Isolation, identification and genome analysis of an avian hepatitis E virus from white-feathered broilers in China.

Avian hepatitis E virus (HEV) is the major causative pathogen of the big liver and spleen disease, hepatitis-splenomegaly syndrome, and hepatic rupture hemorrhage syndrome. Until now, there are 6 different avian HEV genotypes that infect chickens have been reported worldwide. Epidemiologic investigations of the avian HEV demonstrated that avian HEV has been widely spread in China in recent years. In this study, an avian HEV named YT-aHEV was obtained from white-feathered broilers using LMH cells by virus isolation assay in Shandong province, China. The complete genome consists of 6656-nt excluding the poly(A) tail. The isolate was highly similar to the CaHEV strain and segregated into the same branch belonging to avian HEV genotype 3. Indirect immunofluorescence using capsid protein-specific polyclonal antibodies confirmed that YT-aHEV could establish productive infection and replicate stably in LMH cells. Furthermore, an in vivo avian HEV infection model was established successfully in specific pathogen-free chicken embryos by intravenous experiments. In the present study, we demonstrate an avian HEV infection associated with liver lesions of hemorrhage and swelling by LMH cells for the first time in a white-feather broiler flock in China. This research also provides a new diagnosis method for detection of avian HEV, which laid a foundation for the understanding of pathogenicity and molecular biology of this virus for further study.

Correlation between goose circovirus and goose parvovirus with gosling feather loss disease and goose broke feather disease in southern Taiwan.

BACKGROUND: Goslings in several Taiwanese farms experienced gosling feather loss disease (GFL) at 21-35 days and goose broke feather disease (GBF) at 42-60 days. The prevalence ranges from a few birds to 500 cases per field. It is estimated that about 12,000 geese have been infected, the morbidity is 70-80% and the mortality is 20-30%. OBJECTIVES: This study aims to investigate the pathogens that cause GFL and GBF. Focus on the study of the correlation between goose circovirus (GoCV) and goose parvovirus (GPV) with the goose feather loss in southern Taiwan. Furthermore, a phylogenetic tree was established to align the differences between southern and northern Taiwan and compare with virus strains from China and Europe. METHODS: Samples were collected from animal hospitals. Molecular and microscopy diagnostics were used to examine 92 geese. Specific quantitative polymerase chain reaction (Q-PCR) assays are performed to evaluate GPV and GoCV viral loads and simultaneously evaluated the feather loss conditions in geese with the scoring method. RESULTS: High prevalence of GoCV and GPV infection in geese showing signs of GFL and GBF. Inclusion body was detected in the feather follicles and Lieberkühn crypt epithelial cells. The Q-PCR showed the high correlation between feather loss and viruses during 3rd-5th week. However, the infection was not detected using the same test in 60 healthy geese. CONCLUSIONS: Thus, GFL and GBF appear to be significantly closely related to GoCV and GPV. The geese feathers showed increasing recovery after being quarantined and disinfected.

A case of clubbed down syndrome in broilers.

This study presents a case of clubbed down syndrome in conventional broilers. During the first week of life, severe growth retardation was observed in approximately 25% of the flock. The growth-retarded chicks weighed only 45 g and showed a typical feather disorder which was most apparent on their abdomen and was defined in literature as typical for clubbed down syndrome. Necropsies, histology, biochemical analysis of blood and liver samples, serology and different PCR tests were performed in broilers to assess the aetiology of the clinical signs that were present in the affected broiler farm. Because of the suspicion of a possible link with the broiler-breeder farms, different investigations including serology, PCR and feed analysis were also performed on these farms. The results suggest that an accidentally excessive amount of calcium and iron in the feed of broiler-breeders, 3 weeks prior to first clinical signs in broilers, led to the development of clubbed down in the offspring, because of a relative Zn-deficiency in broiler-breeders and an absolute Zn-deficiency in the hatching eggs that were produced during this period. This appeared to be a reversible process as no clinical signs were observed in younger offspring of these broiler-breeders after they had consumed more of the new batch of feed. A potential involvement of Astrovirus could not be completely ruled out. This study demonstrates the importance of correct mineral concentrations in broiler-breeder feed and the impact it can have on the development of the offspring.

