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Schistosoma japonicum: day to day variation in excretion and hatchability of parasite eggs in the domestic pig, Suis suis.

The aim of the present study was to describe the course of an Schistosoma japonicum infection in individual pigs over time, with special regards to fecal egg counts and egg hatchability, emphasizing the extent of variation during and between days. Five specific pathogen free Danish Landrace/Yorkshire/Duroc crossbred male pigs were each given 3500 cercariae intramuscularly. From day 36 to 62 post infection, fecal samples were collected from each pig, morning and evening. Pigs were perfused 62 days post infection. Actual fecal egg counts, miracidial counts, and worm burdens were determined. The trend of fecal egg excretion was adequately described with a third order polynomial and logarithmic link function. Miracidial counts were related directly to morning fecal egg counts through a simple linear function. The study revealed a fairly similar overall pattern of egg excretion for all pigs, showing an increase in egg excretion until week 8, followed by a marked reduction to almost zero. In general, large fluctuations around this trend were seen for all pigs, as revealed in variations in egg counts within the same day as well as between days. However, as revealed by the estimated model lines, these fluctuations are due to random variation in egg density in stools rather than being caused by biologically determined cycles. An exact time for both first occurrence of eggs in feces (days 35-38 post infection) and for peak egg counts (days 44-48 post infection) was predicted for each individual pig. Furthermore, the model revealed that miracidial counts can be related directly to the expected fecal egg counts of the same day through a parameter, which represents hatchability. This hatchability parameter was found to be independent of time post infection for each pig, but large individual differences were seen in hatchability between the pigs.

Animals↗

Maternal thyroid hormones in Japanese quail eggs and their influence on embryonic development.

We addressed the relationship between the thyroid status of hens and the thyroid hormone content of their eggs, as well as the influences of egg hormones on embryonic development. Methods for measuring thyroid hormones in egg yolk were verified by demonstrating consistency in the recovery of yolk thyroid hormones following a methanol/chloroform extraction and in the measurement of thyroid hormones by RIA for a range of hormone concentrations in yolk extracts. Untreated hens produced eggs with yolk thyroxine (T4) concentrations that were low relative to plasma T4, but yolk triiodothyronine (T3) concentrations comparable to those of plasma. Hens dosed twice daily with T4 (1 or 3x the daily thyroid secretion rate, TSR, of T4 per dose) had significantly higher plasma and egg yolk T4 concentrations than did control hens dosed with saline. In general, the T4 concentration of egg yolk varied with the thyroid status of the hen. When the relationship between each hen's plasma T4 and the yolk T4 concentration of her eggs was examined, hens appeared to regulate T4 deposition into yolk at "levels" characteristic of the "levels" of thyroid status produced by the different doses of T4. Embryonic pelvic cartilage, a thyroid hormone-responsive tissue, showed enhanced growth and differentiation in embryos from eggs of hens given the highest dose of T4. Specifically, alkaline phosphatase activity (a marker of differentiation) and pelvic cartilage wet and dry weights were significantly greater in embryos from high T4 eggs (hens on the 3x TSR dose) than those in controls. However, embryos from high T4 eggs did not differ in general body growth (body weight, length, and general morphology) or hatchability compared to controls. In a single T3 experiment, hens were dosed twice daily with 1 microg T3. The embryos from eggs of these hens had accelerated differentiation/maturation of pelvic cartilages (sampled at Day 12) compared to those from control eggs; body growth did not differ from that of controls.

Animals↗

Egg laying in vitro of Echinostoma caproni (Trematoda) in nutritive and nonnutritive media.

