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PubMedDate not supplied

Dot-ELISA for the detection of IgM and IgG antibodies to Schistosoma mansoni worm and egg antigens, associated with egg excretion by patients.

Human schistosomiasis, caused by Schistosoma mansoni, is highly prevalent in Brazil and usually diagnosed by time consuming stool analysis. Serological tests are of limited use in this disease, mainly for epidemiological studies, showing no discrimination between previous contact with the parasite and active infections. In the present study, we standardized and compared a Dot-ELISA for IgM and IgG antibodies against S. mansoni antigens from eggs and worms with a routine IgG and IgM immunofluorescence assay using similar antigens, in the study of sera from 27 patients who had quantified egg stool excretion. The positivity obtained for IgG Dot-ELISA was 96.3% and 88.9% for IgM Dot-ELISA with worm antigen and 92.6% and 90.9% with egg antigen. The IFI presented similar positivities using worm antigen, 92.6% (IgG) and 96.3% (IgM), and lower results with egg antigen, 77.8%(IgG and IgM). The patients studied were divided into two groups according to their egg excretion, with greater positivity of serological tests in higher egg excreters. When comparing the quantitative egg excretion and the serological titers of the patients, we detected a correlation only with IgM Dot-ELISA, with r = 0.552 (p = 0.0127). These data show that Dot-ELISA can be used for the detection of specific antibodies against S. mansoni in sera from suspected patients or in epidemiological studies and, with further purification of egg antigen and larger samples, IgM Dot-ELISA could be a possible tool for rough estimates of parasite burden in epidemiological studies.

Adolescent↗
PubMed1996

The effect of egg yolk sampling on performance parameters and reproductive indices of northern bobwhite quail (Colinus virginianus) eggs.

The sampling of 50 microliters of the yolk (average 1.7% of the total volume) from northern bobwhite quail (Colinus virginianus) eggs on day 0, 6, 12, and 18 of incubation resulted in significantly (P < 0.05) increased embryo mortality in the day 12 and day 18 egg groups. Performance parameters and reproductive indices of the non-sampled eggs and the eggs sampled on day 0, and 6 of incubation did not differ from other reports on C. virginianus. No differences was found in postnatal responsiveness among hatchlings originating from controls and from all the groups of sampled eggs. As demonstrated in this representative galliform species, the technique can be applied for the screening of unembryonated eggs or early-embryogenesis-eggs of wild birds. If female birds are not accessible but their eggs are, information on the reproductive flock can be accessed without stressing adult birds.

Animals↗
PubMed1998

[Respiratory symptoms by oral challenge tests with egg white antigens in egg-allergic children].

One hundred and ninety one subjects showing histories of immediate hypersensitive response to egg white ingestion and/or positive IgE antibody titers specific for egg white were enrolled in double-blind placebo-controlled oral challenge with freeze and dried, heated or heated and ovomucoid-depleted egg white antigens. Seventy seven were male and 114 female, and their ages ranged from 11 month to 10 years 5 month; 118 of them had atopic dermatitis, seven had asthma and 33 had both atopic dermatitis and bronchial asthma and 33 had urticaria. One hundred four children developed 147 positive symptoms including 131 immediate reactions and 16 non-immediate reactions by oral challenge tests. Respiratory symptoms were observed in 25 cases (17%) including cough alone in 12 cases (8.2%), and both wheezing and cough in 13 cases (8.8%). These were all observed as immediate reactions and accompanied with dermal symptoms. Frequency of respiratory symptoms correlated with specific IgE antibody titers for egg white. Heated and ovomucoid-depleted egg white was more hypoallergenic that heated or freeze and dried egg white with respect to respiratory symptoms as well as other symptoms. We concluded that respiratory symptoms were provoked through oral challenges with egg white in a part of egg-allergic children.

Administration, Oral↗
PubMed1988

Determination of lysozyme content in eggs and egg products using SDS-gel electrophoresis.