Comparison of selected indices of internal environment and condition of laying hens kept in furnished cages and in aviaries.

Biochemical and hematological examination of blood and individual assessment of the birds were performed in Lohman Brown laying hens at 45 weeks of age housed in different systems. The biochemical examination revealed higher (p < .01) corticosterone levels, creatine kinase, and aspartate aminotransferase activity and lower (p < .01) levels of lactate, triglycerides, albumin, calcium, and phosphorus in aviary hens compared to hens housed in furnished cages. Hematological examination of hens housed in aviaries revealed higher (p < .05) hematocrit, leukocytes, heterophils, eosinophils, basophils, monocytes, and H/L ratio. Furthermore, hens housed in aviaries had lower (p < .01) body weight than hens in furnished cages, they were worse feathered (p < .001), had more damaged combs (p < .05), and poorer physical condition (p < .01). In contrast, caged hens showed worse (p < .01) feather condition of the wings due to abrasion and claws due to overgrowth. The results have shown that the housing system has a significant impact on the internal environment and condition of birds and that housing in aviaries without taking into account the specifics of such housing may lead to significant stress and disturbance to the welfare of laying hens.

Identification and Characterization of a Distinct Strain of Beak and Feather Disease Virus in Southeast China.

Beak and feather disease virus (BFDV) is an infectious agent responsible for feather degeneration and beak deformation in birds. In March 2017, an epidemic of psittacine beak and feather disease (PBFD) struck a farm in Fuzhou in the Fujian Province of southeast China, resulting in the death of 51 parrots. In this study, the disease was diagnosed and the pathogen was identified by PCR and whole genome sequencing. A distinct BFDV strain was identified and named as the FZ strain. This BFDV strain caused severe disease symptoms and pathological changes characteristic of typical PBFD in parrots, for example, loss of feathers and deformities of the beak and claws, and severe pathological changes in multiple organs of the infected birds. Phylogenetic analysis showed that the FZ strain was more closely related to the strain circulating in New Caledonia than the strains previously reported in China. Nucleotide homology between the FZ strain and other 43 strains of BFDV ranged from 80.0% to 92.0%. Blind passage experiment showed that this strain had limited replication capability in SPF Chicken Embryos and DF-1 Cells. Furthermore, the capsid (Cap) gene of this FZ strain was cloned into the pGEX-4T-1 expression vector to prepare the polyclonal anti-Cap antibody. Western blotting analysis using the anti-Cap antibody further confirmed that the diseased parrots were infected with BFDV. In this study, a PBFD and its pathogen was identified for the first time in Fujian Province of China, suggesting that future surveillance of BFDV should be performed.

A cross-sectional study on feather cover damage in Canadian laying hens in non-cage housing systems.

BACKGROUND: Feather damage (FD) resulting from feather pecking remains a concern in non-cage housing systems for laying hens worldwide. This study aimed to identify bird-, housing-, and management-related factors associated with FD in non-cage housing systems as the egg production sector phases out the conventional cage system in Canada. A survey on housing and management practices was developed and distributed to 122 laying hen farms where 39 respondents provided information on non-cage flocks. Farmers visually assessed 50 birds throughout the barn for FD using a 0-2 scoring scale according to severity. Prevalence of FD was calculated as the percentage of birds with any form of FD (score > 0). Multivariable linear regression modeling was used to identify factors associated with FD prevalence. RESULTS: Six variables were included in the final model and accounted for 64% of the variation in FD between farms. FD prevalence was higher with increasing flock age (0.9% ± 0.29) and when birds were housed in all wire/slatted barns compared with all litter barns (37.6% ± 13.1). Additionally, FD prevalence tended to be higher in barns with manure removal only after depopulation (20.1% ± 10.70). Enrichment also tended to be associated with higher FD (19.1% ± 8.04), possibly indicating that it was provided after FD was observed as a control measure, or, was not efficient in reducing the development of FD. CONCLUSIONS: These findings emphasize the role of litter provision and management (e.g., manure removal effects on air quality), and its potential impact on FD among laying hens in non-cage housing systems in Canada. Further longitudinal and/or intervention studies are needed to assess the potential of the identified factors to function as a management strategy to prevent or reduce FD in non-cage housed laying hens.

