An Unusual Case of Collision Testicular Tumor in a Female DSD Dog.

Collision tumors (CT) consist of two independent neoplasms with distinct neoplastic populations. Disorders of sexual development (DSDs) are characterized by atypical sexual development leading to various abnormalities of the genital tract. Sex reversal (SR) syndromes are a type of DSD characterized by a discrepancy between chromosomal sex and gonadal development (testes/ovaries) and the presence or the absence of the SRY gene. A phenotypically female 8-year-old Jack Russell terrier dog was referred due to anomalous vaginal discharge and non-pruritic cutaneous bilateral symmetrical alopecia on the flanks. During abdominal palpation, a voluminous mass was detected in the left quadrant area, later confirmed by ultrasound. The owner decided to proceed with euthanasia and necropsy. In the abdominal cavity, the left gonad was increased in size, the right one and the uterus were decreased, and the vagina and vulva appeared to be thickened. Histologically, both gonads were revealed to be testes: the left one was affected by a double neoplastic component (sustentacular tumor and interstitial cell tumor), whereas the right gonad showed coarctated seminiferous tubules. PCR amplification of the genes SRY and AMELX revealed the absence of the MSY region of the Y chromosome. To the authors' knowledge, this is the first report describing a case of a testicular collision tumor in a DSD SRY-negative dog.

Reaction of Cornu aspersum Immune System against Different Aelurostrongylus abstrusus Developmental Stages.

Cornu aspersum, the land snail, is recognized as a suitable intermediate host of Aelurostrongylus abstrusus; however, there is little information both on larval development as well as on the intermediate host's immune system reaction to the parasite. The aim of the study was to assess the histological reaction of C. aspersum's immune system against A. abstrusus. Sixty-five snails were provided by a snail farm. Five of them were digested to assess the absence of natural parasitic infections. The remaining sixty were divided into five groups. Three groups of snails were infected with A. abstrusus using by-contact or injection methods; one group was injected only with saline solution and one group was left untreated as the control. The snails of group A were sacrificed and digested on study days 2, 10, and 18; snails of the other groups were collected and examined for histopathological analysis on study days 2, 10, and 18. On study day 2, in the infected snails, several free L1s were observed along with the absence of immune system reactions. On day 10, the L2s elicited an intense reaction in the internal layer of the muscular foot. On day 18, all L3s partially encapsulated by the snail's immune system were observed in the outermost part of the muscular foot, which is near and among the goblet cells. This last finding suggests that L3s could be shed with the snail's mucus and spread in the environment, representing an alternative route of transmission for this feline lungworm.

Comparative genomic analysis of the Dietzia genus: an insight into genomic diversity, and adaptation.

Dietzia strains are widely distributed in the environment, presenting an opportunistic role, and some species have undetermined taxonomic characteristics. Here, we propose the existence of errors in the classification of species in this genus using comparative genomics. We performed ANI, dDDH, pangenome and genomic plasticity analyses better to elucidate the phylogenomic relationships between Dietzia strains. For this, we used 55 genomes of Dietzia downloaded from public databases that were combined with a newly sequenced. Sequence analysis of a phylogenetic tree based on genome similarity comparisons and dDDH, ANI analyses supported grouping different Dietzia species into four distinct groups. The pangenome analysis corroborated the classification of these groups, supporting the idea that some species of Dietzia could be reassigned in a possible classification into three distinct species, each containing less variability than that found within the global pangenome of all strains. Additionally, analysis of genomic plasticity based on groups containing Dietzia strains found differences in the presence and absence of symbiotic Islands and pathogenic islands related to their isolation site. We propose that the comparison of pangenome subsets together with phylogenomic approaches can be used as an alternative for the classification and differentiation of new species of the genus Dietzia.

Does TLS Exist in Canine Mammary Gland Tumours? Preliminary Results in Simple Carcinomas.

