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Evaluation of the protein quality of an isolated soy protein in young men: relative nitrogen requirements and effect of methionine supplementation.

The protein nutritional value of an isolated soy protein (Supro-620) was evaluated in a series of nitrogen balance studies in healthy young male MIT students. Experiment 1 involved giving eight subjects graded intakes of the isolated soy protein while seven additional subjects received egg protein. Mean nitrogen intake required for N balance for the isolated soy protein and the egg protein were not significantly different (p greater than 0.1). In experiment 2, nine subjects received soy and the effects on N balance at various levels of L-methionine supplementation were studied. In experiment 3 eight subjects each were studied at two nitrogen intake levels of isolated soy protein/kg/day with methionine supplementation, and an unsupplemented egg protein period included. Results from experiment 2 and 3 at an 82 mg N/kg/day test level showed that N balance for 1.6% supplementation was significantly below that of unsupplemented egg while 1.1% and unsupplemented soy were indistinguishable from egg. No beneficial effects of methionine supplementation were observed when the test nitrogen intake level was 128 mg N/kg/day. These results indicate that for healthy adults, the isolated soy protein is of high nutritional quality, comparable to that of animal protein sources, and that the methionine content is not limiting for adult protein maintenance.

Adult↗

Digestibility and dietary quality of nonacetylated and acetylated Vicia faba proteins in maintenance. Studies on 15N-labelled adult rats.

N digestibility and the dietary quality of a Vicia faba protein isolate as well as of two protein fractions from this isolate, acetylated and nonacetylated, were estimated in 15N-labelled adult rats. For reference, whole egg, casein and wheat were studied. The diets were fed at maintenance energy intake level. Daily metabolic fecal nitrogen estimated by an isotopic method was 5 mg/100 g body weight in animals receiving whole egg, casein, or Vicia faba proteins. When wheat was included in the diet metabolic fecal nitrogen was increased to 140% of this value. True N digestibility of reference proteins was 91-92% and of Vicia faba proteins 95-97%. Protein quality for maintenance was high and similar for all proteins. Acetylation affected neither N digestibility nor protein quality under maintenance conditions.

Acetylation↗

Effect of induced molting on albumen quality, hatchability, and chick body weight from broiler breeders.

Commercial Cobb broiler breeders were subjected to molting from 55 to 62 wk of age. Incubating eggs were collected before molting and after molting and were stored for 18, 13, 8, or 3 d. Sample eggs were broken to measure albumen Haugh units (HU) and pH. Two hundred twenty-five eggs per treatment, stored for 18, 13, 8, or 3 d, were incubated for 21 d; the hatched chicks were weighed at the end of incubation and again after Day 7 of rearing. As the storage time increased, albumen HU decreased (P < 0.001). At all storage times, HU after molting were higher than those before molting (P < 0.001). Albumen pH increased with storage time (P < 0.001). A molting x storage interaction on pH was observed after 8 d of storage (P = 0.03). Hatchability of eggs increased after hens were molted, if the eggs were stored for a long time (P < 0.001). Body weights of 1-d-old chicks from the eggs of hens before molt were heavier than those from eggs after molting (P < 0.001). Conversely, at 7 d, the chicks from the eggs after molting were significantly heavier than those from eggs before molting (P < 0.001). We concluded that molting was a procedure to improve hatchability and chick juvenile growth. If eggs need to be stored, we recommended that fresh eggs with high HU value be stored rather than those with low HU values.

Aging↗

Effects of elevated egg corticosterone levels on behavior, growth, and immunity of yellow-legged gull (Larus michahellis) chicks.