Rectricectomy With Pygostylectomy for Resolution of Feather Trauma in an Umbrella Cockatoo (Cacatua alba).

Feather trauma is a common problem among pet birds, especially those with trimmed wing feathers, and often affects remiges and rectrices. Damage can become chronic if new feathers remain unprotected by fully formed adjacent feathers. The follicles can become damaged and poorly functional over time. A 6-month-old male umbrella cockatoo (Cacatua alba) was presented for examination because of feather damage that occurred shortly after receiving a wing trim by the owner. Over the next 18 months, various medical, behavioral, and husbandry methods failed to allow normal regrowth of the tail feathers. The bird presented 25 times over the 18-month period for problems associated with broken blood feathers in the tail. Feather imping was ruled out because of the lack of hollow adult shafts to serve as a base. Diagnostic tests included hematologic testing and biochemical analysis, circovirus testing, feather biopsy, and radiographs, results of which were unremarkable. Because of the chronic pain associated with the frequent rectrix damage, a complete rectricectomy was performed. All of the rectrical follicles as well as the pygostyle were surgically removed. Healing was uneventful and the bird has been clinically normal with no further blood feather injury for 24 months after surgery.

Lameness and its relationship with health and production measures in broiler chickens.

The aim of this study was to explore lameness and the associations between lameness and health/production measures of animal welfare in commercial broiler production, using the Welfare Quality® protocol for broilers. A total of 50 flocks were included in the sample and farm visits were conducted for lameness scoring at a mean age of 28.9 days. The percentage of animals (n=7500) in the six different gait score (GS) categories were GS0: 2.53%, GS1: 44.19%, GS2: 33.84%, GS3: 16.32%, GS4: 2.36% and GS5: 0.53%. Production and other welfare data were collected for each flock after slaughter. Higher gait scores were associated with increased hock burn score (P<0.02), increased footpad dermatitis score (P<0.01), reduced bird cleanliness score (P<0.01) and peat litter (P<0.01). Although not statistically significant, there was a tendency for increased flock gait score being associated with wet litter (P=0.07). In addition, condemnations at postmortem inspection were associated with increasing gait scores (P<0.05), indicating that at least a portion of the lameness cases display pathological changes on the carcasses. In conclusion, 19%of the birds showed moderate-to-severe lameness, which was associated with several production or health and welfare observations including feather cleanliness and condemnations as unfit for human consumption at slaughter. Although stocking density and growth rate are already known key factors for lameness, associations of lameness with hock burns, footpad dermatitis and cleanliness of the birds suggest that a suboptimal physical environment (e.g. litter- and air quality) may be detrimental to leg health. Further studies are needed to explore these associations in more detail.

A novel deletion in KRT75L4 mediates the frizzle trait in a Chinese indigenous chicken.

