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Dlx genes integrate positive and negative signals during feather bud development.

In the embryonic chicken skin, feather buds and the intervening interbud tissue form in a reiterated and sequential pattern that is dependent on interactions between the epidermis and dermis. Feather promoting and inhibiting signals such as fibroblast growth factors (FGF) and bone morphogenetic proteins (BMP), respectively, direct the formation of this periodic pattern. However, the transcription factors that mediate the response to these signals and transmit this information to downstream effector genes are largely unknown. Here we have explored the DLX transcription factors as candidate transcriptional mediators downstream of the described feather patterning signals. We show that several Dlx members are expressed in the dermis and epidermis of the developing feather buds and their expression is induced in embryonic chick skin by the ectopic activation of BMP and FGF signaling. Misexpression of Dlx in the chick skin leads to both feather loss and feather bud fusions, suggesting that DLX proteins play a negative as well as a positive role in feather development. Moreover, DLX regulates the expression of NCAM and tenascin, molecules that are important for feather bud initiation as well as bud outgrowth and morphogenesis. Our results suggest that DLX transcription factors serve to integrate and transduce feather patterning signals to downstream effector molecules.

Animals↗

cDermo-1 misexpression induces dense dermis, feathers, and scales.

Reciprocal epithelio-mesenchymal interactions between the prospective epidermis and the underlying dermis are the major driving forces in the development of skin appendages. Feather development is initiated by a still unknown signal from the dermis in feather-forming skin. The morphological response of the ectoderm to this signal is the formation of an epidermal placode, which signals back to the mesenchyme to induce dermal condensations. Together, epidermal and dermal components constitute the outgrowing feather bud. The bHLH transcription factor cDermo-1 is expressed in developing dermis and is the earliest known marker of prospective feather tracts. To test its function during feather development, we forced cDermo-1 expression in embryonic chicken dermis using a retroviral expression vector. In featherless (apteric) regions, cDermo-1 misexpression induced dense, thickened dermis normally observed in feathered skin (pterylae), and leads to the development of regularly spaced and normally shaped ectopic feather buds. In pterylae, cDermo-1 misexpression enhanced feather growth. In hindlimb skin, according to the local skin identity, misexpression of cDermo-1 induced ectopic scale formation. Thus, we show that forced cDermo-1 expression in developing dermis is sufficient to launch the developmental program leading to skin appendage formation. We propose a role of cDermo-1 at the initial stages of feather induction upstream of FGF10.

Animals↗

Methionine and cystine requirements of slow- and fast-feathering male broilers from zero to three weeks of age.

Two experiments were conducted with fast- (Ross x 3F8) and slow- (Ross x 308) feathering broiler males from 0 to 3 wk of age to determine Met and Cys requirements. A corn-soybean meal basal diet was formulated to be deficient in Met and Cys but was adequate in all other nutrients (22.0% CP; 3,050 kcal ME/kg). In experiment 1, diets contained 0.50% dietary Cys with 0.35, 0.40, 0.45, and 0.50% total Met. Feed conversion (FC) of slow- and fast-feathering males improved in a similar manner to 0.50% Met (linear, P < 0.05). Nitrogen retention measured from 20 to 21 d of age optimized at 0.46% Met (quadratic, P < 0.01), regardless of feathering rate. Experiment 2 examined the response to feeding 0.35, 0.40, 0.45, and 0.50% total Cys in diets having total Met at 0.45%. Increasing Cys improved FC that optimized at 0.40% with fast-feathering birds (quadratic, P < 0.01), whereas slow-feathering broilers were not responsive. Nitrogen retention measured from d 20 to 21 did not indicate a difference attributable to feathering but a Cys optimization at 0.43% with both broiler sources. Present experimentation indicates a Met requirement approximating 0.50% is appropriate for broilers 0 to 3 wk of age, regardless of feather rate; however, the estimated Cys requirement for slow-feathering males (0.39%) was less than for fast-feathering (0.44%) males.

Animal Feed↗

A comparison of haemagglutination, haemagglutination inhibition and PCR for the detection of psittacine beak and feather disease virus infection and a comparison of isolates obtained from loriids.

