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In April and November of 2018, multiple commercial laying hen flocks within the same company presented with a sharp increase in mortality and drop in egg production that persisted for several days. These flocks showed striking necropsy lesions consistent with systemic infection and responded to antimicrobial treatment in the feed. Staphylococcus aureus (SA) was the most frequently isolated organism from multiple tissues including comb and wattle lesions, lungs, liver, ovary, spleen, and bone marrow. Given such an uncommon presentation of SA, which is known as a secondary opportunistic pathogen, a challenge study was conducted to evaluate its role in these disease outbreaks. In the present study, laying hens of two ages (22 and 96 wk) were inoculated with SA via three routes: oral gavage, subcutaneous (SC) injection, and intravenous (IV) injection. Both young and old hens in the IV group showed a significant increase in body temperature and drop in body weight; however, the clinical signs observed in the naturally occurring outbreaks were not present. SA was reisolated at multiple time points postchallenge from all challenge groups except the negative control group. While the SC group showed localized necrosis at the injection site, microscopic changes were different from changes observed in birds from the natural outbreaks. Despite observed initial differences in route and age, the SA challenge strain was not capable of reproducing the disease on its own. The results of this study indicate that SA may have played a role in the increased mortality, clinical signs, and necropsy lesions reported with the naturally occurring outbreaks. However, SA should still be considered as a secondary opportunistic pathogen. Other factors that could have caused the initial insult are stress, immunosuppression, or other primary infectious agents. The results of this study may aid veterinary diagnosticians, clinicians, and all poultry professionals to include SA in their differentials list as a secondary opportunistic pathogen in similar cases. This is an uncommon presentation and further field observations and clinical studies are needed to better elucidate the pathogenesis of this disease, which will in turn help to prevent future outbreaks.

Background: Mono-L-aspartyl chlorin e6 (Talaporfin sodium) is a novel photosensitizer, and is currently being used in photodynamic therapy for various malignant tumors in combination with irradiation with a 664 nm laser. An interesting characteristic of Talaporfin sodium is that the skin photosensitivity after injection of this agent disappears faster than any other existing photosensitizers. This study examined the vascular events that occurred postirradiation in the chicken comb as a capillary malformation model after photosensitization with Talaporfin sodium., Materials and Methods: A single intravenous bolus injections of Talaporfin sodium was administered to the chickens, and a 1 cm diameter area of the comb of each animal was irradiated with a 664 nm visible red laser. The gross changes in the chicken combs were recorded for 7-14 days after photodynamic therapy. For the histological examination, HE, PTAH and Azan stained sections were analyzed., Results: All treated chicken combs had blanched after photodynamic therapy. Microscopy demonstrated an absence of erythrocytes and the vessel lumina were obliterated, leaving the normal overlying epidermis completely intact. Concomitantly with selective destruction of the capillaries in the target area, moderate invasion of inflammatory cells and a slight increase in the stroma were observed., Conclusions: In the chicken comb model, photodynamic therapy with Talaporfin sodium effectively achieved selective destruction of the microvasculature while leaving the epidermis intact. Our results strongly suggest that photodynamic therapy with Talaporfin sodium could be a feasible method to treat dermal hypervascular lesions.

Ten-to-14-day-old replacement chickens were vaccinated subcutaneously in the neck with a Mycoplasma gallisepticum bacterin. A few days later, 40% of the flock developed cellulitis of the head. Birds were submitted for diagnostic investigation between 4 and 6 wk of age. Histology revealed lesions in the subcutis consistent with oil-emulsified vaccine/bacterin reaction. No significant bacteria were isolated from the lesions. Birds were treated with 1 mg of methyl prednisol acetate intramuscularly. Improvement was noted as early as 12 hr posttreatment. One week after treatment only 1%-2% of the flock still had facial cellulitis.