BACKGROUND: Highly diversified in morphology and structure, feathers have evolved into various forms. Frizzle feathers, which result from a developmental defect of the feather, are observed in several domestic chicken breeds. The frizzle phenotype is consistent with incomplete dominance of a major gene, but the molecular mechanisms that underlie this phenotype remain obscure. Kirin, a Chinese indigenous chicken breed that originated in the Guangdong province, is famous for its frizzle feathers. The KRT75 gene is considered as the dominant gene responsible for the frizzle trait in several chicken breeds, but this is not the case in the Kirin breed. Thus, the objective of our study was to investigate the genomic region and mutation responsible for this phenotype in this particular breed. RESULTS: A resource population was produced by crossing Kirin and Huaixiang chickens to produce F1 and F2 generations. DNA samples from 75 frizzle feather and normal feather individuals were sequenced with double-digest genotyping by sequencing (dd-GBS). After the detection of 525,561 high-quality variants, a genome-wide association analysis was carried out and the gene responsible for the frizzle phenotype was localized within the type II α-keratin cluster on chromosome 33. Sanger sequencing was used to screen for mutations in the exons of five genes of this type II α-keratin cluster. A 15-bp deletion in exon 3 of KRT75L4 that showed complete segregation with the frizzle phenotype was detected within the F2 population. Transcriptome sequencing demonstrated that KRT75L4 was expressed but that the transcript was shorter in Kirin than in Huaixiang chickens. In addition, by using Sanger sequencing, we were able to confirm that the deletion was in complete linkage with frizzle feathers. CONCLUSIONS: A deletion in the KRT75L4 gene is responsible for the frizzle feather phenotype in the Kirin chicken. The identification of this mutation, which causes a developmental defect of avian integument appendages, will improve our understanding of the mechanisms that are involved in feather formation.

A Retrospective Study of Horner Syndrome in Australian Wild Birds, 2010-2016.

Horner syndrome was identified in 25 of 30 777 avian admissions to Currumbin Wildlife Hospital during 2010-2016. Unilateral ptosis and erection of facial feathers were distinct findings on physical examination and consistent across 9 species. Affected birds were biased toward adults (64%) suffering traumatic injuries (88%). Concurrent injuries requiring treatment were present in 38% of cases, and 76% had additional neurologic deficits. Prognosis for release was poor, with an overall success rate of 32%. Resolution of clinical signs increased to 44% with higher doses of meloxicam and required an average hospitalization of 22 days (range, 3-78 days). Further investigation of the underlying causes of Horner syndrome in birds to provide treatment and prognostic guidelines is warranted.

The effects of a reduced balanced protein diet on litter moisture, pododermatitis and feather condition of female broiler breeders over three generations.

Protein content reduction in broiler breeder diets has been increasingly investigated. However, broiler breeders reared on low protein diets are characterized by a deterioration of the feather condition. Furthermore, polydipsia induced by controlled feed intake increases litter moisture and as a consequence pododermatitis. This project aimed to study the litter moisture, pododermatitis and feather condition of breeders fed with a 25% reduced balanced protein (RP) diet during the rearing and laying period over three successive generations. The experiment started with two treatments for the F0 generation: control (C) group fed with standard C diets and RP group fed with RP diets. The female F0-progeny of each treatment was divided into the two dietary treatments as well, resulting in four treatments for the F1 generation: C/C, C/RP, RP/C and RP/RP (breeder feed in F0/F1 generation). The RP diet fed breeders received on average 10% more feed than C diet fed breeders to achieve the same target BW. The female F1-progeny of each treatment were all fed with C diets which resulted in four treatments for the F2 generation: C/C/C, C/RP/C, RP/C/C and RP/RP/C (breeder feed in F0/F1/F2 generation). Litter moisture, footpad and hock dermatitis were recorded at regular intervals throughout the experimental period in all three generations. For the F0 and F1 generation, the pens of breeders receiving C diets had significantly higher litter moisture than the RP diets fed groups (P<0.05), resulting in an elevated footpad dermatitis occurrence (FDO) (P<0.05). No difference was found in the F2 generation. The feather condition was scored during the laying period for each generation. F0 and F1 breeders reared on the RP diets had poorer feather condition than those receiving the C diets (P<0.05). The C/RP breeders had a significantly poorer feather condition than RP/RP breeders (P<0.05). For the F2 generation, RP/RP/C breeders had a significantly better feather condition compared with the other three groups (P<0.05). The RP/C/C breeders were significantly better feathered than C/C/C breeders (P<0.05). In conclusion, providing RP diets to broiler breeders improved litter condition and hence reduced FDO whereas impaired feather condition. Furthermore, positive transgenerational effects of the maternal RP diets on the feather condition may be inferred, hence potentially altering the welfare status.