Psittacine beak and feather disease (PBFD) is recognized as a threat for endangered psittacine birds in Australia, New Zealand and South Africa. Several diagnostic methods for the detection of beak and feather disease virus (BFDV) infection have been developed but there are few studies comparing the relative merits or sensitivity and specificity of each diagnostic test. In this report, the results of PCR, haemagglutination (HA) and haemagglutination inhibition (HI) testing of diagnostic samples collected from 679 samples from a range of psittacine bird species suspected of being infected with BFDV are summarized and compared. There was a strong agreement (kappa = 0.757; P<0.0001) between PCR and HA testing of feather samples and PCR-negative birds were 12.7 times more likely to have HI antibody than PCR-positive birds. False-positive HA results with titres up to 1:320 were identified in six feather samples that were PCR negative; the haemagglutination detected in these samples was not inhibited by anti-BFDV antisera and was removed by filtration through a 0.22 microm filter. Similarly, one false-negative PCR result was detected in a feather sample that had a high HA titre (>1:40,960) and four false-positive PCR results were detected in a batch of four feather samples. Of 143 birds that were feather PCR positive, only two had detectable HI antibody, and these birds were also feather HA negative, suggesting that they were developing immunity to recent infection. All birds with HI antibody were negative on feather HA testing. The assays confirmed BFDV infection in two endangered swift parrots (Lathamus discolor) and phylogenetic analysis of the sequence data generated from ORF V1 of these isolates provide further evidence of BFDV genotypes clustering in parallel with the Loriidae, Cacatuidae and Psittacidae.

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Permeability of synthetic and feather pillows to live house dust mites and house dust.

BACKGROUND: Previous studies have demonstrated significantly higher house dust mite (HDM) allergen levels from synthetic pillows, compared to feather pillows. Reasons for these differences could be lower permeability of feather pillow coverings to allergen in dust, greater HDM penetration of synthetic pillow covering, or both. OBJECTIVES: To determine the permeability of synthetic and feather pillow coverings to live HDMs and house dust. METHODS: Twenty live adult HDMs were seeded on top of two types of synthetic pillow covering (one standard polyester and one newer polyester/cotton type) and one type of feather pillow coverings with adequate food supply below in sealed culture dishes, kept at 23 degrees C and 70% relative humidity. After 24 and 48 h live HDM numbers remaining on top of the coverings were enumerated microscopically. Three aliquots of fine house dust (each in triplicate) were placed on top of the synthetic and feather pillow coverings, shaken gently for 30 min and penetrated dust was collected and weighed. RESULTS: After 24 h, all 20 HDMs had penetrated the standard synthetic pillow coverings, and no HDMs had penetrated either the feather pillow or the new synthetic pillow coverings after 24 or 48 h. Dust permeability (% of applied dust) for the standard synthetic, new type synthetic and feather pillow coverings were 0.88%, 0.07%, and 0.07%, respectively. This compared to 0.02% for a commercial occlusive pillow cover. CONCLUSIONS: These findings of total permeability of standard synthetic pillow coverings to live HDMs, and their greater permeability to house dust could explain their reported higher HDM allergen levels, compared to feather pillow coverings. Newer types of synthetic pillow coverings are similar to feather pillow coverings in their permeability to live HDMs and house dust.

Allergens↗

Adhesion molecules in skin development: morphogenesis of feather and hair.

Figure 9 summarizes the morphogenetic process of feather and hair. Hair of feathers are formed from a layer of homogeneously distributed mesenchymal cells. The mesenchymal cells start to condense to form foci in response to some unidentified induction signal (Fig. 9B). Several adhesion molecules, including L-CAM, N-CAM, integrin, tenascin, as well as proteoglycan, are involved. These adhesion molecules appear to have different roles in this process, because perturbation with specific antibodies leads to different aborted patterns. Hair or feather follicles then form following cell proliferation and epithelial invagination (Fig. 9C). The dermal papilla is enriched with N-CAM and tenascin, whereas the feather collar (equivalent of hair matrix) is enriched with L-CAM and PDGF receptor. Epithelial cells in the feather collar receive a signal from the dermal papilla and are able to continue to divide. Several growth factors, such as PDGF and EGF, may be involved. As epithelial cells are pushed upwards, they differentiate and keratinize in a cylindrical structure into hair. In feather, another morphogenetic event takes place to form the branched structure. The epithelial cylinder of the feather shaft invaginates to form rows of cells that die to become space and create the secondary branch or barbs (Fig. 9D). N-CAM is enriched in the cells destined to die and appears to form the border of cell groups within which the "death signal" is transmitted. In some, but not all, feathers the same process is repeated, in a way analogous to fractal formation, to form the tertiary branches or the barbules (Fig. 9E). Thus, in each step of the morphogenesis of feather and hair, different adhesion molecules are expressed and are involved in different functions: induction, mesenchymal condensation, epithelial folding, and cell death, depending on different scenarios. We have just begun to elucidate these molecular events.

