Molecular and Phylogenetic Characterization of Novel Papillomaviruses Isolated from Oral and Anogenital Neoplasms of Japanese Macaques (Macaca fuscata).

Papillomaviruses (PVs) are a diverse group of host species-specific DNA viruses, etiologically linked with various benign and malignant neoplasms of cutaneous and mucosal epithelia. Here, we describe the detection and characterization of the first two PVs naturally infecting Japanese macaques (Macaca fuscata), including the determination of their etiological association(s) with the development of original neoplasms. The molecular and phylogenetic analyses were performed on complete genome sequences of Macaca fuscata PV types 1 (MfuPV1) and 2 (MfuPV2), which were completely sequenced in samples of a malignant oral tumor and benign anogenital neoplasm of Japanese macaques, respectively. Subsequently, two type-specific quantitative real-time PCRs were developed to estimate viral loads of MfuPV1 and MfuPV2 and to evaluate their etiological roles. The in silico molecular analyses revealed that both viral genomes encode characteristic PV proteins with conserved functional domains and have a non-coding genomic region with regulatory sequences to regulate and complete the viral life cycle. However, additional experimental evidence is needed to finally confirm the presence and biological functionality of the molecular features of both novel PVs. While MfuPV1, together with PVs identified in other macaques, is classified into the Alphapapillomavirus (Alpha-PV) species 12, MfuPV2 is most likely a representative of the novel viral species within the Alpha-PV genus. Their relatively high viral loads suggest that both PVs are etiologically linked with the development of the original neoplasms.

Papillomavirus infection and squamous cell carcinoma in horses.

Squamous cell carcinoma (SCC) is a common disease that seriously impairs the health and welfare of affected horses and other equids. In humans, almost all cervical carcinomas, a high percentage of anogenital SCCs and a subset of SCCs of the head and neck are caused by high-risk human papillomavirus (hrHPV) infection. Since hrHPV-induced human cancers and equine SCC have similar cytological and histopathological features, it has been hypothesised that equine SCCs could also be induced by papillomaviruses. This review provides an overview of the current evidence for an aetiological association between papillomavirus infections and equine SCCs and SCC precursor lesions. SCC of apparently papillomavirus-unrelated aetiology are also discussed, as are recent advances in equine SCC prophylaxis.

Chemoprevention of spontaneous ovarian cancer in the domestic hen.

The hen is an attractive animal model for in vivo testing of agents that thwart ovarian carcinogenesis because ovarian cancer in the domestic hen features clinical and molecular alterations that are similar to ovarian cancer in humans, including a high incidence of p53 mutations. The objective of the study was to test the potential ovarian cancer chemopreventive effect of the p53 stabilizing compound CP-31398 on hens that spontaneously present the ovarian cancer phenotype. Beginning at 79 wk of age, 576 egg-laying hens (Gallus domesticus) were randomized to diets containing different amounts of CP-31398 for 94 wk, 5 d, comprising a control group (C) (n = 144), which was fed a diet containing 0 ppm (mg/kg) of CP-31398; a low-dose treatment (LDT) group (n = 144), which was fed a diet containing 100 ppm of CP-31398; a moderate-dose treatment (MDT) group (n = 144) which was fed a diet containing 200 ppm of CP-31398; and a high-dose treatment (HDT) group (n = 144), which was fed a diet containing 300 ppm of CP-31398. Hens were killed at 174 wk of age to determine the incidence of ovarian and oviductal adenocarcinomas. Whereas the incidence of localized and metastatic ovarian cancers in the MDT and HDT groups was significantly lower (up to 77%) compared to levels in the C and LDT groups (P < 0.05), the incidence of oviductal cancer was unaffected by CP-31398. CP-31398 appears to be an effective tool for chemoprevention against ovarian malignancies, but does not appear to affect oviductal malignancies.

"TERATOCARCINOMATOSIS" IN AN EMU (DROMAIUS NOVAEHOLLANDIAE).

An adult female emu (Dromaius novaehollandiae) with no previous abnormal clinical signs was found dead in its stall. A postmortem examination revealed carcinomatosis of unknown origin. Histopathology identified the masses as teratomas with malignant transformation by the presence of poorly differentiated tissue and dissemination throughout the coelomic cavity. We propose a new term, "teratocarcinomatosis," for this finding. This is the first case of a reproductive tumor described in an emu and the first case of such marked spread of malignant teratomas in a bird.

Morphological and immunohistochemical observations on leiomyoma of the ventral ligament of the oviduct of the hen.