Neoplastic progression is influenced by the expression of tumour antigens that activate an anti-tumour immune response. Human medical studies show that this body defence is carried out in secondary lymphoid organs (SLOs) but also directly in the tumour through organized cellular aggregates that are called tertiary lymphoid structures (TLSs). However, their occurrence has different meanings in different tumour types. For example, the presence of TLSs in breast cancer is associated with the most aggressive subtypes. This paper aimed to study TLSs in canine mammary simple carcinomas. A morphological assessment of the inflammatory infiltrate was performed on H&E sections of fifty cases. Immunohistochemistry was then carried out to typify the inflammatory cells in the tumour microenvironment. Results showed that, sometimes, inflammatory infiltrates were organized in follicles close to high-grade carcinomas, simulating a lymphoid organization, as in breast cancer. Therefore, we can assume that even in canine mammary tumours, TLSs exist and they are entities to consider due to their presence in the most aggressive histotypes or tumours with a high degree of malignancy.

Nontraumatic Paraureteral Urinoma in a Cat with Urolithiasis.

Urinoma is an encapsulated collection of urine due to a disruption in the collection system of the urinary tract. This condition is rarely reported in veterinary medicine. The aim of this study is to describe the clinical and diagnostic findings of a well-encapsulated paraureteral urinoma associated with urinary tract stones in a critical feline patient. The ultrasound examination of the abdomen revealed a well-defined collection of anechoic fluid containing the left kidney in the retroperitoneal space, while the radiographic examination revealed a loss of soft tissue contrast in the lumbar region. Both techniques confirmed the presence of stones in the bladder. Following the ultrasound-guided drainage of the lesion, the nature of the fluid was also confirmed to be urine. Due to the worsening of the cat's health, to better define the extent and genesis of the lesion, a positive contrast radiographic study was performed by means of an ultrasound-guided percutaneous injection of a contrast medium into the lesion, which highlighted a wide and homogeneous radiopaque area in the left retroperitoneal space. These findings were confirmed with a postmortem examination, corroborating the diagnostic suspicion of urinoma. The percutaneous contrast inoculation performed in this critically ill patient plays a role in the diagnostic process to reach a final diagnosis in cases in which the patient's clinical condition does not allow for performing an excretory positive contrast study.

Penile Varicose Vein in Akita inu, 7-Year-Old Dog: A Clinico-Pathological Study.

Penile varicose veins are a rare lesion described in human medicine. A seven-year-old intact Akita inu male dog presented with a red penile neoformation. It was further referred for a specialist andrological examination. During the visit, ultrasonography of genital apparatus and cytology of the penile lesion were performed. A suspected neoformation of vascular origin was observed on ultrasonography. Cytology was inconclusive as composition of the sample revealed blood elements only. The neoformation was surgically removed and the excised sample was submitted for the histological examination, which revealed an anomalous varicose dilatation of the venous vascular structures. In human medicine, varicose veins are part of the spectrum of chronic venous disease and include spider telangiectasia, reticular veins, and true varicosities. Reports of penile localization of varicose veins is rare in human medicine and, to the best of our knowledge, it has never been reported in veterinary medicine.

Antibacterial Properties of Canine Platelet-Rich Plasma and Other Non-Transfusional Hemo-Components: An in vitro Study.

This in vitro study was carried out to evaluate the potential antibacterial properties of canine non-transfusional hemo-components. Therapeutic formulations commonly used for regenerative medicine purposes (platelet-rich plasma, platelet gel, platelet lysate, fibrin glue), considering both leukocyte-rich and leukocyte-poor formulations, but also platelet-poor plasma and activating substances (thrombin, calcium gluconate), were tested to detect elements with potential antimicrobial properties. The antibacterial effect was tested on different bacterial strains (Staphylococcus aureus subspecies aureus, Staphylococcus cohnii subspecies cohnii, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae subspecies pneumoniae) isolated from canine wounds and classified as susceptible, multidrug-, extensively, and pandrug-resistant bacteria toward a known panel of human and veterinary antibiotics. The evaluation was carried out by agar gel diffusion method (Kirby-Bauer) and micro-inhibition in broth using microplates and spectrophotometer reading. The study findings confirmed the hypothesized antibacterial properties of canine non-transfusional hemo-components. A more effective bacteriostatic effect was found against Gram-negative bacteria, drug-resistant too. The presence of leukocytes or platelets does not appear to be essential for the antibacterial effect. Further studies should be conducted to evaluate the exact mechanism of action of the antimicrobial activity. However, non-transfusional hemo-components could be a useful natural aid in controlling bacterial infections in 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.

