of postmenopausal women participating in a health-care-program. METHODS: Subjects included were 38 healthy postmenopausal women aged between 46 and 60 years, recruited from the Menopause Program of the Madrid City Council. Physical activity, some anthropometric data and dietary information was obtained using a modified version of the dietary history method, which contained a 24-hour-recall and a food frequency questionnaire covering the preceding month as reference period. Dietary quality indexes, including those of the energy provided by macronutrients, alcohol and fatty acids and PUFA + MUFA/SFA and PUFA/SFA ratios were calculated. RESULTS: This group consumed a diet very similar to the traditional Mediterranean diet. Intake of vegetables (415 +/- 165 g/d), fruits (396 +/- 178 g/d) and fish (131 +/- 69 g/d) was high and a wide variety of these products was consumed. Potato and cereal (157 +/- 76 g/d) intake was low. Dairy products, meat, poultry and eggs were only a modest part of this diet. The fat quality (PUFA + MUFA/SFA = 2.26) was satisfactory. CONCLUSION: The diet of this group of postmenopausal women attending a prevention program closely conforms to current nutritional guidelines. Physical activity, body weight and intake of vegetable foods are adequate and may be very useful to counterbalance the increased risk of several pathologies after menopause. However, consumption of carbohydrate rich foods is lower than recommended. Participation in the menopause health-care-program is useful for weight control and dietary advice.

Eating↗

Does nutrition-related stress carry over to spruce budworm, Choristoneura fumiferana (Lepidoptera: Tortricidae) progeny?

Three different patterns of feeding of sixth-instar spruce budworm, Choristoneura fumiferana Clemens were simulated in the laboratory. Larvae were fed artificial diets whose nitrogen and total soluble sugar contents varied according to levels similar to those found in three types of balsam fir, Abies balsamea (L.) Miller foliage (current-year foliage from middle and lower crown sections and one-year-old foliage). The biological performance of offspring was studied according to the nutrition of their parents. Although food quality had no impact on pupal weight of female parents and individual mean egg weight, progeny fitness was influenced by parental nutrition. Old foliage simulated diet, poor in nitrogen, clearly affected the early larval development of progeny, especially the percent of egg hatch and first-instar survival. Lower crown current-year foliage simulated diet, with low total soluble sugar content, reduced the first-instar survival of the progeny. However, the selective pressure exerted by low food qualities on the parental generation and on the early stages of their progenies resulted in C. fumiferana populations having higher tolerance to starvation and higher survival after the diapause period. These results highlighted the potentially direct and indirect effects of C. fumiferana parental nutrition on the next generation. The patterns of feeding of parental generations would appear to affect the quality and size of subsequent populations through several selections on the different life-history stages of both generations.

Animals↗

Quantification of the heat exchange of chicken eggs.

In the incubation process of domestic avian eggs, the development of the embryo is mainly influenced by the physical microenvironment around the egg. Only small spatiotemporal deviations in the optimal incubator air temperature are allowed to optimize hatchability and hatchling quality. The temperature of the embryo depends on 3 factors: (1) the air temperature, (2) the exchange of heat between the egg and its microenvironment and (3) the time-variable heat production of the embryo. Theoretical estimates on the heat exchange between an egg and its physical microenvironment are approximated using equations that assume an approximate spherical shape for eggs. The objective of this research was to determine the heat transfer between the eggshell and its microenvironment and then compare this value to various theoretical estimates. By using experimental data, the overall and the convective heat transfer coefficients were determined as a function of heat production, air humidity, air speed, and air temperature. Heat transfer was not affected by air humidity but solely by air temperature, embryonic heat generation, and air speed and flow around eggs. Also, heat transfer in forced-air incubators occurs mainly by convective heat loss, which is dependent on the speed of airflow. A vertical airflow is more efficient than a horizontal airflow in transferring heat from the egg. We showed that describing an egg as a sphere underestimated convective heat transfer by 33% and was, therefore, too simplistic to accurately assess actual heat transfer from real eggs.

