Ultrasonographic and radiographic determination of egg development of jurarás (Kinosternon scorpioides) in captivity

Kinosternon scorpioides (Linnaeus, 1766), with its common name of jurará, is a quite variable species of turtles, and many different names have been applied to populations throughout its range. Currently, however, four subspecies are considered valid as K. scorpioides arises from southern Panama over most of northern South America and is found in Ecuador, northern Peru, southern Bolivia, northern Argentina, eastern Guyana and Brazil. Thus, an ultrasonographic and radiographic study was performed in order to describe the morphology and development of eggs of 20 female jurará mud turtles K. scorpioides, from September 2005 to August 2006. In the first month, the ovarian cycle was characterized by absence of vitellogenic follicles, atresic follicles or oviduct eggs. From October 2005 to March 2006 on, ultrasonographic scanning allowed to establish the growing vitellogenic follicles. Vitellogenic follicles were observed with spherical to ovoid shapes, with a surrounding echogenic yolk, a nonechogenic albumin layer, and a high echogenic shell. The oviduct eggs were identified by radiography just 180 days after beginning the experiment, when the shell became enough mineralized to impress the radiographic film. This experiment allowed to obtain by means of the 7.5 MHz linear probe images with adequate resolution and penetration for visualization of follicles. Successive ultrasonographic examinations of 20 K. scorpioides females allowed to access initial stages of vitellogenic follicles and oviduct eggs, and radiographic examination revealed to be an easy technique to assess oviduct eggs and to allow evaluation of egg development in jurarás, from 6 months on.

Variability in the fecal egg count and the parasitic burden of hair sheep after grazing in nematode infected paddocks

This study aimed to evaluate the variability in the fecal egg count (FEC) and the parasitic burden of naive hair sheep after grazing in nematode infected paddocks. The research was carried out in Tabasco, Mexico, during two periods (August and December). In each period 32 lambs were grazed for one month on African star grass (Cynodon plectostachyus) contaminated with gastrointestinal parasitic nematodes. FEC, packed cell volume (PCV) and body weight (BW) were recorded. Gastrointestinal worms were recovered at necropsy. Data were analyzed with the MIXED procedure of SAS using a model of repeated measurements over time. A higher number of Haemonchus contortus worms was found in December (2814±838) than in August (1166±305). The opposite occurred with Cooperia curticei (2167±393 and 3638±441, respectively). The FEC and correlation coefficient in respect to the worm burden were higher in December (6516 ± 1599, r=0.83, respectively) than in August (4364±771, r=0.44, respectively). A high variability in resistance-susceptibility to gastrointestinal nematodes (GIN) occurred in Katahdin × Pelibuey lambs after grazing.

Outbreak of infectious laryngotracheitis in large multi-age egg layer chicken flocks in Minas Gerais, Brazil

A recent (November 2010) outbreak of infectious laryngotracheitis (ILT) in a multi-age laying hen facility in Minas Gerais state, Brazil, is described. Previous ILT outbreak in laying hens was only notified in São Paulo state, Brazil, in 2002. In the outbreak described here, the affected population was approximately eight million hens, with flock sizes ranging from 100,000 to 2,900,000 chickens. The average mortality ranged from 1 to 6%, and morbidity was around 90% (most of the twenty seven farms of the area were positive for ILT virus). Three multi-age laying farms from one company were selected for this report. Clinical signs included prostration, dyspnea, conjunctivitis, occasional swelling of the paranasal sinuses and bloody mucous nasal discharge. Severely affected chickens presented with dyspnea, gasping and became cyanotic before death. At necropsy, these chickens had fibrinous exudate blocking the larynx and the lumen of cranial part of the trachea. In addition, conjunctivitis with intense hyperemia, edema and sinuses with caseous exudate were present. On histopathology, there were marked necrosis and desquamation of respiratory ephitelium and conjunctiva with numerous syncytial cells formation and fibrinous exudate. Moderate to marked non suppurative (especially lymphocytes and plasma cells) infiltration in the lamina propria also was observed. Sixteen out of 20 examined chickens, eosinophilic intranuclear inclusion bodies were observed in the syncytial cells. The DNA extracted from larynx and trachea produced positive PCR results for ILT virus (ILTV) DNA using formalin-fixed, paraffin embedded (FFPE) samples. Amplicons from a small region of ICP4 gene were submitted to sequencing and showed 100% identity with ILTV EU104910.1 (USA strain), 99% with ILTV JN596963.1 (Australian strain) and 91% with ILTV JN580316.1 (Gallid herpesvirus 1 CEO vaccine strain) and JN580315.1 (Gallid herpesvirus 1 TCO vaccine strain).

