Experimental Transmission of Walleye Dermal Sarcoma to Yellow Perch

Abstract

Walleye dermal sarcoma was transmitted under experimental conditions to yellow perch Perca flavescens. Fish (20 weeks posthatch) were challenged with cell-free tumor filtrates by topical application on the right flank and then held for observation for 25 weeks in 15°C dechlorinated municipal water. Additional treatment groups included yellow perch challenged with cell-free filtrates of walleye discrete epidermal hyperplasia, yellow perch discrete epidermal hyperplasia, normal yellow perch skin (control), and normal walleye skin (control). Walleye dermal sarcoma was first observed on yellow perch at 20 weeks postexposure, at which time the tumors were small (1–2-mm), slightly raised masses on the right flank of the fish. At the end of the 25-week study, walleye dermal sarcoma was grossly observed in 42% (22 of 53) of the remaining fish and was confirmed by microscopic examination in an additional 29% (9 of 31). Gross and microscopic evaluation of fish at 25 weeks postexposure did not reveal development of lesions in any of the other treatment groups. This study extends the host range of experimentally transmitted walleye dermal sarcoma virus to include the yellow perch.     

Experimental transmission of a dermal sarcoma in fingerling walleyes (Stizostedion vitreum vitreum)

Dermal sarcoma is a benign skin tumor of adult walleyes (Stizostedion vitreum vitreum) with a suspected viral etiology. A laboratory study was initiated to determine if the tumor could be experimentally transmitted by inoculating young walleyes with materials prepared from tumors from adult fish. Eighty walleye fingerlings were divided into four groups of 20 fish each. Two groups were inoculated intramuscularly at 4 months of age either with live tumor cells or with cell-free filtrates of sonicated tumor cells. The two other groups were used as controls and were inoculated either with cultured cells from normal walleye fry or with tissue culture media. Neoplasms, similar to the dermal sarcoma affecting adult walleyes, were observed after 4 months only in fingerlings inoculated with cell-free filtrates of sonicated tumor cells. Like the tumor affecting wild adult walleyes, the transmitted tumors were restricted to the dermis and originated from the superficial surface of scales. They never invaded locally and never metastasized. The transmitted tumors differed from tumors of adult walleyes in their severity and the absence of osteoid. The multicentric origin of transmitted walleye dermal sarcoma suggests that the virus spreads systemically and that tumor cells are polyclonal. This successful transmission of the lesion, along with the presence of C-type virus particles budding from tumor cells in two of seven tumor-bearing fingerlings, supports a retroviral etiology.     

Pathological Changes Associated with Vitamin C Deficiency in Walleyes

Abstract

Walleyes Stizostedion vitreum were fed three experimental diets containing 0, 96, and 190 mg vitamin C/kg for 20 weeks. Retarded growth, increased mortality, and clinical signs of vitamin C deficiency were seen only in the group fed the vitamin-C-free diet. Skeletal deformities, primarily lordosis, were evident in 76% of this group. Pathological changes associated with vitamin C deficiency in walleyes included twisted cartilage of gill filaments and extreme dislocation of vertebrae. Vertebral lesions included focal hemorrhage, compressed spinal cord, and displacement of adjacent kidney and skeletal muscle. Breaking of the isthmus, a clinical sign not previously described for vitamin-C-deficient fish, was the cause of death in 27% of walleyes fed the vitaminC-free diet. Hemorrhagic fins, skin, and eyes were seen occasionally. A diet that was not supplemented with vitamin C but that contained 96 mg vitamin C/kg was adequate for normal growth and prevention of vitamin-deficiency signs in walleye.     

Ether Sensitivity of the Walleye Dermal Sarcoma Virus

Abstract

Current evidence supports the classification of the virus causing dermal sarcoma on walleyes Stizostedion vitreum as a member of the family Retroviridae. One of the characteristics of this group of viruses is the presence of a lipid envelope. In this report we document studies conducted that demonstrate the presence of a lipid envelope on the walleye dermal sarcoma virus.     

Transmission of Walleye Dermal Sarcoma and Lymphocystis via Waterborne Exposure

Abstract

A study was conducted to determine if walleye dermal sarcoma could be experimentally transmitted by waterborne exposure of tumor-free walleyes Stizostedion vitreum to tumor-positive walleyes by cohabitation. Uninfected walleyes were placed in a common raceway with tumor-positive fish for either 5 or 15 d. Direct contact between the two groups of fish was prevented by two screen barriers. The exposed fish were then maintained in the laboratory for a total of 20 weeks. Walleye dermal sarcoma developed in 89% and 71% of the fish exposed for 5 and 15 d, respectively. There was no significant difference (P > 0.05) in incidence of tumors between the two groups of fish exposed for different lengths of time.     