The Feather Model for Chemo- and Radiation Therapy-Induced Tissue Damage.

Chemo- and radiation therapy are the main modalities for cancer treatment. A major limiting factor is their toxicity to normal tissue, thus reducing the dose and duration of the therapy. The hair follicle, gastrointestinal tract, and hematopoietic system are among the target organs that often show side effects in cancer therapy . Although these organs are highly mitotic in common, the molecular mechanism of the damage remains unclear. The feather follicle is a fast-growing mini-organ, which allows observation and manipulation on each follicle individually. As a model system, the feather follicle is advantageous because of the following reasons: (1) its complex structure is regulated by a set of evolutionarily conserved molecular pathways, thus facilitating the effort to dissect the specific signaling events involved; (2) its morphology allows the continuity of normal-perturbed-normal structure in a single feather, thus "recording" the damaging effect of chemo- and radiation therapy; (3) further histological and molecular analysis of the damage response can be performed on each plucked feather; thus, it is not necessary to sacrifice the experimental animal. Here, we describe methods of applying the feather model to study the molecular mechanism of chemo- and radiation therapy-induced tissue damage.

Assessment of the effect of housing on feather damage in laying hens using IR thermography.

Plumage damage represents one of the animal-based measures of laying hens welfare. Damage occurs predominantly due to age, environment and damaging pecking. IR thermography, due to its non-invasiveness, objectivity and repeatability is a promising alternative to feather damage scoring systems such as the system included in the Welfare Quality ® assessment protocol for poultry. The aim of this study was to apply IR thermography for the assessment of feather damage in laying hens kept in two housing systems and to compare the results with feather scoring. At the start of the experiment, 16-week-old laying hens (n=30) were divided into two treatments such as deep litter pen and enriched cage. During 4 months, feather damage was assessed regularly in 2-week intervals. One more single assessment was done nine and a half months after the start of the experiment. The feather damage on four body regions was assessed by scoring and IR thermography: head and neck, back and rump, belly, and underneck and breast. Two variables obtained by IR thermography were used: the difference between the body surface temperature and ambient temperature (ΔTB) and the proportion of featherless areas, which were defined as areas with a temperature >33.5°C. Data were analyzed using a GLM model. The effects of housing, time, region and their interactions on feather damage, measured by the feather scoring and by both IR thermography measures, were all significant (P<0.001). The ΔTB in all assessed regions correlated positively with the feather score. Feather scoring revealed higher damage in enriched cages compared with deep litter pens starting from week 6 of the experiment on the belly and back and rump regions, whereas ΔTB from week 6 in the belly and from week 8 on the back and rump region. The proportion of featherless areas in the belly region differed significantly between the housings from week 8 of the experiment and on the back and rump region from week 12. The IR thermography assessment of the feather damage revealed differences between hens kept in different housing systems in agreement with the feather scoring. In conclusion, it was demonstrated that the IR thermography is a useful tool for the assessment of poultry feather cover quality that is not biased by the subjective component and provides higher precision than feather damage scoring.

Defenses against keratinolytic bacteria in birds living in radioactively contaminated areas.

