Cutaneous transmissible venereal tumor without genital involvement in a prepubertal female dog

An 11-month-old prepubertal crossbreed female dog was presented with multiple nodular lesions disseminated over the cervical, back, flank, and abdominal regions. The lesions were ulcerated and cauliflowerlike, or nodular and subcutaneous, measuring up to 13 cm in diameter. Cytologic preparations of one of the lesions revealed a uniform population of round to oval cells, with lightly basophilic cytoplasm that contained multiple distinct vacuoles. Frequent mitotic figures and occasional lymphocytes were also observed. The cytologic diagnosis was cutaneous transmissible venereal tumor (TVT) in a progressing growth phase. This was confirmed by histologic and immunohistochemical findings. Vaginal TVT was diagnosed later in the dog's mother. TVT is a contagious neoplasm of sexually mature dogs that usually is transmitted by coitus and affects the genital mucosa. To our knowledge, this is the first report of naturally occurring multicentric TVT in a prepubertal female dog and also is unique in its exclusively cutaneous (no mucosal) involvement. We speculate that transmission of neoplastic cells occurred during cohabitation and social/mothering behavior between the dogs. Despite the atypical clinical presentation, response to chemotherapy with vincristine was excellent, leading to complete regression of the neoplasm without relapse after 6 months.     

Immunohistochemical characterization of intraocular metastasis of a canine transmissible venereal tumor

Little has been published on intraocular metastasis of transmissible venereal tumors (TVT) in dogs. This report presents a 4-year-old male Labrador Retriever with a previous history of subcutaneous TVT which underwent total remission after treatment with vincristine. The dog presented with clinical signs of uveitis and increased intraocular pressure (IOP) in both eyes. After enucleation of the left eye, a diagnosis of TVT was made based on morphology, histology and immunohistochemistry (IHC). IHC staining for vimentin, S-100 protein, cytokeratin and HMB45 was performed to differentiate this lesion from TVT, lymphoma, melanoma, carcinomas, neurogenic tumors and fibrosarcoma. The IHC findings supported the diagnosis of TVT for this round cell tumor.     

Evaluation of a genetic assay for canine transmissible venereal tumour diagnosis in Brazil

The canine transmissible venereal tumour (CTVT) is a transmissible cancer that is spread between dogs by the allogeneic transfer of living cancer cells. The infectious agents in CTVT are the living cancer cells themselves, which are transmitted between dogs during coitus. CTVT first arose several thousand years ago and the disease has a global distribution and is frequently observed in dogs from Brazil. We evaluated the utility of a LINE‐MYC quantitative polymerase chain reaction for diagnosis of CTVT cases in Brazil. Our analysis indicated that the LINE‐MYC rearrangement was detectable in all CTVT samples but not in their corresponding hosts. This genetic assay proves to be a useful tool for providing a definitive molecular diagnosis of CTVT, which presents with varying degrees of aggressiveness and invasiveness in different host dogs and can therefore be a diagnostic challenge in some specific cases.     

Clinico-pathological studies on the effect of different anti-neoplastic chemotherapy regimens on transmissible venereal tumours in dogs

Thirty-two dogs affected with transmissible veneral tumour (TVT) were divided into three treatment groups. In group I vincristine sulphate at 0.025 mg/kg body weight, in group II vinblastine sulphate at 0.150 mg/kg body weight, and in group III vinblastine sulphate at 0.100 mg/kg body weight plus methotrexate at 0.35 mg/kg body weight were given intravenously at weekly intervals. Biopsies were performed on days 0, 3, 7 and 14. The tissues were preserved in 10% neutral buffered formalin and processed routinely for haematoxylin and eosin staining. Histopathologically, the untreated TVT was characterized by sheets or bundles of mostly rounded cells having a large, highly basophilic nucleus with a prominent, highly basophilic nucleolus. Both vincristine and vinblastine primarily affected the nuclei of neoplastic cells, causing condensation, karyorrhexis and karyolysis within 3 days of chemotherapy. The regressing tumour mass showed marked infiltration by lymphocytes, lymphoblasts and macrophages by day 7. There was nearly complete regression of the tumour by day 14, as shown by the almost complete loss of neoplastic cells, with fibrous tissue substitution. However, in group III, the changes occurred more slowly and more injections were needed for complete regression. In both groups I and II, 11/12 of the animals responded completely to the chemotherapy within 3 weeks, while in group III, 6/8 of the dogs responded to the treatment by 21–28 days.Abbreviations SD standard deviation - TVT transmissible venereal tumour     

