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Specific domains drive VM32E protein distribution and integration in Drosophila eggshell layers.

A study was made of the localization and assembly of the VM32E protein, a putative vitelline membrane component of the Drosophila eggshell. The results highlight some unique features of this protein compared with the other proteins of the same gene family. At the time of its synthesis (stage 10), the VM32E protein is not detectable in polar follicle cells. However, it is able to move in the extracellular space around the oocyte and, by stage 11 is uniformly distributed in the vitelline membrane. During the terminal stages of oogenesis the VM32E protein is partially released from the vitelline membrane and becomes localized in the endochorion layer also. By analyzing transgenic flies carrying variously truncated VM32E proteins, we could identify the protein domains required for the proper assembly of the VM32E protein in the eggshell. The highly conserved vitelline membrane domain is implicated in the early interactions with other components and is required for cross-linking VM32E protein in the vitelline membrane. The terminal carboxylic domain is necessary for localization to the endochorion layer. Protein with the C-end domain deleted is localized solely to the vitelline membrane and cross-linked only in laid eggs, as occurs for the other vitelline membrane proteins.

Amino Acid Sequence↗

Effect of concentration of mixed-function oxidase on concentration of estrogen, rate of egg lay, eggshell thickness, and plasma calcium in laying hens.

The objective of this study was to investigate the effect of induction or depression of mixed-function oxidases (MFO) by xenobiotics on the peripheral concentration of estrogens, plasma Ca, rate of egg lay, and eggshell thickness in laying hens. In Exp. 1, 100 mg of phenobarbital (PB) administered orally each day for 3 or 7 d reduced concentrations of estrone and estradiol (E2) in serum. In Exp. 2, 25, 50, or 100 mg of PB was administered orally to laying hens for 3 or 7 d. Treatment with 100 mg of PB for 3 d or with 25, 50, or 100 mg for 7 d significantly increased liver:body weight ratios. Microsomal protein increased after 7 d of 50 or 100 mg of PB administration. Feeding PB decreased concentration of E2 and total plasma Ca in a dose- and period-dependent manner. Concentration of E2 was reduced to 10% of control, whereas hepatic cytochrome P-450 increased significantly with dose. The correlation between concentration of E2 and P-450 was negative and significant. Total Ca in the plasma was highly, positively correlated with concentration of E2. Eggshells were thinner from hens treated with 100 mg of PB for both 3 or 7 d than those from control hens. Rate of egg lay was reduced by 100 mg of PB for 7 d. In Exp. 3, .5 mL of CCl4 given orally for 1 d decreased P-450 and increased E2.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Influence of large doses of ascorbic acid on performance, plasma calcium, bone characteristics, and eggshell quality in broilers and Leghorn hens.

Four experiments were conducted using broilers (Experiments 1 and 2) or White Leghorn hens (Experiments 3 and 4) to determine the effects of large doses of dietary ascorbic acid on performance, plasma concentration of total and ionic calcium, bone characteristics, and eggshell quality. A total of 564 male broilers were fed diets containing ascorbic acid (ranging from 0 to 3,000 ppm) from 3 to 7 wk of age. Weight gain and feed conversion were measured, and blood plasma was analyzed for total and ionic calcium. Leg bones (femur, tibia, and metatarsus) were analyzed for bone mineral content, density, and breaking strength. In Experiments 3 and 4, a total of 484 Leghorn hens were fed diets containing ascorbic acid (ranging from 0 to 3,000 ppm) for 4 wk. Egg weight and specific gravity were determined, and plasma and tibiae were analyzed as in Experiments 1 and 2. Results of the broiler experiment (Experiment 2) indicated that plasma ionic calcium was significantly increased (P < .05) in ascorbic-acid-treated birds. Among leg bones examined, femur strength was improved by 16% in birds fed 2,000 ppm of ascorbic acid (Experiment 2). Other bone characteristics were not affected. In the layer experiments (Experiments 3 and 4), egg weight increased up to 5% and egg specific gravity was improved in hens fed 2,000 or 3,000 ppm of ascorbic acid, which also had increased calcium in the blood. Results suggest that large doses of ascorbic acid in the diet influence calcium metabolism, affecting bone and eggshell mineralization in chickens.