Leiomyomas of the ventral ligament (LVLs) of the oviduct from 2-year-old spent layers were examined. These tumours can be present either as single large masses or as multiple smaller nodules. The most common site of origin of the tumours was the centre of the free margin of the ventral ligament, but some small tumours were observed at the insertion of this ligament into the magnum of the oviduct. Most samples were highly vascular and some blood vessels within the tumours had vacuolation of the smooth muscle cells. These findings suggest that the proliferative processes leading to LVLs may include transformation of the blood vessels of the ventral ligament. Immunohistochemically, the tumour cells expressed vimentin, α-smooth muscle actin, desmin and heavy-caldesmon. These avian leiomyomas have been proposed as a model for similar tumours in other species.

Equus caballus papillomavirus-2 (EcPV-2): an infectious cause for equine genital cancer?

REASONS FOR PERFORMING STUDY: The aetiology of genital squamous cell carcinoma (SCC) in horses remains unknown, but the similarity to the disease in man, for which papillomavirus infection has been shown to be a causal factor, requires to be investigated in horses. HYPOTHESIS: One or more novel papillomaviruses cause equine genital SCC and its associated premalignant lesions. METHODS: DNA was extracted from samples of equine genital SCC and performed rolling circle amplification, in order to identify closed circular DNA viral genomes within the samples. The amplified DNA was subcloned and sequenced and the DNA sequence compared to that of other papillomavirus genomes. Using PCR primers developed from these genomic DNA sequences, studies were then carried out in order to identify the frequency at which the viral DNA could be identified in equine genital cancer samples from horses in both the UK, Australia and Austria. Finally, in situ hybridisation using specific probes developed from this DNA sequence were used to confirm the presence of the viral RNA sequences in the neoplastic cells in these lesions. RESULTS: The full length genome of a novel papillomavirus species was characterised from the equine genital SCC tissue and termed Equus caballus papillomavirus-2 (EcPV-2). Viral DNA and RNA was identified in the genital tumour samples, but not in the adjacent histologically normal tissue. EcPV-2 DNA could not be identified in equine ocular or nasal carcinomas or within the scrotal skin or in most smegma samples obtained from tumour-free horses. Sequencing of amplicons, generated from the archived equine genital tumours, identified variations within E1 and E6 on DNA and predicted protein level. CONCLUSIONS: A novel papillomavirus, EcPV-2, is likely to play a causal role in the pathogenesis of equine genital epithelial tumours. POTENTIAL RELEVANCE: Identification of a papillomavirus causal for genital carcinomas in horses may lead to development of a vaccine that could be used to prevent this serious disease in horses. This would be analogous to man, where vaccination against oncogenic papillomavirus species is currently being used to help prevent cervical cancer.

Neoplasms of the genital tract in a Vietnamese potbellied pig.

A 9-year-old, female potbellied pig showed loss of appetite and abdominal distension. After clinical examination and ultrasonography, a tumour was suspected. At laparotomy a large mass was present in the genital tract. Because the mass could not be excised, the pig was euthanized. Pathological examination revealed leiomyoma of the cervix and uterus wall in addition to multifocal adenocarcinomas of the uterus.

Oviduct adenocarcinoma in some species of captive snakes.

This article reports 5 cases of oviduct adenocarcinoma in adult captive snakes from Smithsonian's National Zoological Park. This neoplasm was found in 1 of each of the following species: emerald tree boa (Corallus caninus), Amazonian tree boa (Corallus enydris enydris), Burmese rock python (Python molurus bivittatus), Northern pine snake (Pituophis melanoleucus melanoleucus), and corn snake (Elaphe guttata). Grossly, tan to red firm masses were found within oviducts in 3 cases. In an additional 2 cases, the primary tumor was detected only histologically. Microscopically, neoplasms were papillary, and often extended transmurally. The neoplastic cells were polygonal and organized in acini or cords, with often abundant fibrovascular stroma. Hemorrhages and necrosis were present in all cases. Inflammation, myxomatous material, desmoplasia, and bacteria were often observed. Histologic evidence of metastasis was present in all cases. Solid metastases were seen in all animals except the Northern pine snake and involved several organs including the liver, lung, and heart. Emboli of neoplastic cells were observed in all animals but the Burmese rock python and corn snake.

Tumors of the urogenital system in dogs and cats. Retrospective review of 138 cases.

The aim of the study was to estimate the prevalence and localization of different tumors in the urogenital system in dogs and cats in relation to sex, age and breed of animals. The study was performed on tumors or tissue specimens from tumors of the urinary and genital system obtained during surgery from dogs and cats submitted to the Division of Pathological Anatomy, Department of Clinical Sciences Agricultural University of Warsaw from 1998 to 2005. Most tumors of the urogenital system recognized in the present study derived from dogs (94.20%, 130 cases), and only a few cases were obtained from cats (5.79%). Occurrence and localization of urogenital system tumors in present review is similar to findings reported by other authors. Testicular tumors in males, ovarian lesions in females and urinary bladder tumors in both sexes were most commonly recognized. Older dogs were most often affected, animals with nonmalignant tumors were a bit younger than those with malignant lesions. Any obvious breed predilections were found, but terriers were at increased risk for development of transitional cell carcinoma of the urinary bladder and mixed breed and German shepherd for development of testicular neoplasms.

