Assessment of the impact of direct in vitro PFAS treatment on mouse spermatozoa.

Abstract: Poly- and per-fluoroalkyl substances (PFAS) are synthetic environmentally persistent chemicals. Despite the phaseout of specific PFAS, their inherent stability has resulted in ubiquitous and enduring environmental contamination. PFAS bioaccumulation has been reported globally with omnipresence in most populations wherein they have been associated with a range of negative health effects, including strong associations with increased instances of testicular cancer and reductions in overall semen quality. To elucidate the biological basis of such effects, we employed an acute in vitro exposure model in which the spermatozoa of adult male mice were exposed to a cocktail of PFAS chemicals at environmentally relevant concentrations. We hypothesized that direct PFAS treatment of spermatozoa would induce reactive oxygen species generation and compromise the functional profile and DNA integrity of exposed cells. Despite this, post-exposure functional testing revealed that short-term PFAS exposure (3 h) did not elicit a cytotoxic effect, nor did it overtly influence the functional profile, capacitation rate, or the in vitro fertilization ability of spermatozoa. PFAS treatment of spermatozoa did, however, result in a significant delay in the developmental progression of the day 4 pre-implantation embryos produced in vitro. This developmental delay could not be attributed to a loss of sperm DNA integrity, DNA damage, or elevated levels of intracellular reactive oxygen species. When considered together, the results presented here raise the intriguing prospect that spermatozoa exposed to a short-term PFAS exposure period potentially harbor an alternate stress signal that is delivered to the embryo upon fertilization. Lay summary: PFAS are synthetic chemicals widely used in non-stick cookware, food packaging, and firefighting foam. Such extensive use has led to concerning levels of environmental contamination and reports of associations with a spectrum of negative health outcomes, including testicular cancer and reduced semen quality. To investigate the effects of PFAS on male reproduction, we incubated mouse sperm in a cocktail of nine PFAS at environmentally relevant concentrations before checking for a range of functional outcomes. This treatment strategy was not toxic to the sperm; it did not kill them or reduce their motility, nor did it affect their fertilization capacity. However, we did observe developmental delays among pre-implantation embryos created using PFAS-treated sperm. Such findings raise the intriguing prospect that PFAS-exposed sperm harbor a form of stress signal that they deliver to the embryo upon fertilization.

Half-life of serum anti-Müllerian hormone and changes after gonadectomy in adult female and male dogs with normal and abnormal gonads.

Anti-Müllerian hormone (AMH) is a biomarker for the presence of gonadal tissue. Changes in serum AMH after gonadectomy are not well established, and its serum half-life is unknown in dogs. We measured serum AMH with a validated electro-chemiluminescent immunoassay in adult female (n = 12) and male (n = 7) dogs with normal gonads, as well as in dogs with gonadal pathology (ovarian remnant syndrome, ORS n = 3, testicular tumor [Leydig cell, Sertoli cell, seminoma] n = 3, unilateral abdominal cryptorchid n = 4) on the day of gonadectomy (D0), and on D3, D7, D14 (females and males), and D21, D28 (males only). Males had higher AMH concentrations than females independent of gonadal status (P < 0.001). Dogs with ORS had lower initial AMH (0.45 ± 0.43 ng/ml) than bitches with normal gonads (1.16 ± 0.44 ng/ml; P = 0.027). Cryptorchid dogs had higher initial concentrations (80.57 ± 52.81 ng/ml) than males with normal gonads (7.92 ± 2.45 ng/ml; P = 0.004), and those with testicular tumors (18.63 ± 5.04 ng/ml) were intermediate (P ≥ 0.250). AMH decreased over time (P ≤ 0.012) and was 0.01-0.04 ng/ml by D14 in females and 0.02-0.12 ng/ml by D28 in males. Serum half-life in the whole study population was 2.85 ± 0.51 days and did not differ between groups. In conclusion, serum AMH can differentiate between intact and gonadectomized status of adult dogs by 14 days after ovario(hyster)ectomy in females and by 28 days after surgical castration in males.

Single-cell mRNA sequencing of giant panda (Ailuropoda melanoleuca) seminoma reveals the cellular and molecular characteristics of tumour cells.