Animals↗

Effects of dietary thiouracil on thyroid activity, egg production, and eggshell quality in commercial layers.

The effects of thyroid suppression induced during the rearing period by providing various dietary thiouracil (TU) regimens on plasma thyroxine (T4) concentrations, growth, and subsequent egg production (EP) and eggshell quality were determined in Single Comb White Leghorn chickens. Thiouracil was provided in the feed at levels of 0, .1, and .2% from 0 to 6 wk of age in Experiment 1, and at levels of 0, .05, and .1% from 6 to 16 wk of age in Experiment 2. In both experiments, T4 concentrations were reduced during TU treatment. However, T4 later became elevated at 12 and 20 wk in both dosage level groups in Experiment 1. Additionally, BW and egg weights were suppressed by both TU treatments, and EP was reduced up to Week 23 in the .1% TU-treated birds and through Week 25 in the .2% TU-treated birds. No effects on EP were noted in Experiment 2, but feed consumption (FC) was reduced during Week 6 in birds fed .05% TU and during Weeks 6, 10, and 19 in birds fed .1% TU. Both liver and thyroid weights were increased in .1% TU-treated birds relative to controls at Week 16. Eggshell quality was affected only in Experiment 2, in which birds given .05% TU had a higher relative conductance, or maximum rate of water loss, at Week 38 than 0 and .1% TU dosage levels, and .1% TU-treated birds had a higher breaking strength than 0 and .05% TU-treated birds at Week 22.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

The effect of four levels of ascorbic acid and two levels of calcium on eggshell quality of forced-molted White Leghorn hens.

The present experiment evaluated the effect of four levels of ascorbic acid (0, 100, 250, and 500 ppm) and two levels of calcium (3.0 and 3.5%) on the productivity and eggshell quality of molted Single Comb White Leghorn hens, housed four per 30.5 cm wide x 45.7 cm deep cage. The 4 x 2 factorial treatment combinations were randomly assigned to eight blocks of cages. Egg production increased (P < .06) an average of 5% for the levels of 250 and 500 ppm ascorbic acid. No differences were observed for feed consumption, egg weight, and shell thickness for the different levels of ascorbic acid or calcium in the diet. An increase (P < .01) was observed for specific gravity, as the levels of ascorbic acid increased. Increasing the level of calcium from 3.0 to 3.5% increased (P < .01) specific gravity from 1.073 to 1.075. Shell weight increased (P < .01) with the higher levels of ascorbic acid (250 and 500 ppm). No interactions were observed between the levels of ascorbic acid and calcium. Results suggest that supplementing ascorbic acid to molted laying hens can be beneficial to egg production and eggshell quality.

Animal Feed↗

Eggshell conductance and other functional qualities of ostrich eggs.

Eggshell conductance was measured and eggshell conductance constants were calculated for eggs from ostriches. In addition, egg water, yolk, albumen, shell, and total solids were measured in an effort to determine the maternal investment of ostrich eggs. The results of the study suggest that the optimal incubator humidity for ostrich eggs is less than 25% to allow a 15% loss of initial egg mass during the 45-d incubation period. This low humidity does not preclude increasing humidity during the actual hatching process. In addition, incubation temperatures need to be adjusted to allow a longer development time for the embryo to attain an adequate level of maturity to survive the plateau stage in oxygen consumption or to prevent the use of limited energy of the yolk to survive the anoxia of tucking and internal pipping. The optimal incubation temperature of ostrich eggs appears to be between 36.1 and 36.9 C.

Animals↗

Ultrastructural characteristics of ostrich eggshell: outer shell membrane and the calcified layers.