Air Movements↗

Compressed time and frequency recording of the electrogastrogram by individual wave detection.

Electrogastrography (EGG) is a method for recording and analysis of the bioelectrical activity of the stomach, acquired by cutaneous electrodes. The problem of obtaining an EGG signal as free of artefact as possible has recently found some solutions. Good quality recordings of several hours duration can be acquired, often needed, especially if clinical applications are envisaged. Therefore, a next problem to be considered would be time-compressed presentation of the EGG signal without loss of relevant signal characteristics. Relative amplitude changes can be of clinical value, while frequency variations are traditionally regarded as more important. A method for simple compressed presentation of long term EGG recordings, without losing information about the real amplitude and/or frequency changes or transients, is presented. It makes use of detection of successive wave extrema in an EGG. Successive wave amplitudes and durations are measured and momentaneous frequencies are derived. Then a compressed two-trace plot is generated, representing amplitude and frequency variations with respect to time. The method is virtually insensitive to baseline drift, as only the peak amplitudes are detected and followed. A compression factor with respect to the traditional signal recording (1 cm min(-1) or 2 cm min(-1) paper speed) of 5 up to 15 per trace can be obtained.

Electrodiagnosis↗

The effect of removal of seminal plasma, egg yolk level and season on sperm freezability of canary buck (Capra hircus).

Goat semen is different from that of other domestic species in its limited tolerance to the inclusion of egg yolk in the freezing medium, and this tolerance depends on the presence of enzymes in the seminal plasma that react with egg yolk, producing toxic compounds to the spermatozoa. Moreover, the goat is a seasonal breeder that shows variations in semen quality throughout the year, and those variations may affect semen freezability; hence in freezing protocols, for instance, removal of seminal plasma (washing) yields varying results. This work was designed to study this problem in Canary goats: semen from six males was collected in spring, autumn or winter, washed or non-washed, diluted in a freezing extender with 1.5, 6 or 12% egg yolk, frozen, and thawed after 2 days, 2 or 6 months of cryopreservation. The effect of egg yolk concentration in the freezing extender was far more important than the effect of washing or season on sperm cryosurvival. The quality of frozen-thawed semen tended to improve as egg yolk concentration increased regardless of the effects of season, washing or period of cryopreservation. Washing produced a positive effect on frozen-thawed semen collected during spring or autumn, but the difference decreased as the concentration of yolk increased. However, washing produced a negative effect on frozen-thawed semen collected during winter, diluted with either 6 or 12% egg yolk. There was no apparent seasonal effect on gross measures of sperm production but the seasonal effect was ever present and was reinforced by freezing.

Acrosome↗

Effect of dietary contents of cholecalciferol, 1 alpha,25-dihydroxycholecalciferol and 24,25-dihydroxycholecalciferol on blood concentrations of 25-hydroxycholecalciferol, 1 alpha,25-dihydroxycholecalciferol, total calcium and eggshell quality.

1. Withdrawal of cholecalciferol (D3) supplement from a layers diet drastically reduced blood 25-hydroxycholecalciferol (25-OH-D3), 1 alpha,25-dihydroxycholecalciferol (calcitriol) and egg specific gravity (SG) within two weeks, followed by a decrease in blood total calcium (Ca). 2. Doubling the D3 supplement in the control diet (27.5 micrograms or 1100 IU/kg) almost linearly increased the circulating concentration of 25-OH-D3 without raising the concentration of calcitriol, Ca, or egg SG. 3. Replacing D3 by the optimal concentration of calcitriol (5 micrograms/kg diet) improved egg SG after 21 weeks of treatment without increasing blood calcitriol or total Ca. 4. By itself, 24,25-dihydroxycholecalciferol [24,25-(OH)2D3] was unable to maintain normal blood levels of calcitriol, Ca or egg SG and, when added together with calcitriol in the diet, tended to elevate blood Ca but suppress the beneficial effect of calcitriol on shell quality, with little or no effect on blood calcitriol.