Testicular tumours are zoonoses that can occur in not only human, but other animals, include giant pandas. A middle-aged male giant panda named Fufu was diagnosed with a testicular tumour and underwent surgery to remove the entire left testis. The testis was mainly composed of three substantive parts: normal tissue on the outside, tumour tissue in the middle, and necrosis in the centre. HE stains revealed that the tumour was a seminoma. Single-cell mRNA sequence was applied to characterise cellular states and molecular circuitries of giant panda testicular seminoma. Only germ cell markers expressed in nearly all tumour cells, and the tumour cells appeared to be the same subtype of seminoma cells. We identified four clusters with unique genes expression. They were early apoptosis cells (EAC), inactive cells (IC), active cells subcluster 1 (AC-1) and active cells subcluster 2 (AC-2). We utilised monocle tools and found that IC cells was in the initiation stage, and EAC was one type of terminal stage, suggesting that tumour cells may undergo apoptosis in the future. AC-2 was another type of terminal stage, representing a group of progressive cells. Our study represents the first report to utilise scRNA-seq to characterise the cellular states and molecular circuitries of a giant panda testicular tumour. This investigation proposes CD117 and CD30 as dependable markers for future pathologic diagnosis. Our findings also suggest that CTSV and other genes with unique expression patterns in active and progressive giant panda seminoma cells may act as early prognostic biomarkers.

Occurrence of mammary gland tumours in male dogs and its weak association with development of testicular tumours: a review.

Mammary gland tumours (MGTs) are commonly occurring neoplasms in female dogs. However, rare cases of MGTs in male dogs have been reported for years. Due to the low incidence of MGTs in male dogs in comparison to female dogs, veterinary oncology is mainly focused on mammary neoplasms diagnosed in female dogs and extensive research is conducted in this scientific area. Therefore, there are no sufficient epidemiological data on male dogs and the aetiology of their tumour development is still poorly understood.The aim of this literature review was to present cases of MGTs in male dogs for better understanding the scale of the problem over the years. The analyses of 74 affected male dogs with 92 tumours showed that the majority of MGTs in male dogs were benign tumours (54.3%), especially in form of adenomas, often developed in posterior canine mammary glands (58.1%).The increased number of canine MGTs in male dogs aged 7 -13 years with an age peak at 11 years was noted. The age of affected animals was not related to breed. Mammary gland neoplasms were diagnosed predominately in Crossbreeds (20.2%) followed by Cocker Spaniels (18.9%) and German Shepherds (10.8%).The association between MGT development in male dogs and co-occurrence of testicular tumours (TTs) has been discussed for years. Thus, cases of development of both tumours were included in this study. As a result, only in 12.7% cases of MGTs also history of TTs was described. Therefore, no general association between these tumours should be assumed.

First report of primary testicular leiomyosarcoma in two dogs.

BACKGROUND: Testicular tumours are common in dogs and, among them, interstitial cell tumours, seminomas and sustentacular cell tumours are the most reported. Mesenchymal testicular tumours are rarely reported in humans as in veterinary medicine where only three cases of sarcomas (leiomyomas and leomyosarcomas) have been described in two stallions and in a ram. CASE PRESENTATION: The present cases regarded a 12-year-old mixed-breed dog and a 10-year-old American Staffordshire Terrier that underwent bilateral orchiectomy. Formalin fixed testes were referred for histopathological diagnosis. At gross examination, in one of the testes of both dogs, a white, firm and variably cystic testicular mass, effacing and replacing the testicular parenchyma was detected. Samples were collected from both neoplastic and contralateral testes, routinely processed for histology and serial sections were also examined immunohistochemically with primary antibodies against cytokeratins, vimentin, Von Willebrand factor, inhibin-α, α-smooth muscle actin, smooth muscle myosin and desmin. Histopathological features as well as the immunohistochemical results, positive for vimentin, actin, myosin and desmin, confirmed the mesenchymal origin and the myoid phenotype of both testicular tumours supporting the diagnoses of leiomyosarcoma. CONCLUSIONS: To the authors knowledge these are the first cases of primary testicular sarcoma reported in the canine species. However, even rare, these tumours deserve to be considered in routine diagnosis when a testicular spindle cell tumour is observed. The immunohistochemical panel applied was useful to distinguish the present tumours from undifferentiated Sertoli cell tumours confirming the diagnosis of leiomyosarcoma.

