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Biomedical subjects

Y Nys

Publications and source records attributed to Y Nys.

At least 19 recordsLinked to original sources
PubMed2006

Effects of beak trimming and cage design on laying hen performance and egg quality.

1. Conventional cages are to be replaced by furnished cages or aviary systems to improve the welfare of hens. We compared the performance and egg quality of hens reared in two designs of furnished cages and of two standard cages. We also explored the consequences of the absence of beak trimming when using these designs. 2. Hens (2028) were housed from 18 to 70 weeks of age in 108 standard cages at 6 per cage (60 cmx63.5 cm), in 96 cages at 5 per cage (59.5 cmx55.5 cm) or in two designs of furnished cage at 15 per cage (24 F15M cages and 36 F15P cages made by two manufacturers) which contained equipment varying in size and location (nests, dust baths and perches). Half of the hens were beak-trimmed in each design. 3. Mortality was low in beak-trimmed hens (<5%) but was unacceptably high in non-beak-trimmed hens due to cannibalism (>40%, 516 hens). Mortality was worse in standard cages than in furnished cages. Consequently, hen-housed egg production was significantly lowered in non-beak-trimmed hens. 4. Egg laying in beak-trimmed hens reared in furnished cages occurred mainly in the nest (80 and 84.8% in F15M and F15P) but also in the dust bath (13.3 and 9.4% in F15M and F15P) and in other parts of the cage (6.7 and 5.8% in F15M and F15P). 5. The total percentage of broken (visual observation) and hair-cracked eggs (candling) was high in the furnished cage designs (15.4 and 19.6% in F15M and F15P, respectively) compared with standard cage designs (8.1 and 12.2% in S6 and S5). This was mainly due to hair-cracked eggs, the highest percentages occurring in the nests, especially in the design with a narrow nest and no egg saver (11.1% in F15M compared to 17.6% in F15P) as a consequence of egg accumulation in the cradle and relatively low frequency of manual egg collection. 6. Eggshell quality (index and breaking strength) was only slightly influenced by cage effects so differences in egg breakage were attributable to impacts related to cage design. 7. It is concluded that beak trimming remains the most effective way to prevent cannibalism, although furnished cages with a large group of hens slightly reduced the incidence, and that further development and optimisation of furnished cages is needed to reach egg quality similar to standard cages.

Animal Husbandry↗
PubMed2006

Comparison of eggshell hygiene in two housing systems: standard and furnished cages.

1. The hygienic properties of eggs produced in two designs of furnished cage were investigated and compared to two standard cage designs. 2. At 28, 37, 47 and 58 weeks of age, the proportion of dirty eggs was higher in one of the furnished cage designs while, in the other, it was similar to standard cages. 3. At 27, 33 and 60 weeks, the bacterial load on the eggshell (total aerobic bacteria and enterococci) was higher in furnished cage designs. A seasonal effect was observed with lower counts at 60 weeks (winter) than at 27 weeks (summer). 4. More dirty eggs and a higher bacterial load were observed in eggs laid outside the nests, which suggests egg hygiene in furnished cages could be similar to standard cages if the equipment in furnished cages was improved to enhance nest laying.

Aging↗
PubMed2006

Assessment of dietary zinc requirement of weaned piglets fed diets with or without microbial phytase.

Fifty-four pigs, weaned at 26 days of age at an average body weight of 7.74 kg were used in a 26-day experiment to assess the zinc requirement of piglets, using diets based on maize and soybean meal, with or without microbial phytase. The nine experimental diets were the basal diet containing 33 mg of zinc/kg supplemented with 10, 25, 40, 60 or 80 mg of zinc as sulphate (ZnSO(4), 7H(2)O)/kg and the basal diet supplemented with 0, 10, 25 or 40 mg of zinc as sulphate/kg and 700 units (U) of microbial phytase (Natuphos)/kg. Pigs were fed the basal diet for a 7-day adjustment period prior to the 19-day experimental period. Microbial phytase enhanced plasma alkaline phosphatase (AP) activity, plasma zinc and bone zinc concentrations. These parameters increased linearly with zinc intake, with a similar slope with and without phytase. The response of bone zinc-to-zinc added did not plateau. Without microbial phytase, plasma AP activity and zinc concentration were maximized when dietary zinc reached 86 and 92 mg/kg respectively. With microbial phytase they were maximized when dietary zinc concentration reached 54 and 49 mg/kg respectively. Accounting for a safety margin, the recommended supply of zinc for weaned piglets up to 16 kg fed maize-soybean meal diets supplemented with zinc as sulphate is thus of 100-110 mg/kg diet. This supply may be reduced by around 35 mg if the diet is supplemented with 700 U of microbial phytase.