24,25-Dihydroxyvitamin D 3↗

99mTc ovalbumin labelled eggs for gastric emptying scintigraphy: in-vitro comparison of solid food markers.

BACKGROUND: The reliability of solid phase gastric emptying measurements by scintigraphy requires a marker that remains within the solid component of the test meal, and which is not degraded by the gastric juice throughout the scintigraphic procedure. In Europe, foods are most often labelled with 99mTc rhenium sulfide macrocolloid (RSMC) but this solid phase marker was withdrawn from the market in January 2004. OBJECTIVE: To test other potential solid phase markers and to compare them to the reference marker RSMC. These markers were rhenium sulfide nanocolloid (RSNC), tin fluoride colloid (TFC), phytates and two albumins (Alb and AlbC). All were radiolabelled with 99mTc. METHODS: After quality control, each 99mTc marker was incorporated into the albumin of one egg. Then, egg white and yolk were mixed together, and a well-cooked omelette was prepared. Aliquots of the omelette were incubated with an acidic solution of pepsin at 37 degrees C which mimicked gastric juice. Unbound radioactivity in the supernatant fraction was measured at various times up to 3 h. RESULTS: The radiochemical purity was > 95% for all radiopharmaceuticals. During the in-vitro incubation, the percentage of 99mTc labelled colloids released from the omelette increased continuously: after 3 h, 5% for TFC and RSMC, 8% for phytates, and > 9% for the two albumins and RSNC. CONCLUSION: Considering quality controls and release of 99mTc during in-vitro incubation of the omelette, TFC showed the same behaviour as the reference marker RSMC. Thus, TFC seems to be the best candidate to replace RSMC for the radiolabelling of the solid phase of the gastric emptying test meal.

Eggs↗

Detection of both the normal and mutant alleles in single cells of individuals heterozygous for the sickle cell mutation--prelude to preimplantation diagnosis.

As a preliminary step to preimplantation diagnosis of sickle cell disease in unfertilized eggs or 8-cell embryos of heterozygous parents, we established quality control for detection of the mutant and normal alleles of the beta-haemoglobin gene using single buccal cells. Efficient polymerase chain reaction (PCR) amplification of a 680 base pair sequence of the beta-globin gene spanning the site of the sickle cell mutation was obtained for 79 per cent of single heterozygous cells. In 71 per cent of cases, both alleles were detected. With this current efficiency, we predict that a clinical preimplantation diagnosis at the 8-cell embryo stage could be carried out safely and reliably for a couple at risk of transmitting sickle cell disease to their children.

Alleles↗

Long-term infertility and dominant lethal mutations in male mice treated with adriamycin.

Sperm production and fertility were studied in male mice treated with adriamycin (ADR) at 6 or 8 mg/kg. Testicular sperm production and epididymal sperm counts were markedly reduced after ADR treatment. Gradual recovery of counts occurred, but sperm counts had not reached control levels even more than 1 year after treatment. Epididymal sperm showed treatment-induced morphological abnormalities throughout the experiment; the frequencies of sperm with detached tails and the frequencies of sperm with morphologically abnormal heads remained elevated about 2-3-fold above control. According to the frequency of vaginal plugs, treated male mice mated at control rates with untreated females during the post-treatment sterile period. However, after some fertility was regained the fertilization rate (calculated as the fraction of eggs, flushed from the oviduct 2 days after mating, that had been fertilized and had cleaved) was markedly reduced and remained depressed for the remainder of the experiment. The fertilization rate reached only 0.29 at 23-32 weeks after 8 mg/kg ADR and 0.76 at 16-23 weeks after 6 mg/kg ADR; both values were significantly below the control value of 0.94. Dominant lethal mutations in the zygotes flushed from the oviduct were measured in culture by the loss of the zygote's ability to develop to a stage characterized by trophectoderm outgrowths and formation of an inner cell mass. The frequencies of dominant lethal mutations detected in vitro were 1.7 or 7.4% after 6 mg/kg, and 32 or 40% after 8 mg/kg ADR; each value was calculated in two different ways, with 3 of these 4 values significantly different from zero. We conclude that even after mice regain fertility following ADR exposure, the level of fertility remains permanently subnormal as evidenced by a lack of fertilization of eggs that is probably due to the decreased quantity and quality of spermatozoa produced. Furthermore, ADR can induce genetic damage in stem spermatogonia, which can be transmitted through fertile spermatozoa. Thus, there may be a genetic risk to the offspring of cancer patients treated with ADR chemotherapy, but at present we are unable to quantitate that risk.