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The pathology of psittacine beak and feather disease.

Psittacine beak and feather disease is characterised by loss of feathers, abnormally shaped feathers and overgrowth and irregularity of the surface of the beak. The disease occurs in a number of psittacine species including the Sulphur-crested Cockatoo, Lovebirds , Budgerigars and Galahs . The abnormal appearance of feathers and beak is due to a dystrophic process within the epidermis of the feather and beak. The process consists of epidermal cell necrosis, epidermal hyperplasia and hyperkeratosis. Many of the feather abnormalities are due to retention of a hyperkeratotic feather sheath. A characteristic microscopic finding is the presence of macrophages containing purple intracytoplasmic inclusions in affected epidermis and feather pulp. The inclusions consist of aggregates of particles 17 to 22 nm in diameter. Similar but smaller inclusions occur in epidermal cells. In addition, non-suppurative inflammation occurs in the feather pulp. The findings are suggestive of a viral infection.

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Quail-duck chimeras reveal spatiotemporal plasticity in molecular and histogenic programs of cranial feather development.

The avian feather complex represents a vivid example of how a developmental module composed of highly integrated molecular and histogenic programs can become rapidly elaborated during the course of evolution. Mechanisms that facilitate this evolutionary diversification may involve the maintenance of plasticity in developmental processes that underlie feather morphogenesis. Feathers arise as discrete buds of mesenchyme and epithelium, which are two embryonic tissues that respectively form dermis and epidermis of the integument. Epithelial-mesenchymal signaling interactions generate feather buds that are neatly arrayed in space and time. The dermis provides spatiotemporal patterning information to the epidermis but precise cellular and molecular mechanisms for generating species-specific differences in feather pattern remain obscure. In the present study, we exploit the quail-duck chimeric system to test the extent to which the dermis regulates the expression of genes required for feather development. Quail and duck have distinct feather patterns and divergent growth rates, and we exchange pre-migratory neural crest cells destined to form the craniofacial dermis between them. We find that donor dermis induces host epidermis to form feather buds according to the spatial pattern and timetable of the donor species by altering the expression of members and targets of the Bone Morphogenetic Protein, Sonic Hedgehog and Delta/Notch pathways. Overall, we demonstrate that there is a great deal of spatiotemporal plasticity inherent in the molecular and histogenic programs of feather development, a property that may have played a generative and regulatory role throughout the evolution of birds.

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Gradients of homeoproteins in developing feather buds.

Homeoproteins are functionally involved in pattern formation. Recently, homeoproteins have been shown to be distributed in a graded fashion in developing limb buds. Here we examine the expression of homeoproteins in chicken feather development by immunocytochemical localization. We find that XlHbox 1 antigen is present in cell nuclei and is distributed in a gradient in the mesoderm of developing feather buds, with strongest expression in the anterior-proximal region. The gradient is most obvious in feather buds from the mid-trunk level. Feather buds from the scapular level express very high levels of XlHbox 1 and feather buds from the caudal region express no XlHbox 1, suggesting that a broad gradient along the body axis is superimposed on a smaller gradient within each individual feather bud. Feather ectoderm also expresses XlHbox 1 antigen but without an obvious graded pattern. Another homeoprotein, Hox 5.2, is also expressed in developing feather buds in a graded way, and its distribution pattern is partially complementary to that of XlHbox 1. These observations suggest that homeoproteins may be involved in setting up the anteroposterior polarity of cell fields at different levels, first for the body axis, then for the limb axis and finally for the feather axis.

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Effect of feather meal as a source of valine for lactating sows.