MicroRNA as epigenetic regulators of canine cryptorchidism.

Cryptorchidism, the failed descent of one or both testes into the scrotum, is a common developmental disorder in male dogs. Cryptorchidism may affect canine fertility, reducing the quality of the semen, and may promote spermatic cord torsion and onset of neoplasia. MicroRNAs (miRNAs) are epigenetic regulators of gene expression and their dysregulation is associated with disorders of spermatogenesis and testis neoplasia. The present study aimed at investigating the expression of miRNAs in formalin-fixed, paraffin-embedded (FFPE) canine retained testes and testes affected by seminoma, and at integrating miRNAs to their target genes. Forty testicular FFPE specimens from 30 dogs were included - 10 scrotal and 10 contralateral retained from 10 unilateral cryptorchid dogs; 10 tumoral testes affected by seminoma from non-cryptorchid dogs; 10 scrotal normal testes from non-cryptorchid dogs included as the control. The expression level of three miRNAs, namely miR-302c-3p, miR-302a-3p, and miR-371-3p, associated with testicular disorders, were quantified using RT-qPCR. The comparative analysis demonstrated that the level of miR-302a-3p and miR-371a-3p were quantifiable exclusively in control testes. The expression level of miR-302c-3p was higher in the control than in the other groups; its expression decreased in retained testes compared to scrotal testes and testes with seminoma. Gene Ontology analysis pointed out that these miRNAs may be involved in the modulation of estrogen and thyroid hormone signaling pathways. In conclusion, this study demonstrated that miRNAs are dysregulated in canine cryptorchid and seminoma-affected testes compared to control tissues, confirming the pivotal role of miRNAs in cryptorchidism.

Metastatic Sertoli cell tumour in a captive giant anteater (Myrmecophaga tridactyla).

There are a few studies on diseases of anteaters, but reports on reproductive lesions and neoplasms of these animals are scarce. This is the first report of a case of metastatic Sertoli cell tumour in a giant anteater (Myrmecophaga tridactyla). The animal had renal lesions associated with impaired renal function as indicated by serum biochemistry. Histopathological and immunohistochemical examinations provided a conclusive diagnosis of Sertoli cell tumour with metastasis to the liver, kidneys and lymph nodes.

A rare case of ocular and testicular T-cell lymphoma in a hermaphrodite koi carp (Cyprinus carpio Linnaeus 1758): clinical, histopathological, and immunohistochemical study.

BACKGROUND: Lymphatic neoplasia may occur in various types, such as lymphoma, lymphosarcoma, lympholeukemia, and plasmacytoid leukemia. Lymphoma, defined as a malignant tumour of lymphoid tissue, has been found in a number of fish families including Esocidae and Salmonidae. However, the occurrence of lymphoma is rare in those belonging to the Cyprinidae. A final diagnosis of ocular and testicular T-cell lymphoma in the present study was based on the clinical signs, morphology, and texture of the tumour masses in the macroscopic and microscopic examinations. In addition, histopathological and immunohistochemical findings corresponded to T-cell lymphoma characteristics. CASE PRESENTATION: A 2-year-old hermaphrodite koi carp (Cyprinus carpio Linnaeus 1758) with a large ocular mass and severe exophthalmia in the right eye was referred to the Ornamental Fish Clinic in October 2020. Under anesthesia, enucleation was performed. 57 days after enucleation of the right eye, exophthalmia in the left eye was discovered. 221 days after surgery, the fish was discovered to be dead. At necropsy, a large soft tissue mass attached to the left testis was discovered. There were also small whitish nodules on the surface of the liver. Histopathology revealed a hypercellular ocular mass with scant connective tissue. The sections also revealed multifocal hemorrhages, round to ovoid neoplastic cells, mild-to-moderate anisokaryosis and anisocytosis, and mitotic figures. Basophilic neoplastic cells were found in blood vessels within the testicular mass, raising the possibility of systemic spread. The liver showed microscopic metastasis with morphologic similarities to the ocular and testicular tumors. The neoplastic cells infiltrating the left and right eyes as well as the testicular mass were immunohistochemically positive for CD3 but negative for CD20. The masses were diagnosed as T-cell lymphoma based on histopathological and immunohistochemical findings. CONCLUSIONS: This case report provides the first evidence of clinical, histopathological, morphological, and immunohistochemical findings of an ocular and testicular T-cell lymphoma in a hermaphrodite koi carp (Cyprinus carpio) in Iran.