Ocular involvement of transmissible venereal tumor in a dog

A case of metastatic transmissible venereal tumor (TVT), with lesions on the penis, conjunctiva, buccal mucosa and skin (lips and trunk), is presented in this case report. The clinical picture is described along with the cytological and histopathological features of the tumor leading to definitive diagnosis of TVT. Possible reasons for the unusual metastatic behavior of TVT and full recovery of the dog after chemotherapy are discussed.     

Mitosis and apoptosis in canine cutaneous histiocytoma and transmissible venereal tumour

Cell proliferation and apoptosis in canine cutaneous histiocytomas and transmissible venereal tumours were examined in twenty cases. The Ki-67 immunohistochemistry and Tunel methods were used to detect mitotic activity and apoptosis, respectively. The number of Ki-67 immunoreactive cells was 11.65 (+/- 1.1706) in canine cutaneous histiocytomas and 17 (+/- 2.1751) in transmissible venereal tumours. The mean values of apoptotic cells for canine cutaneous histiocytomas and transmissible venereal tumours were 13.25 (+/- 1.8758) and 8.52 (+/- 1.1007), respectively. It was considered that mitotic activity and apoptotic indices were useful in differentiation of canine cutaneous histiocytomas and transmissible venereal tumours. The correlation values for canine cutaneous histiocytomas and transmissible venereal tumours were 0.359 (+/- 0.330) and -0.232 (+/- 0.344), respectively. No significant (P > 0.05) correlation was found between mitosis and apoptosis in these two tumour types.     

The distribution of C-bands in canine transmissible venereal tumor cells

Colchicine-arrested metaphase preparations, derived from canine transmissible venereal tumor (TVT) cells grown in culture, were characterized based on the occurrence and distribution of constitutive heterochromatin. Analysis of the data with regard to the distribution of C-bands in the pericentric, interstitial and telomeric segments revealed a nonrandom distribution along the arms of many chromosomes with the bulk of the bands occurring in the centromeric region. The frequency of C-banded regions differed from those reported for normal dog cells and both primary and transplanted tumors. These results suggest that similar nondifferentially-stained TVT karyotypes do not necessarily exhibit identical distribution of constitutive heterochromatin.     

Ultrastructural characteristics of canine transmissible venereal tumor at various stages of growth and regression.

Ultrastructural study of canine transmissible tumors in developing, mature, and regressing stages from 6 dogs revealed the presence of healthy and degenerating tumor cells in all neoplasms. The total number of neoplastic cells seemed to decrease, and the number of degenerating neoplastic cells seemed to increase in mature tumors. Macrophages, lymphocytes, and plasma cells infiltrated mature and regressing tumors. Alteratons in degenerating tumor cells consisted mainly of cytoplasmic changes in early stages and of both nuclear and cytoplasmic changes in cells in which degeneration was more advanced. Amounts of endoplasmic reticulum and ribosomes were decreased. There were swelling and vacuolation of mitochondria. Nuclear chromatin was clumped along the nuclear envelope, and the perinuclear space was widened. Degenerating cells often contained membrane-bound granules and clusters. Lamellar complexes were observed in tumor cells from 2 dogs. Virus particles were not seen.     