Eggs of vertebrates contain steroid hormones of maternal origin that may influence offspring performance. Recently, it has been shown that glucocorticoids, which are the main hormones mediating the stress response in vertebrates, are transmitted from the mother to the egg in birds. In addition, mothers with experimentally elevated corticosterone levels lay eggs with larger concentrations of the hormone, which produce slow growing offspring with high activity of the hypothalamo-adrenal axis under acute stress. However, the effects and function of transfer of maternal corticosterone to the eggs are largely unknown. In the present study, we injected corticosterone in freshly laid eggs of yellow-legged gulls (Larus michahellis), thus increasing the concentration of the hormone within its natural range of variation, and analyzed the effect of manipulation on behavioral, morphological, and immune traits of the offspring in the wild. Eggs injected with corticosterone had similar hatching success to controls, but hatched later. Mass loss during incubation was greater for corticosterone-treated eggs, except for the last laid ones. Corticosterone injection reduced rate and loudness of late embryonic vocalizations and the intensity of chick begging display. Tonic immobility response, reflecting innate fearfulness, was unaffected by hormone treatment. Elevated egg corticosterone concentrations depressed T-cell-mediated immunity but had no detectable effects on humoral immune response to a novel antigen, viability at day 10, or growth. Present results suggest that egg corticosterone can affect the behavior and immunity of offspring in birds and disclose a mechanism mediating early maternal effects whereby stress experienced by females may negatively translate to offspring phenotypic quality.

Animals↗

Nutritional quality of prebreeding diet influences breeding performance of the Florida scrub-jay.

Food supplementation studies of breeding birds have traditionally concentrated on energetic constraints on breeding performance. It is only recently that the nutritional quality of the prebreeding diet has also been considered influential. We examined the importance of specific nutrients in the prebreeding diet of the Florida scrub-jay ( Aphelocoma coerulescens). Birds were provided with one of two supplements (rich in protein and fat or rich in fat only) prior to breeding in 2000 and 2001 and their breeding performance, in relation to unsupplemented (control) birds, was examined. Birds receiving both supplements significantly advanced laying in both years, and increased clutch size in 2000 but not in 2001. Laying date explained variation in clutch size in birds on dietary supplements. Egg mass and volume declined with laying order, irrespective of dietary treatment, but birds on the high fat, high protein diet laid heavier third eggs than controls and this was independent of laying date. Laboratory analysis of 14 abandoned and unhatched eggs revealed that as egg mass increased so did the absolute amount of protein and water while fat content remained relatively fixed. Using these relationships between the masses of egg components and fresh egg mass, we calculated that heavier third eggs laid by birds on high fat and high protein, compared with those laid by controls, contained more water that may be fundamental to chick growth and survival. This is the first demonstration for an avian species that nutritional quality of prebreeding diet can simultaneously influence laying date, clutch size, and egg size and composition.

Animal Feed↗

Effect of epididymis handling conditions on the quality of ram spermatozoa recovered post-mortem.

Post-mortem spermatozoa recovery is an important technique for obtaining germplasm reserves from genetically valuable animals or endangered species. However, there are many factors that influence the outcome of this technique. We have studied the effect of the interval between animal's death and sperm recovery (0, 24 or 48 h) on the quality and freezability of ram spermatozoa from cauda epididymidis. Storage temperature of epididymis (room temperature or 5 degrees C) was also analysed. Spermatozoa were diluted with Tes-Tris-Fructose solution supplemented with egg yolk (10%) and glycerol (4%), and frozen using a programmable biofreezer (-20 degrees C/min). Pre-freeze and post-thaw sperm samples showed viable spermatozoa up to 48 h after the animal's death, although their quality declined significantly as post-mortem storage time increased. Epididymis sperm stored at 5 degrees C showed better motility and a lower percentage of abnormal forms than epididymis stored at room temperature after 24 and 48 h. The fertilizing ability of cauda epididymis ram spermatozoa obtained at 0 and 24h after the animal's death is similar to that of ejaculated spermatozoa. Therefore, a good protocol for post-mortem semen collection in rams when epididymal spermatozoa cannot be collected immediately, is to preserve the epididymis at 5 degrees C and process the samples in the first 24h after the animal's death.

Acrosome↗

Post-thaw bovine spermatozoal quality estimated from fresh samples.