Multiple Sex Cord-stromal Tumors in a Standardbred Stallion Testis.

A 12-year-old Standardbred stallion presented with a 5-month history of a growing mass in the left testis as well as an overall decrease in left testicular size. Palpation and ultrasonography of the left testis revealed a firm, hypoechoic, clearly delineated soft tissue mass in the craniolateral portion of the testis that measured 2.5 × 2.3 × 1.9 cm. Two smaller, hypoechoic regions also were visible ultrasonographically in the left testis, suggesting the presence of multifocal/multicentric neoplasia. The affected testis was very small (testicular volume of 40.3 cm3). The right testis was significantly larger (144.3 cm3), and the parenchyma was ultrasonographically normal. Due to the concern that these findings could indicate the presence of a more aggressive tumor type, unilateral orchiectomy was performed. Multiple soft tissue masses were identified grossly, and histopathologic evaluation identified the larger mass as a Sertoli cell tumor and the two smaller masses as mixed sex cord-stromal tumors with Sertoli cell and Leydig cell differentiation. To our knowledge, this the first report of concurrent Sertoli cell and mixed sex cord-stromal tumors in a single descended equine testis.

Streptococcus canis prostatitis and endocarditis with thromboembolism in a dog with sertoli cell tumour in a cryptic testis and prostatic squamous metaplasia.

We describe an unusual case of prostatitis caused by Streptococcus canis evolving to endocarditis and splenic, renal, and cerebral thromboembolism in a dog, associated with a Sertoli cell tumour in a cryptic testis and diffuse prostatic squamous metaplasia. A nine-year-old, intact male, mixed-breed dog was presented to a veterinary teaching hospital with abdominal pain and prostration. Physical examination and abdominal ultrasonography revealed an atrophic right testicle located in the subcutaneous tissue. The left testicle was in the abdominal cavity with increased dimensions and irregular contours. Complete blood count analysis showed marked neutrophilic leukocytosis and thrombocytopenia. After clinical worsening, euthanasia was performed, and the dog was submitted to post-mortem examination. The main gross findings included testicular malposition with one cryptic and one ectopic testis, enlarged prostate with purulent content, distension of the urinary bladder with cloudy urine, vegetative valvular endocarditis in the mitral valve, and spleen and renal infarcts. Histological examination showed a Sertoli cell tumour in the abdominal testis, diffuse prostatic squamous metaplasia with marked keratinization associated with bacterial prostatitis, fibrinonecrotic cystitis, bacterial endocarditis with marked myxomatous degeneration in the mitral valve, and splenic, renal, and cerebral thromboembolism. Microbiological analysis identified Streptococcus canis in the prostate and mitral valve. Sertoli cell tumour of cryptic testis increases oestrogen production and leads to squamous metaplasia of the prostate, which should be considered as predisposing factors for ascending S. canis infection from the urogenital tract to the prostate. Then, haematogenous spread of S. canis from the prostate to mitral valve cause endocarditis and subsequent thromboembolism and infarcts, all decisive to poor prognosis in this case.

Diagnosis of Seminoma With Metastasis to the Brain and Kidneys in a Moluccan Eclectus (Eclectus roratus) Using Advanced Contrast Based Imaging.

