Squamous cell carcinoma in a tuatara (Sphenodon punctatus)

CASE HISTORY: A mature male tuatara was presented with a history of recurrent cloacal prolapse. CLINICAL AND PATHOLOGICAL FINDINGS: The prolapsed tissue included a 12 mm diameter mass, which histologically showed cords and nodules of neoplastic epithelial cells. DIAGNOSIS: The mass was diagnosed as a squamous cell carcinoma with ulceration of the overlying stratified squamous epithelium and diffuse inflammation of the surrounding dermis. CLINICAL RELEVANCE: This case presented a rare opportunity to investigate squamous cell carcinoma in a long-lived lower vertebrate species. Future recurrence or metastasis of the mass may add useful information to the current base of knowledge of the behaviour of malignant neoplasms in reptiles.     

Health screening for a translocation of captive-reared tuatara (Sphenodon punctatus) to an island refuge

AIMS: To screen tuatara undergoing translocation from a captive crèche to an island refuge for evidence of health and known diseases, and apply basic epidemiological techniques to assess the significance of disease test results.

METHODS: Tuatara (n=353) were physically examined and samples were taken from a random selection (n=30) for estimated white cell counts, screening for haemoparasites, and culture for Salmonella, Yersinia, Aeromonas and Campylobacter spp. Direct faecal smears were carried out on-site, and faecal floats were later performed to assess levels of endoparasitism with helminths and protozoa (n=69). Modified Ziehl-Neelsen staining was used to screen faecal smears, and positive specimens were further screened using an immunofluorescence antibody (IFA) test for Cryptosporidium oocysts.

RESULTS: There was no evidence of external parasites on any of the animals examined and only one animal had a gross abnormality. All estimated white cell counts were in the range 2.8– 17.5 × 10⁹/L. No haemoparasites were observed. There were no enteric pathogens cultured, indicating the intestinal carriage of these bacteria in the tuatara was 9.4%. Of the 69 individual faecal samples examined, 12 (17%) had unidentified coccidial oocysts, 21 (30%) had nematode ova of various kinds, and 12 (17%) had intestinal carriage of motile protozoa consistent with Trichomonas spp and another unidentified organism. Nineteen (28%) tuatara had acid-fast oocysts present; however, IFA staining failed to detect any Cryptosporidium oocysts.

CONCLUSIONS: Our understanding of the diversity of gastrointestinal endoparasites affecting tuatara is inadequate as many of the parasite ova seen could not be identified. This is the first record of tuatara as a host for Trichomonas spp of protozoa in the gastrointestinal tract.     

Presence of antibodies to Salmonella in tuatara (Sphenodon punctatus) sera

Colonisation of a host by pathogenic microorganisms is a near constant threat to the health of all vertebrates and most species have evolved an efficient adaptive immune response which produces antibodies following exposure to a specific antigen. The strength of this response can be influenced by many factors including sex and season. Tuatara are exposed to Salmonella through contact with infected skinks and soil; however, no gastrointestinal colonisation of tuatara with Salmonella has been found. Using Western blot and flow cytometry we have demonstrated that tuatara possess antibodies which recognise Salmonella antigens, but many of these antibodies are not specific and are cross-reactive with two closely related and ubiquitous bacteria, Escherichia coli and Citrobacter koseri. Our study describes the anti-Salmonella immune responses in tuatara and will help to inform decisions around maintaining wildlife health, as well as providing important insights into the role and development of adaptive immunity in reptilian species.     

T cell function in tuatara (Sphenodon punctatus)

Tuatara are the sole survivors of an entire order of reptiles that thrived during the age of the dinosaurs. Therefore, knowledge of their physiology is critical to understanding the phylogeny of reptiles. Previous studies of the immune system of the tuatara did not assess T cell function. We analyzed T cell function among six captive tuatara by assessing concanavalin A (Con A), phytohemagglutinin (PHA) and mixed lymphocyte reaction (MLR) induced T cell proliferation. Peripheral blood mononuclear cells from six out of six and four out of four tuatara tested exhibited significant proliferative responses to Con A and PHA, respectively, as measured by an MTT reduction assay. A lower level of proliferation was detected in an MLR. However, Con A activated lymphocytes were not cytotoxic for a xenogeneic murine mastocytoma cell line (P815).     