6-Phytase↗
PubMed2005

Changes in eggshell mechanical properties, crystallographic texture and in matrix proteins induced by moult in hens.

The effect of moult on eggshell mechanical properties, on composition and concentrations of organic matrix components and on eggshell microstructure was investigated. The observed changes were studied to understand the role of organic matrix and eggshell microstructure in eggshell strength. Moult was induced by zinc oxide (20 g zinc/kg diet) in 53 ISA Brown laying hens at 78 weeks of age. No difference was observed for egg or eggshell weights after moult. In contrast, moult improved the shell breaking strength (28.09 vs 33.71 N). After moult, there was a decrease in the average size of calcite crystals composing the eggshell and in their heterogeneity, whereas crystal orientation remained basically the same. After moulting, the total protein concentration in eggshell increased slightly. The comparisons of SDS-PAGE profiles of the organic matrix constituents extracted before and after moulting showed changes in staining intensity of certain bands. After moult, bands associated with main proteins specific to eggshell formation (OC-116 and OC-17) showed higher staining intensity, while the intensity of the egg white proteins (ovotransferrin, ovalbumin and lysozyme) decreased. ELISA confirmed the decrease in ovotransferrin after moult. Its concentration was inversely correlated with breaking strength before moult. These observations suggest that changes in eggshell crystal size could be due to changes in organic matrix composition. These changes may provide a mechanism for the improvement in shell solidity after moulting.

Animals↗
PubMed2003

Supplementation of a corn-soybean meal diet with manganese, copper, and zinc from organic or inorganic sources improves eggshell quality in aged laying hens.

An experiment was conducted to evaluate the effects on eggshell quality of supplementing the diet of laying hens with a combination of Zn, Mn, and Cu from organic or inorganic sources. Three groups of hens, aged 32, 60, and 69 wk were fed a corn-soybean basal diet containing 32.6 mg/kg Zn, 24.7 mg/kg Mn, and 4.95 mg/kg Cu or a basal diet supplemented with 30-30-5 and 60-60-10 mg/kg of Zn, Mn, and Cu, respectively. Addition of Zn, Mn, and Cu in combination increased their concentrations in egg yolk and slightly decreased egg weight during some sampling from aged hens. The Zn, Cu, and Mn dietary supplementation did not affect the following measures of eggshell quality: percentage eggshell, eggshell index (shell weight per unit surface area), and eggshell stiffness. However, there was a small increase in elastic modulus of the eggs from one group of the older hens due to supplementation with Zn, Mn, and Cu. More consistently we observed improved breaking strength and fracture toughness (resistance to fracture) in the two oldest groups of hens at 1, 5, and 9 wk after introduction of the experimental diets. The effect was observed regardless of the source of trace elements.

Aging↗
PubMed2003

Purification of ovocalyxin-32, a novel chicken eggshell matrix protein.

The eggshell is a highly ordered structure resulting from the deposition of calcium carbonate and an organic matrix from the acellular uterine fluid. Characterization of the individual matrix components is necessary to determine their influence upon calcite crystal shape, size, and orientation during eggshell calcification. We have purified and sequenced a novel 32-kDa protein, ovocalyxin-32 (OCX-32), which is present at high levels in the uterine fluid during the terminal phase of eggshell formation, and is localized predominantly in the outer eggshell. Database searches identified expressed sequence tags (ESTs) whose alignment yielded the complete cDNA. OCX-32 protein possesses limited identity (32%) to two unrelated proteins: latexin, a carboxypeptidase inhibitor expressed in rat cerebral cortex and mast cells, and to a skin protein that is encoded by a retinoic acid receptor-responsive gene, TIG1. The timing of OCX-32 secretion into the uterine fluid suggests that it may play a role in the termination of mineral deposition.