Egg laying in vitro was studied in Echinostoma caproni adults placed in 10 ml of nutritive or nonnutritive media for 48 h in petri-dish cultures maintained at 37 degrees C in an atmosphere containing 7.6% CO2. Maximal egg laying occurred within 24 h in the defined medium RPMI 1640. Egg laying was significantly greater in this medium than in McCoy's or Locke's solution. Eggs released into the RPMI medium were capable of producing miracidia that were infective to Biomphalaria glabrata snails. Fried and Huffman (1996) referred to a technique used to obtain eggs of Echinostoma caproni in the defined medium RPMI 1640, but details of the study were not given. No information is available on egg laying of echinostomes in vitro. Such information could contribute to a better understanding of egg release in digeneans and would also be helpful in the acquisition of eggs for biology and chemistry studies. Current techniques used to obtain echinostome eggs involve worm homogenization, teasing of eggs from the worms' uteri, or recovery of eggs from feces (see Idris and Fried 1996 for details). The purpose of this communication is to report on an efficient procedure for the acquisition of eggs of E. caproni after the placement of adult worms in the defined medium RPMI 1640. E. caproni adults were grown in ICR mice for either 17 (young worms) or 112 days (old worms) as described previously (Ursone and Fried 1995a). Worms were removed from the small intestines and rinsed rapidly in three changes of sterile Locke's solution containing penicillin (200 IU/ml) and streptomycin (200 micrograms/ml; Fried and Contos 1973). Worms were placed in culture media within 30 min of their removal from hosts. Nutritive media consisted of RPMI 1640 and McCoy's medium (Sigma, St. Louis, Mo.). Non-nutritive media consisted of Locke's or Locke's 1:1 (Ursone and Fried 1995b). All media contained antibiotics as described above.

Animals↗

High-DHA eggs: feasibility as a means to enhance circulating DHA in mother and infant.

Dietary DHA enhances infant attention and visual development. Because the DHA content of red blood cells and plasma lipids varies approximately threefold in pregnancy, maternal DHA status may influence subsequent infant function. It would be feasible to study the effects of higher maternal DHA intake on infant development if dietary intake of DHA could be increased by a reliable means. This study was designed to determine whether women provided with one dozen high-DHA hen eggs (135 mg DHA/egg) would consume the eggs and have higher blood DHA levels than women consuming ordinary eggs (18 mg DHA/egg). The study was a randomized, double-masked comparison of the effect of eggs with different concentrations of DHA on intake and blood lipid DHA content of women and their infants. A third nonrandomized group ate few eggs. In this study, DHA intake reported from eggs was eightfold higher in the high-DHA egg group compared to the ordinary egg group. Including all groups, DHA intake ranged from 0 to 284 mg/d. In this intake range, maternal blood lipid DHA content at enrollment best predicted DHA content at delivery, accounting for 36.5 and 51.7% of the variance in ordinary and high-DHA egg intake groups, respectively. The high-DHA vs. ordinary egg groups had similar maternal and cord blood lipid DHA, but there was a positive relationship between maternal plasma phospholipid DHA and daily DHA intake from eggs controlled for study duration (r = 0.278, P = 0.048). DHA intake and birth weight were also correlated (r = 0.299, P = 0.041). High-DHA eggs were well accepted and increased DHA intake. Other benefits of DHA intake during pregnancy were also suggested.

Adolescent↗

Experimental control of the site of embryonic axis formation in Xenopus laevis eggs centrifuged before first cleavage.

In Xenopus laevis, the dorsal structures normally develop from regions of the egg opposite the side of sperm entry. Gravity is known to affect this topographic relationship in eggs inclined obliquely from their normal vertical orientation in the period before first cleavage. This effect has been explored in detail, making use of low-speed centrifugation (10-50 g) for short durations (4 min). Eggs were immobilized in gelatin and oriented with their animal-vegetal axes 90 degrees to the force vector, with the sperm entry point (SEP) side of the egg either toward or away from the center of the rotor. It has been found that the egg shows three distinct periods of response to centrifugal force in the interval from fertilization to first cleavage: Prior to 0.4 (40% of the first cleavage interval), the egg is very sensitive to centrifugal force and develops dorsal structures from its centrifugal side, regardless of the position of the SEP in the centrifugal field. Thus, the dorsal structures of the embryo are reversed from normal in eggs centrifuged with the SEP away from the center of the rotor. In the period 0.4 to 0.7, the egg is still very sensitive to centrifugal force and develops dorsal structures from its centripetal side, regardless of the position of the SEP in the centrifugal field. Thus, the dorsal structures of the embryo are reversed from normal in eggs centrifuged with the SEP toward the center of the rotor. In the period 0.7-1.0, the egg becomes increasingly resistant to centrifugal force and forms dorsal structures at the normal position opposite the SEP side. This resistance can be overcome in some egg clutches by 50 g centrifugation followed by prolonged 90 degrees off-axis inclination at 1g. Midway in the second cell cycle, there is a brief period of sensitivity to centrifugal force. These These results are discussed in terms of the types of cytoplasmic rearrangements occurring in the egg at different times of the cell cycle, and in terms of the process of cytoplasmic localization of determinants of dorsal axial development.