Genetic polymorphism of fifteen microsatellite loci in Brazilian (blue-egg Caipira) chickens

The purpose of this study was to investigate the genetic polymorphism of fifteen microsatellites loci in Brazilian (blue-egg Caipira) chickens. Samples were collected from 100 blue eggs of Caipira chickens from rural properties in the city of Dois Lajeados, RS. After DNA extraction, the fragments related to molecular markers LEI0248, LEI0221, LEI0214, LEI0192, LEI0217, LEI0254, LEI0194, LEI0212, MCW0371, ADL0278, LEI0234, MCW0183, MCW0216, MCW0330 and MCW0081 were obtained by polymerase chain reaction (PCR). The statistical analysis were carried out with the softwares ARLEQUIN 3.5 version and CERVUS 3.0.3 version. The allelic and genotypic frequencies, deviations from Hardy-Weinberg equilibrium, estimates of observed (HO) and expected (HE) heterozygosity and polymorphic information content (PIC) were obtained for each marker locus. A total of 186 alleles from 15 loci were obtained, with sizes ranging of 83 to 490 base pairs. The medium number of alleles was 12.4, the HE was 0.76±0.14 and HO was 0.49±0.21 and PIC was 0.706. The first conclusion is that the microsatellites used are polymorphic and can be used to genetic studies in chickens. The second is that the "Caipira" chicken (blue eggs) population investigated has a great genic variability, which makes than an important source of genetic resources for future animal breeding programs.

Membrane properties of structurally modified ceramides : effects on lipid lateral distribution and sphingomyelin-interactions in artificial bilayer membranes

Ceramides comprise a class of sphingolipids that exist only in small amounts in cellular membranes, but which have been associated with important roles in cellular signaling processes. The influences that ceramides have on the physical properties of bilayer membranes reach from altered thermodynamical behavior to significant impacts on the molecular order and lateral distribution of membrane lipids. Along with the idea that the membrane physical state could influence the physiological state of a cell, the membrane properties of ceramides have gained increasing interest. Therefore, membrane phenomena related to ceramides have become a subject of intense study both in cellular as well as in artificial membranes. Artificial bilayers, the so called model membranes, are substantially simpler in terms of contents and spatio-temporal variation than actual cellular membranes, and can be used to give detailed information about the properties of individual lipid species in different environments. This thesis focuses on investigating how the different parts of the ceramide molecule, i.e., the N-linked acyl chain, the long-chain sphingoid base and the membrane-water interface region, govern the interactions and lateral distribution of these lipids in bilayer membranes. With the emphasis on ceramide/sphingomyelin(SM)-interactions, the relevance of the size of the SMhead group for the interaction was also studied. Ceramides with methylbranched N-linked acyl chains, varying length sphingoid bases, or methylated 2N (amide-nitrogen) and 3O (C3-hydroxyl) at the interface region, as well as SMs with decreased head group size, were synthesized and their bilayer properties studied by calorimetric and fluorescence spectroscopic techniques. In brief, the results showed that the packing of the ceramide acyl chains was more sensitive to methyl-branching in the mid part than in the distal end of the N-linked chain, and that disrupting the interfacial structure at the amide-nitrogen, as opposed to the C3-hydroxyl, had greater effect on the interlipid interactions of ceramides. Interestingly, it appeared that the bilayer properties of ceramides could be more sensitive to small alterations in the length of the long-chain base than what was previously reported for the N-linked acyl chain. Furthermore, the data indicated that the SM-head group does not strongly influence the interactions between SMs and ceramides. The results in this thesis illustrate the pivotal role of some essential parts of the ceramide molecules in determining their bilayer properties. The thesis provides increased understanding of the molecular aspects of ceramides that possibly affect their functions in biological membranes, and could relate to distinct effects on cell physiology.