Animals↗

Avian embryonic stem cells.

Blastodermal cells derived from the area pellucida of a stage X (EG&K) embryo have the potential to contribute to the somatic tissues and the germ line when reintroduced into a stage X (EG&K) recipient embryo. This chapter describes a method to culture chicken embryonic stem (cES) cells derived from blastodermal cells. Within the first week of culture, the cells change their morphology; they become smaller with a large nucleus and a prominent nucleolus. The cES cells remain chromosomally normal and can be cultured for extended periods. They can be modified genetically using standard electroporation procedures and, after injection into a recipient embryo, can contribute to all somatic tissues. Using a surrogate shell culture system, the injected embryos can be manipulated and visualized easily throughout incubation. We have generated high-grade chimeras by compromising the recipient embryos and maintaining the ES cells in stage X (EG&K) recipients for a few days at 15 degrees before incubating them at 37.5 degrees. The cES system provides a novel experimental paradigm for the investigation of developmental and physiological mechanisms in the chicken.

Animals↗

Solid-state fermentation of cornmeal with the basidiomycete Hericium erinaceum for degrading starch and upgrading nutritional value.

The ability of the basidiomycete Hericium erinaceum to degrade starch and upgrade nutritional value of cornmeal during solid-state fermentation was studied. On the basal medium which consisted of cornmeal and salt solution, H. erinaceum produced a strong alpha-amylase on the 15th day after inoculation, which resulted in a 52% degradation of the starch. By supplementation with 5-15 g soybean meal per 100 g cornmeal the alpha-amylase activity and degradation rate of starch was raised significantly (P < 0.01). Prolongation of fermentation time from 15 to 30 days did not increase significantly the degradation rate of starch, though the alpha-amylase activity reached its maximum value of 179 U/g on the 20th day after inoculation. Under conditions close to the theoretical optimum fermentation conditions, that was after 25 days at 25 degrees C in the medium with added 15 g soybean meal per 100 g cornmeal, the starch content in the product decreased from 63% to 22% (P < 0.001) and protein content increased from 12% to 17% (P < 0.01). In the protein in the product, the lysine content was increased from 36 to 56 mg/ g and tryptophan from 9 to 13 mg/g. Using egg protein as a standard, an evaluation on the protein quality of the fermented product showed that it was superior to that of the nonfermented control and to other cereals, was close to that of soybean and chicken, but was inferior to that of milk and red meats.

Basidiomycota↗

Comparison of protective media and freezing techniques for cryopreservation of human semen.

