John P. Sundberg

Cutaneous Fibropapillomas and Renal Myxofibroma in a Green Turtle, Chelonia mydas

A debilitated 7 kg juvenile green turtle (Chelonia mydas mydas) with multiple ulcerated and infected cutaneous fibropapillomas was clinically evaluated and found to have a nonregenerative anemia, hypoproteinemia, hypoalbuminemia and several electrolyte abnormalities. Surgery was performed to remove the larger tumors. The turtle did not eat post surgically, and an attempt was made to place a pharyngostomy tube utilizing endoscopy. Edematous esophageal papillae, the angulation of the gastroesophageal junction, and a S-shaped configuration of the esophagous prevented successful placement of the tube. The animal was found dead the next day and necropsied. Multiple large white firm nodules were seen bulging from both kidneys. Microscopic examination of the nodules resulted in a diagnosis of renal myxofibroma.

Cloning and characterization of an equine cutaneous papillomavirus

Equine papillomaviruses (EqPV) from naturally occurring cases of cutaneous papillomatosis in several ponies and one horse were isolated, cloned, and characterized. Group specific papillomavirus structural antigens were detected in sections of the papillomas by the peroxidase-antiperoxidase technique, and virions were observed in the in the nuclei of cells in the stratum granulosum and corneum. Negatively stained virions purified from papilloma homogenates by isopycnic CsCl centrifugation were 55 nm in diameter and had typical papillomavirus morphology. The entire viral genomes of two separate isolates were cloned at a single BamHI site into pBR322. A detailed restriction map of the viral genome is presented. Using nick-translated subgenomic fragments of BPV-1 as probes in Southern blot hybridizations, the organization of the EqPV genome was established. Southern blot analysis under various conditions of stringency revealed that EqPV shares relatively more homology with the late region of the BPV-1 genome and with the E2 region of the HPV-1 genome than with other parts of the same viral DNAs. Papillomavirus-specific sequences were found in papillomas from other anatomic sites using the EqPV DNA as a probe in Southern blot hybridizations. Genomes detected in DNA from penile papillomas had a different restriction pattern and hybridized to the EqPV probe only under nonstringent conditions.

GENITAL PAPILLOMATOSIS IN SPERM WHALE BULLS

Examination of 31 male sperm whales (Physeter catodon) caught off the western coast of Iceland revealed three cases of genital papillomatosis involving the unsheathed penis. One subadult and two sexually mature bulls were affected. Gross lesions resembled papillomas common in terrestrial mammalian species. Transmission electron microscopy of these lesions revealed nonenveloped intranuclear virus particles 28–40 nm in diameter and round to hexagonal in shape. In two cases immunoperoxidase staining was negative for group-specific papillomavirus antigen. These findings indicate that the spectrum of animal species affected with virus-associated genital papillomatosis includes at least one globally distributed species of the order Cetacea (whales, dolphins, and porpoises).

Cross-Hybridization and Relationships of Various Papillomavirus DNAs at Different Degrees of Stringency

Cloned DNAs of 21 different papillomaviruses which naturally infect mammals and one bird papillomavirus were compared for relative homology by Southern blot hybridization. Blots were carried out under low (Tm-40 degrees), medium (Tm-33 degrees), and high (Tm-22 degrees) stringency conditions. At higher stringency, human papillomaviruses cross-hybridized with each other reflecting species-specific similarities. Bovine papillomavirus types 1, 2, 5, European elk papillomavirus, and deer papillomavirus also cross-hybridized at higher stringencies probably reflecting the association of these viruses with fibroblast-prolific lesions. The hybridization data presented here may be useful in future classification attempts. They are also useful as a guide in the selection of papillomavirus DNA probes for analysis of extracts from warts and tumors.

Venereal Papilloma and Papillomavirus in a Colobus Monkey (Colobus guereza)

A papilloma on the penis of a colobus monkey was found to contain papilloma-virus group specific antigens by immunohistochemical analysis and virus-like particles in the nuclei of epithelial cells by transmission electron microscopy. In low-stringency Southern blot hybridizations, DNA from the lesion annealed with human papillomavirus 11 DNA, but not with the DNAs of 13 other papillomaviruses. Using human papillomavirus 11 DNA as a probe in Southern blot hybridizations, DNA from the penile papilloma was shown to contain a supercoiled DNA approximately 8 kilobases in size. This represents the first demonstration of a papillomavirus-associated venereal lesion in a nonhuman primate.