Squamous cell carcinoma of the paranasal sinuses in two horses

This case report describes the history, clinical findings, diagnostic imaging (including computed tomography), gross pathology and histopathology of 2 cases of squamous cell carcinoma in the paranasal sinuses. Radiography of the skull is a useful tool to detect a mass in the paranasal sinuses, but the position, extent and character of the lesion can be better visualised with computed tomography.     

Squamous cell carcinoma associated with a periorbital mass in a veiled chameleon (Chamaeleo calyptratus)

This report describes a squamous cell carcinoma in a 1-year-old female veiled chameleon (Chamaeleo calyptratus). The lesion developed as a small (1 by 1 mm) left periocular discoloration of a scale never involving the eye. The mass was first diagnosed as an abscess, increased in size (4 by 8 by 3 mm), and recurred after two surgical resections combined with antibiotic therapy. Poor nutritional condition and egg production by the chameleon complicated management of this condition. The mass was removed surgically a third time at which point histopathologic evaluation revealed a locally invasive squamous cell carcinoma. Bacterial culture of the mass isolated a pure culture of Pseudomonas aeruginosa. Ceftazidime was administered at 20 mg/kg IM every 48 h for 20 days. The animal died 3 months later from complications during an ovariohysterectomy for pregnancy toxemia and oviduct inertia. Necropsy showed no local recurrence or metastasis of the tumor.     

Failure to detect Salmonella species in a population of wild tuatara (Sphenodon punctatus)

AIM: To assess the prevalence of faecal excretion of Salmonella serovars by wild tuatara (Sphenodon punctatus) on Stephens Island, New Zealand. METHODS: One hundred cloacal swabs obtained as part of health-screening for the translocation of adult tuatara from Stephens Island were subjected to general aerobic culture and enrichment, and cultured specifically for Salmonella spp. RESULTS: No Salmonella spp were cultured from any of the cloacal samples, which suggests that, at the 95% confidence interval, the maximum prevalence of tuatara in the island population that were shedding Salmonella spp not detected by our sample size was 1.5%. Mixed bacteria were grown from the 70 cloacal swabs cultured aerobically. A predominant organism was evident in 30 cultures, and these were identified as Hafnia alvei type 1 (n=16) and type 2 (n=7), Corynebacterium spp (n=4), Klebsiella oxytoca (n=2), and Moraxella spp (n=1). CONCLUSIONS: The absence of intestinal carriage of Salmonella spp by the tuatara sampled in this study may indicate either lack of exposure, or an innate resistance to intestinal colonisation in tuatara. The significance of the other bacteria cultured as potential pathogens to the tuatara and as zoonotic risks is also uncertain. Wildlife managers should screen translocated reptiles for Salmonella spp, and thereby avoid exposing wild and managed populations to infection.     

Health screening for a translocation of captive-reared tuatara (Sphenodon punctatus) to an island refuge

AIMS: To screen tuatara undergoing translocation from a captive crèche to an island refuge for evidence of health and known diseases, and apply basic epidemiological techniques to assess the significance of disease test results. METHODS: Tuatara (n=353) were physically examined and samples were taken from a random selection (n=30) for estimated white cell counts, screening for haemoparasites, and culture for Salmonella, Yersinia, Aeromonas and Campylobacter spp. Direct faecal smears were carried out on-site, and faecal floats were later performed to assess levels of endoparasitism with helminths and protozoa (n=69). Modified Ziehl-Neelsen staining was used to screen faecal smears, and positive specimens were further screened using an immunofluorescence antibody (IFA) test for Cryptosporidium oocysts. RESULTS: There was no evidence of external parasites on any of the animals examined and only one animal had a gross abnormality. All estimated white cell counts were in the range 2.8- 17.5 x 10(9)/L. No haemoparasites were observed. There were no enteric pathogens cultured, indicating the intestinal carriage of these bacteria in the tuatara was <9.4%. Of the 69 individual faecal samples examined, 12 (17%) had unidentified coccidial oocysts, 21 (30%) had nematode ova of various kinds, and 12 (17%) had intestinal carriage of motile protozoa consistent with Trichomonas spp and another unidentified organism. Nineteen (28%) tuatara had acid-fast oocysts present; however, IFA staining failed to detect any Cryptosporidium oocysts. CONCLUSIONS: Our understanding of the diversity of gastrointestinal endoparasites affecting tuatara is inadequate as many of the parasite ova seen could not be identified. This is the first record of tuatara as a host for Trichomonas spp of protozoa in the gastrointestinal tract.     