Genetic interactions involving major genes at the dwarf locus in egg-type chickens

Three egg-type stocks segregating dwarf (dw) and bantam (dwB) genes in female progeny were produced from the same 18 heterozygous (dwB/dw) sires used to inseminate dams of three different genotypes: normal (dw+), dwarf (dw) and bantam (dwB) dams. The heritability of 8-week body weight estimated from full-sibs of the same phenotype of progeny was 0.40, and that estimated from paternal half-sibs of the same phenotype (dwarf or bantam), and from the same genotype of dam was 0.38. Therefore, maternal and non-additive effects within genotypic classes of dam made little contribution to the genetic variance for 8-week body weight among their progeny. The interaction of sires (S) with genotypes (dw+, dw and dwB) of dam (G) was significant at the 5% level. This indicates that the rankings of the sires within each one of the three genotypes of dam were not the same, probably due to non-additive genetic variation among genotypes of dams. The evidence indicated that in general the genes from individual sires combined differently with each type of dam (G). Those genes which combined well with the genes from normal (dw+) dams combined poorly with both the genes from the dwarf (dw) and the genes from the bantam (dwB) dams. The interaction of sires (S) with phenotypes (dwarf and bantam) of progeny (P) was significant at the 10% level. The results indicated a probable gene x genotype interaction for 8-week weight between genes at the dwarf locus (dw and dwB) and the background genotype (single and/or polygenes). The correlation among paternal half-sibs was influenced more by the S x G than by the S x P interaction, but the effects tended to be cumulative

Synthesis of stocks and phenotypic effects of dwarf and bantam sex-linked major genes in egg-type chickens

The present study describes the production of stocks segregating dwarf (dw), bantam (dwB) and normal (dw+) alleles, as well as the characters, shank length, adult body weight, age at sexual maturity and egg production. Heterozygous K dw+/k dwB sires were mated to normal (dw+) dams to produce stock D6.a, and mated to dwB females to produce stock D6.b. Stock D4.a came from mating F1 heterozygous dwB dw sires to dwarf Leghorns. In a third series of matings, 7/8 Sebright and 1/8 dw-Leghorn dwB dw sires were crossed to three groups of dams of different genotypes. The progeny of the normal (dw+), dwarf (dw), and bantam (dwB) dams were designated as stocks D4.b, D4.c and D4.d, respectively. The dw+ dams were White Leghorn strain cross females. The difference between the rate of laying of normal (69.7%) and their bantam sisters (68.6%) was not statistically significant when the average 32-week body weight of the dw+ sisters was 1,897 g. However, when the 32-week body weight of the normal daughters from the same sires and smaller dams was around 1,646 g, the difference between the rate of laying of the normal (78.1%) and their bantam sisters (75.9%) was significant (P < 0.05). The dwB gene may have a similar but smaller effect on the rate of egg laying than its dwarf allele. The difference between sexual maturity of normal and bantam daughters of either the largest or the smallest dams was not statistically significant, even though the smallest dwB pullets were in average 2.9 days older at first egg. The use of shank length combined with adult body weight allowed a precise discrimination between bantams and dwarfs

Egg size, yolk mass extrusion and hatching behavior in two cryptic speciesof Anastrepha fraterculus (Wiedemann) (Diptera, Tephritidae)

Variations in egg length were observed for two populations of cryptic species of Anastrepha fraterculus (Wiedemann). The eggs of type I flies were smaller than those of type II individuals. For both types, in regard to yolk mass extrusion, four classes of embryos were detected. Class 1: embryos that extrude masses at both extremities; class 2: embryos in which extrusion occurs only at the anterior pole; class 3: embryos that eliminate mass only at the posterior pole, and class 4: embryos that do not extrude any mass. Embryo class frequencies were similar for populations belonging to the same type, but different between types. Individual females may produce eggs from different embryo classes, but for any given female the pattern remains constant during a long period of oviposition. Variation in size of the extruded masses was similar for both populations. Individual females produced embryos with a small range of mass diameters, and different females produced masses of different mean size. However, individual mass size remained constant during oviposition. The results suggest the existence of genetic components involved in the control of this unusual process. Larvae of both types presented, just before eclosion, similar unusual behaviors: they ingest the anterior extruded mass, rotate 180°, absorb the posterior mass and eclose near the posterior pole. Data show that cryptic A. fraterculus type I and type II differs in regard to egg size as well as to the phenomenon of yolk mass extrusion