A cutaneous mycosis caused by Candida albicans that involved the combs and less frequently the wattles, facial skin, ear lobes, and neck of male broiler breeders is described. Roosters were 35 wk old and housed with hens in two conventional broiler breeder houses on a farm in western North Carolina. Morbidity was approximately 10% in one house and less than 2% in the other house. Mortality and flock fertility were not affected. Three birds from the most affected house were examined. All birds had white adherent material on their combs that presented as crusty patches or lighter diffuse areas. Often, lesions were roughly circular or had a defined margin. Small black scabs were present in a few lesions. Similar but less extensive lesions were located on the wattles, facial skin, ear lobes, and rictus. In one bird, lesions extended down the neck, and they were accompanied by hyperemia and feather loss. Hyperkeratosis with little to no inflammation and intralesional fungi occurring as yeast and pseudohyphae were seen microscopically. High numbers of C. albicans were isolated and identified from the lesions.

Objective: To define the optimal treatment parameters of chlorophyll derivative IV (CPD4) in photodynamic therapy(PDT) of port wine stains(PWS) by observing the morphological changes of chicken combs., Methods: 102 animals were divided into 6 groups at random, including 3 control groups(given no treatment, given light without photosensitizer or given photosensitizer without light) and 3 experimental groups treated with different CPD4 dosages(5, 7.5, 10, 12.5, 15 mg/kg body weight), power density(50, 100, 150, 200 mW/cm2), energy density (15, 30, 45, 60, 120, 135, 180 J/cm2) and different intervals between CPD4 injection and irradiation (immediately, 5, 10, 20, 30, 60, 120, 180, 240 minutes). The main emitting peak of light source was at 650 nm, and CPD4 was injected intravenously, then the appearance of chicken comb was recorded everyday. The prepared sections were stained with HE. The histological changes of chicken comb were observed by light microscope., Results: The control animals showed no significant comb changes, while the PDT could cause blanching of the chicken comb in the experimental group. The histologic changes included damage of erythrocytes and endothelial cells, vascular rupture, hemorrhage or disappearance, denaturation of perivascular collagen fiber and local necrosis of chicken combs. The treatment parameters which caused the most obvious blanching of the chicken comb were 7.5-10 mg/kg body weight of CPD4 dosage, 100-150 mW/cm2 of power density, 60-90 J/cm2 of energy density, and the intervals between CPD4 injection and irradiation were 0-20 minutes., Conclusion: CPD4 in PDT can cause selective damage of dermal capillaries in chicken comb and blanching of the chicken comb with optimal treatment parameters. The results will offer the basis for further investigation of clinic treatment parameters.

Two experiments were conducted to induce the delayed footpad reaction (DFR) to killed Staphylococcus aureus antigen. In Experiment 1, tracheal, cloacal, and choanal swabs were collected from chickens prior to sensitization with S. aureus to determine the carrier status of S. aureus. The second experiment compared the DFR to the delayed wattle reaction (DWR). Chickens were subjected to single or multiple sensitizations in the neck with S. aureus antigen between 4 and 6 wk of age. One week later, birds were challenged with S. aureus either in the right footpad or wattle. The left footpad or wattle was injected with PBS. The thicknesses of the footpad or the wattle were measured up to 96 h postchallenge. The recoveries of S. aureus from the choanal slit and trachea were significantly higher than that of the cloaca (P < 0.001). Birds of Experiment 1 showed a significant DFR (P < 0.0001) following intradermal challenge with killed S. aureus that was sustained through 48 h postchallenge with no difference in the DFR between carrier and noncarrier birds. In Experiment 2, the thicknesses of the footpad and wattle were significantly increased following challenge with S. aureus (P < 0.0001), with the footpad showing a greater response than the wattle (P < 0.001). Three sensitizing dosages, as compared to two dosages, resulted in a less pronounced DFR and DWR (P < 0.02). These results indicate that the DFR can be used as a delayed reaction model in the study of staphylococcosis in poultry.