Lysozyme can be separated commercially from other egg components and is used as a natural, antibiotically active protein. A method for the detection of de-lysozymed egg products using SDS-polyacrylamide gel electrophoresis (PAGE) is described. The method can also be used for lysozyme quantification. Lysozyme concentrations in egg products are tentatively assigned to three classes, based on the lysozyme contents found in intact eggs. The results obtained from a series of egg products on the market allow two groups to be distinguished. The first group has values for average lysozyme content, standard deviation and total protein content very close to the values found for intact eggs, and the second group clearly shows a lower lysozyme content but a total protein content comparable with intact eggs.

Animals↗
PubMed1997

D values of Salmonella enteritidis isolates and quality attributes of shell eggs and liquid whole eggs treated with irradiation.

Irradiation sensitivity of five Salmonella enteritidis isolates inoculated either on the surface or inside of whole shell eggs were determined. The shell eggs were irradiated at doses of 0, 0.5, 1.0, and 1.5 kGy. A minimal dose of 0.5 kGy was sufficient to eliminate all the isolates from the surface of whole eggs; however, the same isolates were more resistant to irradiation when present inside the eggs. The ATCC 13076 isolate was significantly more sensitive to irradiation, with a D value of 0.32 kGy, than the other four isolates from animal origin. Irradiation D values of the latter ranged from 0.39 to 0.41 kGy. Liquid whole eggs were also inoculated (2.4 x 10(6) cells per milliliter) with two S. enteritidis isolates and were heat-treated at 50 C for 0, 20, 40, or 60 min followed by irradiation at 0, 0.25, 0.5, 0.75, or 1.0 kGy. The results indicate that mild heating prior to irradiation was ineffective in reducing the irradiation D values. However, on the basis of the D values obtained, an irradiation dose of 1.5 kGy should be sufficient to reduce Salmonella counts by approximately 4 log10 in both whole shell and liquid eggs. Results also indicate that color and thermal characteristics of the whole or liquid eggs were unaffected by a 1.5-kGy dose of irradiation.

Animals↗
PubMed1985

Allergens in the white and yolk of hen's egg. A study of IgE binding by egg proteins.

The radioallergosorbent test (RAST) was used to compare the IgE binding of egg white and yolk, and allergenic proteins were detected by immunoelectrotransfer ('Western blotting'). The main allergens were found in egg white, but for a large proportion of the egg-sensitive patients, yolk contained specific IgE-binding constituents. For blood sera from 36 patients, there was a positive correlation between the results of RAST for egg white and for yolk. Lysozyme was found to be an allergen for some patients. The effect of heating on the allergenicity of egg white was examined and the allergenicity of hen egg white was compared with that of a duck egg. The allergens in yolk were associated with each of the three yolk fractions, and several of the proteins in the low-density lipoprotein fraction bound IgE.

Adolescent↗
PubMed1975

In vitro fertilization of rat and mouse eggs by ejaculated sperm and the effect of energy sources on in vitro fertilization of rat eggs.

In vitro fertilization of rat and mouse eggs by ejaculated or epididymal spermatozoa in chemically defined media was studied. Penetration rates by ejaculated sperm was very low (0 to 8%) in the rat, but 11 to 41% of eggs were penetrated by ejaculated sperm in the mouse. The optimal concentration of sperm for in vitro fertilization appears to be similar whether ejaculated or epididymal sperm were used. The time of sperm penetration in the mouse eggs, however, was delayed for one-half to one hour when ejaculated sperm were used. The importance of sodium pyruvate, sodium lactate and glucose in the medium containing bovine serum albumin for in vitro fertilization of rat eggs was examined. When rat eggs in cumulus clot were exposed to epididymal sperm preincubated for five hours, the presence of sodium pyruvate, sodium lactate and glucose was found to play an important role. When exposed to non-incubated epididymal sperm sodium pyruvate could be omitted without much decline of the fertilization rate. When the denuded eggs were exposed to non-incubated sperm, penetration rates were very low (0 and 5%) in the absence of pyruvate. It appears that although lactate, pyruvate and glucose are all important for in vitro fertilization of rat eggs, pyruvate can be supplied by the follicular cells surrounding the eggs.