Squamous cell carcinoma in a digit of the hind limb with systemic metastasis in a 17-year-old female koala

We encountered a case of cutaneous squamous cell carcinoma (SCC) in a 17-year-old female koala at a zoo. A fragile, papillary, elevated mass was found on the third digit of the right hind limb. SCC was identified histopathologically: squamous cell-like polygonal tumor cells showed a nest-like growth pattern with epidermal down growth, central keratinization and necrotic foci, and invaded dermal connective tissues. Metastatic lesions were observed in various organs, including the lung and axillary lymph node: in the lung, multiple metastatic foci similar to the primary lesion, and in the axillary lymph node, individual polygonal tumor cells infiltrated the sinusoids. Immunohistochemistry revealed that the tumor cells were positive for proliferating cell nuclear antigen, which exhibited 32–33% of labeling indices in the tumor cells. To our knowledge, this is the first report of a case of SCC in a digit of a koala.     

Squamous cell carcinoma of the nasal planum in cats and dogs

The purpose of this article is to review the therapeutic options available for the treatment of squamous cell carcinoma of the nasal planum in cats and dogs. The techniques of complete and partial nasal planum resection in the cat are described in detail. Surgical treatment offers the greatest chance of cure, although several options are available for early, less invasive lesions.     

Long-term outcome of treatment of a squamous cell carcinoma of the foot by amputation of the distal limb in a pony

This report describes a rare case of squamous cell carcinoma (SCC) in a 10-year-old Shetland pony. The pony was presented for evaluation of a chronic, ulcerating mass of the foot associated with a nonweightbearing lameness of the right forelimb. Foot radiographs revealed an aggressive bone lesion with severe osteolysis of the distal phalanx. Amputation of the digit was performed under general anaesthesia at the level of the metacarpophalangeal joint using a palmar flap technique and a transfixation cast for protection of the stump. Avascular necrosis, infection and dehiscence of the stump occurred 3 weeks later and a second amputation was performed at the level of the proximal third of the third metacarpal bone. Histopathology revealed a squamous cell carcinoma. After healing of the stump, a prosthesis was fitted to the limb for improved ambulation of the pony. Two years after the amputation, telephone follow-up with the referring veterinarian and the owner, revealed that the pony was healthy and able to go out daily in a paddock with its prosthesis. SCC represents an unusual indication for limb amputation. Successful outcome is rarely reported in horses.     

Detection of Paranannizziopsis australasiensis in tuatara (Sphenodon punctatus) using fungal culture and a generic fungal PCR

AIMS: To describe the methods used at the Animal Health Laboratory (AHL, Ministry for Primary Industries) to identify Paranannizziopsis australasiensis.

METHODS: Skin biopsy samples from two adult male tuatara were submitted to the AHL in March 2014. Approximately half of each sample was processed for fungal culture and incubated on mycobiotic agar containing cycloheximide at 30°C. Following morphological examination of the culture products, DNA was extracted from suspect colonies. PCR was used to amplify the internal transcribed spacer (ITS) region of fungal rRNA using primers ITS1 and ITS4. Positive amplicons were subjected to DNA sequencing and the results were compared to published sequences. In addition, DNA was extracted from the remaining skin samples and the same PCR was carried out to compare the results.

RESULTS: After 7 days of incubation, colonies morphologically resembling P. australasiensis were observed. DNA extracted from these isolates tested positive for P. australasiensis by PCR and DNA sequencing. Samples of DNA extracted directly from the infected skin samples tested negative for P. australasiensis using the generic fungal PCR.

CONCLUSIONS AND CLINICAL RELEVANCE: Isolation and identification of P. australasiensis was carried out using a combination of fungal culture and molecular testing available at AHL. Results were available in significantly less time than in the past, when isolates had to be sent overseas. PCR and sequencing of fungal isolates is a valuable tool for identification of species that have few, if any, unique macroscopic or microscopic features to aid identification. Further sampling from captive and wild New Zealand reptiles will provide important information on the epidemiology of P. australasiensis, and the conservation and management implications for tuatara and other native reptile species.     

Corneal squamous cell carcinoma in a Border Collie

A 6-year-old, female, spayed Border Collie was presented to the Unit of Comparative Ophthalmology at the Animal Health Trust with a 6-month history of a progressive nonpainful opacity of the left cornea. A keratectomy was performed and the tissue submitted for histopathology. The diagnosis was squamous cell carcinoma. There has been no recurrence of the neoplasm to date (5 months). Canine corneal squamous cell carcinoma (SCC) has not been reported previously in the UK.     