Background: Spontaneous animal models of human autoimmune diseases provide the means to study the very first pathogenetic events, which is not possible in their human counterparts. This is particularly true for connective tissue diseases in which clinical symptoms become manifest only after a long and still obscure course of immunologic, inflammatory, and fibrotic processes. University of California at Davis line 200 chickens (UCD-200) develop a hereditary scleroderma-like disease resembling the entire spectrum of human systemic sclerosis, such as early endothelial cell damage, severe lymphocytic infiltration, and accumulation of collagen in skin and internal organs., Materials and Methods: In the present study, we investigated mRNA levels of alpha 1(I), alpha 2(I), alpha 1(II), alpha 1(III), alpha 1(VI), alpha 2(VI), and alpha 3(VI) procollagen and GAPDH using digoxigenin-labeled antisense probes in a nonradioactive ribonuclease protection assay (RPA). We analyzed tissue samples from comb, esophagus, heart, lung, and liver of UCD-200 chickens at different stages of the disease, and healthy UCD-058 chickens., Results: During the early inflammatory stage of the disease, the ratios of procollagen types VI/I and types VI/III increased 7-fold in comb tissue, followed by a 3-fold elevation in type I procollagen transcripts in the late acute stage. In the chronic stage, alpha 1(III) procollagen message was increased 2-fold. Additionally, hybridization with the 180 bp alpha 2(I) antisense probe resulted in two bands of 180 bp and 115 bp, respectively, in the RPA. The ratio of these two previously undescribed bands changes in the early stage of the disease both in comb and esophagus., Conclusions: In an animal model with a spontaneous scleroderma-like disease we found a characteristic, sequential increase in type VI, type I, and type III procollagen transcripts, and we found evidence for the presence and altered ratio of two mRNA variants of alpha 2(I) procollagen, possibly caused by alternative splicing. Comparative analysis of alpha 2(I) procollagen variants in early stages of avian scleroderma and human SSc might provide answers to unresolved questions concerning the molecular basis for generalized fibrosis in scleroderma.

Comb necrosis with leg weakness was seen in 41-day-old female layer breeder chickens. This disease occurred in three flocks at a breeder farm, but not in other flocks of growing chickens and broiler breeder hens at the same farm. The disease started in 35-day-old chicks in three flocks. The morbidity of comb necrosis was 10% and that of leg weakness was 3%. Characteristic gross lesions of affected chickens were swelling and necrosis of the whole comb. Histologically, liquefactive necrosis of epidermal epithelial cells with hyperplasia, vesicle formation in the epidermis, congestion, and hemorrhages with fibrinous thrombi of underlying dermis in the comb were noted. In mature comb lesions, the epidermis showed eosinophilic necrosis (scab formation). In the livers, multiple fibrinous thrombi were present in the sinusoids and there was necrosis of hepatic cells. Staphylococcus aureus and Pasturella spp. were isolated from comb lesions. There were no significant lesions causing leg weakness.

A flock of commercial Single Comb White Leghorn laying hens, diagnosed as having fatty Liver Hemorrhagic Syndrome (FLHS), was surveyed to ascertain the relationship between comb color and selected FLHS characteristics. Twenty-eight hens with and without pale combs were selected from 14 cages for paired comparisons. Hens with pale combs had a higher (P less than or equal to .05) liver score and relative liver weight than hens with normal combs. Differences were not detected in BW, comb weight, relative comb weight, liver fat and plasma estrogen concentration. In flocks diagnosed with FLHS, comb appearance is associated with a higher incidence of FLHS.

Favus, or avian ringworm, was diagnosed in a backyard flock of game chickens from which Microsporum gallinae was isolated. Infected birds had white crusts on the comb and on the skin of the head and neck. Histopathological lesions included hyperkeratosis of the skin epithelium with invasion of the stratum corneum by fungal mycelia, acanthosis, acantholysis, and hydropic degeneration of cells in the stratum spinosum. The underlying dermis was infiltrated by mononuclear cells and contained lymphoid foci. Daily topical treatment with miconazole was applied in the field and in the laboratory, with apparent success.

Drug induced immunosuppression of chicken immune response was studied in F1 hybrids of the CB and IC inbred lines. In tuberculin reaction complete inhibition of wattle swelling was induced by the administration of methotrexate, colcemid (1 mg/KBW), and 6-mercaptopurine. The cellular infiltration was substantially reduced in these cases. Cyclophosphamide and colcemid (0.1 mg/KBW) reduced partially the wattle swelling but had no apparent effect on the cellular infiltration. Acetinomycin D did not affect in measurable degree the wattle swelling. The histologic picture was in this case the same as in the control animals. The same drug administration schedule had less pronounced effect on anti-HSA antibody production. No anti-HSA antibody was found after the 500 mg/animal doses of 6-mercaptopurine. Significant reduction of anti-HSA titres was found after 50 mg/animal doses of 6-mercaptopurine, colcemid (1 mg/KBW), 25 MG/KBW or cyclophosphamide and after the methotrexate treatment.