Effects of Water Temperature on Experimental Transmission of Dermal Sarcoma in Fingerling Walleyes

Abstract

The effects of temperature on experimental transmission of dermal sarcoma, a spontaneous tumor of adult walleyes Stizostedion vitreum to fingerling walleyes was determined. Test temperatures were 10, 15, and 20°C. Three-month-old walleyes were inoculated intramuscularly with a cell-free filtrate from a dermal sarcoma collected from an adult walleye in the spring of the year. Tumors were first grossly visible in fish held at 15°C at 8 weeks postinoculation. At 12 weeks postinoculation, all fish were euthanized and examined for presence of tumors. Tumor transmission was most successful at 15°C, followed by that at 20°C; many fish maintained at these temperatures had grossly visible tumors. Although the majority offish held at 10°C also had tumors, tumors in these fish developed to a lesser degree than those observed in fish held at the higher temperatures.     

The cannabinoid receptors system in horses: Tissue distribution and cellular identification in skin

BackgroundThe endocannabinoid system (ECS) is composed of cannabinoid receptors type 1 (CBR1) and type 2 (CBR2), cannabinoid‐based ligands (endogenous chemically synthesized phytocannabinoids), and endogenous enzymes controlling their concentrations. Cannabinoid receptors (CBRs) have been identified in invertebrates and in almost all vertebrate species in the central and peripheral nervous system as well as in immune cells, where they control neuroimmune homeostasis. In humans, rodents, dogs, and cats, CBRs expression has been confirmed in the skin, and their expression and tissue distribution become disordered in pathological conditions. Cannabinoid receptors may be a possible therapeutic target in skin diseases.ObjectivesTo characterize the distribution and cellular expression of CBRs in the skin of horses under normal conditions.AnimalsFifteen healthy horses.MethodsUsing full‐thickness skin punch biopsy samples, skin‐derived primary epidermal keratinocytes and dermal‐derived cells, we performed analysis of Cnr1 and Cnr2 genes using real‐time PCR and CBR1 and CBR2 protein expression by confocal microscopy and Western blotting.ResultsNormal equine skin, including equine epidermal keratinocytes and dermal fibroblast‐like cells, all exhibited constant gene and protein expression of CBRs.Conclusions and Clinical ImportanceOur results represent a starting point for developing and translating new veterinary medicine‐based pharmacotherapies using ECS as a possible target.     

Serum Lysozyme Levels in Paddlefish and Walleye

Abstract

The turbidimetric assay of lysozyme activity in serum revealed ranges of 0.0–27.8 units/mL in walleyes Stizostedion vitreum (spawners and nonspawners) and 11.1–238.9 units/mL in paddlefish Polyodon spathula (spawners and juveniles). A unit of lysozyme activity was defined as the amount of the sample causing a decrease in absorbance of 0.001 /min. Lysozyme activity was significantly different (P < 0.05) in the two species. This first demonstration of lysozyme in the sera of paddlefish and walleye is further evidence of the occurrence of this nonspecific immune factor, which has been ubiquitous in fishes investigated to now.     

Thiamine and fatty acid content of walleye tissue from three southern U.S. reservoirs

We determined the thiamine concentration in egg, muscle, and liver tissues of walleyes Sander vitreus and the fatty acid content of walleye eggs from three southern U.S. reservoirs. In two Tennessee reservoirs (Dale Hollow and Center Hill), in which there were alewives Alosa pseudoharengus in the forage base, natural recruitment of walleyes was not occurring; by contrast in Lake James Reservoir, North Carolina, where there were no alewives, the walleye population was sustained via natural recruitment. Female walleye tissues were collected and assayed for thiamine (vitamin B1) and fatty acid content. Thiamine pyrophosphate was found to be the predominant form of thiamine in walleye eggs. In 2000, mean total egg thiamine concentrations were similar among Center Hill, Dale Hollow, and Lake James reservoirs (2.13, 3.14, and 2.77 nmol thiamine/g, respectively). Egg thiamine concentration increased as maternal muscle (r2 = 0.73) and liver (r2 = 0.68) thiamine concentration increased. Walleye egg thiamine does not appear to be connected to poor natural reproduction in Tennessee walleyes. Threadfin shad Dorosoma petenense, which are found in all three reservoirs, had higher thiaminase activity than alewives. Six fatty acids differed among the walleye eggs for the three reservoirs. Two were physiologically important fatty acids, arachidonic acid (20:4[n-6]) and docosahexaenoic acid (22:6[n-3]), which are important eicosanoid precursors involved in the regulation of biological functions, such as immune response and reproduction.     

