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PubMed1997

Evaluation of chromatin condensation in human spermatozoa: a flow cytometric assay using acridine orange staining.

The quality of sperm chromatin is an important factor in fertilization and is especially critical where one spermatozoon is artificially selected for fertilizing an egg (as in intracytoplasmic sperm injection). In this study, flow cytometry after staining of human spermatozoa with Acridine Orange was used to study chromatin structure. A method is described for estimating the percentage of cells in a human sperm sample that have completed epididymal maturation in regard to chromatin condensation. Of the 121 samples of the semen that were examined, nine contained a higher percentage of hypocondensed spermatozoa and six samples contained elevated amounts of hypercondensed spermatozoa. In addition to aberrancies in chromatin condensation other defects showed up as satellite populations of spermatozoa with higher than normal ratios of red/green fluorescence after Acridine Orange staining. Such defects were found in 15 semen samples. The use of swim-up and Percoll gradient centrifugation methods was shown to improve the percentage of spermatozoa with normal chromatin structure in some samples with poor initial quality.

Acridine Orange↗
PubMed2004

Influence of brood, vent screening, and time of year on honey bee (Hymenoptera: Apidae) pollination and fruit quality of greenhouse tomatoes.

Greenhouse tomatoes, Lycopersicon esculentum Miller (Solanaceae), are autogamous, but facilitated pollination results in increased fruit size and set. Previous research examining honey bee pollination in greenhouse tomato crops established that fruit quality resulting from honey bee visitation is often comparable to bumble bees (Bombus spp.) and significantly better than in flowers that receive no facilitated pollination. However, management alternatives have not been studied to improve tomato fruit quality when honey bees are the only pollination option available for the high-value greenhouse industry. We investigated whether the quantity of brood (eggs, larvae, and pupae) in a honey bee colony in the winter and screening on greenhouse vents in the summer would encourage honey bee foraging on tomato flowers. We also established the influence of time of year on the potential for honey bees to be effective pollinating agents. We constructed small honey bee colonies full of naive forager bees with either two frames of brood ("brood colonies") or two empty frames ("no-brood") and compared total fruit set and the number of tomato seeds resulting from fruit potentially visited by honey bees in each of these treatments to bagged flowers that received no facilitated pollination. There was no significant difference in the quality of fruit resulting from honey bees from "brood" and "no-brood" colonies. However, these fruits produced significantly more seeds than bagged flowers restricted from facilitated pollination. Honey bees from brood and no-brood colonies also resulted in 98% fruit set compared with 80% fruit set in bagged flowers that received no facilitated pollination. During the summer, the number of seeds per fruit did not differ significantly between unbagged flowers potentially visited by honey bees in screened greenhouses and unscreened greenhouses and bagged flowers that received no facilitated pollination. However, time of year did have a significant influence on the quality of fruit produced by honey bees compared with flowers that received no facilitated pollination, because no difference in seed number was observed between the treatments after mid-April. The results from this study demonstrate that the management of brood levels and vent screening cannot be used to improve the quality of fruit resulting from honey bee pollination and that honey bees can be a feasible greenhouse pollination alternative only during the winter.

Agriculture↗
PubMed1992

Comparative sensitivity of gametes and early developmental stages of a sea urchin species (Echinometra mathaei) and a bivalve species (Isognomon californicum) during metal exposures.

Bioassays were developed using sperm of a sea urchin (Echinometra mathaei), and sperm, embryos, and larvae of a bivalve species (Isognomon californicum). Sea urchin spawning was restricted to only a few months of the year and viability of sperm throughout the year varied from 5 to 75%. Sea urchin fertilization assays were affected by temporal variation in sperm viability. Spawning in this bivalve species occurs year-round and there is little temporal variation in sperm viability. Since stringent sperm:egg ratios are not required for the bivalve embryo and larval assays, these were less affected by variation in gamete quality. The relative sensitivity of the various assays were compared during exposure to three different metal pollutants: cadmium, copper, and tributyltin. Gametes and embryos were relatively resistant to cadmium toxicity, but larvae were very sensitive. With copper and tributyltin, sea urchin and bivalve fertilization assays were the least sensitive; and bivalve growth assays were the most sensitive, followed closely by the bivalve embryo assays. On the basis of sensitivity, ease and time required to conduct the assay, and salinity tolerance, the bivalve embryo assay was recommended as the overall single most reliable toxicity bioassay. However a multispecies, multidimensional approach using sperm fertilization assays as well as embryo assays should be employed, perhaps in a hierarchal manner. Larval growth assays were deemed too tedious and time consuming to be used routinely, but due to their high sensitivity, should still be considered as a valuable comparative tool.