A virulent fowlpox virus was isolated from a natural outbreak of the disease in the Sudan. The virus was used for experimental transmission of the disease to susceptible chickens. Scarification and intravenous methods were used to infect one-month-old chickens. Scarification induced typical pox lesions at 5-7 days postinoculation, whereas intravenous injection induced only swelling of combs or wattles, at 10-11 days postinoculation, although neutralizing antibodies against fowlpox virus were detected 2, 3, and 4 weeks postinoculation. The histopathological features of the pox lesions obtained in both natural and experimental infection were described.

The first diagnosis of favus in chickens in Costa Rica was made in a 1-year-old fighting cock that had lesions surrounding the comb. The etiologic agent was isolated and identified as Microsporum gallinae. The rooster recovered during topical treatment with tolnaftate and oral treatment with griseofulvin.

Domestic fowl were found to be hypersensitive to a second injection of highly purified mineral oil if they had been primed 3 weeks previously with oil in the presence of mycobacteria. This response did not appear to be related to impurities originally present in the oil or introduced into it during sterilization, and did not follow priming with mycobacteria in vegetable oil or in saline. The reaction at the challenge site was prolonged and developed a characteristic histology identical to that found at the priming site. It is postualted that the hypersensitivity seen has an immunological basis and that birds are able to respond to antigenic determinants formed by long chain aliphatic hydrocarbons.

Posthatching differentiation of arteriovenous anastomoses (AVAs) and the effect of heat exposure (38 degrees C) and cold acclimation (2-3 degrees C) on AVA density were studied in naked skin areas (eyelids, comb, wattles) of chickens. The AVAs were identifiable in the newly hatched chick, but they were extremely simple. The number of cell layers in the wall of the AVAs increased from two at hatching to four to five in the 5-month-old chickens. The density of the AVAs increased significantly during posthatching maturation. Cold acclimation of the chickens increased the density of AVAs by a factor of 2.1-3.2. Daily heat exposure also increased the density of AVAs significantly in the eyelids but had no effect in the other skin areas. Heat and cold had only minor effect on the vascularity of the skin; the only significant change recorded was a slight increase in the number of blood vessels in the wattles of the cold-acclimated chickens.

Studies were undertaken to determine the presence of lesions and immuno-fluorescent (IF) antigen of Marek's disease (MD) in the comb, wattles and skin overlaying the shank of MD affected birds. Lesions were more common in the comb than in the wattles and shanks, and were characterized by degeneration of epithelial lining cells and accumulation of lympho-reticular cells around the blood vessels of the dermis. Diffuse infiltration of these cells replacing varialbe areas of dermis were also seen in some cases. Immunofluorescent antigen was mostly confined to epidermal epithelium and was detected in comb from majority of cases, and the frequency of detection of IF antigen was similar to the presence of MD lesions in these tissues. This study indicates that the presence of lesions and viral antigen (If) of MD in these cutaneous appendages might play a significant role in epidemiology and diagnosis of this disease.

Egg production dropped after hens were inoculated with fowlpox virus originally isolated from a natural mild infection. The drop started from the 2nd week and continued to the 5th week after inoculation. All birds developed focal lesions at the site of inoculation, and some developed secondary lesions. The drop in individual birds was related to the severity of lesions. Birds challenged with fowlpox virus fifteen months after vaccination with pigeon pox vaccine were susceptible.

Marek's disease was diagnosed as the cause of prolonged high mortality in two commercial flocks of white leghorn layers by gross and microscopic examination of affected tissues and by immunofluorescent study of live tumor cells using anti-Marek's disease tumor-associated surface antigen, anti-immunoglobulin M, and monoclonal antibodies. The disease was characterized by swollen, necrotic, lymphomatous combs and mortality that rose above normal at 28 weeks of age, peaked at 35-36 weeks, and returned to normal by 45-46 weeks.