Animals↗
PubMed1979

In vitro fertilization of two species of deer mouse eggs by homologous or heterologous sperm and penetration of laboratory mouse eggs by deer mouse sperm.

Newly ovulated eggs from immature deer mice (Peromyscus maniculatus and P. polionotus) and mature laboratory mice (Mus musculus) treated with PMSG and HCG were inseminated in vitro with spermatozoa recovered from the cauda epididymidis of mature males. The time required for capacitation of deer mouse sperm in culture was estimated to be about two to five hours based on the dispersal of sperm agglutination and increase of sperm motility. The rate of sperm penetration through the zona pellucida of deer mouse eggs by homologous or heterologous sperm was relatively high (72-91%) but that of laboratory mouse eggs by deer mouse sperm was low (20-21%). After penetration through the zona pellucida, a high proportion of deer mouse eggs (79-93%) were fertilized by homologous or heterologous deer mouse sperm but no laboratory mouse eggs were fertilized by sperm of two species of deer mice. The zona pellucida was dissolved in a higher proportion of laboratory mouse eggs cultured with P. maniculatus (45%) than with P. polionotus sperm (3.4%), but this did not happen by incubation of deer mouse eggs with homologous or heterologous sperm. It seems that there is little difference in sperm penetration and fertilization between these two closely related species of deer mice but the reactions between the mouse eggs and deer mouse sperm are quite different.

Animals↗
PubMed2002

Cytochalasin-D retards sperm incorporation deep into the egg cytoplasm but not membrane fusion with the egg plasma membrane.

The fertilization process is impaired when spermatozoa are previously incubated with Cytochalasin-D (Cyt-D). Although this fact reveals the participation of polymerized actin in fertilization, the specific event obstructed by Cyt-D treatment has not been determined. To identify this event, we capacitated guinea pig spermatozoa in minimal capacitating medium with pyruvate and lactate (MCM-PL) with Cyt-D, to inseminate hamster zona pellucida (ZP)-free eggs. Cyt-D (70 microM) decreased F-actin relative concentration in capacitated spermatozoa to a larger extent than in spermatozoa incubated under control conditions. Cyt-D also cancelled the F-actin increase normally observed in acrosome-reacted cells, and decreased the number of these cells with normal F-actin localization at the equatorial zone. Insemination of eggs with Cyt-D treated spermatozoa did not change early fertilization events such as the egg cortical reaction (CR), membranes fusion, and egg F-actin new localization, but clearly retarded, by 16 hr, spermatozoa incorporation deep into the egg cytoplasm, and decondensation of egg metaphase II chromosomes. These results show that actin polymerization is necessary for spermatozoa incorporation deep into the egg cytoplasm, but not for plasma membrane fusion nor egg activation early steps.

Acrosome Reaction↗
PubMed1998

Trace elements in egg contents and egg shells of slider turtles (Trachemys scripta) from the Savannah River Site.

In this paper we examine the levels of trace elements in the egg contents and egg shells of slider turtles (Trachemys scripta) from the Savannah River Site, near Aiken, South Carolina. Trace elements have seldom been examined in the tissues or eggs of reptiles, although some turtles and large snakes occupy a high trophic level. Lead, mercury, cadmium, selenium, chromium, and manganese levels were examined in one egg and its egg shell collected from each of 16 females that laid in late May and June 1996. We were interested in determining background levels, whether certain metals were sequestered in the egg shells, and whether levels were higher in contents or shells. Concentrations were higher in egg contents than in shells for lead, mercury, and selenium, while chromium was higher in the shell. There were no differences for cadmium and manganese. Compared to eggs from other reptiles, levels in slider turtles were generally similar for cadmium and selenium, lower for chromium and lead, and higher for manganese.

Animals↗
PubMed1988

Identification of a secondary sperm receptor in the mouse egg zona pellucida: role in maintenance of binding of acrosome-reacted sperm to eggs.