Spontaneous lesions in the reproductive tract and mammary gland of female non-human primates.

Because of their close phylogenic relationship with humans, the use of non-human primates (NHP) as experimental subjects has a long history in biomedical research. Although research topics have shifted focus and species used have changed, NHP remain vital as models in basic and applied research. While there is a wealth of information available on the spontaneous lesions of NHP, most of this information is fragmented, dated, or narrow in focus, often limited to single case reports. This review attempts to integrate this information to illustrate and enumerate the spectrum of spontaneous pathology of the reproductive tract and mammary gland of NHP. Although not the focus of this review, steroid-related changes are inextricably linked to these tissues, and brief consideration is given to this subject as well.

Steroid receptors in canine and human female genital tract tumours with smooth muscle differentiation.

The expression of oestrogen receptor-alpha (ERalpha) and progesterone receptor (PR) was examined in 32 canine genital tract tumours diagnosed as smooth muscle tumours (benign or malignant, pure or mixed). The immunohistochemical expression of calponin was used to assess the smooth muscle differentiation of the tumours. Nineteen human uterine leiomyomas were also examined. Calponin expression was detected in 89.3% of canine and 100% of human genital tract tumours diagnosed as leiomyomas, as well as in the majority of other tumours examined (canine or human, genital or extragenital, benign or malignant) with the exception of canine negative control tumours (cutaneous fibroma and hepatoid gland adenoma). ERalpha was found in 56.3% of canine and 52.6% of human leiomyomas, while PR was found in 84.4% of canine and 94.7% of human tumours. These results indicate that calponin is a good marker for differentiating neoplasia of the canine genital system of uncertain origin, as in human patients. They also show that canine tumours with smooth muscle differentiation of the genital tract of the bitch express steroid hormone receptors, a finding that opens up the possibility of hormone therapy.

Spontaneously occurring fibroid tumors of the laying hen oviduct.

Spontaneously occurring benign uterine leiomyomas (fibroids) are the most common tumors of reproductive-age women. It is estimated that more than 70% of all women will develop uterine fibroids, and the presence of these tumors is a primary cause of hysterectomies. Research into the causes and treatment of uterine fibroids is hampered by a lack of reliable animal models for the disease. Leiomyomas that appear to be outwardly similar to human uterine fibroid tumors are known to occur on the oviducts of laying hens over 2 yr of age. The objective of this study was to characterize these tumors and compare them to human uterine fibroids to determine the suitability of the aging hen as a model system for the study of the disease. In this study, hens at 5 yr of age were examined for the presence of oviduct-associated fibroid tumors. Tumors were found attached to the internal surface of the oviduct, embedded in the oviduct wall, or attached to the exterior of the magnum and isthmus. Tumor and normal oviduct samples were frozen or fixed in formalin for histological analyses or immunohistochemistry for estrogen and progesterone receptors, proliferating cell nuclear antigen and Bcl-2 protein expression. Human uterine fibroid samples were acquired and evaluated compared with hen oviduct fibroids. The results indicate that laying hen fibroid tumors are similar to human fibroid tumors with respect to estrogen and progesterone receptors, localized cellular proliferation, and expression of the Bcl-2 protein.

Reproductive oncology.

The actual incidence of reproductive tract tumors is difficult to ascertain, presumably because a significant percentage of dogs and cats are neutered, particularly in the United States. Data from European countries, where elective neutering is less common, provides insights. Several studies document the relative frequency of reproductive tumors. Larger studies of the more common tumors, e.g., testicular tumors, provide a substantial amount of information on the clinical presentation and response to treatment of dogs, but overall there is less information available on cats. Numerous case reports document the less commonly reported tumors and will be included in the following discussion.

Genital tract smooth muscle tumors are common in zoo felids but are not associated with melengestrol acetate contraceptive treatment.

In a survey of gynecologic lesions in female zoo felids conducted to determine if the widely used progestin contraceptive melengestrol acetate (MGA) had adverse effects, numerous leiomyomas and leiomyosarcomas were detected. This current study aimed to characterize the morphologic features of these tumors, determine their prevalence, and assess if MGA was a risk factor for their genesis. Genital tracts from 219 zoo felids representing 23 species were evaluated, and leiomyomas were detected in 24% of the felids. Leiomyomas were often multiple and occurred in the myometrium, ovary, or adjacent broad ligament. The risk of developing leiomyomas increased with age, but MGA treatment or parity had no effect. Five other felids had leiomyosarcomas. Leiomyosarcomas were distinguished from poorly demarcated leiomyomas by the presence of local invasion, metastasis, and cellular atypia, but necrosis and mitotic rate were not distinguishing criteria. Four of five felids with leiomyosarcomas had been treated with MGA. These results indicate that leiomyomas are common spontaneous lesions in the genital tracts of zoo felids and their genesis is not linked to MGA exposure. Whether progression to malignancy is promoted by MGA warrants further investigation.