Bidimensional and Contrast-Enhanced Ultrasonography of the Spleen in Dogs Affected by Leishmaniosis.

Canine leishmaniosis (CanL) is responsible for splenic pathological changes. The main features detectable from ultrasound examination are splenomegaly and diffuse alterations of the echostructure. The study aimed to highlight whether these ultrasound changes are related to the severity of the disease or to a modification of splenic microvascularization that can be detected in vivo through contrast-enhanced ultrasonography (CEUS). Twenty-five adult dogs tested for CanL were enrolled in this prospective, controlled study and staged according to LeishVet guidelines. Bidimensional ultrasonography revealed that splenomegaly was seen in 50% of the affected dogs, and diffuse parenchymal changes were seen in more than 60% of dogs with splenomegaly, showing a positive correlation with severity of the disease; therefore, splenomegaly could be of prognostic significance. CEUS showed that a persistent heterogeneous distribution pattern appeared only in spleens with diffuse echostructure alterations. The evaluation of quantitative CEUS parameters regarding the volume and velocity of flow in three regions of interest did not show differences between affected and control dogs. Diffuse spleen microvascular modifications evidenced by CEUS were reported for the first time in dogs with CanL. In endemic areas, CanL could be included in the differential diagnoses list when detecting splenic alterations in dogs.

Immune Cells and Immunoglobulin Expression in the Mammary Gland Tumors of Dog.

Inflammatory cells have a role in tumor progression and have prognostic and therapeutic potential. The immunohistochemical expression for Mast Cell Tryptase, Macrophage Marker, CD79a, IgA, IgM and IgG on 43 cases of canine mammary gland lesions was analyzed. In hyperplasia, a few B cells (BCs) and Tumor-Associated Macrophages (TAMs) were observed, while the number of Tumor-Associated Mast Cells (TAMCs) was the highest. In the peritumoral stroma of malignant lesions, low number of TAMCs and a high number of TAMAs and BCs were present. Immune cells of each type were always lower in the intratumoral than peritumoral stroma. Positivity to CD79a was also detected in the epithelial cells of simple and micropapillay carcinomas. Immunoglobulin reactivity was mainly located in the epithelial cells where an intense positivity to IgA and IgG and a weak positivity for IgM were detectable. On the basis of our preliminary results and literature data, we suggest that such cells and molecules could be directly involved in the biology of canine mammary gland tumors. In breast cancer, stromal inflammatory cells and cancer derived immunoglobulins have been correlated with the progression, malignancy and poor prognosis of the tumor. The results herein reported show that the dog's mammary gland epithelium also expresses immunoglobulins, and they mostly show a direct relationship with the infiltration of macrophages. In addition, this study shows that the infiltration of mast cells, B-cells and macrophages varies depending on the degree of malignancy of neoplasia.

Endometrial Cytology During the Different Phases of the Estrous Cycle in Jennies: New Evidences.