An experiment was conducted to evaluate feather meal as a source of Val in lactating sow diets. Sows (five farrowing groups; mean parity = 2.34) were allotted to one of two dietary treatments on the basis of ancestry, parity, and weight and date of d 110 of gestation. The treatment diets included 1) corn-soybean meal lactation diet (n = 40) or 2) corn-soybean meal lactation diet with 2.5% feather meal (n = 39). The diets were formulated on an equal Lys basis. All litters were adjusted to 10 pigs within 24 h after farrowing, and all sows weaned at least nine pigs. Sows were bled at 110 d of gestation and at weaning, and serum urea N was determined. Backfat thickness was determined ultrasonically at 110 d of gestation and at weaning. Serum urea N and backfat thickness at d 110 of gestation were used as covariates for serum urea N and backfat thickness at weaning, respectively. The litter response criteria (weaning weight, litter weight gain, and percentage survival) were not affected (P > .10) by feather meal. The sow response criteria (weaning weight, weight loss per day, weaning backfat thickness, change in backfat thickness, ADFI, and days to estrus) were not affected (P > .10) by feather meal. Sows fed feather meal had increased (P < .01) serum urea N and tended (P = .15) to have decreased sow weaning weight. Following the initial analysis of the data, the data set was split into two groups: 1) sows with litters gaining less than 2.17 kg/d (n = 19 and 20 for control and feather meal diets, respectively) and 2) sows with litters gaining more than 2.17 kg/d (n = 21 and 19 for control and feather meal diets, respectively). These two groups were analyzed separately. In sows with litters gaining less than 2.17 kg/d, the litter and sow criteria were not affected (P > .10) by treatment. In sows with litters gaining more than 2.17 kg/d, sow weaning weight was decreased (P < .04) and sow weight loss (P < .02) and serum urea N (P < .01) were increased in sows fed feather meal. Feather meal (as a source of Val) did not improve litter weight gain, but it increased serum urea N.

Animal Feed↗

Profiles of pulp infiltrating lymphocytes at various times throughout feather regeneration in Smyth line chickens with vitiligo.

Smyth line (SL) chickens develop a spontaneous, autoimmune, posthatch loss of pigment cells (vitiligo) in regenerating feather tissue. Smyth line vitiligo (SLV) is associated with lymphocyte infiltrations prior to and throughout the development of the disorder. It was the purpose of this study to determine the type, relative amounts, and proportions of pulp-infiltrating lymphocytes at various times throughout the growth of regenerating feathers. Feathers were plucked from 8-week-old chickens with and without SLV. Feather pulp cell suspensions were prepared when the regenerating feathers were 2, 3, 4, and 6 weeks of age. Cells were fluorescently labeled using a panel of mouse monoclonal antibodies specific for chicken lymphocytes. Both T and B cells infiltrated the feather pulp of chickens with SLV. T cell levels remained elevated throughout the 6 weeks of feather growth, while B cell levels steadily declined to control levels over the same time. The pulp-infiltrating cells were primarily T cells with an alphabeta T cell receptor expressing the Vbeta1 gene (TCR2+). The ratio between CD4+ and CD8+ cells was 1.42 and 0.75 in 2- and 6-week-old regenerating feathers from chickens with autoimmune SLV, respectively. In non-vitiliginous chickens this ratio was always near 1. These data suggest that TCR2+ T cells play an important role in SLV. CD4+ cells may play a recruiting/activating role, whereas CD8+ cells may have cytotoxic activity specifically directed against melanocytes. Additionally, this is the first report demonstrating the infiltration of B cells into the feather pulp of vitiliginous chickens. These B cells may directly/indirectly contribute to melanocyte destruction in SLV.

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Horse hooves and bird feathers: Two model systems for studying the structure and development of highly adapted integumentary accessory organs--the role of the dermo-epidermal interface for the micro-architecture of complex epidermal structures.