A 20-year-old male Moluccan eclectus (Eclectus roratus) was presented with a history of falling off the perch, ataxia, unilateral blindness, hyporexia, and progressive lethargy. The eclectus was found to have a leukocytosis (52 × 103 cells/µL), characterized by an absolute and relative heterophilia (42.6 × 103 cells/µL and 82%, respectively), relative lymphopenia (18%), elevated bile acids (88 µmol/L), and elevated triglycerides (236 mg/dL). Radiographic images revealed a mass effect within the caudal coelom. After 13 days of outpatient supportive care, the patient was rechecked and had normalized bile acids (<35 µmol/L), static triglycerides (232 mg/ dL), and hyperuricemia (18.6 mg/dL). Computed tomography was performed antemortem with and without iodinated contrast agent, 4 mL/kg IV over 2 minutes, and a mass was found associated with the left kidney. Due to worsening neurologic signs and involvement with surrounding structures, the owner elected euthanasia. Before the postmortem examination, a postmortem intravascular contrast agent was used to provide a more thorough visualization of internal anatomical structures, including left renal vasculature disruption, mass vasculature, caudal coelomic organ displacement, and increased irregular optic chiasm radiodensity. Postmortem, a 4 × 4.1 × 5.1-cm white to tan mass was identified. Histopathology confirmed a seminoma with metastasis to the kidneys and optic chiasm. Seminomas have been described in avian species; however, seminoma metastasis in an eclectus parrot confirmed by histopathology has not been reported, and optic chiasm metastasis of a seminoma has not been described in any avian species. This report describes postmortem computed tomographic angiography of metastatic seminoma in a Moluccan eclectus with metastasis to the optic chiasm.

Widespread metastasis of a spermatocytic seminoma with concomitant hepatic peliosis in a Southern African hedgehog (Atelerix frontalis).

ABSTRACT: A six-year-old intact male Southern African hedgehog (Atelerix frontalis) presented with a history of chronic mild to moderate weight loss, and sub-acute hind limb ataxia that progressed to complete paralysis, at which point the hedgehog was euthanised. At autopsy, a large multinodular pale mass had completely replaced the left testicle and transcoelomically metastasised to the diaphragm and the peri-renal area, from where it then invaded the vertebral column and spinal cord. Multifocal, irregular to round, well-demarcated, blood-filled, proliferative lesions were also present in the hepatic parenchyma. Histological analysis of both the testis and metastatic lesions revealed diffuse sheets of neoplastic cells with moderate pale cytoplasm, large irregular to round nuclei and mostly one prominent magenta nucleolus, consistent with metastatic seminoma. The neoplastic cells were negative for periodic acid-Schiff (PAS) stain and positive for CD117 by immunohistochemistry (IHC). Taken together with the morphology of the neoplastic cells and the advanced age of the animal, this is suggestive of a spermatocytic seminoma. Histological analysis of the liver revealed multifocal lesions consisting of large anastomosing blood-filled spaces bordered by compressed hepatocytes, consistent with hepatic peliosis. This is the first report of a neoplasm in the Southern African hedgehog (Atelerix frontalis), the first report of a metastatic seminoma in a hedgehog, together with diagnosis of spermatocytic subtype, and the first report of a hedgehog with concomitant hepatic peliosis.

Spontaneous neoplasms in harbour porpoises Phocoena phocoena.

Harbour porpoises are widely distributed in the North Atlantic and represent the most abundant cetacean species in the North and Baltic Seas. Spontaneous neoplasms are relatively rarely reported in cetaceans, and only little is known about neoplasia in harbour porpoises. Thus, archival material was reviewed for spontaneous neoplasms in harbour porpoises recorded during post-mortem examinations between 1999 and 2018. Neoplasms were identified in 7 adult porpoises: 6 animals originating from the North and Baltic Seas and investigated as part of German and Dutch systematic health monitoring programs, and 1 porpoise from Greenlandic waters. The tumours were of different histogenetic origins and further characterised by histology and immunohistochemistry. One individual had a neoplasia in the digestive tract (adenocarcinoma, n = 1); 4 animals, in the genital tract (Sertoli cell tumour, n = 1; genital leiomyoma/fibroleiomyoma, n = 3); and 2 porpoises, in endocrine organs (adrenal adenoma, n = 2). This is the first report of an adenocarcinoma in the liver, a testicular Sertoli cell tumour and adrenocortical adenomas in harbour porpoises. The cause of the tumorigenesis in examined cases remains undetermined. The involvement of endogenous factors, including mutation of cell cycle regulating genes, such as the tumour-suppressor gene p53, cannot be ruled out. The aetiopathogenetic significance of exogenous factors, such as infectious agents like liver flukes or anthropogenic factors, including persistent organic pollutants, should be the subject of future investigations.

Dependence of the Ki67 Labelling Index of Selected Canine Tumours on Patient Age, Sex and Tumour Size.