Since in the mare and other animal species such as bitches and cats, the endometrial cell pattern varies depending on the phase of the estrous cycle, the aim of this study was to describe and quantify the endometrial cytological (EC) findings in cycling jennies. EC of eight nonpregnant jennies by cytobrush (CB) at diestrus (day 1 and day 14) and estrous (day 21) were evaluated. All slides were stained with Wright´s stain and microscopically examined at both 400× and 1000× magnification. Seven high-power fields (400×) were assessed in each smear and the endometrial epithelial cells and neutrophils (PMNs) were counted. Endometrial epithelial cells were classified as intact, distorted or fragmented and, on the basis of the presence of dense groups, in monolayer or single clusters. Cytoplasmic characteristics, such as vacuolation or streaming and size, form, position of nuclear characteristics, including karyorrhexis, were recorded. Background aspect, as clear, proteinaceous, or debris, was also considered. In general, sampling by CB provided a yield of cells and clumped endometrial epithelial cells in many smears, being more abundant in estrus than early and late diestrus. Individual endometrial epithelial cells, during estrous, presented a columnar morphology, ciliated or not ciliated and basal nuclei. During diestrus phase, endometrial epithelial cells presented a more cuboidal ciliated or not ciliated morphology. Moderate amount of proteinacious material and red blood cells (RBC) was also observed. Non variation in the percentage of PMNs during diestrus was obtained, but lower and segmented PMNs in CB smears were shown in estrous. This study provides new insights on the physiological changes of endometrial epithelial cells in cycling jennies during the estrus cycle. The CB technique represents a suitable and adequate method for endometrial evaluation, taking into account cytological and/or cytopathological purposes also in jennies.

Atypical Multibacterial Granulomatous Myositis in a Horse: First Report in Italy.

Infectious causes of myositis are reported relatively uncommonly in horses. Among them, bacterial causes include Streptococcus equi subsp. zooepidemicus, Actinobacillus equuli, Fusobacterium spp. Staphylococcus spp, and Corynebacterium pseudotuberculosis. Infection can be spread to muscles via haematogenous or extension from skin lesions. Parasitic myositis has also been documented. In this report, a 12 year-old Italian Quarter Horse mare presented with diffuse subcutaneous nodules and masses ranging from 2 × 3 to 5 × 20 cm in size, and adherent to subcutis and muscles that were first macroscopically and cytologically diagnosed as pyogranulomas. Subsequently, histological, molecular, bacteriological, and biochemical investigations were performed. All the data obtained allowed to diagnose a severe and diffuse multibacterial granulomatous myositis caused by Corynebacterium pseudotuberculosis and Corynebacterium amycolatum. Following the therapy and an initial disappearance of most of the lesions together with a general improvement of the mare, the clinical condition deteriorated, and new nodules appeared. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and PCR techniques revealed the presence of bacteria as Glutamicibacter creatinolyticus and Dietzia spp. To the authors' knowledge, this case report represents the first description of multibacterial granulomatous myositis due to Corynebacterium pseudotuberculosis, Corynebacterium amycolatum, Glutamicibacter creatinolyticus, and Dietzia spp. in a horse reared in Italy.

Complete genome analysis of Glutamicibacter creatinolyticus from mare abscess and comparative genomics provide insight of diversity and adaptation for Glutamicibacter.

Bacteria of the genusGlutamicibacterare considered ubiquitous because they can be found in soil, water and air. They have already been isolated from different habitats, including different types of soil, clinical samples, cheese and plants. Glutamicibacter creatinolyticus is a Gram-positive bacterium important to various biotechnological processes, however, as a pathogen it is associated to urinary tract infections and bacteremia. Recently,Glutamicibacter creatinolyticusLGCM 259 was isolated from a mare, which displayed several diffuse subcutaneous nodules with heavy vascularization. In this study, sequencing, genomic analysis ofG. creatinolyticusLGCM 259 and comparative analyseswere performedamong 4representatives of different members of genusfromdifferent habitats, available in the NCBI database. The LGCM 259 strain's genome carries important factors of bacterial virulence that are essential in cell viability, virulence, and pathogenicity. Genomic islands were predicted for 4 members of genusGlutamicibacter,showing ahigh number of GEIs,which may reflect a high interspecific diversity and a possible adaptive mechanism responsible for the survival of each species in its specific niche. Furthermore,G. creatinolyticusLGCM 259 sharessyntenicregions, albeit with a considerable loss of genes, in relation to the other species. In addition,G. creatinolyticusLGCM 259 presentsresistancegenes to 6 differentclasses ofantibiotics and heavy metals, such as: copper, arsenic, chromium and cobalt-zinc-cadmium.Comparative genomicsanalysescouldcontribute to the identification of mobile genetic elements particular to the speciesG. creatinolyticuscompared to other members of genus. The presence of specific regions inG. creatinolyticuscould be indicative of their rolesin host adaptation, virulence, and the characterization ofastrain that affects animals.