The ultrastructure of the eggshell of the domestic hen has been well researched and structural studies of other avian species, such as the ostrich, often base their interpretation of egg shell structure on that of the chicken. In the ostrich, lowered hatchability and hatching trauma may be due to shell ultrastructural abnormalities. In the present study the ultrastructure of the calcified portion, and the outer shell membrane (OSM), of domesticated ostrich eggshells was investigated using standard electron microscopic techniques. Transmission and scanning electron microscopy studies demonstrated intimate contact between cup-shaped structures present on the OSM and the mammillary layer of the calcified portion of the shell. The initial calcium carbonate growth of the calcified shell was of a dendritic nature with nucleation sites on the surface of the cup's contents. The dendritic growth gave way to a more randomly-orientated, smaller crystallite growth structure, which changed in form as it neared the vertical crystal layer (VCL). The VCL is described as being both amorphous and 'crumbly' depending on the plane of fracture. These observations suggest that firstly, initial calcification is contained within the cups and is then directed outwards to form the shell and that secondly, the VCL may contain an evolutionary, calcified cuticular layer. These observations serve as a baseline for studies investigating the effect of shell structure and strength on hatchling trauma and the influence of maternal diet.

Animals↗

Eggshell membrane structure and penetration by Salmonella typhimurium.

Eggshell membrane was removed from 10 broiler hatching eggs at approximately monthly intervals through the productive life of a commercial flock. A piece of membrane (2 by 2 cm) was used to cover an opening in an apparatus designed to test penetration by Salmonella Typhimurium. The membrane was placed between two chambers as the only means of liquid transfer. The chamber above the membrane was filled with a suspension of Salmonella Typhimurium cells. Sampling of the bottom chamber provided cultural evidence of the penetration by a marker strain of Salmonella Typhimurium. Samples were drawn at 15 min and at 1, 3, 5, 7, 12, 24, and 48 h. Following the cultural penetration experiment, the same pieces of membrane were removed and stained for microscopic examination. Membrane structure was examined with the use of a confocal laser scanning microscope. Each image consisted of a composite of eight 1-microm optical slices showing the fibers making up the outer surface of the membrane. These images were transferred to an image analysis software package that allowed the measurement of the interfiber area. No clear correlation could be made between the average size of all interfiber areas or the total interfiber area measured in the outer 8 microm of the outer membrane and the ability of Salmonella Typhimurium to penetrate the same piece of eggshell membrane.

Animals↗

Detection of the mgtC gene in multidrug-resistant Salmonella sp. based on isolation of chicken eggshell swabs from traditional Surabaya markets.

BACKGROUND: The virulence of Salmonella sp. is increased by the presence of the mgtC gene, which allows the bacteria to survive in environments with low magnesium levels, such as inside macrophages. Salmonella sp. found on eggshells when they show resistance to three or more classes of antibiotics can be classified as multidrug-resistant (MDR) bacteria. AIM: This study aimed to identify the presence of Salmonella sp. MDR and the mgtC gene in chicken eggshell swabs from traditional markets in Surabaya. METHODS: Swab samples were collected from 160 eggs (80 from layer chickens and 80 from free-range chickens) at 10 traditional markets in Surabaya, Indonesia. Isolation and identification were performed using culture media, including Salmonella Shigella Agar, Gram staining, Triple Sugar Iron Agar, Sulfide Indole Motility, Simmons Citrate Agar, Methyl Red and Voges Proskauer, and Urea Agar. Antibiotic sensitivity testing was performed using the disc diffusion method on Mueller-Hinton Agar. Molecular detection of the mgtC gene was performed using polymerase chain reaction. RESULTS: The results showed that 16.87% (27/160) of the samples were detected positive for Salmonella sp. All Salmonella sp. isolates (27) were resistant to Erythromycin (100%). Resistance was also found to Ampicillin (77.77%, 21/27), Tetracycline (29.62%, 8/27), and Ciprofloxacin (18.51%, 5/27). No resistance to chloramphenicol was observed. In addition, eight of the 27 isolates (29.62%) were classified as Salmonella sp. MDR. The Salmonella sp. MDR isolates also carried the mgtC gene at 87.5% (7/8). CONCLUSION: These findings demonstrate the potential global public health threat posed by MDR Salmonella sp. with the mgtC gene, emphasizing the importance of monitoring and controlling antibiotic resistance in humans and animals.

Animals↗

Salmonella typhimurium penetration through the eggshell of hatching eggs.