Animals↗
PubMed2004

Embryonic exposure to oestrogen causes eggshell thinning and altered shell gland carbonic anhydrase expression in the domestic hen.

Eggshell thinning among wild birds has been an environmental concern for almost half a century. Although the mechanisms for contaminant-induced eggshell thinning are not fully understood, it is generally conceived to originate from exposure of the laying adult female. Here we show that eggshell thinning in the domestic hen is induced by embryonic exposure to the synthetic oestrogen ethynyloestradiol. Previously we reported that exposure of quail embryos to ethynyloestradiol caused histological changes and disrupted localization of carbonic anhydrase in the shell gland in the adult birds, implying a functional disturbance in the shell gland. The objective of this study was to examine whether in ovo exposure to ethynyloestradiol can affect eggshell formation and quality in the domestic hen. When examined at 32 weeks of age, hens exposed to ethynyloestradiol in ovo (20 ng/g egg) produced eggs with thinner eggshells and reduced strength (measured as resistance to deformation) compared with the controls. These changes remained 14 weeks later, confirming a persistent lesion. Ethynyloestradiol also caused a decrease in the number of shell gland capillaries and in the frequency of shell gland capillaries with carbonic anhydrase activity. These data suggested that a disrupted carbonic anhydrase expression was involved in the mechanism for the oestrogen-induced eggshell thinning found in this study. The results support our hypothesis that eggshell thinning in avian wildlife can result from a structural and functional malformation in the shell gland, induced by xeno-oestrogen exposure during embryonic development.

Animals↗
PubMedDate not supplied

Effect of skin colour and selected physical characteristics on Schistosoma mansoni dependent morbidity.

The effect of the colour group on the morbidity due to Schistosoma mansoni was examined in two endemic areas situated in the State of Minas Gerais, Brazil. Of the 2773 eligible inhabitants, 1971 (71.1%) participated in the study: 545 (27.6%) were classified as white, 719 (36.5%) as intermediate and 707 (35.9%) as black. For each colour group, signs and symptoms of individuals who eliminated S.mansoni eggs (cases) were compared to those who did not present eggs in the faeces (controls). The odds ratios were adjusted by age, gender, previous treatment for schistosomiasis, endemic area and quality of the household. There was no evidence of a modifier effect of colour on diarrhea, bloody faeces or abdominal pain. A modifier effect of colour on hepatomegaly was evident among those heaviest infected (> or = 400 epg): the adjusted odds ratios for palpable liver at the middle clavicular and the middle sternal lines were smaller among blacks (5.4 and 6.5, respectively) and higher among whites (10.6 and 12.9) and intermediates (10.4 and 10.1, respectively). These results point out the existence of some degree of protection against hepatomegaly among blacks heaviest infected in the studied areas.

Brazil↗
PubMed1995

Factors affecting the developmental stage of embryos recovered on day 7 from superovulated dairy cattle.