Role of exfoliative cytology in the diagnosis of canine transmissible venereal tumour

The objectives of this study were to evaluate the role of exfoliative cytology in the diagnosis of canine transmissible venereal tumour and to improve the success rate of surgical excision of this tumour. The technique was used to screen 360 dogs and, at the time of surgery, 34 clinical cases. Seventy-five per cent of the cases detected by screening were described as early stage disease in comparison with 23 per cent of cases reported by owners. The value of exfoliative cytology at the time of surgery to determine tumour cell removal was demonstrated by the reduction in local tumour recurrence from 22 per cent to 8 per cent. The technique is, therefore, recommended for use in screening dogs for canine transmissible venereal tumour and in determining the extent of surgical removal. However, more work is needed to assess its applicability in practice by determining its sensitivity, specificity and predictive values of a positive and negative test answer.     

Incidence of drug resistance and transmissible R factors in strains of E. coli isolated from faeces of healthy pigs

Two hundred and twenty-six strains of E. coli were isolated from faeces of 107 pigs at different ages and without clinical signs of infectious diseases. The resistance of the strains to sulphonamide, tetracycline, streptomycin, chloramphenicol, ampicillin, and nalidixic acid was determined. In 74 % of the animals the predominant E. coli flora was found to be resistant to one or more of the drugs mentioned. Fifty-three % of the strains were resistant. Multiple resistance was predominant among resistant strains (67 %). R factors transmissible to a sensitive strain of E. coli K12 W3132 were demonstrated in 28 %. The proportion of resistant strains was largest in young animals (0–14 weeks) accounting for 65 % of the strains isolated, as compared to 43 % of strains from pigs and sows (6 months or more). The incidence of resistance to sulphonamide, tetracycline, and streptomycin was high, whereas most of the strains were sensitive to ampicillin and chloramphenicol. All strains were sensitive to nalidixic acid.The incidence of resistance to antibiotics in a population of pigs to whom these drugs are not fed but applied as therapeutic agents solely seems rather high. When based on clinical findings only, the value is therefore questionable of sulphonamide, tetracycline and streptomycin treatment of infectious diseases caused by E. coli.     

Prevalence of Salmonella spp. in Cane Toads (Bufo marinus) from Grenada,West Indies,and their Antimicrobial Susceptibility

Cloacal swabs and caecal contents sampled from 58 cane toads (Bufo marinus) in St George’s parish, Grenada, during a 7‐month period in 2011 were examined by an enrichment and selective culture method for presence of Salmonella spp. Twenty‐four (41%) toads were positive for Salmonella spp. of which eight were Salmonella enterica serovar Javiana, and eight were S. enterica serovar Rubislaw. The other serovars were as follows: Montevideo, 6; Arechavaleta, 1; and serovar: IV:43:‐:‐, 1. The high frequency of isolation of serovar Javiana, an emerging human pathogen associated with several outbreaks in the recent years in the eastern United States, suggests a possible role for cane toads in transmission of this serovar. Although S. Rubislaw has been isolated from lizards, bats and cases of some human infections, there is no report of its carriage by cane toads, and in such high frequency. The rate of carriage of S. Montevideo, a cause for human foodborne outbreaks around the world was also over 10% in the 58 toads sampled in this study. The antimicrobial drug susceptibility tests against amoxicillin‐clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, imipenem, nalidixic acid, streptomycin, tetracycline and trimethoprim‐sulfamethoxazole showed that drug resistance is minimal and is of little concern. Antimicrobial resistance was limited to ampicillin and amoxicillin‐clavulanic acid in one isolate of S. Javiana and one isolate of S. Rubislaw. This is the first report of isolation and antimicrobial susceptibilities of various Salmonella serovars not identified previously in cane toads in Grenada, West Indies.     

Risk factors of ciguatera in the French West Indies in Saint-Barthélémy, Saint-Martin and Anguilla