Molecular cloning and partial characterization of a parrot papillomavirus

The genome of a papillomavirus isolated from a cutaneous lesion on the head of an African grey parrot (Psittacus erithacus timneh) was cloned into pBR322, and a restriction map was prepared. Several short portions of the DNA were sequenced allowing the genome to be aligned with HPV-la. In Southern blot hybridizations, under conditions of low and medium stringency (Tm-40 and -33 degrees), but not at a higher stringency, this viral DNA annealed weakly with only 1 of 17 mammalian papillomavirus genomes tested. Furthermore, PePV DNA hybridized with the DNA from the European chaffinch only at low stringency, indicating that it represents a unique avian papillomavirus.

Characterization of papillomaviruses isolated from cutaneous fibromas of white-tailed deer and mule deer.

Abstract Deer fibromavirus (DFV), a member of the papillomavirus genus, was isolated from cases of fibromatosis in white-tailed deer (Odocoileus virginianus) and mule deer (Odocoileus hemionus). SDS-polyacrylamide gel electrophoresis analysis of virions indicated no molecular weight differences in the major structural protein. DFV agglutinated mouse erythrocytes and the reaction could be inhibited by both DFV-specific and BPV-1-specific antisera. Although analysis of restriction endonuclease digestion products indicated minor differences in cleavage patterns, the DFV DNAs were indistinguishable by liquid phase hybridization and restriction enzyme cleavage maps indicated most of the sites in common. Comparison of DFV DNA to BPV-1 and BPV-2 DNA under stringent liquid-phase hybridization conditions indicated that 3 to 9% DNA sequence homology could be detected between the genomes of these viruses. Blot-transfer hybridization revealed that DFV DNA reannealed to the same restriction fragments of BPV-2 DNA under stringent conditions that DNA from other papillomaviruses hybridize to under nonstringent conditions.

Extrachromosomal deer fibromavirus DNA in deer fibromas and virus-transformed mouse cells

The non-virus-producing fibromatous portions of five deer fibromas were examined for deer fibromavirus (DFV) DNA sequences. Liquid-phase hybridization revealed 100 to 330 copies per cell of the virus genome. Southern blot analysis of undigested deer tumor DNA preparations indicated that most of the DFV DNA was present as monomeric, unintegrated genomes; however, restriction enzyme digestion patterns suggest a small population of resistant DFV sequences. DFV DNA was also present in virus-transformed NIH/3T3 mouse cells as multiple, extrachromosomal genomes.

Cloning and molecular characterization of an oral papillomavirus of domestic rabbits

DNA obtained from New Zealand white rabbit oral papillomas was analyzed for the presence of papillomavirus DNA. The viral genome was cloned as three separate subclones, which were each mapped and oriented with respect to one another. Comparisons with other papillomavirus DNAs by Southern blot hybridization under various conditions of stringency revealed a strong area of conservation among the DNAs of the rabbit oral papillomavirus (ROPV) and CRPV, HPV-1a, HPV-16, and BPV-5, but not with 12 other papillomavirus DNAs. This region, which spans the junction of the presumptive E2 and L2 open reading frames of ROPV, was sequenced and compared to other known papillomavirus sequences. These analyses revealed a high degree of DNA homology in the C-terminal E2 and N-terminal L2 regions between ROPV and both HPV-1a and CRPV. The homology with HPV-16 was limited to the L2 open reading frame. The predicted amino acid sequences of each region were also compared and bore out the same conclusions. In addition, no E5 open reading frame was detected in the ROPV sequence.

Cryptosporidium in a Wild Cottontail Rabbit (Sylvilagus floridanus)

Michael J. Ryan, Battelle Columbus Laboratories, 505 King Avenue, Columbus, Ohio 43201: John P. Sundberg, Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Illinois, Urbana, Illinois 61801, USA: Richard J. Sauerschell, Sterling-Winthrop Research Institute, Rensselaer, New York 11 144, USA; and Kenneth s. Todd, Jr., Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Illinois, Urbana, Illinois 61801, USA