Animals↗
PubMed2002

Influence of the microstructure on the shell strength of eggs laid by hens of different ages.

1. The eggshell is a bioceramic material constructed of columnar calcite crystals preferentially oriented with their c-axis perpendicular to the shell surface. 2. The influence of microstructure (crystal size, shape and crystallographic orientation of crystal grains) on the mechanical properties of eggshells (shell strength) was investigated using eggs from hens of different ages. 3. There was a strong correlation between crystallographic texture and the strength of the eggshell in the case of eggs laid by young hens. The strength of eggshells increased as the preferential orientation of the crystals constituting the eggshell decreased. 4. By comparing two age populations, the effect of hen age on eggshell properties was evaluated. In general, eggshells from aged hens had a lower breaking strength (less than half that of those laid by young hens) and showed a greater variability in their structural properties such as thickness, grain morphology and crystallographic texture. 5. Texture analysis revealed that shells from eggs laid by aged hens have two preferred crystal orientations, after (001) and (104), compared with mainly one, after (001), in eggs laid by young hens. 6. These observed changes in eggshell properties could be due to changes in the organic matrix of the eggshell associated with ageing of the hens.

Age Factors↗
PubMed2001

Ovocalyxin-32, a novel chicken eggshell matrix protein. isolation, amino acid sequencing, cloning, and immunocytochemical localization.

The eggshell is a highly ordered structure resulting from the deposition of calcium carbonate concomitantly with an organic matrix upon the eggshell membranes. Mineralization takes place in an acellular uterine fluid, which contains the ionic and matrix precursors of the eggshell. We have identified a novel 32-kDa protein, ovocalyxin-32, which is expressed at high levels in the uterine and isthmus regions of the oviduct, and concentrated in the eggshell. Sequencing of peptides derived from the purified protein allowed expressed sequence tag sequences to be identified that were assembled to yield a full-length composite sequence whose conceptual translation product contained the complete amino acid sequence of ovocalyxin-32. Data base searches revealed that ovocalyxin-32 has limited identity (32%) to two unrelated proteins: latexin, a carboxypeptidase inhibitor expressed in the rat cerebral cortex and mast cells, and a skin protein, which is encoded by a retinoic acid receptor-responsive gene, TIG1. High level expression of ovocalyxin-32 was limited to the isthmus and uterus tissue, where immunocytochemistry at the light and electron microscope levels demonstrated that ovocalyxin-32 is secreted by surface epithelial cells. In the eggshell, ovocalyxin-32 localizes to the outer palisade layer, the vertical crystal layer, and the cuticle of the eggshell, in agreement with its demonstration by Western blotting at high levels in the uterine fluid during the termination phase of eggshell formation. Ovocalyxin-32 is therefore identified as a novel protein synthesized in the distal oviduct where hen eggshell formation occurs.

Amino Acid Sequence↗
PubMed2001

Ovotransferrin is a matrix protein of the hen eggshell membranes and basal calcified layer.

The eggshell is an highly ordered structure deposited in the distal oviduct and composed of calcium carbonate and an organic matrix which is believed to influence its fabric. We have identified ovotransferrin as an 80 kDa matrix protein observed at high concentration in the uterine fluid at the initial stage of shell mineralization, by N-terminal sequencing and western blotting using monoclonal and polyclonal antibodies. It is present in extracts from demineralized eggshell and was localized by immunofluorescence in the eggshell membranes and mammillae, which are the sites of calcite nucleation. Northern blotting and RT-PCR demonstrated that ovotransferrin message was expressed in the proximal oviduct (magnum and white isthmus), and at a lower magnitude in the distal oviduct (red isthmus and uterus). Ovotransferrin was revealed by immunofluorescence in the tubular gland cells of the uterus. Calcium carbonate crystals grown in vitro in the presence of purified ovotransferrin showed large modifications of the calcite morphology. These observations and its presence in eggshell and membranes suggest a dual role for ovotransferrin, as a protein influencing nucleation and growth of calcite crystals and as a bacteriostatic filter to reinforce its inhibition of Salmonella growth in egg albumen.