The objective of this retrospective study was to evaluate the factors influencing developmental stage of bovine embryos recovered from superovulated dairy cattle 7 d after estrus. From 217 superovulated dairy cows and heifers, 2,211 eggs were recovered, of which 1,495 were classified as transferable embryos based on morphological evaluation of developmental stage and quality. From the evaluated embryos, 1,429 were non-surgically transferred to recipients to produce 623 calves. The transferable embryos were classified into five developmental stages and four quality grades. The least-developed transferable embryos tended to be classified into poorer quality grades. A multifactorial statistical model was used to analyze whether the following factors were associated with the developmental stage and quality grade of the embryos: donor breed, parity, gonadotropin preparation, embryo sex, insemination bull, embryologist (the person evaluating the embryo), year, and season of recovery. Among these factors, only the embryologist and the donor animal accounted for significant variation in embryo development. It was concluded that the developmental stage of embryos recovered at d 7 from superovulated cattle, when evaluated by simple morphological criteria, was correlated with the embryo's quality and was affected by the donor animal but in this study not by the embryo sex, donor breed and parity, gonadotropin preparation, and insemination bull used. The embryo's quality grading was influenced by the embryologist. Consequently, sexing of an embryo recovered from superovulated cattle is not possible by simple morphological evaluation of the embryo's developmental stage.

Animals↗
PubMed1991

The effect of indomethacin on eggshell quality.

The objective of the present study was to determine whether indomethacin, an inhibitor of prostaglandin synthesis, could improve shell quality in birds laying a high incidence (greater than 15%) of soft-shelled (SS) and shell-less (SL) eggs and in birds laying exclusively hard-shelled (HS) eggs. Treatments consisted of two injection types, indomethacin or oil (controls), and two injection times, 4 h or 16 h postentrance of the egg into the uterus. Indomethacin delayed oviposition in both HS and SS or SL egg layers, with indomethacin being more effective in delaying oviposition when injected at 16 h as compared with 4 h postentrance of an egg into the uterus. Indomethacin increased shell thickness of the first egg laid following injection in the SS or SL groups but not in the HS groups. Egg weight was not affected by indomethacin injection. It was concluded that indomethacin delayed oviposition and prevented the premature expulsion of some SS or SL eggs. This delay in oviposition resulted in thicker shells of the first egg laid postinjection.

Animals↗
PubMed1982

Constancy of net protein intake in rats on self-selection regimen with proteins of various qualities.

Voluntary intake of proteins of various qualities in relation to dietary protein utilization was investigated in growing and adult rats. The rats were given two diets, one containing high protein and the other no protein, and were allowed to self-select protein and energy intakes freely from both diets. The results showed that total food intake (protein diet plus protein-free diet) and body weight gain were similar among five proteins tested. However, the amount of protein consumed by the growing rats per 100 g of total food intake (i.e., dietary protein level) was different depending upon the protein qualities, that is, wheat gluten (WG) 44 g, casein (CA) 30 g, soy protein (SP) 21 g, lactalbumin (LA) 19 g, and amino acid mixture simulating egg protein (AA) 11 g. Net protein utilization (NPU), estimated as the proportion of protein intake that is retained in the body, was as follows: WG, 20%; CA, 33%; SP, 44%; LA, 50%; and AA, 74%. From the above figures, net dietary protein value, which is a measure of utilizable protein in the diet, was calculated by multiplying the dietary protein level by NPU. In contrast to the difference in protein intake, net dietary protein value was quite constant in spite of large differences in the dietary protein quality, being 8 to 10%. A similar relationship between protein intake and protein utilization was obtained also with adult rats, except that the net dietary protein value was smaller in adult rats than in growing rats. These results may suggest that the animals can regulate the intake of dietary protein to keep the amount of protein available for the body constant.

Amino Acids↗
PubMed2006

Solar ultraviolet-B radiation alters the attractiveness of Arabidopsis plants to diamondback moths (Plutella xylostella L.): impacts on oviposition and involvement of the jasmonic acid pathway.