Animals↗

Improved methods for isolating DNA from Ostertagia ostertagi eggs in cattle feces.

A multiplex PCR assay for differentiating strongyle eggs from cattle has recently been described; however, the egg disruption and DNA extraction procedures, though effective, are inadequate for large studies or clinical application. The purpose of this research was to evaluate methods for disrupting trichostrongyle eggs, then assess commercial kits for extracting egg DNA using Ostertagia ostertagi as a model species. Egg disruption procedures tested included probe sonication, bath sonication, bead beating, boiling, microwaving, proteinase K/SDS digestion, freezing, and various combinations of the above with the incorporation of sodium dodecyl sulfate. These procedures were evaluated in conjunction with four commercial DNA extraction kits: DNA Stool mini kit and DNeasy Plant kit (Qiagen), Fast DNA kit (QBiogene), and the MAP extraction kit (Tetracore). Results showed that egg disruption was best accomplished with the bead beater and ceramic beads, resulting in 100% disruption within 1min. When DNA extraction was preceded by the isolation of eggs from feces, all procedures except the Fast DNA kit produced PCR-ready DNA from at least two eggs. The DNeasy Plant kit allowed consistent detection of DNA released from one egg. Due to the morphological similarities among trichostrongyle eggs in ruminants, strongyle eggs in equids, and hookworm eggs, the methods described herein may have broad application to other nematodes.

Animals↗

Faecal examination of Fasciola eggs fixed with formalin solution using the beads technique.

Faecal examination for Fasciola eggs which were preserved in 5% and 10% formalin and TAF solutions for 6 years was carried out using the Beads technique. The morphological characters and the distinctive yellowish color of Fasciola eggs were retained and appeared similar to those of fresh eggs. The mean egg recovery rate of eggs in TAF solution was significantly lower than the others (P < 0.01). The egg recovery rates from 5% and 10% formalin samples were 57.2% and 60.8%, respectively, and these values were similar to the value of 58.8% for fresh Fasciola eggs in cattle faeces. For Paramphistomum eggs, preserved with 5% formalin, the mean egg recovery rate was 64.7%. Fasciola eggs and Paramphistomum eggs were easily identified by their morphological characters in this experiment. The present data demonstrated that preservation with 5% formalin solution is an acceptable means of storing faecal samples for later examination of Fasciola and Paramphistomum eggs using the Beads technique and presumably by other fluke sedimentation methods.

Animals↗

Ascaris egg profiles in human faeces: biological and epidemiological implications.

Since 2 morphological forms (fertilized and unfertilized) of egg can be produced by Ascaris, infected humans can release in their faeces fertilized eggs only (FEO), unfertilized eggs only (UEO) or both fertilized and unfertilized eggs (FUE) (designated herein as the 3 different egg profiles). Epidemiologically, fertilized eggs are of significance as they enable effective transmission of the parasite. This study, for the first time, characterizes the Ascaris egg profiles in human faeces in an endemic region of China, explores possible host- and parasite-factors related to these profiles, and discusses the biological and epidemiological implications of the findings. The 3 egg profiles were recorded throughout the study period of 2 years, and the overall percentages of people with FEO, FUE and UEO profiles were approximately 41-47%, 32-42% and 17-21%, respectively. The overall number of unfertilized eggs for the entire population accounted for approximately 6-9% of all eggs excreted. The different Ascaris egg profiles showed no correlation to host gender, but they did relate to age and worm burden of the host and to the sex ratio and developmental status of the parasite. While an annual universal anthelmintic treatment resulted in some fluctuation in the values of individual egg profiles, the general features of these profiles remained similar throughout the study period. The findings of this study should have significant implications for understanding transmission patterns of Ascaris and for the implementation of control measures against ascariasis in endemic regions.