Cell proliferation is a fundamental criterion in the assessment of malignant progression of many tumours and an essential parameter in several grading schemes. However, proliferation may be dependent on patient age and other variables, as shown in normal tissues, cultured cells and human neoplasms. We thus hypothesized that age or other patient or tumour-related parameters might generally affect proliferation in canine neoplasms, which might be of value for optimizing prognostic algorithms. We performed linear regression analyses to associate age, sex and tumour size with digitally quantified immunohistochemical Ki67 labelling indices (Ki67-LIs) of 495 canine tumours, including cutaneous mast cell tumours (MCTs, n = 70), soft tissue sarcomas (n = 61), plasmacytomas (n = 86), trichoblastomas (n = 62) and perianal gland adenomas (PGAs, n = 95) as well as testicular interstitial (n = 65) and Sertoli cell tumours (n = 56). In MCTs, the Ki67-LI increased 1.13-fold per year of age (P <0.05) in bitches but not in males. Conversely, in PGAs it rose 1.10-fold per year in males (P <0.05) while it decreased 0.95-fold in bitches (P = 0.37). Only in MCTs and PGAs was the Ki67-LI associated with tumour size, albeit in oppositional directions (MCTs: 1.26-fold per cm diameter, P <0.01; PGAs: 0.76-fold, P <0.01). No correlations were found in the other tumour types. The few sex-dependent correlations with patient age and tumour size established here indicate highly tumour-type specific mechanisms, but the diagnostic consequences are uncertain.

Medullary bone in male budgerigars (Melopsittacus undulatus) with testicular neoplasms.

Medullary bone is a calcium-rich, labile bone normally occurring in female birds with each egg-laying cycle. The stimulus for formation of medullary bone is, in part, the cyclic increase in serum estrogens produced by preovulatory ovarian follicles. Increased bone density due to formation of medullary bone, particularly in pneumatic bones, has been termed polyostotic hyperostosis, even if physiologic. This study investigated the formation of medullary bone in nonpneumatic (femur) and pneumatic (humerus) bones in sexually mature male budgerigars submitted for autopsy. Of the 21 sexually mature male budgerigars submitted for autopsy, 7 (33%) had medullary bone in 1 or more bones examined. All 7 male budgerigars with medullary bone had a testicular neoplasm, which was morphologically consistent with a testicular sustentacular cell tumor, seminoma, or interstitial cell tumor. Medullary bone was not present in the 14 cases with other diseases. Medullary bone formation in pneumatic and nonpneumatic bones can occur in male budgerigars with testicular neoplasms. Radiographic increases in medullary bone density, particularly in the humerus, could provide antemortem indication of testicular neoplasia in male budgerigars.

Outcome of dogs with bone marrow suppression secondary to Sertoli cell tumour.

Sertoli cell tumours are one of the most common canine testicular neoplasia. These tumours are significantly more likely to arise in cryptorchid dogs and are often functional, oestrogen-secreting tumours which can lead to fatal myelotoxicity. The goal of this study was to describe the outcome of dogs with oestrogen-induced bone marrow suppression secondary to Sertoli cell tumours in seven client-owned dogs. Medical records from April 1, 2011 through April 1, 2021 were reviewed to identify dogs that underwent surgical management of a Sertoli cell tumour with documented bone marrow suppression. Overall, 5/7 dogs required transfusion of blood products peri-operatively. Cases 1 and 6 received a transfusion of packed red blood cells (RBC) prior to surgery and case 5 required a transfusion of whole blood. Case 1 also required a transfusion of platelets before surgery. Post-operatively, cases 1 and 2 received packed RBC's and case 6 received two transfusions of whole blood. Case 3 required transfusions of both fresh frozen plasma and platelets post-operatively. All dogs survived to discharge and 6/7 dogs had documented improvement in haematopoietic values. Two dogs remained chronically thrombocytopenic. The median hospital stay was 4 days. One dog died within 4 weeks of surgery from worsening pancytopenia. Survival for greater than 1 year was documented in 4/7 dogs, and one dog was lost to follow-up 4 months post-operatively. One dog remained severely pancytopenic 4 weeks post-operatively and received oral lithium treatment. Improvements in all blood cell lines were observed within the 4 weeks and resolution of pancytopenia within 6 weeks. Historically, the prognosis for dogs with bone marrow suppression secondary to Sertoli cell tumours was guarded to poor. This report documented improved outcomes for dogs that underwent surgery, including one dog that received lithium chloride as treatment for Sertoli cell tumour-induced bone marrow suppression.