Comparisons were made among flow cytometrically quantified populations of fresh and cryopreserved bovine spermatozoa, tri-stained with fluorophores rhodamine 123 (R123), 5- (and 6-) carboxy-4',5'-dimethylfluorescein diacetate (CMFDA), and propidium iodide (PI), and analyzed by dual parameter flow cytometry. The purpose was to find parameters in fresh semen samples that were potentially predictive of frozen sample parameters so that bulls with marginal cryopreservation capacity could be identified. Fresh and cryopreserved aliquots of semen from two sets of six bulls were semen processed in either milk (bulls 1-6) or egg yolk citrate (bulls 11-16). Membrane-damaged red (PI) and intact green (CMFDA + R123) populations were evaluated as percentages of 10,000-cell samples or numbers per straw. In milk, gated central subsets of membrane-damaged sperm cells in fresh samples and moribund cells in post-thaw samples were significantly correlated for sample percentages (r = 0.90, P = 0.014) and cell numbers per straw (r = 0.94, P = 0.006). In egg yolk citrate, fresh and frozen membrane-damaged populations were correlated (percentages: r = 0.81, P = 0.048; numbers: r = 0.88, P = 0.019). Additionally, post-thaw motility estimated by a photographic method was correlated with the number of sperm cells per straw in the intact central green subset (r = 0.98, P = 0.0006). These findings suggest that partitioning red and gree populations into smaller, central subset populations reveals significant relationships between fresh and cryopreserved bull ejaculates. The proportion of membrane-damaged spermatozoa in fresh semen seems to be predictive of the proportion of post-thaw membrane-damaged or moribund spermatozoa. The data consistently showed ejaculates of bulls 1 and 12 as having the greatest cryopreservation potentials, and bulls 4 and 16 the least. Thus, flow cytometric evaluation of fresh semen may be useful for identifying young sires with relatively poor fertilizing potential.

Analysis of Variance↗

Periodical cicadas use light for oviposition site selection.

Organisms use incomplete information from local experience to assess the suitability of potential habitat sites over a wide range of spatial and temporal scales. Although ecologists have long recognized the importance of spatial scales in habitat selection, few studies have investigated the temporal scales of habitat selection. In particular, cues in the immediate environment may commonly provide indirect information about future habitat quality. In periodical cicadas (Magicicada spp.), oviposition site selection represents a very long-term habitat choice. Adult female cicadas insert eggs into tree branches during a few weeks in the summer of emergence, but their oviposition choices determine the underground habitats of root-feeding nymphs over the following 13 or 17 years. Here, field experiments are used to show that female cicadas use the local light environment of host trees during the summer of emergence to select long-term host trees. Light environments may also influence oviposition microsite selection within hosts, suggesting a potential behavioural mechanism for associating solar cues with host trees. In contrast, experimental nutrient enrichment of host trees did not influence cicada oviposition densities. These findings suggest that the light environments around host trees may provide a robust predictor of host tree quality in the near future. This habitat selection may influence the spatial distribution of several cicada-mediated ecological processes in eastern North American forests.

Animals↗

An analysis of semen quality and sperm function in cases of oligozoospermia.

This study is based upon an analysis of 27 severely oligozoospermic men, in whom defects in sperm motility and morphologic defects were also evident, and 35 control subjects of proven fertility. Semen samples recovered from these men were assessed by conventional criteria, time-exposure photomicrography, and the zona-free hamster egg penetration test. Evidence of severely defective sperm function was obtained for the oligozoospermic group, which gave a mean +/- standard error (SE) fertilization rate of 2.8% +/- 1.5, compared with 44.0% +/- 3.4 for the normal fertile control subjects. Multivariate discriminant analysis was used for selection of those parameters of semen quality that could most accurately identify the 70% of oligozoospermic samples that failed to exhibit any fertilizing potential in the hamster egg assay. This analysis correctly identified 89% of such samples, on the basis of seven discriminating variables, of which the most significant were percentage of motile sperm, the log of sperm density, and a log transformation of the concentration of progressive spermatozoa.