Adolescent↗

Oocyte proteomics: localisation of mouse zona pellucida protein 3 to the plasma membrane of ovulated mouse eggs.

In order to gain a deeper understanding of the molecular underpinnings of sperm-egg interaction and early development, we have used two-dimensional (2D) electrophoresis, avidin blotting and tandem mass spectrometry to identify, clone and characterise abundant molecules from the mouse egg proteome. Two-dimensional avidin blots of biotinylated zona-free eggs revealed an abundant approximately 75-kDa surface-labelled heterogeneous protein possessing a staining pattern similar to that of the zona pellucida glycoprotein, mouse ZP3 (mZP3). In light of this observation, we investigated whether mZP3 specifically localises to the plasma membrane of mature eggs. Zona pellucidae of immature mouse oocytes and mature eggs were removed using acid Tyrode's solution, chymotrypsin or mechanical shearing. Indirect immunofluorescence using the mZP3 monoclonal antibody (mAb) IE-10 demonstrated strong continuous staining over the entire surface of immature oocytes and weak microvillar staining on ovulated eggs, regardless of the method of zona removal. Interestingly, in mature eggs, increased fluorescence intensity was observed following artificial activation and fertilisation, whereas little to no fluorescence was observed in degenerated eggs. The surface localisation of ZP3 on mature eggs was supported by the finding that the IE-10 mAb immunoprecipitated an approximate 75-kDa protein from lysates of biotinylated zona-free eggs. To further investigate the specificity of the localisation of mZP3 to the oolemma, indirect immunofluorescence was performed using the IE-10 mAb on both CV-1 and CHO cells transfected with full-length recombinant mZP3 (re-mZP3). Plasma membrane targeting of the expressed re-mZP3 protein was observed in both cell lines. The membrane association of re-mZP3 was confirmed by the finding that biotinylated re-mZP3 (approximately 75 kDa) is immunoprecipitated from the hydrophobic phase of Triton X-114 extracts of transfected cells following phase partitioning. Immunoprecipitation assays also demonstrated that surface re-mZP3 was released from transfected CV-1 in a time-dependent manner. These results demonstrate that ZP3 is specifically associated with the surface of mature eggs and its subsequent release from the cell surface may represent one mechanism by which ZP3 is secreted. Furthermore, the increase in ZP3 surface expression following fertilisation suggests that ZP3 may have a functional role during sperm-oolemma binding and fusion. These results also validate the usefulness of using the 2D proteomic approach to identify and characterise egg-surface proteins.

Animals↗

Effects of temperature and humidity on the development of eggs of Toxocara canis under laboratory conditions.

The influence of temperature and humidity on the survival and development of Toxocara canis eggs in an in vitro model system was investigated. Two soil samples were inoculated with T. canis eggs and maintained at 3% and 50% humidity and temperatures of 19-24 degrees C. Nine soil samples were inoculated with T. canis eggs of which three samples were kept at 4 degrees C with humidities at 3%, 15%, and 30%; three were maintained at 21 degrees C and three more were incubated at 34 degrees C, and at the same three humidity levels. Samples were monitored every 7 days for a total of 2 months, for the presence and development of eggs. With increasing temperature, the number of eggs undergoing development increased (P<0.01); the number of deformed eggs decreased, the number of infective eggs increased (P<0.01), and egg maturation was accelerated. A decrease in the survival of infective eggs occurred at 34 degrees C. An increase in humidity produced a rise in the number of developed eggs at all three temperatures (P<0.01). This study suggests that elevated temperatures accelerated the development as well as the degradation of eggs of T. canis, whereas the range in humidity was directly correlated with egg development.