Atypical cytomorphologic description of a seminoma in a rabbit.

Seminoma is reported as a rare disease associated with specific micro-morphologic findings. In the present report, we describe the case of a testicular seminoma in a 7-year-old Holland Lop rabbit, in which the cytology presented an atypical pattern. Upon presentation, the left testicle was severely enlarged, and the patient also had a history of radiation therapy for a previously diagnosed thymoma. Following excision and histopathology of the abnormal organ, results showed a mixed intratubular-diffuse pattern with evidence of torsion. Moreover, cytology revealed a cohesive pattern with multiple malignancy criteria. To our knowledge, this is the first published report of a seminoma with these cytologic features.

Immunohistochemical insights into a hidden pathology: Canine cryptorchidism.

Cryptorchidism is a common disorder in the canine population with some aspects still unclear. Although the bilateral condition is known to lead to fertility problems and predisposition to testicular cancer, the neoplastic risk for scrotal testis in unilateral cryptorchid dog is controversial. Therefore, the therapeutic approach to the canine unilateral cryptorchid is arbitrary so far. This study aimed to investigate precancerous testicular lesions, such as immaturity and atrophy, and compare them in scrotal and undescended testes using an in-depth diagnostic analysis based on immunophenotypic patterns. With this purpose, 26 adult male dogs of different ages and breeds, affected by unilateral or bilateral cryptorchidism were enrolled. After surgical removal, testes were examined immunohistochemically to assess their positivity for specific markers of the canine foetal/neonatal period, that is vimentin (VIM), cytokeratin (CK), desmin (DES), inhibin-α (INH), and anti-Müllerian hormone (AMH) in Sertoli cells, and placental alkaline phosphatase (PLAP) in germ cells. Except for the ubiquitous VIM, all the markers were more expressed in neoplastic gonads compared to healthy ones (P < 0.05). Similarly, testes detected with Sertoli cell-only tubules as well as with Sertoli cells hyperplasia showed higher expression than gonads without such alterations for CK, DES, AMH and PLAP, and for CK and DES, respectively (P < 0.05). The same trend was observed in undescended respect to scrotal testes even though their positivity was significant only for DES, INH and AMH (P < 0.05). Immunohistochemical positivity found in scrotal testes of unilateral cryptorchid dogs, even in absence of detectable anatomical abnormalities, was suggestive of precancerous lesions. Despite the limited sample size, this study could help to clarify the predisposition to neoplastic development in normally descended testes. These markers expression in adult life could allow identifying the early stages of the testicular carcinogenesis process besides suggesting a precautionary bilateral surgical approach in unilateral cryptorchid dogs.

A case of neoplastic synchronism in a dog.

INTRODUCTION: Synchronous primary tumors are considered severe, comorbid conditions in people representing neoplasm that develop independently and concomitantly. A diagnosis of synchronous tumors was made in a dog, underlying the difficulties to reach it without the aid of multiple diagnostic techniques aimed to demonstrate the simultaneous coexistence of different tumor types. MATERIALS AND METHODS: A 7-year-old male Boxer dog presented several tumors located on the skin of the left hind limb and the scrotal region. Moreover, additional tumors in the testicles, after palpation and ultrasound examination, were detected. Following diagnostic results, the cutaneous tumor, scrotum, and testes were surgically removed. RESULTS: Pathological investigations revealed the presence of five different tumors: a cutaneous mast cell tumor; a scrotal melanocytoma; three testicular neoplasms (Sertoli Sustentacular cell tumor, seminoma, and interstitial Leydig cell tumor). CONCLUSIONS: The present report describes a neoplastic synchronism due to the presence of five different primary tumors in a dog and, for the first time the presence of a collision testicular tumor together with other non-testicular primary tumors. The occasional finding underlines the importance of the knowledge of such conditions in the process of decision-making and in carrying out all the proper diagnostic procedures for a correct diagnosis and clinical staging.