OBJECTIVE: To evaluate the influence of cryopreservation medium and freezing-thawing techniques on human sperm motility and morphology. STUDY DESIGN: 63 semen samples were obtained from 39 donors to the artificial insemination programme. Possible effects of the sperm dilution with cryomedium on the motility were examined 10 min after exposure of 24 high initial quality semen samples to TEST-yolk ¿zwitterion-citrate-egg yolk extender containing TES [N-Tris (hydroxymethyl) methylaminoethane sulfonic acid] and Tris [(hydroxymethyl) aminomethane]¿ and human sperm preservation medium (HSPM). Post-thaw sperm motility from 24 frozen semen samples was examined comparing the cryoprotective efficacy of TEST-yolk and HSPM following different freezing techniques (vapour freezing, fast programmable freezing and slow programmable freezing). The relationship of sperm morphology to the effects of freezing was investigated on 39 semen samples following different freezing techniques. Post-thaw sperm motility from 39 frozen semen samples was compared among three groups divided according to the percentage of morphologically normal cells (<40, 40-50 and >50%) in fresh semen. RESULTS: Exposure of spermatozoa to cryomedia for 10 min at room temperature significantly reduced motility in TEST-yolk treatment group for 9% and in HSPM treatment group for 18% (P<0.01). The recovery of motile sperms (mean+/-standard deviation) was 49+/-15.7, 43+/-15.2 and 52+/-16.8% when TEST-yolk was used and 34+/-17.8, 32+/-18.2 and 50+/-13.6% when HSPM was used as a cryopreservative following vapour freezing, and fast and slow programmable freezing, respectively. Following vapour freezing and also following fast programmable freezing, the recovery of motile sperm was significantly higher (P<0.05) after addition of TEST-yolk medium than after addition of HSPM. Post-thaw motility of the sperm cryopreserved in HSPM showed significant differences (P<0.05) after three different freezing techniques. The recovery of motile sperms was 57+/-26.4, 38+/-8.6 and 38+/-17.3% in groups with >50, 40-50 and <40% morphologically normal cells, respectively. The percentage of morphologically normal spermatozoa was reduced 8% after vapour freezing and 6 and 3% after fast and slow programmable freezing, respectively. The results were statistically analysed using SAS/STAT software. CONCLUSIONS: Slow programmable freezing was superior to vapour freezing and fast programmable freezing as a method for sperm cryopreservation. However, none of these methods of freezing had discernible effects on sperm morphology. Motility of spermatozoa decreased due to the exposure of semen to cryomedium. TEST-yolk was a superior cryomedium to HSPM. Fresh semen with more than 50% of morphologically normal cells showed the best recovery of motile cells after freezing and thawing.

Buffers↗

Detection of sodium chloride in cured salmon roe by SW-NIR spectroscopy.

Salt and moisture content of cured salmon roe (ikura) was determined using short-wavelength-near-infrared (SW-NIR) reflectance spectroscopy (600-1100 nm). SW-NIR can be used to measure chloride species. Calibrations for salt in bulk samples of high-quality roe (R(2) = 0.904, SEP = 0.421%, RPD = 3.21) and average-quality roe (R(2) = 0.711, SEP = 1.13%, RPD = 1.81), crushed eggs (R(2) = 0.857, SEP = 0.509%, RPD = 2.62), and individual eggs (R(2) = 0.731, SEP = 0.698%, RPD = 1.98) were developed using partial least squares (PLS) regression models. The heterogeneous distribution of lipid and moisture in the individual eggs limit the sensitivity of this method; however, this method provides a rapid nondestructive method for high-value food products where destructive testing is expensive or impractical and for process control applications.

Animals↗

Characteristics of oviposition patterns of turkey hens and the influence of different wavelengths of light.

1. Characteristics of individual oviposition patterns of turkey breeder hens were determined for 160 d of photostimulation with fluorescent blue, green or red light and with incandescent light. 2. In all treatments most eggs were laid during the afternoon and evening. The maximum frequency occurred at 14.00 to 15.00 h (8 to 9 h after light onset); about 15% of all eggs were laid in the dark period. 3. The hourly oviposition distribution remained unchanged and was specific to individual hens throughout the study period. However, it was highly variable between hens, ranging from highly dispersed to tightly clustered. 4. The total number of clutches per hen did not differ between treatments and eggs were laid in relatively short clutches; 65% were of two eggs or fewer, 30% of three to 5 eggs and only 5% of 6 eggs or more. 5. The oviposition time of the first egg in a clutch was very variable; however, oviposition time became more clustered and uniform as the position of the egg in a clutch increased. 6. Mean time of lay, mean interval between eggs and total lag time were not influenced by light treatment. Mean interval was about 27 h for all eggs and was not influenced by clutch size. 7. Light quality at the intensity used in this study did not appear to play an important role in oviposition patterns of turkey hens.

Animals↗