Fertilization↗

Egg globulins in rapid virus diagnosis.

Egg-derived antibodies specific for a range of human viruses are now available commercially. These globulins are prepared by inoculating chickens with the relevant virus then harvesting antibodies from egg yolks. Reagents for influenza A and B, parainfluenza 1 and 3, adenovirus group antigen and rotavirus were tested. The successful use of these reagents on tissue culture and clinical material is described as well as their quality assessment. The advantages of this type of antibody are discussed.

Adenoviridae↗

Combined roles of contact stimulant and deterrents in assessment of host-plant quality by ovipositing zebra swallowtail butterflies.

Zebra swallowtail (Eurytides marcellus) butterflies are stimulated to oviposit by a single compound, 3-caffeoyl-muco-quinic acid (1). Analysis of the aqueous extracts of the leaves of the host. Asimina triloba, showed that they contained stimulant 1, its isomer (2), and the flavonoids rutin (3) and nicotiflorine (4) as major components. We compared the concentrations of compounds 1-4 in terminal leaves (TL) and expanded leaves (EL) of the host plants at four different times throughout the growing season. In spring, the concentration of 1 was highest in TLs, and flavonoids were not detectable or present at low levels. As the season progressed, however, the concentrations of flavonoids increased, reached maxima by late summer, and then decreased as the plants started senescing. There were also significant differences in the concentrations of these compounds between TLs and ELs. In a choice assay with model leaves, we tested equivalent amounts of post-dichloromethane aqueous extracts made in spring (May) and in fall (September). September extracts received significantly fewer approaches and eggs. In greenhouse experiments with potted A. triloba plants, the butterflies chose some leaves to lay eggs, while others were rejected or ignored. Analyses showed that the concentrations of compound 1 were not significantly different in the three kinds of leaves. The flavonoids (3 and 4), however, were significantly higher in the leaves that were ignored. Multiple-choice tests using model plants suggested that concentrations of both flavonoids and stimulant were important in assessing host suitability. There was a gradual decrease in approaches as the concentration of 1 decreased. Higher amounts of flavonoids deterred egg laying even in the presence of high concentrations of stimulant 1. At lower concentrations of 1, the addition of low doses of flavonoids deterred egg laying. Thus, the results suggest that the butterflies use both qualitative and quantitative information about these compounds to assess host quality. This behavior may have evolved to take advantage of seasonal variation in the chemistry of their host, A. triloba.

Animals↗

Genetic regulation of egg and embryo survival.

In both mice and humans, 15-50% of embryos die during the preimplantation period from mechanisms that are largely unknown. Two major criteria predict preimplantation embryo quality, the rate of development and the degree of fragmentation. We review evidence that both of these criteria have a genetic basis. Rate of development and subsequent embryo survival are controlled by a gene, Ped, we discovered in the mouse. Although progress is being made in the search for the human homologue of the mouse Ped gene, it has not yet been identified. Fragmentation, observed in both mouse and human embryos, is probably the result of apoptosis. We analysed transcription of two genes that regulate apoptosis, bcl-2 and bax, and found that both are transcribed in mouse and human preimplantation embryos. Overall, the literature reviewed and new data presented in this paper support the concept that there is a genetic basis for preimplantation egg and embryo survival.

Animals↗

Effects of sodium azide on sea urchin embryos and gametes.