Solar ultraviolet-B radiation (UV-B) can have large impacts on the interactions between plants and herbivorous insects. Several studies have documented effects of UV-B-induced changes in plant tissue quality on the feeding performance of insect larvae. In contrast, the effects of UV-B-induced plant responses on the behavior of adult insects have received little attention. We carried out a series of field and glasshouse experiments using the model plant Arabidopsis thaliana L. and the crucifer-specialist insect Plutella xylostella L. (diamondback moth) to investigate the effects of UV-B on natural herbivory and plant-insect interactions. Natural herbivory under field conditions was less severe on plants exposed to ambient UV-B than on plants grown under filters that attenuated the UV-B component of solar radiation. This reduced herbivory could not be accounted for by effects of UV-B on larval feeding preference and performance, as P. xylostella caterpillars did not respond to changes in plant quality induced by UV-B. In contrast, at the adult stage, the insects presented clear behavioral responses: P. xylostella moths deposited significantly more eggs on plants grown under attenuated UV-B levels than on plants exposed to ambient UV-B. The deterring effect of UV-B exposure on insect oviposition was absent in jar1-1, a mutant with impaired jasmonic acid (JA) sensitivity, but it was conserved in mutants with altered ethylene signaling. The jar1-1 mutant also presented reduced levels of UV-absorbing phenolic compounds than the other genotypes that we tested. Our results suggest that variations in UV-B exposure under natural conditions can have significant effects on insect herbivory by altering plant traits that female adults use as sources of information during the process of host selection for oviposition. These effects of natural UV-B on plant quality appear to be mediated by activation of signaling circuits in which the defense-related hormone JA plays a functional role.

Age Factors↗
PubMed1999

Prospecting enhances breeding success of first-time breeders in the great cormorant, Phalacrocorax carbo sinensis.

In many species of colonial seabirds, young birds visit colonies in the years before they start breeding. This prospecting behaviour may allow them to obtain information that could enhance their future breeding success. We examined the reproductive consequences of prospecting behaviour in the colonial great cormorant, and found support for this idea. New breeders that had been prospecting actively in the previous year obtained breeding sites of higher quality (i.e. closer to sites where conspecifics had fledged young in the previous year) and had higher breeding success than those that had been less active. Prospecting occurred mostly late in the breeding season, and coincided with the time when the majority of the eggs had hatched but before the chicks started fledging, that is, when breeding success in the colony reflected habitat suitability. These results are thus consistent with the use of conspecific reproductive performance as a cue for the quality of a breeding habitat as expected from the 'performance-based conspecific attraction hypothesis'. Copyright 1999 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed1997

Effects of an oxygen-enriched environment on the survival of turkey embryos between twenty-five and twenty-eight days of age.

The hypothesis was tested that increased partial pressure of oxygen during the plateau (25 to 26 d of incubation for turkeys) and paranatal (27 to 28 d of incubation) stages of incubation may increase survival rates of turkeys from selected genetic lines. Partial pressure of oxygen inside the incubator cabinet was increased to 171 + 3 mm Hg of the barometric pressure during the plateau stage in oxygen consumption and compared to ambient oxygen (152 + 3 mm Hg). Turkey embryos from genetic lines selected for egg production (E) or growth (F) were compared to their respective randombred controls. These genetic lines have previously been shown to differ in egg weight, eggshell conductance, length of incubation period, embryonic gluconeogenesis, and survival rates during late incubation. Blood, liver, heart, and pipping muscle samples were obtained prior to pipping, at internal pipping and external pipping, and at hatching. The blood was analyzed for glucose concentration and the remaining tissues were assayed for glycogen concentrations. Survival rates were determined on approximately 2,200 eggs in each of three independent trials of the experiment. Interactions of oxygen treatment and genetic line were observed for embryonic survival, heart growth, and hepatic glycogen content. The data suggest that the response to increased oxygen tension in selected genetic lines has been diminished. It was concluded that embryos have been altered metabolically by genetic selection and the concomitant increase in mortality of selected lines during the plateau and paranatal stages is not simply the result of shell quality and hypoxia.

Aging↗
PubMed2001

[Implementation of a new electronic patient record in surgery].