Animals↗

Probability of an egg cracking during packing can be predicted using a simple non-destructive acoustic test.

1. The aim of this investigation was to test the predictive power of the dynamic stiffness measurement to identify eggs which are most likely to crack under field conditions. 2. A representative sample of eggs (n = 1660) was collected from the front of the cages in a commercial battery unit. Egg weight, % damping and dynamic stiffness (Kdyn) were recorded using an acoustic crack detection device. Intact eggs were marked and replaced in the front of the cages. These eggs were subsequently passed through online collection, grading and packing machinery, along with a volume of unmarked eggs. At the end of packing the acoustic test was repeated on the marked eggs, and these were subsequently categorised as being either intact (0) or cracked (1). 3. A logistic regression of the probability of cracking vs Kdyn revealed that as the Kdyn measurement decreases below 15,000 N/m there is a rapid increase in the probability that an egg will crack during routine handling. 4. Additional variables (visit, egg weight,% damping and position in the house (battery [1 to 7], side [1, 2] and tier [1 to 8]) were also fitted to the model but only egg weight, visit and tier effects significantly improved the model fit. 5. This study confirms that the dynamic stiffness measurement can predict the probability of an egg cracking in the field and with high precision. As this measurement also has a high heritability, it could be incorporated into breeding programmes, where it would offer an excellent method to improve eggshell quality and reduce the incidence of cracked eggs.

Acoustics↗

A model for persistency of egg production.

The objectives of our study were to propose a new definition for persistency of egg production and to develop a mathematical model to describe the egg production curve, one that includes a new measure for persistency, based on the proposed definition, for use as a selection criterion to improve total egg production. Persistency of egg production is an important determining factor for total egg production. Hens with the same total production, however, can exhibit different egg production curves because of differences in persistency. We propose a new definition for persistency of egg production: the number of weeks during which a level of constant production is maintained. No egg production model exists that includes a measure of persistency in terms of duration of time or that allows this measure of persistency to be derived from model parameters. It was necessary, therefore, to develop a new model to describe an egg production curve for a flock: equation [see text] and for an individual: equation [see text] where y(t) = egg production at time t, t1 and t2 = times at transition, r = duration of transition, y(p) = level of constant production, b4 = rate of decline in production, and P = persistency of constant production. These parameters measure directly the important biological characteristics of an egg production curve. To illustrate the model, six data sets were used: two from flocks (one pullet flock and one hen flock) and four from two pairs of individuals. The proposed definition of persistency should be important for genetic selection because it might be desirable to select for increased persistency. The novel approach to the definition and measure of persistency presented here should provide a better understanding of the relationship between the new measure of persistency and other characteristics of egg production.

Animals↗

Effect of dietary folic acid supplementation on egg folate content and the performance and folate status of two strains of laying hens.

Enrichment of eggs with folate is possible when dietary folic acid levels are increased. However, development of optimal strategies for the production of folate-enriched eggs requires knowledge as to differences due to strain of bird and a greater understanding of the factors limiting egg folate deposition. To this end, a study was designed to determine the response of two leghorn strains that differ in production performance. Hyline W36 and W98 hens (n = 6 per diet) received a barley-based ration containing 0, 2, 4, 8, 16, 32, 64, or 128 mg/kg of crystalline folic acid for 21 d. Response criteria included production parameters, measures of blood folate status, and egg folate content. Significant (P < 0.05) main effects of folate supplementation were observed for egg folate content and plasma folate, which increased, and homocysteine concentrations, which decreased with supplementation; performance, however, was not affected. The Hyline W98 strain had significantly (P < 0.05) higher total egg and yolk weights and feed consumption when compared with the W36. Significant (P < 0.05) ration x strain interactions were observed for egg and yolk weight, egg folate content, and plasma homocysteine. The higher egg mass producing strain, Hyline W98, benefited from increased folic acid through a reduction in plasma homocysteine concentrations, suggesting that this strain has a higher requirement for folate than the W36 strain. Overall, egg folate content is maximized when crystalline folic acid is supplemented to the diet at 2 mg/kg or higher. Higher levels of egg folate are not achieved due to the saturation of the precursor pool for egg folate deposition.