Histopathological studies on neoplasms in the genital tract of female donkeys in Egypt.

The genital tracts of 165 female donkeys were collected and examined with special concern to neoplasms. Ovarian Neoplasms were found in 12 cases (7.72%). A granulosa cell tumor was found in one case (0.61%). A cavernous haemangioma was found in 11 cases (6.67%). Uterine neoplasms were found in 14 cases (8.84%). Endometrial polyps and fibroleiomyoma were found in one case (0.61%) each. Leiomyoma was found in 2 cases (1.21%). Cavernous haemangioma was found in 10 cases (6.06%), and cervical polyps in 5 cases (3.03%).

Common metastatic carcinoma of California sea lions (Zalophus californianus): evidence of genital origin and association with novel gammaherpesvirus.

Tissues from 10 adult California sea lions (Zalophus californianus, seven females and three males) that had metastatic carcinoma in sublumbar area lymph nodes were examined histologically. A distinctive epithelial proliferative lesion interpreted as intraepithelial neoplasia was found in genital tracts of all ten animals; in vagina (5/7), cervix (7/7), uterus (3/7), penis (3/3) and prepuce (3/3). Intraepithelial neoplasia closely resembled metastatic carcinomas and was directly contiguous with invasive carcinoma in one animal. Rare eosinophilic intranuclear inclusion bodies were found in penile and preputial intraepithelial neoplasia (one animal), cervical intraepithelial neoplasia (one animal), invasive cervical carcinoma (one animal) and metastatic carcinoma (two animals). Electron microscopic examination of tissues from two sea lions (one with intraepithelial neoplasia and one with metastatic carcinoma) demonstrated viral particles consistent with a herpesvirus. An immunohistochemical stain for the latent membrane protein of Epstein-Barr virus was positive in intraepithelial neoplasia in one sea lion. Herpesvirus DNA sequences were detected by consensus primer polymerase chain reaction (PCR) in metastatic carcinomas from all four sea lions from which unfixed tumor samples were available. Results of sequencing were consistent with a novel gammaherpesvirus in the genus Rhadinovirus. DNA extracted from the four metastatic carcinomas also was tested for papillomavirus by Southern blot and PCR with consensus papillomavirus primers; all samples were negative by both methods. These findings support the genital origin of the sea lion carcinoma and implicate a novel gammaherpesvirus as a possible cause.

Caprine genital leiomyosarcoma.

An aged Saanen doe was euthanized following repeated severe hemorrhage from the vulva. Necropsy examination revealed mural thickening of tubular genitalia with firm, protruding intralumenal masses containing blood-filled cavitations, and effacement of normal cervical architecture. Histologically, uterine and cervical masses comprised a variably dense population of mildly pleomorphic spindle cells forming interlacing streams supported by variably dense collagenous stroma. Immunoperoxidase staining of neoplastic cells was positive for muscle-specific actin, supporting the diagnosis of low-grade leiomyosarcoma. Months later, the doe's twin was likewise euthanized due to persistent bleeding from the vulva associated with a large vulvar mass having histopathologic features similar to those of the previous case. The clinical, gross, and histologic findings are similar to five cases of caprine genital leiomyosarcoma identified in retrospectively analyzed case material. Analysis of caprine tumor accessions over 20 years demonstrated a significantly higher incidence of genital leiomyosarcoma within the Saanen breed.

Spontaneous tumours in tree shrews (Tupaia belangeri): population studies.

In a necropsy series at the German Primate Centre, 63 of 1132 tree shrews were found to have tumours. The youngest affected animal was < 6 months old, but the incidence of tumours increased strikingly beyond 2 years of age; of 17 animals aged > 8 years, no less than 14 (82.4%) were affected. The sites of neoplasia were the genital system (22 cases), haematopoietic system (18 cases of malignant lymphoma, many involving the digestive tract), lungs (14 cases), integument (five cases), digestive tract (three cases other than those with malignant lymphoma) and urinary bladder (one case).

Neoplasia of the female reproductive tract.

Granulosa cell tumors of the ovary and SCC and melanoma of the external genitalia are the most common neoplasms in the mare. Tumors of the tubular reproductive tract and the mammary gland are rare.