This study was undertaken to demonstrate penetration of Salmonella typhimurium through the eggshell of newly laid broiler hatching eggs. Eggs were challenged either by lightly spraying the bacteria over the blunt end of the egg or by contact with contaminated dry nest litter. Exposure time for both groups was 10 minutes; afterward, all eggs were disinfected and incubated 19 days under normal conditions. Chorioallantoic membranes and yolk sacs were cultured in brain-heart infusion broth on day 19 to demonstrate penetration. Isolation of the bacteria from chorioallantoic membranes and yolk sacs, respectively, were as follows: sprayed group: 100% and 83%; contact group: 59% and 29%. These results showed that although water enhanced S. typhimurium penetration, its presence on the eggshell is not essential for penetration to occur.

Allantois↗

Egg dipping in hydrogen peroxide solution to eliminate Salmonella typhimurium from eggshell membranes.

The effectiveness of egg dipping in a 6% hydrogen peroxide solution to eliminate Salmonella typhimurium from eggshell membranes was evaluated. The first step was to assess the water uptake from broiler hatching eggs dipped in tap water once or twice using a positive-pressure differential method in order to determine the best method to use in the disinfection trials. Double dipping increased water uptake by 86% more than single dipping. Dipping S. typhimurium-contaminated eggs twice in a 6% hydrogen peroxide solution reduced the average number of organisms in eggshell membranes by 95% and the number of S. typhimurium-positive eggs by 55% compared with the infected untreated group. Dipping the eggs in a 6% hydrogen peroxide solution did not adversely affect hatchability.

Allantois↗

Experimental reduction of eggshell conductance during incubation. I. effect on the susceptibility to ascites syndrome.

Two hundred-four fertile broiler chicken eggs were obtained from a commercial source and divided into three equal groups. On day 1 of incubation, 68 eggs were selected randomly and four strips of vinyl tape were applied to the shell below the air cell (tape-day 1) to reduce eggshell conductance. This procedure was repeated with an additional group of 68 eggs on day 14 (tape-day 14). Sixty-eight eggs were incubated without treatment, as controls. One week after hatch, 20 chickens from each treatment group and control group were placed into a hypobaric chamber (simulated altitude of 2500 m) for 5 weeks. The remaining chickens in each group were maintained under normobaric conditions. The hematocrit and the mean frontal resultant electrical vector (MRV) of the heart were measured following 1 week, 3 weeks, and 5 weeks of hypobaric or normobaric exposure. Surviving chickens were euthanized at the end of 5 weeks. The weight ratio of right ventricle to left ventricle plus septum (the hypertrophy index [HI]) and the cardiac index, the HI divided by body weight, were determined. All mortality during the study was subjectively scored for the presence of ascites syndrome lesions. The percentage of chickens dying during, or exhibiting ascites syndrome at the completion of, the 5-week hypobaric exposure was 16.7%, 66.7%, and 58% for control, tape-day 1, and tape-day 14 treatments, respectively. MRV values of birds following hypobaric exposure were significantly different between treatment and control groups of the hypobaric exposure and between the two tape treatments. These results suggest that reducing conductance of the eggshell during incubation significantly potentiates the development of ascites syndrome in the broiler chicken.

Aging↗

Eggshell ultrastructure in four mosquito genera (Diptera, culicidae).

Eggshell ultrastructure in 8 species of mosquitoes from the genera Aedes, Anopheles, Culex, and Toxorhynchites is compared using transmission electron microscopy. A 3-layered organization is present. The vitelline envelope is of similar character in all genera--homogeneous, rather electrondense, and with no substructure. The endochorion always includes a lamellate layer (with at least one lamella) and, except in Toxorhynchites, tubercles of varying size and shape. The exochorion is a thin layer covering the outside of the tubercles, sometimes like a sheet, but more often weblike. The ventral side of the Anopheles egg lacks exochorion entirely. The endo- and exochorion in Toxorhynchites are fused and contain numerous large empty spaces. Ultrastructural differences were found to be greater between different strains than between different species. It is suggested that species-specific characters should be chosen only after study of populations from all parts of species' ranges. Possible functional trends in the eggshells are discussed, as well as the importance of integrating ecological studies with morphology to understand how environmental and other factors act upon eggs. Egg characters suitable for phylogenetic analysis are suggested.

Aedes↗

Structure of synthetic peptide analogues of an eggshell protein of Schistosoma mansoni.