Regression of Dermal Sarcoma in Adult Walleyes

Abstract

Dermal sarcoma is a benign skin tumor of walleyes Stizostedion vitreum, and a seasonal prevalence for this condition has been observed in feral fish. This study was conducted to determine if dermal sarcomas regress. Evidence of dermal sarcoma regression was documented on individually identified adult walleyes. Degree of regression was variable, ranging from none to complete. This report provides the first definitive evidence that dermal sarcoma does regress seasonally from spring to summer.     

Investigation into the pathogenesis of Atrophic Rhinitis in pigs

Summary

In two groups of swine herds, herds with and without clinical AR the presence of Atrophic Rhinitis (AR) correlated with the presence of toxinogenic Pasteurella multocida (PM) and not with the Bordetella bronchiseptica (BB) infection.

Six BB‐ and eighteen PM‐strains have been investigated for AR pathogenicity. Broth cultures were injected intradermally in guinea‐pigs (GPST) orintranasally in 3‐week‐old colostrum deprived specific pathogen free (SPF) piglets.

The average atrophy of the ventral conchae (A VC) correlated with the GPST in 4BB‐ and 7 PM‐strains. One BB‐ and 2 PM‐strains were qualified as doubtful, the others as non‐AR pathogenic. With AR pathogenic BB‐ and PM‐strains clinical AR could be induced in 3‐ and 6‐week‐old piglets. AVC lesions (gradation> 1) could be induced with BB in piglets of 6 and with pathogenic PM in 16‐week‐old piglets. Six of seven AR pathogenic PM‐strains resembled Carter‐type D and one resembled type A. No significance was found between AR pathogenicity and somatic serotypes.

Intranasal instillations of cell‐free broth culture filtrates of AR pathogenic PM‐strains also caused AR in piglets. These filtrates also caused lethality in piglets and in mice lethalitytest (MLT) and induced a positive GPST. After heating the pathogenic effects of the filtrates disappeared. The name AR toxin has been introduced for this thermolabile, haemorrhagic dermonecrotic (HDNT) fraction of the AR inducing filtrates. The severity of the AR lesions depended on the amount of the AR toxin intranasally instilled in pigs.

Cross protecting antibodies obtained in rabbits against the AR toxins of two PM strains could be demonstrated by a toxin neutralisation test in the MLT and the GPST.

Broth cultures were injected intradermally in guinea‐pigs (GPST) or intranasally in 3‐week‐old colostrum deprived specific pathogen free (SPF) piglets.     

Micro-parasites of the ruminant abomasum

Abstract

CASE HISTORY: Nodular lesions were found on the skin of two immature brown kiwi (Apteryx mantelli) less than 6 months of age living freely on Ponui Island off the North Island of New Zealand. The lesions were observed during routine external examination undertaken as a part of the management of other research projects, one in 2006 and the other in 2011. Apart from the skin lesions, both birds showed no signs of illness and the lesions resolved spontaneously over a 2-month period.

PATHOLOGICAL FINDINGS: The first case showed several 3-mm diameter firm, brown nodules located on the skin below the hock of both legs. The second case had a single multinodular mass that measured 7×20 mm, on the base of the bill. A portion of the mass and scab samples were collected for diagnosis. Histological examination of the nodules revealed severe ballooning degeneration of keratinocytes and epithelial hyperplasia. Round eosinophilic structures resembling avipoxvirus (APV) intracytoplasmic inclusion bodies (Bollinger bodies) were observed in the layers of keratinocytes. In deeper layers of the epidermis, there was evidence of secondary bacterial growth and inflammation.

DIAGNOSIS: DNA was extracted from tissue samples and subjected to PCR analysis. Avipoxvirus 4b core protein gene was detected in both samples by PCR. Bootstrap analysis of APV 4b core protein gene revealed that APV isolates from two kiwi comprised two different subclades. One isolate displayed 100% sequence homology to subclade B1, and the other presented 100% sequence homology to subclade A3.

CLINICAL RELEVANCE: This study confirmed that kiwi are susceptible to APV infection and that at least two different strains of APV are present in the population examined. Since there is no information on the origin, virulence, or prevalence of APV in kiwi, a seroprevalence study would be useful to elucidate the degree of exposure and immune response to the disease. This would allow a more informed approach to risk management of the disease in wild and captive populations.     