Sodium azide (SA) was tested on sea urchin embryos and gametes (Paracentrotus lividus). Developing embryos were exposed to SA (10(-6) to 10(-3) M) up to pluteus larval stage, or for shorter intervals before or after hatching. Developmental defects in SA-exposed embryos consisted mainly of gut abnormalities, without any detectable differences between pre- or post-hatch-exposed embryos. SA-induced damage to gut was exerted during gastrulation, as evident by lectin binding of extracellular matrix. No mitotic damage was observed in SA-exposed embryos, nor could pH-related variations be detected in SA-induced embryotoxicity at pH's ranging from 8 to 6. Concurrently, no effect ensued in the exposure of unfertilized eggs to SA (10(-5) to 10(-2) M) both in terms of fertilization success and of offspring quality. When sperm were suspended in filtered seawater at pH's ranging from 8 to 6, and SA levels ranging from 10(-5) to 10(-2) M, fertilization success of SA-exposed sperm appeared to be modulated by pH, by displaying three distinct dose-response trends at pH 8, 7, or 6. The consequences of sperm pretreatment on offspring quality failed to show any significant SA-induced changes on larval malformations or mortality, while confirming the previously reported pH-induced increase of developmental defects in the offspring of acid-exposed sperm (Pagano et al.: Teratogenesis Carcinogen Mutagen 5:113-121, 1985).

Animals↗

Altered nuclear activation parameters of rat sperm treated in vitro with chromatin-damaging agents.

Standard semen measures do not assess the genetic integrity of sperm. A human sperm activation assay (HSAA) has proven very useful for assessing sperm quality and predicting pregnancy outcome. The HSAA involves incubating permeabilized sperm in cytoplasmic extracts of Xenopus laevis frog eggs. The extracts activate sperm nuclei, which undergo chromatin decondensation, DNA synthesis, and chromatin recondensation, mimicking events that occur after fertilization in vivo. However, no animal model sperm activation assay has been reported. We hypothesize that sperm activation assays will be useful for studying molecular mechanisms of sperm DNA repair by egg cytoplasm and for screening sperm for damaged DNA. Thus, the objectives of this study were to develop an in vitro rat sperm activation assay (RSAA) using cytoplasmic extracts of X. laevis frog eggs and to determine how chemically damaging the sperm chromatin would affect two sperm activation parameters, chromatin decondensation and DNA synthesis. We incubated demembranated rat sperm in a cytoplasmic extract of X. laevis frog eggs supplemented with tritiated thymidine triphosphate ([3H]TTP). The activated sperm nuclei underwent chromatin decondensation and DNA synthesis. Decondensation kinetics were examined using image analysis to measure the size of the sperm nuclei as they decondensed. DNA synthesis kinetics were examined using autoradiography of incorporated [3H]TTP. To investigate how chemical damage affects nuclear activation, we treated rat sperm in vitro with ethylene glycolbis(sulfosuccinimidyl-succinate; SEGS), a reversible crosslinking agent, or hydroxylamine (HA), a DNA base modifier. Treatment with SEGS blocked decondensation in a dose-dependent manner. In contrast, treatment with HA enhanced decondensation, induced gross chromatin abnormalities, and increased [3H]TTP incorporation into activated sperm nuclei, responses consistent with an attempt by the egg cytoplasm to repair DNA damage. These results suggest that the RSAA may be useful for detecting damaged sperm chromatin as a result of toxicant exposure.

Animals↗

Differences in induced volatile emissions among rice varieties result in differential attraction and parasitism of Nilaparvata lugens eggs by the parasitoid Anagrus nilaparvatae in the field.

We compared the volatiles of JA-treated plants of six rice varieties and then determined, in the laboratory and field, if they differed in attractiveness to Anagrus nilaparavate Pand et Wang, an egg parasitoid of rice planthoppers. Analyses of volatiles revealed significant differences among varieties, both in total quantity and quality of the blends emitted. On the basis of these differences, the six varieties could be roughly divided into three groups. In a Y-tube olfactometer, female wasps preferred odors from two groups. These preferences corresponded to observed parasitism rates in a field experiment. A comparison of the volatiles with results from behavioral assays and field experiments indicates that the quality (composition) of the blends is more important for attraction than the total amount emitted. The results imply that the foraging success of natural enemies of pests can be enhanced by breeding for crop varieties that release specific volatiles.