Extracts of Ascaris suum egg and adult worm share similar immunosuppressive properties

Adult Ascaris suum body extract (Asc) prepared from male and female worms (with stored eggs) down-regulates the specific immune response of DBA/2 mice to ovalbumin (OA) and preferentially stimulates a Th2 response to its own components, which is responsible for the suppression of the OA-specific Th1 response. Here, we investigated the participation of soluble extracts prepared from male or female worms or from eggs (E-Asc) in these immunological events. Extracts from either sex (1 mg/animal) or E-Asc (0.35 or 1 mg protein/animal) suppressed the delayed-type hypersensitivity (DTH) reaction (60-85%), proliferative response (50-70%), IL-2 and IFN-gamma secretion (below detection threshold) and IgG1 antibody production (70-90%) of DBA/2 mice to OA. A dose of 0.1 mg E-Asc/animal did not change DTH or proliferation, but was as effective as 0.35 mg in suppressing IL-2 and IFN-gamma, and OA-specific IgG1 antibodies. Lymph node cells from DBA/2 mice injected with Asc (1 mg/animal) or a high dose of E-Asc (1 mg protein/animal) secreted IL-4 upon in vitro stimulation with concanavalin A. As previously demonstrated for Asc, the cytokine profile obtained with the E-Asc was dose dependent and changed towards Th1 when a low dose (0.1 mg protein/animal) was used. Taken together, these results suggest that adult worms of either sex and eggs induce the same type of T cell response and share similar immunosuppressive properties.

Toxicity to sea urchin egg development of the quinone fraction obtained from Auxemma oncocalyx

Auxemma oncocalyx Taub. belongs to the Boraginaceae family and is native to the Brazilian northeast where it is known as "pau-branco". We investigated the ability of the water soluble fraction isolated from the heartwood of A. oncocalyx to inhibit sea urchin egg development. This fraction contains about 80% oncocalyxone A (quinone fraction), a compound known to possess strong cytotoxic and antitumor activities. In fact, the quinone fraction inhibited cleavage in a dose-dependent manner [IC50 of 18.4 (12.4-27.2) µg/ml, N = 6], and destroyed the embryos in the blastula stage [IC50 of 16.2 (13.7-19.2) µg/ml, N = 6]. We suggest that this activity is due to the presence of oncocalyxone A. In fact, these quinones present in A. oncocalyx extract have strong toxicity related to their antimitotic activity.

Pressure-assisted cold denaturation of hen egg white lysozyme: the influence of co-solvents probed by hydrogen exchange nuclear magnetic resonance

COSY proton nuclear magnetic resonance was used to measure the exchange rates of amide protons of hen egg white lysozyme (HEWL) in the pressure-assisted cold-denatured state and in the heat-denatured state. After dissolving lysozyme in deuterium oxide buffer, labile protons exchange for deuterons in such a way that exposed protons are substituted rapidly, whereas "protected" protons within structured parts of the protein are substituted slowly. The exchange rates k obs were determined for HEWL under heat treatment (80ºC) and under high pressure conditions at low temperature (3.75 kbar, -13ºC). Moreover, the influence of co-solvents (sorbitol, urea) on the exchange rate was examined under pressure-assisted cold denaturation conditions, and the corresponding protection factors, P, were determined. The exchange kinetics upon heat treatment was found to be a two-step process with initial slow exchange followed by a fast one, showing residual protection in the slow-exchange state and P-factors in the random-coil-like range for the final temperature-denatured state. Addition of sorbitol (500 mM) led to an increase of P-factors for the pressure-assisted cold denatured state, but not for the heat-denatured state. The presence of 2 M urea resulted in a drastic decrease of the P-factors of the pressure-assisted cold denatured state. For both types of co-solvents, the effect they exert appears to be cooperative, i.e., no particular regions within the protein can be identified with significantly diverse changes of P-factors.