Animal Feed↗

Effect of inorganic versus organic selenium on hen production and egg selenium concentration.

A 28-d experiment using 288 Hy-Line W-36 laying hens was conducted to compare sodium selenite (SS) with Se-enriched yeast (SY). The Se from SS or SY was supplemented into a corn-soybean meal basal diet at 0, 0.15, 0.30, 0.60, or 3.00 ppm, and the basal diet was formulated to provide 0.82% lysine and 2,950 kcal/kg of ME. Each treatment was replicated 4 times with 2 cages of 4 hens per cage in each replicate. Hen production was assessed daily, and 2 eggs per replicate were collected every 4 d for whole-egg Se analysis. Albumen quality was assessed at 2 egg storage temperatures (7.2 vs. 22.2 degrees C) with the eggs collected on d 24 and 28, respectively. The percentage of dirty and cracked eggs was greater (P < 0.04) in hens fed SY than in those fed SS. Percentage hen-day production was not affected (P > 0.05) by diet. Albumen quality of eggs stored at 22.2 degrees C was improved (P < 0.04) in eggs from hens fed SS, but there was no difference (P > 0.05) in albumen quality of eggs stored at 7.2 degrees C. Egg weight was linearly increased (P < 0.01) by SY. Whole-egg Se levels were linearly increased (P < 0.01) as dietary Se level increased for both sources of Se, but eggs from hens fed SY had higher (P < 0.01) Se concentrations than those fed SS. The results from this experiment indicate that percentage hen-day production is not affected by Se source, and that SY increases egg Se concentrations more than SS.

Animal Feed↗

Specific serum IgE in the diagnosis of egg and milk allergy in adults.

Levels of specific serum IgE to cow's milk, whole hen's egg, egg white, and egg yolk were compared to the outcome of double-blind, placebo-controlled food challenge (DBPCFC) with fresh egg and/or milk in 21 adults with a case history of immediate hypersensitivity to egg and/or milk. Specific serum IgE was measured by four different commercially available tests and by an inhouse Maxisorp RAST using freshly prepared food extracts. Sensitivities and negative predictive accuracies were generally high with egg white and milk, but low with egg yolk. Specificities and positive predictive accuracies were low for all allergens and tests. Changing the cutoff levels did not improve the ability of the tests to predict clinical allergy. Among commercially available test allergens, egg white gave the most consistent results in levels and class scores, and the highest degree of concordance with DBPCFC, whereas egg yolk and milk varied more. Applying freshly prepared food extracts in Maxisorp RAST did not improve diagnostic value. Measuring specific serum IgE levels in control subjects tolerant to egg/milk showed that false positive reactions occurred frequently among patients with another food allergy and atopic dermatitis, whereas most tests were likely to be negative in pollen-allergic and nonallergic volunteers. In conclusion, specific IgE measurements with egg white and milk were useful for exclusion of symptomatic hypersensitivity to egg and milk in patients with a positive history, whereas DBPCFC is still mandatory in patients with positive history and positive test. Measuring egg-yolk-specific IgE or using freshly prepared food extracts for specific IgE measurements added no further diagnostic information. The rate of clinically insignificant positive test results seems to be influenced by the prevalence of other food allergies and/or atopic dermatitis in the population under study.

Adolescent↗

Eradication of Arizona hinshawii from artificially infected turtle eggs.