Use of contrast-enhanced ultrasound for assessment of nodular lymphoid hyperplasia (NLH) in canine spleen.

BACKGROUND: Nodular lymphoid hyperplasia (NLH) is one of the most common non-neoplastic splenic lesions in dogs, especially in old ones, showing a splenic enlargement. More recent studies have been focused on Contrast Enhanced Ultrasonography (CEUS) analysis of the spleen for establishing normal perfusion patterns and blood pool phase peculiarities of focal lesions. The aim of the study was to evaluate the qualitative and quantitative CEUS analysis of the canine splenic NLH, characterizing the CEUS pattern of this pathology on 20 clinical cases. RESULTS: A prospective, observational study was performed using a system equipped with contrast-tuned imaging technology. Mechanical Index was set from 0.08 to 0.11; the contrast medium was a second generation contrast medium composed of sulphur hexafluoride encapsulated of a shell of phospholipids (SonoVue®). Qualitative and quantitative assessment of the enhancement pattern of splenic NLH were performed. Cytology and histology identified 20 splenic NLH. All of the benign hyperplastic lesions assessed were isoechoic with a homogeneous pattern than the surrounding normal spleen, during the wash-in phase (10-20 s) of the CEUS exam. Before finishing the wash-in phase, 20-45 s from the contrast medium inoculation, 19/20 benign nodules became markedly hypoechoic to the adjacent spleen. Sensitivity of hypoechoic pattern for NLH was 95%. CONCLUSIONS: These findings should prove useful in the evaluation of focal splenic masses in dogs. Since enhancement and perfusion patterns of NLH seem to coincide with some neoplastic lesions of the spleen previously reported, in clinical practice attention must be paid to the final diagnosis of canine splenic lesions using only the CEUS exam.

Quantitative evaluation of canine urinary bladder transitional cell carcinoma using contrast-enhanced ultrasonography.

BACKGROUND: In veterinary medicine, contrast-enhanced ultrasonography allowed the accurate quantification of liver, splenic and kidney vascularization in healthy dogs and the differentiation between malignant and benign hepatic, renal, and splenic nodules in dogs and cats based on perfusion patterns. The utility of contrast-enhanced ultrasonography in other applications is still under study. The aim of this study was to develop diagnostic criteria by contrast-enhanced ultrasonography in 8 client-owned adult dogs affected by urinary bladder transitional cell carcinoma with definitive diagnosis made by cytopathologic evaluation after suction biopsy. The contrast enhancement pattern and the quantification of blood flow parameters of this tumor were reported. RESULTS: Examinations with B-mode, Doppler ultrasonography and contrast-enhanced ultrasonography were performed in all not sedated dogs. Assessments of bladder masses and bladder wall infiltration were performed. Each dog received 2 bolus injections of sulfur hexafluoride during the contrast-enhanced ultrasonography. Quantitative analysis of the contrast-enhanced ultrasonography images were performed. For each dog, one region of interest was manually drawn around the entire tumor. Software analysis of contrast-enhanced time-intensity curves was used to identify peak enhancement, time to peak enhancement, regional blood volume, regional blood flow, and mean transit time. Contrast-enhanced ultrasonography showed an avid enhancement of the tumour tissue, with a heterogeneous or homogeneous pattern. The exam also showed the loss of planes between the lesion and the muscular layer. The presence of vascularized tissue through the bladder wall confirms the infiltrative feature of the tumour. Post-processing quantitative analysis showed a time-intensity curve with a rapid wash-in, a low level of signal intensity and a slow wash-out. CONCLUSIONS: Contrast-enhanced ultrasonography provided useful clinical information and defined a vascular enhancement patterns and calculated parameters associated with TCC. It may be a useful, noninvasive and reproducible tool for detecting these tumors in dogs.

A case of diprosopiasis in Trachemys scripta scripta.