Accessory organs of the integument are locally modified parts of the potentially feather-bearing skin in birds (e.g., the rhamphotheca, claws, or scales), and of the potentially hairy skin in mammals (e.g., the rhinarium, nails, claws, or hooves). These special parts of the integument are characterised by a modified structure of their epidermal, dermal and subcutaneous layers. The developmental processes of these various integumentary structures in birds and mammals show both similarities and differences. For example, the development of the specialised epidermal structures of both feathers and the hoof capsule is influenced by the local three-dimensional configuration of the dermis. However, in feathers, in contrast to hooves, the arrangement of the corneous cells is only partially a direct result of the particular arrangement and shape of the dermal surface of the papillary body. Whereas the diameter of the feather papilla, as well as the number, length, and width of dermal ridges on the surface of the feather papilla influence the three-dimensional architecture of the feather rami, there is no apparent direct correlation between the dermo-epidermal interface and the development of the highly ordered architecture of the radii and hamuli in the feather vane. In order to elucidate this morphogenic problem and the problem of locally different processes of keratinisation and cornification, the structure and development of feathers in birds are compared to those of the hoof capsule in horses. The equine hoof is the most complex mammalian integumentary structure, which is determined directly by the dermal surface of the papillary body. Perspectives for further research on the development of modified integumentary structures, such as the role of the dermal microangioarchitecture and the selective adhesion and various differentiation pathways of epidermal cells, are discussed.

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Possible control mechanisms of feather follicle movement in the pectoral tract of the chicken.

Feather follicle movement control was studied on feathers of the pectoral tract in the anaesthetized chicken. Dissection of nerves leading to the follicles showed that their origin was at least partially in the sympathetic ganglia. Reflectoric ruffling of feathers could be obtained after adequate stimulation. Intravenous injections of drugs in doses which influenced the circulation gave the following results: Adrenaline caused erection of feathers which was abolished by phentolamine. Noradrenaline caused up and down movements after injection only at high doses. Similar results were obtained by placing of skin pieces in saline containing the drugs. The reaction to drugs was typical always for a given group of feathers. Killing of birds and anaesthesia caused general erection of feathers which was not abolished by phentolamine. The findings suggest that adrenergic synapses are involved in feather follicle movement control, but at least another mechanism (CNS) regulates feather follicle movement.

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Estimating the latitudinal origins of migratory birds using hydrogen and sulfur stable isotopes in feathers: influence of marine prey base.

Hydrogen stable isotope analysis of feathers is an important tool for estimating the natal or breeding latitudes of nearctic-neotropical migratory birds. This method is based on the latitudinal variation of hydrogen stable isotope ratios in precipitation in North America (deltaD(p)) and the inheritance of this variation in newly formed feathers (deltaD(f)). We hypothesized that the typically strong relationship between deltaD(p) and deltaD(f) would be decoupled in birds that forage in marine food webs because marine waters have relatively high deltaD values compared to deltaD values for local precipitation. Birds that forage on marine prey bases should also have feathers with high delta(34)S values, since delta(34)S values for marine sulfate are generally higher than delta(34)S values in terrestrial systems. To examine this potential marine effect on feather stable isotope ratios, we measured deltaD and delta(34)S in the feathers of nine different species of raptors from both inland and coastal locations across North America. Feathers from coastal bird-eating raptors had consistently higher deltaD and delta(34)S values than feathers from inland birds. Birds that had high delta(34)S values also deviated strongly from the typical relationship between deltaD(p) and deltaD(f). We recommend measuring both sulfur and hydrogen stable isotope ratios in feathers when some members of a migrant population could potentially forage in marine habitats. We suggest using a practical cutoff of delta(34)S >10 per thousand to remove marine-foraging birds from a migrant sample when using stable isotopes of hydrogen to estimate the latitudinal origins of migrants because high deltaD(f) values for marine-foraging birds could potentially distort estimates of origins.

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Avian feather development: relationships between morphogenesis and keratinization.

Morphogenesis and expression of the alpha and beta keratin polypeptides are controlled by epidermal-dermal interactions during development of avian skin derivatives. We have examined the relationship between morphogenesis of the embryonic feather and expression of the feather alpha and beta keratins by routine histology, indirect-immunofluorescence, and SDS-PAGE. Initially beta keratins are expressed only in the feather sheath. Following barb ridge morphogenesis beta keratins can be detected in the barb ridge, coincident with the differentiation of barb ridge cells into eight distinct morphological types. Beta keratinization occurs in gradients; from feather apex to base, and from periphery of the barb ridge to the interior. The onset of beta keratinization in the barb ridges is paralleled by an increase in the major feather beta keratin polypeptides, as detected by SDS-PAGE. The alpha keratins are present in both the periderm and feather sheath at early stages of feather development, but become greatly reduced after hatching, when the down feather emerges from the sheath.