Turtle eggs, 24 h old, were infected with Arizona hinshawii and treated 48 h later with gentamicin sulfate (Garasol; Shering Corp., Allantown, N.J.) by pressure differential egg dip treatment to ascertain the concentration of this reagent required to eradicate this pathogen from eggs. Infected eggs treated with 1,000 or 1,500 micrograms of gentamicin per ml of dip solution eliminated detectable A. hinshawii from eggs as determined by testing shells and embryo-yolk homogenates of 12-day-old eggs and the gastrointestinal tracts, kidneys, livers and gall bladders, and yolks of 50-day-old embryos. Treated eggs produced hatchlings which did not excrete detectable A. hinshawii at 72 h or 30 days after hatching, nor was this organism recovered from the visceral organs of these hatchlings when necropsied 30 days after hatching. Bacteriological assays on infected nontreated eggs showed that greater than 70% of the eggs harbored A. hinshawii, and eggs in this group produced hatchlings which actively excreted and harbored A. hinshawii. Eggs not infected or treated also produced turtles which excreted and systemically carried A. hinshawii and Salmonella spp. though not at the same level as did the turtles produced from infected, nontreated eggs.

Animals↗

Inactivation of viable Ascaris eggs by reagents during enumeration.

Various reagents commonly used to enumerate viable helminth eggs from wastewater and sludge were evaluated for their potential to inactivate Ascaris eggs under typical laboratory conditions. Two methods were used to enumerate indigenous Ascaris eggs from sludge samples. All steps in the methods were the same except that in method I a phase extraction step with acid-alcohol (35% ethanol in 0.1 N H(2)SO(4)) and diethyl ether was used whereas in method II the extraction step was avoided by pouring the sample through a 38-microm-mesh stainless steel sieve that retained the eggs. The concentration of eggs and their viability were lower in the samples processed by method I than in the samples processed by method II by an average of 48 and 70%, respectively. A second set of experiments was performed using pure solutions of Ascaris suum eggs to elucidate the effect of the individual reagents and relevant combination of reagents on the eggs. The percentages of viable eggs in samples treated with acid-alcohol alone and in combination with diethyl ether or ethyl acetate were 52, 27, and 4%, respectively, whereas in the rest of the samples the viability was about 80%. Neither the acid nor the diethyl ether alone caused any decrease in egg viability. Thus, the observed inactivation was attributed primarily to the 35% ethanol content of the acid-alcohol solution. Inactivation of the eggs was prevented by limiting the direct exposure to the extraction reagents to 30 min and diluting the residual concentration of acid-alcohol in the sample by a factor of 100 before incubation. Also, the viability of the eggs was maintained if the acid-alcohol solution was replaced with an acetoacetic buffer. None of the reagents used for the flotation step of the sample cleaning procedure (ZnSO(4), MgSO(4), and NaCl) or during incubation (0.1 N H(2)SO(4) and 0.5% formalin) inactivated the Ascaris eggs under the conditions studied.

Animals↗

Association of Salmonella enterica serovar enteritidis yafD with resistance to chicken egg albumen.

Salmonella enterica serovar Enteritidis is a major cause of food-borne diseases in industrialized countries. The incidence of S. enterica serovar Enteritidis infections has increased substantially in recent decades, and S. enterica serovar Enteritidis is now one of the leading serovars of Salmonella in the United States. A unique epidemiological characteristic of S. enterica serovar Enteritidis is its association with chicken shell eggs, since approximately 80% of all human gastrointestinal diseases can be traced to contaminated egg products. Eggs are contaminated when bacteria from reproductive tissues of infected hens are packaged into the eggs and persist inside the hostile egg albumen environment. Therefore, resistance to egg albumen is an important aspect in the transmission of S. enterica serovar Enteritidis. We identified a gene, yafD from S. enterica serovar Enteritidis, whose overexpression conferred upon S. enterica serovar Typhimurium enhanced resistance to egg albumen, while disruption of this gene in S. enterica serovar Enteritidis rendered the organism more susceptible to egg albumen. YafD is homologous to members of an exonuclease-endonuclease-phosphatase family, including some enzymes involved in DNA repair. Furthermore, we discovered that egg albumen has nuclease activities and uses both circular and linear DNA as substrates. We propose that YafD provides a survival advantage to S. enterica serovar Enteritidis in eggs by repairing DNA damage caused by egg albumen and that it may be one of the biologic determinants that contribute to the epidemiological association of S. enterica serovar Enteritidis with egg products.

Albumins↗