INTRODUCTION: The increasing amount of clinical data, intensified interest of patients in medical information, medical quality management and the recent cost explosion in health care systems have forced medical institutions to improve their strategy in handling medical data. In the orthopedic department (3,600 surgeries, 75 beds, 14,000 consultations) software application for comprehensive patient data management has been developed. METHOD: When implementing the electronic patient history following criteria were evaluated: 1. software evaluation, 2. implementation, 3. work flow, 4. data security/system stability. RESULTS: In the first phase the functional character was defined. Implementation required 3 months after parametrization. The expense amounted to 130,000 DM (30 clients). The training requirements were one afternoon for the secretaries and a 2-h session for the residents. The access speed on medically relevant data averaged under 3 s. The average saving in working hours was approximately 5 h/week for the secretaries and 4 h/week for the residents. The saving in paper amounted to 36,000 sheets/year. In 3 operational years there were 3 server breakdowns. CONCLUSIONS: Evaluation of the saving on working hours showed that such a system can amortize within a year. The latest improvements in hardware and software technology made the electronic medical record with integrated quality-control practicable without massive expenditure. The system supplies an extensive platform of information for patient treatment and an instrument to evaluate the efficiency of therapy strategies independent of the clinical field.

Database Management Systems↗
PubMed2006

Expression of vasa (vas)-related genes in germ cells and specific interference with gene functions by double-stranded RNA in the monogenean, Neobenedenia girellae.

Neobenedenia girellae, a monogenean, is an important pathogen in marine cultured fish such as yellowtail and amberjack. An effective control method is required but none has yet been established. Aiming to establish a new control method by interfering with the gametogenesis of N. girellae, we focused on vasa (vas)-related genes that are expressed exclusively in the germline granules in Drosophila, Caenorhabditis elegans and other animals. Three vas-related genes (N. girellae vasa-like gene, Ngvlg1, Ngvlg2 and Ngvlg3) were isolated by PCR and Ngvlg1 and Ngvlg2 were shown to be expressed only in germ cells. We demonstrated that introduction of double-stranded Ngvlg1 or Ngvlg2 RNA by soaking resulted in partial or complete loss of germ cells. Moreover, the hatching rate of eggs from animals showing partial loss of germ cells decreased significantly. These results suggest that Ngvlg1 and Ngvlg2 are essential genes for germ cell quantity and quality. The possibility that a new control method can be developed by controlling gametogenesis of N. girellae was proven, because sterilised N. girellae could be produced.

Amino Acid Sequence↗
PubMed1999

Prospective approaches to avoid flock fertility problems: predictive assessment of sperm function traits in poultry.

This paper discusses why it is important to evaluate males as individuals and how advances made in understanding and measurement of sperm function can be used to improve reproductive efficiency in poultry. Commercial turkey breeding relies on pooling semen from multiple toms. It generally is assumed that sperm in good quality semen from all toms are equally fecund. (Fecund is defined, for males, as an individual whose semen contains a majority of sperm with the potential of producing fertilized eggs, which includes success at all steps in the fertilization process: sperm movement, storage in the hens' sperm storage tubules, binding and penetrating the perivitelline layer, and fertilization.) However, when DNA fingerprinting was used to determine paternity efficiency after pooling ejaculates from seven or more toms, it was found that 18 of 26 males produced very few, or no, offspring. In addition, the traditional measures of poultry semen quality: semen volume, sperm concentration, sperm viability, and subjective motility assessment, were poor predictors of paternity. In recent years, a concentrated effort has been made to develop and evaluate methods that quantify sperm function in poultry. Methods to measure some of these traits are reviewed: sperm motility, sperm storage in the hen, and sperm binding and penetration of the ovum. Data supporting use of these tools for managing flock fertility from the male perspective are explored.

Animal Husbandry↗
PubMed1991

Effects of selection for growth and selection diet on eggshell quality and embryonic development in Japanese quail.