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Altered proteoglycan synthesis disrupts feather pattern formation in chick embryonic skin.

We have tested the role of proteoglycans in the development of feather pattern by culturing 7-day-old embryonic chick skins on medium containing para-nitrophenyl-beta-D-xyloside (2 mM). Xylosides compete with core proteins of proteoglycans by acting as exogenous acceptors for the synthesis of glycosaminoglycans leading to the synthesis of under- or unglycosylated core proteins and free glycosaminoglycans. We have demonstrated that xyloside treatment alters the structure of the proteoglycans synthesized by embryonic skin and disrupts the feather pattern. The altered pattern is seen as the fusion of individual feather rudiments. Fusion can occur diagonally, and in an anteroposterior and mediolateral direction. The effect induced by the disruption of proteoglycan structure takes place during the first 24 hr of culture during which time all the rows of feather rudiments are being established. The effect is reversible if the skins are returned to control medium after 24 hr but not after 48 hr of treatment with xyloside. Once established during the first 24 hr the feather pattern can only be slightly distorted by the xyloside treatment. The results are interpreted to mean that proteoglycans play a developmental role in the establishment of the feather pattern but not in its maintenance, suggesting that the two processes are under different developmental control. The altered feather pattern obtained by disrupting proteoglycan structure is highly similar to that obtained when skins are cultured in the presence of antibodies to L-CAM (W.J. Gallin, C.-M., Chuong, L.H. Finkel, and G.M. Edelman (1986), Proc. Natl. Acad. Sci. USA 83, 8235-8239). This observation suggests that there may be a functional relationship between the extracellular matrix and cell adhesion molecules in the establishment of feather pattern.

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Preen gland function in layer fowls: factors affecting morphology and feather lipid levels.

The effects of age and external factors on preen gland morphology and feather lipid concentrations in layer fowl (Gallus domesticus) have not previously been described. This work examines how floor substrate, whether a bird is feather pecked or not, and age influence both gross and histological morphology. Furthermore, the effects of age, beak trimming and floor substrate on feather lipid concentration (mg lipid per g feathers) are investigated. Neither absolute nor relative preen gland weight were reliably affected by floor substrate or feather pecked status. However, these parameters were strongly affected by changes in bird age and related body weight changes. The microstructures of the gland also increased consistently with age. Feather lipid concentration generally increased with age. Feather lipid was strongly influenced by the presence or absence of a dustbathing substrate. Of all the parameters tested here, the age and growth of the bird had the greatest effects on preen gland development. Feather lipid concentration is also influenced by age, and by the presence or absence of a dustbathing substrate.

Aging↗

Novel applications of feather tip extracts from MDV-infected chickens; diagnosis of commercial broilers, whole genome separation by PFGE and synchronic mucosal infection.

Marek's disease virus (MDV) productive replication occurs in the feather follicle epithelium and the feather tips are valuable both for research and disease diagnosis. Three novel applications of feather tip extracts are described now: (A). As a source of DNA for amplifying either MDV and/or ALV-J. In two clinical situations a marked advantage was obtained compared to blood and organs; in broiler breeder flocks with a mixed MDV and ALV-J infection, and in young broilers with neurological Marek's disease (MD). (B). Separation of the large ( approximately 200 kbp) MDV genome directly from the infected chickens. Using pulsed field gel electrophoresis, the DNA extracted from tumors or feather tips was separated and hybridized to a 132 bp tandem repeat MDV probe. Compared to 2/55 polymerase chain reaction (PCR) positive tumor samples, 15/61 feather tip extracts contained whole MDV genomes. (C). Experimental MDV infection was induced by the mucosal route by dripping feather tip extract to the eye and mouth of the bird. That attempted to reproduce the native infection process, however the use of extracts, instead of dry feather dust was a compromise, aimed to synchronize the infection. In one trial, tumors were induced 6 weeks after dripping day-old broilers, while in another, feather tips were PCR positive 16 days after dripping of 2-month-old layers.

Animals↗