Amino Acid Sequence↗
PubMed2000

Identification and localization of lysozyme as a component of eggshell membranes and eggshell matrix.

The avian eggshell is a composite biomaterial composed of non-calcifying eggshell membranes and the overlying calcified shell matrix. The calcified shell forms in a uterine fluid where the concentration of different protein species varies between the initial, rapid calcification and terminal phases of eggshell deposition. The role of these avian eggshell matrix proteins during shell formation is poorly understood. The properties of the individual components must be determined in order to gain insight into their function during eggshell mineralization. In this study, we have identified lysozyme as a component of the uterine fluid by microsequencing, and used western blotting, immunofluorescence and colloidal-gold immunocytochemistry to document its localization in the eggshell membranes and the shell matrix. Furthermore, Northern blotting and RT-PCR indicates that there is a gradient to the expression of lysozyme message by different regions of the oviduct, with significant albeit low levels expressed in the isthmus and uterus. Lysozyme protein is abundant in the limiting membrane that circumscribes the egg white and forms the innermost layer of the shell membranes. It is also present in the shell membranes, and in the matrix of the calcified shell. Calcite crystals grown in the presence of purified hen lysozyme exhibited altered crystal morphology. Therefore, in addition to its well-known anti-microbial properties that could add to the protective function of the eggshell during embryonic development, shell matrix lysozyme may also be a structural protein which in soluble form influences calcium carbonate deposition during calcification.

Animals↗
PubMed2000

Extraction and quantification by ELISA of eggshell organic matrix proteins (ovocleidin-17, ovalbumin, ovotransferrin) in shell from young and old hens.

The eggshell matrix is mainly composed of proteins that are thought to influence shell formation and calcification and, thus, modify the resulting properties of the shell. We investigated the potential of some of these proteins as biomarkers of eggshell quality by developing a competitive indirect ELISA for quantifying ovotransferrin, ovalbumin, and ovocleidin-17 in eggshell extract. Eggshell fragments were demineralized in acetic acid (20%) and freeze-dried. The micro-extraction yield was markedly increased (>50%) when Tween 20 was added to the subsequent extraction and dialysis milieus. Microplates were coated with ovotransferrin and ovalbumin in a 0.1M carbonate-bicarbonate buffer, but ovocleidin-17 was fixed with acetone (-20 C, 20 min). Optimal dilutions of the monoclonal (ovotransferrin) and polyclonal (ovalbumin and ovocleidin-17) antibodies were 1/3,000, 1/25,000 and 1/4,000, respectively. The inhibition curves were optimized by preincubating the antibodies and proteins overnight. The intraassay coefficient (<5%), parallelism of the standards and samples curves, and recovery (101%) were satisfactory for ovotransferrin. Measurements of ovalbumin were less precise because of higher interassay variation and differences between the slopes of standard and sample inhibition curves. Ovocleidin-17 assays showed similar slopes for standard and eggshell extracts. Although the total protein in soluble matrix extracts was not affected by age, the concentrations of these proteins were higher in eggshell extracts from older hens compared with those from young hens: 1.98x for ovotransferrin, 1.86x for ovalbumin, and 1.58x for ovocleidin-17. The quantification of specific eggshell matrix proteins in shell of differing quality is, therefore, a promising tool for analyzing the origin of eggshell faults and may provide useful information for breeding programs.

Aging↗
PubMed2000

The effect of avian uterine fluid on the growth behavior of calcite crystals.