Microorganisms have shaped the evolution of a variety of defense mechanisms against pathogenic infections. Radioactivity modifies bacterial communities and, therefore, bird hosts breeding in contaminated areas are expected to adapt to the new bacterial environment. We tested this hypothesis in populations of barn swallows (Hirundo rustica) from a gradient of background radiation levels at Chernobyl and uncontaminated controls from Denmark. Investment in defenses against keratinolytic bacteria was measured from feather structure (i.e., susceptibility to degradation) and uropygial secretions. We studied degradability of tail feathers from areas varying in contamination in laboratory experiments using incubation of feathers with a feather-degrading bacterium, Bacillus licheniformis, followed by measurement of the amount of keratin digested. The size of uropygial glands and secretion amounts were quantified, followed by antimicrobial tests against B. licheniformis and quantification of wear of feathers. Feathers of males, but not of females, from highly contaminated areas degraded at a lower rate than those from medium and low contamination areas. However, feathers of both sexes from the Danish populations showed little evidence of degradation. Individual barn swallows from the more contaminated areas of Ukraine produced the largest uropygial secretions with higher antimicrobial activity, although wear of feathers did not differ among males from different populations. In Denmark, swallows produced smaller quantities of uropygial secretion with lower antimicrobial activity, which was similar to swallow populations from uncontaminated areas in Ukraine. Therefore, barn swallows breeding in contaminated areas invested more in all defenses against keratinolytic bacteria than in uncontaminated areas of Ukraine and Denmark, although they had similar levels of feather wear. Strong natural selection exerted by radioactivity may have selected for individuals with higher defense capacity against bacterial infections during the 30 years since the Chernobyl disaster.

The effects of eggshell temperature fluctuations during incubation on welfare status and gait score of broilers.

The aim of the current study was to determine the effects of different eggshell temperatures (EST); low (33.3 to 36.7°C), control (37.8 to 38.2°C), and high (38.9 to 40.0°C) during 10 to 18 days of incubation on welfare status including foot pad dermatitis (FPD), hock dermatitis (HD) and feathering status, and gait score in broilers. Score 2, 4, and 5 of FPD were found to be similar among the treatment groups, whereas a score of 3 was found to be higher in the control and high EST groups (27.7% and 29.2%) compred to the low EST group (16.9%). The eggshell temperature fluctuations were significantly affected the incidence of HD, whereas broiler sex did not. All of the broilers in the high EST group had HD with various scores, while a percentage of 21.1% and 6.9% of broilers had the score 1 of HD in the low and control EST groups, respectively. Feathering status showed a difference between body parts including wing, neck, back, and vent and also a general mean score of broilers from low EST treatment had the highest score for feathering. A higher incidence of gait score was observed in broilers from the control EST treatment than low and high EST groups. This can be attributed to a higher live weight of broilers from the control EST group. On the other hand, the incidence of a gait score of 3 and 4 was found for broilers from control and high EST treatment groups. Male and female broilers from the high EST group had the higher gait score. In conclusion, gait score and welfare status of broilers were affected by fluctuations in EST between 10 and 18 days of incubation.

Associations between and development of welfare indicators in organic layers.

The retail market share of organic eggs in Denmark is high, and the consumers expect high animal welfare standards in the organic production. Documentation of animal welfare is important, however, knowledge about the associations between animal-based welfare indicators is limited. The aims of the study were to investigate the associations between selected welfare indicators at two ages (peak and end of lay), and to examine the development with age of the chosen welfare indicators. The chosen welfare indicators were Ascaridia galli (roundworm) infection, Heterakis sp. (caecal worm) infection, keel bone damages, back feathering, body feathering, foot damages, comb colour and wounds on the body. An observational study with 12 organic egg farms was conducted in 2012 and 2013 with a total of 214 hens assessed individually at the peak and the end of lay. Insufficient data were obtained on helminth infection at the peak of lay. At the end of lay, all helminth infected hens were positive for A. galli, and only three of them had in addition a Heterakis sp. infection. Foot damages, pale combs and wounds on the body occurred at frequencies <5% and were therefore, together with the prevalence of Heterakis sp. infection, left out of the analysis of associations. A graphical model was used to analyse the associations between the remaining clinical welfare indicators, A. galli infection, housing systems and age of the hens at end of lay. A. galli infection was only directly associated with back feathering at end of lay (P=0.011) with an increased incidence of A. galli infection in hens with good back feathering. Between the two visits, the prevalence of hens with keel bone damages increased (P<0.001), and the plumage condition deteriorated (P<0.001), whereas the number of hens with plantar abscess (P=0.037) and pale combs (P=0.020) decreased. No significant differences were found for other foot damages or for skin damage. In conclusion, back feathering at end of lay provided information about a possible helminth infection, but this is not a useful indicator in daily on-farm management. In addition, evidence was found that the deterioration of the plumage condition with age was not only due to accumulation of damage over time.