The peptide (Gly-L-Tyr-L-Asp-L-Lys-L-Tyr)6, referred to as F4-6, was synthesized as a model for a schistosome eggshell protein in which the Gly-Tyr-Asp-Lys-Tyr consensus sequence is repeated over 40 times. Analysis by CD, Fourier transform infrared spectroscopy, potentiometric and spectrophotomertric titrations, NMR, and molecular modeling suggests that F4-6 forms some type of left-handed structure. A likely possibility appears to be a left-handed alpha-helix stabilized by Lysi-Aspi +4 salt bridges and possibly Aspi-Tyri +4 hydrogen bonding and Tyr-Tyr interactions. Spectroscopic studies of a number of F4-6 analogues support this conclusion. For example, substitution of D-Ala for Gly produces a peptide with enhanced left-handed helical spectral characteristics, whereas an L-Ala substitution results in a peptide with minimal structure. These studies suggest that the F4 protein from Schistosoma mansoni may be the first example of a naturally occurring protein devoid of proline and carbohydrate that forms a left-handed helix composed of L-amino acids, although alternative forms of other left-handed structures have yet to be rigorously excluded.

Amino Acid Sequence↗

Molecular architecture of helicoidal proteinaceous eggshells.

I will not attempt to summarize this chapter, which is already a summary. I merely wish to point out that experimental and theoretical evidence to date clearly suggests that antiparallel beta-pleated sheet dictates self-assembly in helicoidal proteinaceous eggshells. Molecular details of this process have started to become clear after the development of the specific, most probably correct, protein structural models in the case of the silkmoths, where amino acid information is available and with the help of several experimental techniques. However, for people seeking universal mechanisms the picture should still be far from complete. Several analogous systems should be studied before providing final answers.

Amino Acid Sequence↗

Influence of chlorinated hydrocarbons, Hg2+ and methyl-Hg+ on steroid hormone receptors from eggshell gland mucosa of domestic fowls and ducks.

A series of chlorinated hydrocarbons were investigated for their ability to inhibit the binding of tritiated progesterone, dexamethasone and testosterone to their respective cytoplasmic receptors, prepared from the eggshell gland mucosa of the domestic fowl. o,p'-DDE and toxaphene were most potent in inhibiting the binding of progesterone and testosterone to their receptors, while p,p'-DDE was most potent in inhibiting the binding of dexamethasone. The effect of the latter compound on dexamethasone binding was biphasic. There were significant differences between ducks and domestic fowls in the dissociation constant (KD) and maximal number of binding sites (Bmax) for several of the steroid hormones. No effect of steroid binding was observed in the p,p'-DDE-treated ducks that were laying thin-shelled eggs. Some metals were also tested for their ability to inhibit progesterone binding. Hg2+ was most potent (IC50 = 5.8 x 10(-5) M) in this respect, while Cd2+, Pb2+ and Al3+ inhibited the binding only when administered in very high concentrations (10(-3) M). Me-Hg+ was less potent than Hg2+. It is concluded that chlorinated hydrocarbon pesticides may interact with several different steroid hormone receptors. These findings are discussed in relation to some reproductive and other hormonal disturbances observed in birds and mammals.

Animals↗

Salmon Eggshell Protein Expression: A Marker for Environmental Estrogens.

: A liver complementary DNA expression library from Atlantic salmon (Salmo salar) pretreated with estradiol-17beta (E2) was constructed and screened with antibodies raised against salmon eggshell (zona radiata) proteins. Two clones, SalZr2_19 and SalZr2_23 were sequenced and shown to encode proteins of approximately 50 kDa. SalZr2_23 contains 12 octamer sequence lpqr/kpa/vq repeats also found in SalZr2_19, but only twice. Alignment reveals that the two salmon sequences are similar to piscine zona radiata proteins and mammalian zona pellucida proteins. Several transcripts ranging from 2.3 to 12 kb appeared in liver extracts of E2-treated fish. Juvenile fish treated with E2 or 4-nonylphenol showed strong induction of zona radiata protein. The use of egg envelope transcriptional and translational induction in male or juvenile fish as a biological marker of environmental estrogens is discussed.

Journal Article↗