An epidemiological study on ciguatera fish poisoning in the French West Indies (St-Barthelemy, St-Martin and Anguilla) was conducted during the years 1985-1986. The investigation on intoxications shows a non seasonal significant prevalence. Though it was difficult to list the cases, the morbidity seems to vary between 7 and 30 per thousand. A study of fish toxicity was realised; 46 different species were tested (individually or by pools) by the mosquito bio-test. Observations of intoxications give the following results: High risk species: Caranx bartholomaei, C. lugubris, Seriola dumerili, Lutjanus apodus, L. jocu, Pristipomoides macrophtalmus, Gymnothorax funebris, G. moringa, Scomberomorus cavalla, S. regalis, Mycteroperca venenosa, M. tigris, Epinephelus morio, Sphyraena barracuda. Intermediate species: Caranx latus, C. ruber, Lachnolaimus maximus, Lutjanus analis, L. buccanella, L. griseus, Malacanthus plumieri, Scomberomorus maculatus. Low risk species: Balistes vetula, Alectis ciliaris, Haemulon album, Bodianus rufus, Halichoeres radiatus, Priacanthus arenatus, Alphestes afer. Many species are involved in the toxic food chain. New ones have been identified, but it is difficult to determine the toxic level range. A cartography is presented but no place is free of risk. A research of Gambierdiscus toxicus, the causal agent, on algal surface from dead corals was conducted around St-Barthelemy and St-Martin. The dinoflagellate is found in low or medium populations all around the islands with no difference between North and South. There is a maximal activity during the spring. A model of the epidemiology of the ciguatera in the area is proposed.     

Growth and morphological characteristics of canine venereal tumor cells in vitro

Canine venereal tumor cells were grown in monolayer and tumor tissue fragments were maintained in vitro for 56 days in medium-199. Attachment and cell replication were evident within 17 hours and monolayer was obtained by the tenth day. Growth at first comprised mainly round cells with abundance of cytoplasm and vesicular nuclei, and a few spindle-shaped cells. Later cells became more elongated and like fibroblasts. Cell degeneration started by the 17th day and most were degenerated by the 24th day. Necrosis and depletion of cell population were prominent in tumor explants during the first 2 weeks. Distinct cell multiplication was evident by the 21st day and tumor fragments were repopulated with cells resembling the original tumor by the 56th day. Two distinct morphological cell types were seen: small cells with vesicular, round to oval nuclei and acidophilic cytoplasm; and the large cells with large hyperchromatic nuclei and acidophilic cytoplasm.     

Cytology of canine cutaneous round cell tumors. Mast cell tumor, histiocytoma, lymphosarcoma and transmissible venereal tumor.

Sixty-four canine cutaneous round cell tumors were divided into 25 mast cell tumors, 15 histiocytomas, nine cutaneous lymphosarcomas and 15 transmissible venereal tumors. The final diagnosis was made from cytologic, clinical and histologic findings. Cytologic features were significantly distinctive in mast cell tumor, transmissible venereal tumor, and most cases of histiocytoma and lymphosarcoma to allow a diagnostic opinion. This opinion was supported by subsequent histologic examination. In some instances cytology was considered essential in rendering a diagnostic opinion even though histology was available.     

Prevalence,Serovars and Antimicrobial Susceptibility of Salmonella spp. from Wild and Domestic Green Iguanas (Iguana iguana) in Grenada,West Indies

Cloacal swabs from 62 green iguanas (Iguana iguana), including 47 wild and 15 domestic ones from five parishes of Grenada, were sampled during a 4‐month period of January to April 2013 and examined by enrichment and selective culture for the presence of Salmonella spp. Fifty‐five per cent of the animals were positive, and eight serovars of Salmonella were isolated. The most common serovar was Rubislaw (58.8%), a serovar found recently in many cane toads in Grenada, followed by Oranienburg (14.7%), a serovar that has been causing serious human disease outbreaks in Japan. Serovar IV:48:g,z₅₁:‐ (formerly, S. Marina) highly invasive and known for serious infections in children in the United States, constituted 11.8% of the isolates, all of them being from domestic green iguanas. Salmonella Newport, a serovar recently found in a blue land crab in Grenada, comprised 11.8% of the isolates from the green iguanas. The remaining four less frequent serovars included S. Javiana and S. Glostrup. Antimicrobial susceptibility tests conducted by a disc diffusion method against amoxicillin–clavulanic acid, ampicillin, cefotaxime, ceftazidime, ciprofloxacin, enrofloxacin, gentamicin, nalidixic acid, streptomycin, tetracycline and trimethoprim–sulfamethoxazole showed that drug resistance is minimal, with intermediate susceptibility, mainly to streptomycin, tetracycline and cefotaxime. This is the first report of isolation and antimicrobial susceptibilities of various Salmonella serovars from wild and domestic green iguanas in Grenada, West Indies.     