Piroxicam and carboplatin as a combination treatment of canine oral non-tonsillar squamous cell carcinoma: a pilot study and a literature review of a canine model of human head and neck squamous cell carcinoma

Results of the treatment with a combination of carboplatin and piroxicam in seven dogs with advanced non‐tonsillar oral squamous cell carcinoma (SCC) were retrospectively analysed. This multi‐agent protocol was well tolerated by all dogs and resulted in a complete regression of the tumour without additional surgery in four of seven patients. Additional surgery was necessary to remove a metastatic lymph node in one dog and residual tumour in a second dog, which achieved a partial response following medical therapy. Median follow‐up for all the dogs was 534 days, while the time‐to‐recurrence, time‐to‐progression and overall survival for this group of patients have not yet been reached. Our study, although limited in number of animals, suggests that this multiagent approach is a useful treatment option for oral non‐tonsillarSCC in dogs and warrants wider application.     

Transitional cell carcinoma metastatic to the eye in a collared peccary (Tayassu tajacu)

A 15-year-old female collared peccary (Tayassu tajacu) was presented for ophthalmic examination following sudden onset of blindness. Bilateral retinal detachment was diagnosed, neoplasia suspected, and euthanasia performed. Widespread tumor dissemination was apparent at autopsy, and transitional cell carcinoma was diagnosed histologically. The tumor was identified as arising from the ovary. Epidemiologic features of this case are discussed.     

Detection of a novel herpesvirus associated with squamous cell carcinoma in a free-ranging Blanding’s turtle

The spread of both infectious and noninfectious diseases through wildlife populations is of increasing concern. Neoplastic diseases are rarely associated with population-level impacts in wildlife; however, impacts on individual health can be severe and might reflect deteriorating environmental conditions. An adult male free-ranging Blanding’s turtle (Emydoidea blandingii) originally captured in 2005 and deemed healthy, was recaptured in 2018 with a 1 × 1.5 cm intra-oral broad-based right mandibular mass. An excisional biopsy was performed, and histopathology revealed squamous cell carcinoma (SCC). Consensus herpesvirus PCR identified a novel herpesvirus (proposed name Emydoidea herpesvirus 2 [EBHV-2]) within the tumor. EBHV-2 shares 85% sequence homology with Terrapene herpesvirus 2 (TerHV-2), a herpesvirus linked to fibropapillomas in eastern box turtles (Terrapene carolina carolina). Virus-associated fibropapillomas have been identified in multiple marine turtle species and have had debilitating effects on their populations, but to date, virus-associated SCCs are rarely reported.     

Dermatomycosis caused by Paranannizziopsis australasiensis in five tuatara (Sphenodon punctatus) and a coastal bearded dragon (Pogona barbata) in a zoological collection in New Zealand

CASE HISTORY: Health monitoring of tuatara (Sphenodon punctatus) at Auckland Zoo between 2001 and 2009 showed that 58/93 tuatara had been affected by dermatitis of unknown origin. From 2011 onwards, cases of suspected fungal dermatitis underwent extensive diagnostic investigations.

CLINCAL FINDINGS: Six cases of dermatomycosis were attributed to Paranannizziopsis australasiensis, five in tuatara and one in a coastal bearded dragon (Pogona barbata). Cases presented typically as raised, yellow to brown encrustations on the skin. Severe cases progressed to necrotising ulcerative dermatitis, and in the bearded dragon to fatal systemic mycosis. Following topical and systemic treatments, lesions resolved in all five tuatara.

LABORATORY FINDINGS: Histopathological examination of skin biopsy samples revealed dermatitis with intralesional septate branching hyphae. Fungal culture yielded isolates morphologically resembling Chrysosporium species, and isolates were submitted for molecular confirmation and sequencing of DNA.

DIAGNOSIS: All six cases were confirmed as dermatitis due to infection with P. australasiensis, on the basis of fungal culture and DNA sequencing of isolates.

CLINICAL RELEVANCE: These are the first reported cases of dermatomycosis associated with P. australasiensis infection in tuatara, and the first cases in which systemic therapeutic agents have been used in the treatment of such disease. Tuatara at the Auckland Zoo are now routinely examined every 3 months and tissue samples from any lesions sent for histopathology and fungal culture. Further work to elucidate the epidemiology and significance of P. australasiensis infections in reptiles in New Zealand is important for both welfare and conservation purposes.