Polymelia and Syndactyly in a Swainson's Hawk (Buteo swainsoni).

A hatch-year Swainson's Hawk (Buteo swainsoni) recovered from Modoc County, California, US, on 12 August 2012 had malformations of the rear limbs consisting of bilateral polymelia and syndactyly. We describe the malformations and evaluate potential causes. Postmortem examination revealed varus rotation of both femurs and abnormal appendages originating from the distal medial surface of the tibiotarsi with two nonfunctional digits on the right leg and one digit on the left leg. There was syndactyly between digits III and IV of both feet. Avian pox viral dermatitis was present on the skin of the ventral abdomen. A definitive cause of the skeletal malformations was not identified.

Specific characteristics of the aviary housing system affect plumage condition, mortality and production in laying hens.

Feather pecking and high mortality levels are significant welfare problems in non-cage housing systems for laying hens. The aim of this study was to identify husbandry-related risk factors for feather damage, mortality, and egg laying performance in laying hens housed in the multi-tier non-cage housing systems known as aviaries. Factors tested included type of system flooring, degree of red mite infestation, and access to free-range areas. Information on housing characteristics, management, and performance in Belgian aviaries (N=47 flocks) were obtained from a questionnaire, farm records, and farm visits. Plumage condition and pecking wounds were scored in 50 randomly selected 60-week-old hens per flock. Associations between plumage condition, wounds, performance, mortality, and possible risk factors were investigated using a linear model with a stepwise model selection procedure. Many flocks exhibited a poor plumage condition and a high prevalence of wounds, with considerable variation between flocks. Better plumage condition was found in wire mesh aviaries (P<0.001), in aviaries with no red mite infestation (P=0.004), and in free-range systems (P=0.011) compared to plastic slatted aviaries, in houses with red mite infestations, and those without a free-range area. Furthermore, hens in aviaries with wire mesh flooring had fewer wounds on the back (P=0.006) and vent (P=0.009), reduced mortality (P=0.003), and a better laying performance (P=0.013) as compared to hens in aviaries with plastic slatted flooring. Flocks with better feather cover had lower levels of mortality (P<0.001). Red mite infestations were more common in plastic slatted aviaries (P=0.043). Other risk factors associated with plumage condition were genotype, number of diet changes, and the presence of nest perches. Wire mesh flooring in particular seems to have several health, welfare, and performance benefits in comparison to plastic slats, possibly related to decreased feather pecking, better hygiene, and fewer red mite infestations. This suggests that adjustments to the aviary housing design may further improve laying hen welfare and performance.

Expression of fluorescent proteins within the repeat long region of the Marek's disease virus genome allows direct identification of infected cells while retaining full pathogenicity.

Marek's disease virus (MDV) is a lymphotropic alphaherpesvirus and causes Marek's disease (MD) in chickens. RLORF4 is an MDV-specific gene located in the repeat long (RL) regions of the genome and is directly involved in attenuation. In this report, we generated recombinant (r)MDVs in which eGFP or mRFP was inserted in-frame of the 3' end of the RLORF4 gene. In vitro growth was unaffected and infected cells could be identified by using fluorescent microscopy. Interestingly, though inserted in-frame with RLORF4, eGFP and mRFP were expressed alone, confirming mRNA expression and splicing within the RL of MDV is complex. In vivo, rMDVs expressing mRFP or eGFP caused tumors similar to wild-type MDV. Fluorescent protein expression could be seen in spleen, tumor, and feather follicle epithelial cells. These results show that expression of fluorescent proteins within the RL region results in fluorescent rMDVs that still maintains full pathogenicity in the chicken.