Pegylated liposomal doxorubicin as a chemotherapeutic agent for treatment of canine transmissible venereal tumor in murine models

The effectiveness of Doxil as a new chemotherapeutic agent against canine transmissible venereal tumor was evaluated, using NOD/ SCID and CD1-nu xenograft mouse models and the response between the two mouse strains was compared. Samples of xenografted venereal tumor were inoculated SC into 20 six week-old NOD/SCID mice and 20 six week-old CD1-nu mice. Seven weeks later, tumor-bearing mice were divided into treatment and control groups. Treatment group was injected with Doxil (6 mg/kg, IP, as a single injection). Control group was injected with buffered saline (0.75cc, IP). Tumor size was determined by caliper measurements and tumor response was assessed according to standard criteria. In both strains there was a significant decrease in tumor size in response to Doxil treatment (P<0.0001). In CD1-nu eight out of nine tumors (88%) responded to the treatment, and in 2 cases complete remission was observed. In NOD/SCID group response to the treatment was seen in eight out of ten tumors (80%) but none regressed fully. Response to the treatment was statistically equal in both strains even though the apoptotic rate, confirmed by TUNEL staining, was higher in NOD/SCID than in CD-1-nu (8.65% and 0.7%, respectively) and tumor infiltrating cells were different: eosinophils in NOD/SCID and CD45R-positive B lymphocytes, and plasma cells in CD-1-nu. In untreated CD1-nu mice, tumor progress was slower than in NOD/SCID. Our results indicate that Doxil is effective against CTVT in mouse xenograft models.     

Risk factors for Campylobacter colonization in broiler flocks in Japan

The present study was conducted to estimate the prevalence of and to identify the risk factors for Campylobacter colonization in broiler flocks in Japan. Campylobacter colonization status in flock was evaluated by culturing pooled caecal excrement from 124 broiler flocks. Potential exposure to risk factors was evaluated with a questionnaire for the broiler producers. Odds ratios (ORs) were calculated using a multivariable logistic regression analysis. The prevalence of Campylobacter-positive flocks was 43.5% (upper and lower limits of 95% confidence interval (CI(95%) ): 34.8, 52.3). Multivariable logistic regression model identified two variables as risk factors for Campylobacter colonization. The ORs of Campylobacter colonization were higher in flocks in western Japan (OR=2.68; CI(95%) : 1.04, 6.91) than in eastern Japan, and in flocks supplied with undisinfected drinking water (OR=7.41; CI(95%) : 3.11, 17.66) than in those supplied with disinfected drinking water. These findings indicate that water may play an important role in Campylobacter colonization in broiler flocks in Japan and the use of disinfected water may reduce the risk of Campylobacter colonization.     

Computer tomographic imaging in 4 dogs with primary nasal canine transmissible venereal tumor and differing cellular phenotype

Primary nasal canine transmissible venereal tumor (CTVT) without genital affection is uncommon. The aim of this report was to describe the primary nasal CTVT findings and CT staging in 4 dogs with different cytological phenotypes. Three male dogs and 1 bitch were evaluated for their chronic histories of sneezing, snoring, mucopurulent nasal discharge and nasal deformation. Cytological examination of nasal secretions suggested CTVT, confirmed by histopathological examination and LINE‐1/c‐myc. Males had the plasmacytoid phenotype of CTVT, and the bitch had the lymphocytoid phenotype. CTVT were staged based on the CT findings using modified Adams staging system. The bitch was classified as stage 1, 2 males were classified as stage 3 and 1 male as stage 4. All dogs had a complete tumoral remission after chemotherapy. Plasmacytoid phenotype was identified in cases with most important damage of the nasal cavity. However, the cytological type did not affect the response to chemotherapy.