Isolation and Characterization of Ovomucin - a Bioactive Agent of Egg White

The hen’s egg is a source of new life. Therefore, it contains many biologically active compounds. In addition to being a very nutritious food and also commonly used in the food industry due to its many techno-functional properties, the egg can serve as a source of compounds used as nutra-, pharmaand cosmeceuticals. One such interesting compound is ovomucin, an egg white protein responsible for the gel-like properties of thick egg white. Previous studies have indicated that ovomucin and ovomucin-derived peptides have several different bioactive properties. The objectives of the present study were to develop isolation methods for ovomucin, to characterize the structure of ovomucin, to compare various egg fractions as sources of ovomucin, to study the effects of various dissolving methods for ovomucin, and to investigate the bioactive properties of ovomucin and ovomucin-derived peptides. A simple and rapid method for crude ovomucin separation was developed. By using this method crude ovomucin was isolated within hours, compared to the 1-2 days (including a dialysis step) needed when using several other methods. Structural characterization revealed that ovomucin is composed of two subunits, α- and β-ovomucin, as egg white protein formerly called α1-ovomucin seemed to be ovostatin. However, it might be possible that ovostatin is associated within β- and α-ovomucin. This interaction could even have some effect on the physical nature of various egg white layers. Although filtration by-product fraction was a very prominent source of both crude and β-ovomucin, process development has reduced its amount so significantly that it has no practical meaning anymore. Thus, the commercial liquid egg white is probably the best option, especially if it generally contains amounts of β-ovomucin as high as were found in these studies. Crude ovomucin was dissolved both by using physical and enzymic methods. Although sonication was the most effective physical method for ovomucin solubilisation, colloid milling seemed to be a very promising alternative. A milk-like, smooth and opaque crude ovomucin suspension was attained by using a colloid mill. The dissolved ovomucin fractions were further tested for bioactive properties, and it was found that three dissolving methods tested produced moderate antiviral activity against Newcastle disease virus, namely colloid milling, enzymatic hydrolysis and a combination of sonicaton and enzymatic hydrolysis. Moreover, trypsin-digested crude ovomucin was found to have moderate antiviral activity against avian influenza virus: both subtype H5 and H7.

INFLUENCE OF ASCORBIC ACID ON EGG QUALITY

The egg quality may be influenced by some nutrients added to laying hens diets and among these nutrients, the ascorbic acid is included. According to some researches, this acid, besides of improving the quality, it can increase the egg production and weight. In addition, it reduces the quantity of cracked eggs and food intake. In order to contribute with more information, this experiment was developed. Concentrations of 100; 150 and 200mg/kg of ascorbic acid were added to rations of laying hens during 60 days, with the objective of determining the influence of ascorbic acid on the quality of eggs kept under room temperature conditions (26.7 ± 2.2°C and 60 ± 5% moisture) for a period of 28 days. A total of 1440 eggs were used, being 720 of white shell and 720 of brown shell. Egg quality was determined by Haugh unit and yolk index, every 7 days. The obtained results allowed to conclude that ascorbic acid, independent on the level of supplementation, was not enough to decrease.

Physicochemical study of CaCO3 from egg shells

Calcium carbonate, a pharmaceutical excipient, is widely used as diluent in solid dosage forms. It is also used as a base for medicinal and dental preparations, a buffering and dissolution aid for dispersible tablets, a food additive and a calcium supplement. Egg shells are a rich source of mineral salts, mainly calcium carbonate, which corresponds to about 94% of the shell. Layer farms produce large amounts of shells, whose final disposal poses a challenge from the environmental standpoint. This work was designed to evaluate the physicochemical and thermal properties of calcium carbonate obtained from egg shells. The findings indicated that calcium carbonate from egg shells can be used as an alternative pharmaceutical excipient.

Preparation of powdered egg yolk using a mini spray dryer

Powdered egg is used as an emulsifying agent in emulsion formulations. It is an excellent source of high quality protein, of which the yolk contains 44% and the egg white 56%. Spray drying is a widely applied method for drying aqueous or organic solutions and emulsions in the chemical and food industries. Spray drying can be used to preserve food or simply as a rapid drying method. The objective of this work was to study the viability of obtaining powdered egg yolk powder using a Büchi B-190 Mini Spray Dryer. The egg yolk protein was evaluated by the semi-micro Kjeldahl method. It was concluded that the use of the Büchi B-190 Mini Spray Dryer to produce powdered egg yolk is perfectly feasible.