Consistent detection of bovine papillomavirus in lesions, intact skin and peripheral blood mononuclear cells of horses affected by hoof canker

Reasons for performing the study: Equine hoof canker is a chronic proliferative pododermatitis of as yet unknown aetiology. Like equine sarcoid disease, canker is a therapy‐resistant disorder characterised by hyperkeratosis, acanthosis and a marked tendency to recur. Hypothesis: There is an association of sarcoid‐inducing bovine papillomaviruses of types 1 and 2 (BPV‐1, BPV‐2) with hoof canker disease. Methods: Using PCR‐based techniques, we assessed canker tissue, intact skin and/or peripheral blood mononuclear cells (PBMCs) of 25 canker‐affected horses for the presence of sarcoid‐associated BPV‐1 and ‐2. Results: Conventional PCR revealed BPV‐1/‐2 DNA in 24/24 canker, 12/13 skin and 10/11 PBMC DNA isolates. Using inverse PCR, full‐length BPV episomes were detected in 1/5 canker specimens. Sequencing of viral early and late genes amplified from canker, intact skin and PBMC DNA of 2 cases revealed an overall identity of 98% to BPV‐1. Viral DNA loads amounted to ≤16 copies per cell in canker tissue and intact skin, and to ≤0.35 copies per PBMC, as determined by quantitative PCR. Using RT‐PCR, the viral major oncogene E5 was shown to be transcribed in 2/4 canker tissue specimens and 5/7 PBMC isolates. Immunocapture PCR from 7 canker and 6 skin extract supernatants revealed capsomere‐associated viral DNA in one canker and one skin sample. Hoof tissue, skin and PBMCs collected from 13 individuals with no signs of canker or BPV‐related malignancies scored negative throughout the experiments. Conclusion: These findings suggest that the observed presence of BPV‐1/‐2 in canker‐affected horses is not coincidental but indicative of an active contribution to hoof canker disease. Potential relevance: The use of antivirals and/or immune modulators may help improving canker therapy.     

Alterations of epidermal proliferation and cytokeratin expression in skin biopsies from heavy draught horses with chronic pastern dermatitis

We report the historical, clinical and histopathological characteristics of skin lesions in biopsies from 37 heavy draught horses with chronic pastern dermatitis. The skin lesions were divided into four macroscopic groups: scaling (group I, n=5), hyperkeratotic and hyperplastic plaque-like lesions (group II, n=14), nodular skin masses (group III, n=16) and verrucous skin lesions (group IV, n=2). The principal histological findings were hyperkeratosis and epidermal hyperplasia. There was a gradual increase in epidermal hyperplasia from groups I to IV, suggesting that the lesions represent different stages of disease. In all cases, there was perivascular dermatitis dominated by T lymphocytes with an increase in MHC class II-positive dendritic-like cells. Immunohistochemical labelling for cytokeratins CK5/6(4), CK10 and CK14 indicated a change in their expression pattern. This correlated with the degree of epidermal hyperplasia, indicating abnormal differentiation of keratinocytes. There was a statistically significant correlation between the severity of skin lesions and several other factors including increasing age, increasing cannon circumference, prominence of anatomical structures such as fetlock tufts of hairs, ergots and chestnuts, and bulges in the fetlock region.     

Intralesional bovine papillomavirus DNA loads reflect severity of equine sarcoid disease

Reasons for performing study: Sarcoids are nonmetastasising, yet locally aggressive skin tumours that constitute the most frequent neoplasm in equids. Infection by bovine papillomaviruses types 1 and 2 (BPV‐1, BPV‐2) has been recognised as major causative factor in sarcoid pathogenesis, but a possible correlation of intralesional virus load with disease severity has not been established thus far. Hypothesis: Given the pathogenic role of BPV‐1 and BPV‐2 in sarcoid disease, we suggest that intralesional viral DNA concentration may reflect the degree of affection. Methods: Severity of disease was addressed by recording the tumour growth kinetics, lesion number and tumour type for 37 sarcoid‐bearing horses and one donkey. Viral load was estimated via quantitative real‐time PCR (qPCR) of the E2, E5, L1 and L2 genes from the BPV‐1/‐2 genome for one randomly selected lesion per horse and correlated with disease severity. Results: Quantitative PCR against E2 identified viral DNA concentrations ranging from 0–556 copies/tumour cell. Of 16 horses affected by quiescent, slowly growing single tumours or multiple mild‐type lesions, 15 showed a viral load up to 1.4 copies per cell. In stark contrast, all equids (22/22) bearing rapidly growing and/or multiple aggressive sarcoids had a viral load between 3 and 569 copies per cell. Consistent results were obtained with qPCR against E5, L1 and L2. Conclusions: While tumours of the same clinical type carried variable virus load, confirming that viral titre does not determine clinical appearance, we identified a highly significant correlation between intralesional viral load and disease severity. Potential relevance: The rapid determination of BPV viral load will give a reliable marker for disease severity and may also be considered when establishing a therapeutic strategy.     