Eggshell formation takes place on the eggshell membrane in an acellular medium, the uterine fluid that contains the inorganic minerals and precursors of the organic matrix. The high degree of eggshell structure could be due to an interaction between calcium carbonate and the organic matrix. The aim of this study was to demonstrate such an interaction by measuring the effect in vitro of uterine fluid collected at various phases of shell formation on precipitation kinetics, size, and morphology of calcite crystals. The SDS-PAGE profiles of the organic constituents differed between the different phases of eggshell formation. The predominant constituents were ovalbumin and ovotransferrin at the initial phase and lysozyme, ovocleidin-17, ovocalyxin-32, 36- and 21-kDa bands, and ovocleidin-116 at the growth phase. These proteins were numerous in the terminal phase and showed an increased staining of the 32- and 66-kDa bands and appearance of very low molecular weight bands. The precipitation lag time was shortened in proportion to the protein concentration at the initial stage. The effect was observed with a lower magnitude in the presence of constituents of growth and terminal phases. Crystal size was smaller in the presence of constituents from the three stages compared with the control. Components from the initial phase induced the formation of twinned crystals and of rounded corners in the rhombohedric crystals. The presence of components from the growth and terminal phases strongly modified the morphology of the calcite crystals. The majority of the corners became rough and developed curved faces. These observations confirm the interaction of the uterine fluid with calcite and its contribution to eggshell structure.

Animals↗
PubMed1999

Molecular cloning and ultrastructural localization of the core protein of an eggshell matrix proteoglycan, ovocleidin-116.

The role of avian eggshell matrix proteins in shell formation is poorly understood. This calcitic biomaterial forms in a uterine fluid where the protein composition varies during the initial, calcification, and terminal phases of eggshell deposition. A specific antibody was raised to a 116-kDa protein, which is most abundant in uterine fluid during active eggshell calcification. This antiserum was used to expression screen a bacteriophage cDNA library prepared using mRNA extracted from pooled uterine tissue harvested at the midpoint of eggshell calcification. Plasmids containing inserts of differing 5'-lengths were isolated with a maximum cDNA sequence of 2.4 kilobases. Northern blotting and reverse transcriptase-polymerase chain reaction demonstrated that the 2. 35-kilobase message was expressed in a uterine-specific manner. The hypothetical translational product from the open reading frame corresponded to a novel 80-kDa protein, which we have named ovocleidin-116. After removal of the predicted signal peptide, its N-terminal sequence corresponded almost exactly with that determined from direct microsequencing of the 116-kDa uterine protein (this work) and with that previously determined for the core protein of a 120-kDa eggshell dermatan sulfate proteoglycan (Corrino, D. A., Rodriguez, J. P., and Caplan, A. I. (1997) Connect. Tissue Res. 36, 175-193). Ultrastructural colloidal gold immunocytochemistry of ovocleidin-116 demonstrated its presence in the organic matrix, in small vesicles found throughout the mineralized palisade layer, and the calcium reserve assembly of the mammillary layer. Ovocleidin-116 thus is a candidate molecule for the regulation of calcite growth during eggshell calcification.

Amino Acid Sequence↗
PubMed1999

Features of eggshell formation in guinea fowl: kinetics of shell deposition, uterine protein secretion and uterine histology.

1. Rate of calcium carbonate deposition, duration of eggshell formation, organic composition of the uterine fluid, morphology of the egg shells and histochemistry of the uterus were studied in guinea fowl to analyse the origin of such thick, strong egg shells. 2. The egg shell was linearly deposited from 6.4 h to 21.8 h after the oviposition of the previous egg. The rate of egg shell deposition was similar to that in laying hens. However, the duration of linear shell deposition was increased by 2.1 h relative to that in hens. This explained the increased egg shell weight observed in the guinea fowl. 3. Intervals between oviposition of intra-clutch eggs were 24 h throughout the laying period. Ovulation occurred just after oviposition of the previous egg in the guinea fowl, as previously observed in hens but the duration of egg white protein deposition, of plumping and of initiation of shell mineralisation were all 1.5 h shorter than in domestic hen. 4. Uterine fluid can only be collected during the growth and terminal phase of shell formation. The electrophoretic profiles of the uterine fluid differed between phases and were somewhat different from those previously observed in the hen. Ovalbumin and ovocleidin-17 were both present in the uterine fluid and also in egg shell extract. Ovocleidin-17 was predominant during the growth phase. 5. The histology of the uterus differed slightly in guinea fowl compared to hens. Ovocleidin and ovalbumin are both secreted by the tubular glands. 6. Examination of radial ultrathin sections of eggshell showed, above the mammillary layer, intricate interlacing of adjacent exospherite in guinea fowl in contrast to the continuous columnar microstructure in hens. 7. The kinetics of egg shell deposition largely explains the increased egg shell weight of guinea fowl. The organic matrix proteins may be associated with the contrast between the structural organisation of the guinea fowl egg shell and that of the hen egg shell.