During fertilization in mice, acrosome-intact sperm bind via plasma membrane overlying their head to a glycoprotein, called ZP3, present in the egg extracellular coat or zona pellucida. Bound sperm then undergo the acrosome reaction, which results in exposure of inner acrosomal membrane, penetrate through the zona pellucida, and fuse with egg plasma membrane. Thus, in the normal course of events, acrosome-reacted sperm must remain bound to eggs, despite loss of plasma membrane from the anterior region of the head and exposure of inner acrosomal membrane. Here, we examined maintenance of binding of sperm to the zona pellucida following the acrosome reaction. We found that polyclonal antisera and monoclonal antibodies directed against ZP2, another zona pellucida glycoprotein, did not affect initial binding of sperm to eggs, but inhibited maintenance of binding of sperm that had undergone the acrosome reaction on the zona pellucida. On the other hand, polyclonal antisera and monoclonal antibodies directed against ZP3 did not affect either initial binding of acrosome-intact sperm to eggs or maintenance of binding following the acrosome reaction. We also found that soybean trypsin inhibitor, a protein reported to prevent binding of mouse sperm to eggs, did not affect initial binding of sperm to eggs, but, like antibodies directed against ZP2, inhibited maintenance of binding of sperm that had undergone the acrosome reaction on the zona pellucida. These and other observations suggest that ZP2 serves as a secondary receptor for sperm during the fertilization process in mice and that maintenance of binding of acrosome-reacted sperm to eggs may involve a sperm, trypsin-like proteinase.

Acrosome↗
PubMed1996

Lectin staining of trichostrongylid nematode eggs of sheep: rapid identification of Haemonchus contortus eggs with peanut agglutinin.

The binding of 10 different lectins to the surface of trichostrongylid nematode eggs of sheep were investigated. Osage orange seed agglutinin bound to eggs from Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis, jack bean agglutinin to H. contortus and O. circumcincta eggs and peanut agglutinin to H. contortus eggs only. Staining of egg mixtures with fluorescein-labelled peanut agglutinin showed that the lectin correctly identified the percentage of H. contortus eggs present. Identification of eggs by lectin staining correlated well with results from larval differentiation. The findings suggest that peanut agglutinin can be used in routine diagnostic parasitology for the rapid identification of H. contortus eggs.

Animals↗
PubMed1981

Isolation of a glycopeptide fraction from the surface of the sea urchin egg that inhibits sperm-egg binding and fertilization.

The role of cell surface glycoproteins of the sea urchin egg in binding sperm has been examined by studying the biological activity of glycopeptides derived from these glycoproteins. Glycopeptides were produced from egg surface glycoproteins by Pronase digestion. After fractionation by gel filtration the glycopeptides were tested for their ability to inhibit the binding of sperm to eggs, presumably by competing with the egg surface glycoproteins for binding sites on the sperm. One glycopeptide fraction with an apparent molecular weight of approximately 6,000 was found to be a potent inhibitor of sperm-egg binding, as well as fertilization, even at nanomolar concentrations. This activity was heat stable and exerted its effect against the sperm and not the egg. Experiments with a radiolabeled form of the glycopeptide fraction directly demonstrated that at least one component of it bound to sperm. Specific binding of the radiolabeled glycopeptide occurred only to acrosome-reacted sperm. Because the isolated glycopeptide fraction has many of the characteristics that one would expect of a biologically active fragment of an egg surface receptor for sperm, these findings are consistent with the idea that one or more glycoconjugates on the surface of the egg are involved in sperm binding.

Acrosome↗
PubMed1994

Endogenous interleukin 12 (IL-12) regulates granuloma formation induced by eggs of Schistosoma mansoni and exogenous IL-12 both inhibits and prophylactically immunizes against egg pathology.