The development of multiple cutaneous inverted papilloma following ovariohysterectomy in a dog

Abstract

CASE HISTORY: Ovariohysterectomy was performed on an adult Cavalier King Charles Spaniel. The skin that had been clipped for surgery was noticed to be erythematous 8 days later.

CLINICAL AND PATHOLOGICAL FINDINGS: Poorly defined patches containing multiple papules were visible bilaterally within the clipped skin. These became larger over the following 2 weeks, and samples were collected for histology. Seven days later, the lesions were multiple raised masses, up to 5 cm in diameter. Histology revealed numerous cup-shaped epidermal proliferations extending into the dermis. The presence of keratinocytes with increased quantities of blue-grey cytoplasm, and koilocytosis suggested papillomavirul infection; Canis familiaris papillomavirus (CfPV-2) DNA was amplified from two separate samples. Complete regression was observed 8 weeks after the lesions had been initially observed.

DIAGNOSIS: Multiple inverted papilloma confined to skin that had been clipped for surgery.

CLINICAL RELEVANCE: This is the first time that the development of canine cutaneous papillomas has been associated with surgery. The nature of the association between surgery and development of the papillomas is uncertain. However, it is possible that damage to superficial skin could promote the formation of papillomas. This is the first identification of CfPV-2 in New Zealand.     

Characterization of a novel papillomavirus species (ZcPV1) from two California sea lions (Zalophus californianus)

A seven-year old California sea lion (Zalophus californianus) presented with focally extensive, bilaterally symmetric, proliferative axillary skin lesions and preputial lesions. A second California sea lion in the same population presented with similar proliferative lesions on the underside of the tail. Histopathology revealed epidermal hyperplasia with severe hyperkeratosis, with proliferating keratinocytes forming broad, branching pegs that extended into the dermis. Pan-papillomaviral consensus PCR was used to obtain initial E1 sequence template and the complete genome was determined using a combination of rolling circle amplification and specific-primer PCR. Analysis revealed a novel papillomavirus, Zalophus californianus papillomavirus 1 (ZcPV1), with seven open reading frames encoding five early proteins (E6, E7, E1, E2 and E4) and two late proteins (L1 and L2). Phylogenetic analysis revealed that (ZcPV1) is most closely related to Equine papillomavirus 1 (EcPV1) in the genus Zetapapillomavirus, and Canine papillomaviruses 3 and 4 (CPV3, CPV4) in the genus Chipapillomavirus. The lesions regressed without intervention over a period of several months.     

Detection of feline herpes virus 1 via polymerase chain reaction and immunohistochemistry in cats with ulcerative facial dermatitis, eosinophilic granuloma complex reaction patterns and mosquito bite hypersensitivity

Ulcerative dermatitis caused by feline herpes virus 1 (FHV-1) is an uncommon disease characterized by cutaneous ulcers secondary to epidermal, adnexal and dermal necrosis. Differential diagnoses for FHV-1 lesions include, but are not limited to, mosquito bite hypersensitivity and eosinophilic granuloma complex. Histopathological diagnosis of FHV-1 dermatitis is based on the detection of the intranuclear inclusion bodies. In cases where intranuclear inclusions are missing but clinical and histological findings are compatible with FHV-1 dermatitis, immunohistochemistry (IHC) and PCRs have been used. In this retrospective study, we evaluated the presence of FHV-1 by IHC and PCR in skin biopsies and compared the results of the two tests. Sixty-four skin biopsy specimens from cats with compatible lesions were reviewed and tested via PCR and IHC for evidence of FHV-1. Polymerase chain reaction was positive in 12 of 64 biopsies; PCR and IHC were positive only in two of 64 biopsies, and these cases were considered true positive cases. The higher number of PCR-positive cases was possibly attributed to amplification of viral DNA from a live attenuated vaccination, but a previous FHV-1 infection with subsequent amplification of latently inserted FHV-1 could not be excluded. If clinical signs and histopathology suggest FHV-1 infection in the absence of typical inclusion bodies, IHC is the preferred diagnostic test; PCR may be useful for initial screening, but due to false positives is not sufficient for a definitive diagnosis.