Animals↗
PubMed1999

Organic matrix composition and ultrastructure of eggshell: a comparative study.

1. The avian eggshell is a biomineralised composite ceramic consisting of calcium carbonate embedded in an organic matrix. Matrix components are supposed to be involved in the control of mineralisation, crystallographic texture and biomechanical properties of eggshell. 2. The structure and eggshell matrix composition of various domesticated bird species were compared to gain insight into the universality of the eggshell mineralisation process. 3. The SDS-PAGE profiles of soluble eggshell matrix were specific within groups of birds (a: laying hen, breeder hen, quail, pheasant and possibly turkey; b: guinea fowl; c: duck and goose) but some of the protein bands were common to all groups. 4. Analogies between species were confirmed by Western blotting using hen protein antibodies. Ovocleidin-17 (OC-17) and ovalbumin were revealed in all species (except quail for OC-17). Lysozyme was present only in hen eggshell. Another egg white protein: ovotransferrin showed a positive signal in hens, turkey and quail. Osteopontin was observed in laying and breeder hens and quail. 5. Different proteoglycans were localised to discrete regions within the eggshell. Dermatan sulphate was observed within the matrix of the calcified shell of all species except quail which contained chondroitin-6-sulfate. Keratan sulphate was observed in mammillary bodies of breeder and laying hen, quail, pheasant and turkey while chondroitin sulphate was also present in guinea fowl and duck. 6. The general structural organisation of the different avian eggshells was similar but specific differences were observed in the ultrastructure of the mammillary layer. Species of the same taxonomic family could be grouped according to their structural analogies: breeder hen, turkey and pheasant resembled that of the domestic fowl. Guinea fowl was unique. Goose and duck were quite similar with large and confluent mammillary bodies. 7. Some matrix components are therefore common to eggshells of various species but more information is needed to relate differences in matrix composition between taxonomic groups with differences in ultrastructure.

Animals↗
PubMed1999

Effect of dietary zinc content and sources on the growth, body zinc deposition and retention, zinc excretion and immune response in chickens.

1. In areas of intensive animal production heavy metals such as zinc (Zn), which is present at high concentrations in poultry excreta in relation to plant requirements, may be at the origin of soil phytotoxicity This study was conducted in order to determine the effect of decreasing dietary Zn content on growth, plasma, tibia and whole body Zn concentrations, immune function, enzyme activity, Zn body retention and Zn concentration in excreta in broilers. 2. Two experiments were carried out using 160 and 80 1-day-old chicks. Broilers received diets with increased Zn contents of 20 to 190 mg/kg. In experiment 1, two sources of zinc methionine were compared to zinc sulphate. 3. A dietary Zn concentration of 45 mg/kg was sufficient to obtain normal broiler performance at 21 d of age. 4. Tibia and plasma Zn concentrations increased linearly with Zn dietary content and reached a plateau at 75 mg/kg, whereas the whole body Zn was saturated when the dietary Zn content was 90 mg/kg. 5. Antibody titres in response to SRBC injection and plasma alkaline phosphatase activity were not affected by dietary zinc concentration. 6. When the dietary Zn content was decreased from 190 to 65 mg/kg, body Zn retention was increased from 8% to 20% and Zn concentration in broiler manure was reduced by 75%. 7. Zn sources had no effect on the parameters measured in this study. 8. A nutritional approach, that is by lowering dietary Zn supplementation may reduce the risks of phytotoxicity in the soil resulting from excessive Zn concentration in manure.

Animal Feed↗