Morbidity in humans infected with Schistosoma mansoni results primarily from the deposition of parasite eggs in portal areas where they induce a granulomatous response. In mice infected with this helminth granuloma formation is a CD4+ T helper (Th) cell-dependent process that is associated with a strong Th2 cytokine response which appears to evolve through a Th0 phase. In this report, we asked whether endogenously synthesized or exogenously induced interferon (IFN)gamma through its suppression of Th2 cell expansion exerts a regulatory role on egg pathology. Depletion of IFN-gamma or natural killer cells resulted in a marked enhancement of granuloma formation around intravenously injected eggs and was associated with increased Th2 and decreased Th1 and interleukin (IL)12 mRNA expression. Similar changes occurred when egg-injected mice were treated with neutralizing monoclonal antibodies specific for IL-12 indicating a role for this cytokine in the regulation of the granulomatous response. In contrast, treatment with exogenous rIL-12 profoundly inhibited primary granuloma formation while increasing IFN-gamma, IL-2, IL-10, and IL-12 pulmonary mRNA levels and suppressing IL-4, IL-5, IL-6, and IL-13 mRNA expression. Cytokine depletion studies indicated that the effects of IL-12 could be attributed primarily to increased IFN-gamma. Importantly, IL-12 also inhibited secondary granuloma formation in mice presensitized with eggs demonstrating a role for the cytokine in reversing established Th2-type responses. Moreover, mice sensitized with eggs in combination with IL-12 to precommit them toward a Th1 response developed only minimal granulomas upon subsequent egg challenge. The latter findings suggest that simultaneous vaccination with antigen plus IL-12 may provide a strategy for the prevention of schistosome egg pathology as well as other diseases stemming from Th2 cytokine production.

Animals↗
PubMed2005

The significance of background odour for an egg parasitoid to detect plants with host eggs.

Scots pine has been shown to produce a volatile bouquet that attracts egg parasitoids in response to oviposition of the herbivorous sawfly Diprion pini. Previous analyses of headspace volatiles of oviposition-induced pine twigs revealed only quantitative changes; in particular, the sesquiterpene (E)-beta-farnesene was emitted in significantly higher quantities by oviposition-induced pine. Here we investigated whether (E)-beta-farnesene attracted the egg parasitoid Chrysonotomyia ruforum. We tested the behavioural response of C. ruforum females to different concentrations of (E)-beta-farnesene. Egg parasitoids did not respond to this sesquiterpene at either concentration tested. However, they did respond significantly to (E)-beta-farnesene when this compound was offered in combination with the volatile blend emitted from pine twigs without eggs. This response was dependent on the applied concentration of (E)-beta-farnesene. Further bioassays with other components [(E)-beta-caryophyllene, delta-cadinene] of the odour blend of pine were conducted in combination with the volatile blend from egg-free pine as background odour. None of the compounds tested against the background of odour from an egg-free pine twig were attractive to the egg parasitoid. These results suggest that the egg parasitoids responded specifically to (E)-beta-farnesene, but only when this compound was experienced in the 'right' context, i.e. when contrasted with a background odour of non-oviposition-induced pine volatiles.

Animals↗
PubMed1998

Daily mean number of eggs laid by the southern cattle tick (Acari: Ixodidae) compared with mean egg mass weight.

Among several methods used to estimate the efficacy of an acaricide is the Drummond adult test, based on formulas which permit the assessment of the reproductive capacity of treated female ticks in comparison with controls. A constant value of 20,000 is normally used in these formulas, because this figure approximates the number of larvae that will hatch from 1 g of eggs. In laboratory studies conducted on large numbers of ticks, egg counting is a time-consuming and fastidious operation. Therefore, in quantitative studies, sampling techniques are more convenient. The use of the weight of egg masses laid by ixodids is adequate because eggs congregate in clusters. Experimental results have demonstrated that the analysis of the mean daily weight of egg aliquots in comparison with actual counts, from the 1st to the 20th d of oviposition, did not permit rejection of the hypothesis that 20,000 would be a representative number of eggs contained in 1 g. When aliquots were weighed only on the 20th or the 30th d of the oviposition-incubation period, and the results were analyzed in comparison with egg counts, the value of 20,000 no longer was reliable to express the number of Boophilus microplus (Canestrini) eggs contained in 1 g.