This short communication describes a case of diprosopiasis in Trachemys scripta scripta imported from Florida and farmed for about 4 months by a private owner in Palermo, Sicily, Italy. The water turtle showed the morphological and radiological features characterizing such deformity. This communication aims to advance the knowledge of the reptile's congenital anomalies and suggests the need for more detailed investigations to better understand its pathogenesis.

Feline lungworm Oslerus rostratus (Strongylida: Filaridae) in Italy: first case report and histopathological findings.

Oslerus rostratus syn. Anafilaroides rostratus (Strongylida: Filaroididae) is a metastrongyloid transmitted by snails, which localizes in peri-bronchial tissues and in the lung parenchyma of wild as well as domestic cats. In Europe, this nematode has been reported only on two occasions, being diagnosed in cats from Majorca Island and in northern Spain. Here, we describe a case of O. rostratus infection in a necropsied 4-year-old cat in Sicily (southern Italy). At the inspection of lungs, slender and greyish nematodes (four females and two males) were found embedded in the peri-bronchial tissues and in the bronchial walls. Parasites were morphological and molecularly identified as O. rostratus, with their 18S sequences being identical among them and showing a high homology (99%) with those available in public databases. At the histology, nematodes were encapsulated in a pseudo-cystic formation surrounded by an interstitial inflammatory process and fibrous tissue. Lung lesions were mainly represented by peri-luminal fibrosis, hyperplasia and hypertrophy of the bronchial mucosa and glands, respectively. This first record of O. rostratus infection from Italy indicates that this parasite should be included in the differential diagnosis of feline of lungworm infection.

Adult of Dirofilaria repens in a dog with recurrent multiple subcutaneous nodular lesions.

Subcutaneous dirofilariosis is a zoonosis parasitizing the dog, transmitted by Culicidae mosquitoes. An increasing number of new cases have been reported both in dogs and in humans all over the world. The aim of this work was to describe an uncommon case of recurrent multiple subcutaneous nodules by Dirofilaria repens in a 2-year-old dog. Surgical excision of two nodules on both hind limbs revealed a single thread-like live nematode and multiple parasite fragments, respectively. The parasite was morphologically identified as an adult female of D. repens. Haematoxylin-eosin tissue sections of nodules revealed a diffuse pyogranulomatous angiocentric dermatitis with an intense eosinophilic infiltrate throughout the dermis. One month post-surgery, a new nodular lesion was detected on the right thigh and resolved spontaneously in a few weeks. During the following 3 months, two other nodules were briefly noted both on the right hip and on the left side of the chest. The dog was periodically treated with milbemycin oxime and praziquantel, starting a week before the appearance of the first nodule. Microfilariae were not detected neither in the bloodstream nor in fine-needle aspirates at any time, and the diagnosis was made only after surgery. Practitioners should consider dirofilariosis in the differential diagnosis of subcutaneous swellings, especially if they have a recurrent behaviour, previously described in humans but not in dogs.

Mycobacterium tuberculosis in a red-crowned parakeet (Cyanoramphus novaezelandiae).

An adult female red-crowned parakeet (Cyanoramphus novaezelandieae) was presented for necropsy and histopathologic evaluation. The bird had died after exhibiting lameness, weight loss, respiratory signs, and hemoptysis. Postmortem radiographs revealed lesions in the diaphysis of the left femur and soft-tissue opacities in the lungs. At necropsy, the muscles of the left femur were pale and swollen, white-yellow small nodules were visible in lungs and air sacs, and the liver and other coelomic organs appeared pale. On histologic examination, areas of necrosis in the lungs were extensively infiltrated with acid-fast positive bacilli surrounded by macrophages, epithelioid cells, and giant cells. Acid-fast bacilli were also present in the left leg muscle and in granulomas in the liver, kidneys, and intestine. Fungal hyphae associated with a Splendore-Hoeppli phenomenon were visible in the left leg muscle. Mycobacterium tuberculosis was confirmed by polymerase chain reaction testing performed on muscle samples. Results were indicative of infection with M tuberculosis complicated by mycotic myositis. The disease in avian species is of zoonotic importance, and infected birds may be a useful sentinel for human infection.