Decreased hatchability in Japanese quail following selection for growth and relative improvements in hatchability following selection during goitrogen treatment were investigated. An unselected quail line (Line C) and lines selected for high 4-wk body weight while being fed diets containing either 20% CP and .2% thiouracil (TU) (Line T), or 28% CP and no TU (Lines P and H-CD) were used. Egg weight loss between 0 and 4, and 0 and 14 days of incubation, hatchability, and developmental stage of embryonic mortalities were determined in Experiment 1. Length of the incubation period, and percentages of body water and dry body weight were determined for 10- and 14-day embryos and 18-day-old chicks in Experiment 2. Eggshell quality was determined in Experiment 3. Hatchability was lower in selected than in unselected quail due to increased early and late embryonic mortality. Egg weight loss during incubation was greater in Line P than in Line C eggs. Weight loss, measured across lines, was lower in eggs that hatched than in those in which embryos died early. Percentage body water was higher in Line C than in Lines P and T at 14 and 18 days of incubation; however, percentage body water in Line T was lower than Line P at 14 days. Percentage of dry body weight was greater in Lines P and T than in Line C at 14 and 18 days. Eggshell thickness was greater in Line H-CD when compared with Line C. Increases in eggshell permeability occurring during selection for growth were associated with increased embryonic mortality and decreased hatchability.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed2006

Assessment of gamete quality for the eastern oyster (Crassostrea virginica) by use of fluorescent dyes.

Evaluation of sperm motility is the single most widely used parameter to determine semen quality in mammals and aquatic species. While a good indicator for fresh sperm viability, post-thaw motility is not always effective at predicting fertilizing ability. Techniques using fluorescent dyes can assess functionality of mammalian sperm, but have not been widely applied in aquatic organisms. The eastern oyster Crassostrea virginica is an important mollusk in the United States, and cryopreservation protocols have been developed to preserve sperm and larvae to assist research and hatchery production. In this study, protocols were developed to assess sperm cell membrane integrity and mitochondrial function by flow cytometry and to assess viability of eggs by fluorescence microscopy. The fluorescent dyes SYBR 14 and propidium iodide (PI) (to assess membrane integrity) and rhodamine 123 (R123) (to assess mitochondrial membrane potential) were used to evaluate the quality of thawed oyster sperm previously cryopreserved with different cryoprotectant and thawing treatments. Membrane integrity results were correlated with motility of thawed sperm and mitochondrial membrane potential with fertilizing ability. Fluorescein diacetate (FDA) was used to assess cytotoxicity of cryoprotectant solutions and post-thaw damage to oyster eggs. The results indicated that membrane integrity (P=0.004) and thawing treatments (P=0.04), and mitochondrial membrane potential (P=0.0015) were correlated with motility. Fertilizing ability was correlated with cryoprotectant treatments (P=0.0258) and with mitochondrial membrane potential (P=0.001). The dye FDA was useful in indicating structural integrity of fresh and thawed eggs. Exposure of eggs, without freezing, to dimethyl sulfoxide yielded higher percentages of stained eggs and fertilization rate than did exposure to propylene glycol (P=0.002). Thawed eggs were not stained with FDA (<1%) and larvae were not produced.

Animals↗
PubMed2004

A first-generation microsatellite linkage map of the Japanese quail.

A linkage map of the Japanese quail (Coturnix japonica) genome was constructed based upon segregation analysis of 72 microsatellite loci in 433 F(2) progeny of 10 half-sib families obtained from a cross between two quail lines of different genetic origins. One line was selected for long duration of tonic immobility, a behavioural trait related to fearfulness, while the other was selected based on early egg production. Fifty-eight of the markers were resolved into 12 autosomal linkage groups and a Z chromosome-specific linkage group, while the remaining 14 markers were unlinked. The linkage groups range from 8 cM (two markers) to 206 cM (16 markers) and cover a total map distance of 576 cM with an average spacing of 10 cM between loci. Through comparative mapping with chicken (Gallus gallus) using orthologous markers, we were able to assign linkage groups CJA01, CJA02, CJA05, CJA06, CJA14 and CJA27 to chromosomes. This map, which is the first in quail based solely on microsatellites, is a major step towards the development of a quality molecular genetic map for this valuable species. It will provide an important framework for further genetic mapping and the identification of quantitative trait loci controlling egg production and fear-related behavioural traits in quail.

Animals↗