Animals↗
PubMed2006

Divergent selection on feather pecking behavior in laying hens has caused differences between lines in egg production, egg quality, and feed efficiency.

The correlated changes in egg production, egg quality, and feed efficiency (FE) due to selection for feather pecking (FP) were investigated by analyzing the data from an experiment including 2 divergently selected lines and a control line. The experiment was conducted with hens from 42 to 46 wk (hatch 1) and 39 to 43 wk (hatch 2) of age in the fifth generation of selection. The number of FP bouts per hour in the low FP line (LFP) was lower than the high FP line (HFP; 0.38 vs. 2.01), and total plumage score in line LFP was better than in line HFP (16.9 vs. 11.6). During the 4 wk, egg number and egg mass in line LFP were higher than those in HFP (24.4 vs. 18.3 and 1,223 vs. 1,132 g, respectively). On the other hand, line HFP had greater egg weight (60.7 vs. 59.2 g), albumen height (73.0 vs. 64.9 in Haugh units), shell thickness (38.1 vs. 37.0 mm), and yolk percentage (30.6 vs. 29.5%) than the LFP line. The control line was intermediate for those traits. The residual feed consumption (RFC) was highest in line HFP, lowest in line LFP, and intermediate in line C. Partial regressions of feed consumption (FC) on BW gain and egg mass were not significantly different among the 3 lines, whereas a significant difference in regression on metabolic BW (32.6 g/d in line LFP, 38.0 g/d in control line, and 43.4 g/d in line HFP) was observed. In addition, there was a negative regression of FC per day on plumage score (-1.73 g). The adjustment for plumage score accounted for 60% of the difference between regressions on metabolic BW in lines LFP and HFP. These results indicated that selection for FP has led to a change in egg production, egg quality, and FE. The better FE in line LFP resulted from a lower requirement for maintenance energy. The later was partly accounted for by a better plumage cover.

Animal Nutritional Physiological Phenomena↗
PubMed1995

Mouse sperm-egg plasma membrane interactions: analysis of roles of egg integrins and the mouse sperm homologue of PH-30 (fertilin) beta.

The guinea pig sperm protein, PH-30 (also known as fertilin), is postulated to participate in the interaction between the sperm and egg plasma membranes. The beta subunit of guinea pig PH-30 (gpPH-30 beta) contains a domain with homology to disintegrins, snake venom proteins that bind to integrins via an integrin-binding domain containing the tripeptide RGD. This raises the question of whether an egg integrin serves as a receptor for PH-30. Although mouse eggs express integrin subunits, their role in mouse fertilization is unresolved. Therefore, we examined fertilization for two different hallmarks of integrin function, namely, dependence of ligand binding on divalent cations and the ability to inhibit ligand binding with RGD peptides. We demonstrate that sperm binding to zona pellucida-free eggs is supported by Ca2+, Mg2+, or Mn2+. Ca2+ was necessary and sufficient for sperm-egg fusion, with 2.5 mM Ca2+ being the most effective concentration. In addition, fertilization could be partially inhibited with various RGD peptides, which caused a decrease in sperm-egg fusion by 30-58%. This partial inhibition of fusion with RGD peptides prompted the cloning of the mouse homologue of gpPH-30 beta (hereafter referred to as mPH-30 beta) to determine if it possessed the tripeptide RGD or a different amino acid sequence in its disintegrin domain. mPH-30 beta, which is expressed during meiotic and post-meiotic phases of spermatogenesis, shares significant similarities to gpPH-30 beta throughout the length of the molecule, from the signal sequence to the cytoplasmic tail. The full-length deduced amino acid sequence of mPH-30 beta. The disintegrin domain of mPH-30 beta has the tripeptide QDE (instead of RGD) in its cell recognition region. Peptides containing this QDE sequence decrease the binding and fusion of sperm with zona pellucida-free eggs by approximately 70%, suggesting that the disintegrin domain of mPH-30 beta participates in the interaction between sperm and egg membranes.

ADAM Proteins↗