Animals↗

Liquid chromatographic determination of fluoroquinolones in egg albumen and egg yolk of laying hens using fluorometric detection.

A liquid chromatographic method was developed for the determination of ciprofloxacin, enrofloxacin, and sarafloxacin at 10-200 ppb in both egg yolk and egg albumen of laying hens. Egg yolk or albumen was acidified with 1 M phosphoric acid followed by deproteination with acetonitrile and centrifugation. The supernate was pipetted out, and the remaining protein pellet was extracted three times with acetonitrile. The supernates were combined and concentrated at 50 degrees C to <0.7 mL. The final volume was adjusted to 2 mL with 0.02 M potassium phosphate buffer, pH 2.5. Separation of the analytes was achieved using reversed-phase HPLC with fluorometric detection. The recoveries were >80% and coefficients of variation <20%. After validation, the method was applied for use in a national survey for fluoroquinolones in table eggs. Of the 276 eggs assayed, none was found positive for fluoroquinolones. The findings suggest that illegal use of fluoroquinolones in laying hens is not widespread.

Animals↗

Organic matrix morphology and distribution in the palisade layer of eggshells sampled at selected periods during lay.

1. 1 cm2 pieces of eggshells from a commercial battery flock were plasma etched to remove the outer shell membranes. 2. They were decalcified using EDTA (200 g/l, pH 6.9 to 7.0) in paraformaldehyde (20 g/l) and 25% gluteraldehyde (20 ml in 0.98 l) in phosphate buffer, then prepared for light and transmission electron microscopy. 3. Light microscopy revealed a differential distribution of matrix material within all 3 regions of the palisade layer at the beginning of lay. 4. Transmission electron microscopy revealed a more even distribution of matrix at the beginning of lay, although morphological differences were observed. At the end of lay all 3 regions showed an increase in % matrix and vesicles/10 cm2 of micrograph compared to the middle and beginning of lay periods. 5. It is hypothesised that matrix vesicles are involved in the regulation of the physiochemical environment within the forming eggshell and that the decline in shell quality associated with the end of lay is related to a concomitant change in matrix quality.

Animals↗

The effect of female age and ovarian reserve on pregnancy rate in male infertility: treatment of azoospermia with sperm retrieval and intracytoplasmic sperm injection.

Factors other than spermatozoa could be the major determinant of the success of assisted reproduction treatment in cases of male infertility. Our aim was to evaluate the effect of the wife's age and ovarian reserve on assisted reproduction success rates in the most severe type of male infertility, i.e. azoospermia. A total of 249 consecutive couples suffering from male infertility caused by azoospermia underwent microsurgical epididymal sperm aspiration (MESA) or testicular sperm extraction (TESE) with intracytoplasmic sperm injection (ICSI). Of these men, 186 had irreparable obstructive azoospermia, and 63 had non-obstructive azoospermia due to testicular failure. Neither the pathology, the source, the quantity, nor the quality of spermatozoa had any effect on fertilization or pregnancy rates. Maternal age and ovarian reserve (number of eggs) had no effect on fertilization or embryo cleavage, but did dramatically affect the embryo implantation, pregnancy and delivery rates. Wives of azoospermic men who were in their 20s had a 46% live delivery rate per cycle, wives aged 30-36 years had a 34% live delivery rate per cycle, wives aged 37-39 years had a 13% live delivery rate per cycle, and wives > or = 40 years had only a 4% live delivery rate per cycle. The number of eggs retrieved also affected pregnancy and delivery rate, but to a lesser extent than age. In virtually all cases of obstructive azoospermia, and in 62% of cases with non-obstructive azoospermia caused by germinal failure, sufficient spermatozoa could be retrieved to perform ICSI, with normal fertilization and embryo cleavage. However, the pregnancy rate and the live delivery rate were dependent strictly on the age of the wife, and on her ovarian reserve. Unfortunately, exaggerated claims of high pregnancy rates can thus easily be made by manipulating, in a very simple way, selection for female factors.

Adult↗