Muscle Pathology in Free-Ranging Stranded Cetaceans.

Despite the profound impact that skeletal muscle disorders may pose for the daily activities of wild terrestrial and marine mammals, such conditions have been rarely described in cetaceans. In this study, the authors aimed to determine the nature and prevalence of skeletal muscle lesions in small and large odontocetes and mysticetes ( n = 153) from 19 different species. A macroscopic evaluation of the epaxial muscle mass and a histologic examination of the longissimus dorsi muscle were performed in all cases. The only macroscopically evident change was variable degrees of atrophy of the epaxial muscles ( longissimus dorsi, multifidus, spinalis) in emaciated specimens. The histopathological study revealed single or combined morphological changes in 91.5% of the cases. These changes included the following: degenerative lesions (75.2%), muscle atrophy (37.9%), chronic myopathic changes (25.5%), parasitic infestation (9.2%), and myositis (1.9%). The skeletal muscle is easily sampled during a necropsy and provides essential microscopic information that reflects both local and systemic conditions. Thus, skeletal muscle should be systematically sampled, processed, and examined in all stranded cetaceans.

Biliary cirrhosis caused by Campula spp. in a dolphin and four porpoises.

Biliary cirrhosis produced by Campula spp. is described in 1 striped dolphin Stenella coeruleoalba and 4 harbour porpoises Phocoena phocoena. The hepatic lesions consisted of severe proliferation of fibrous connective tissue with loss of the lobular pattern, nodular regeneration of the hepatic tissue, bile duct hyperplasia and severe inflammatory infiltrate composed of eosinophils, macrophages, lymphocytes and plasma cells. These lesions were associated with severe infestation by Campula spp. Although inflammatory and degenerative hepatic lesions are frequently found in stranded dolphins, biliary cirrhosis has not been previously reported in cetaceans. Massive infestation by these parasites should be included as a cause of hepatic failure resulting in stranding of marine mammals.

Exuberant mucometra associated with atresia of the cervix in a queen.

This short communication reports the clinical, ultrasonographic and histopathological findings in a cat with atresia of the uterine cervix and mucometra. After 6 months of continuous oestrous behaviour, a remarkable abdominal enlargement was observed in a 14-year-old queen. A presumptive diagnosis of mucometra was concluded after the ultrasound evaluation and based on clinical signs and blood analyses. Ovariohysterectomy revealed a notable symmetrical distension (4-5 cm in diameter) of both uterine horns that were filled with fluid (690 ml); microbiological analyses confirmed the aseptic nature of the uterine fluid. Ovarian follicular cysts and cystic subsurface epithelial structures, >1.5 cm in diameter, were present in both ovaries and no corpora lutea were observed. Gross and microscopic evaluation of the uterus confirmed the development of cystic endometrial hyperplasia and the absence of an internal cervical os. The endometrial hyperplasia and mucometra could have developed as a consequence of repeated oestrogenic stimulation.

Weissella ceti sp. nov., isolated from beaked whales (Mesoplodon bidens).

During an investigation into the microbiota of beaked whales (Mesoplodon bidens), nine isolates were obtained from different organs of four animals. The isolates were Gram-positive-staining, catalase-negative, short rod-shaped or coccoid organisms. A phylogenetic analysis based on 16S rRNA gene sequences of these isolates allocated them to the genus Weissella, showing 96.3 % and 96.0 % 16S rRNA gene sequence similarity with Weissella viridescens NRIC 1536(T)and Weissella minor NRIC 1625(T), respectively. On the basis of phenotypic, physiological and phylogenetic evidence, it is proposed that the new isolates from whales represent a novel species of the genus Weissella, Weissella ceti sp. nov. The type strain of Weissella ceti is 1119-1A-09(T) ( = CECT 7719(T) = CCUG 59653(T)).

Congenital biventricular cardiac diverticula in a dog.

Multiple diverticula of the right and left cardiac ventricular walls were incidental findings at necropsy in a juvenile, male, mixed-breed dog with no clinical history of heart disease. Each saccular diverticulum had a narrow communication into the corresponding ventricular chamber. Histologically, the diverticular walls consisted mainly of collagen without elastin fibers and atrophic cardiomyocytes. No inflammation or other lesions were associated with the diverticula or the rest of the myocardium. Based on the history and pathologic findings, a diagnosis of congenital biventricular fibrous diverticula was made.

Morphological and immunohistochemical characterization of spontaneous mammary gland tumors in the guinea pig (Cavia porcellus).

Ten spontaneous mammary gland tumors affecting guinea pigs (GP) were analyzed histologically and immunohistochemically. Histologically, 3 were benign (2 simple adenomas and 1 benign mixed tumor) and 7 were malignant (1 simple solid carcinoma and 6 simple tubulopapillary carcinomas). Immunohistochemical data revealed the glandular immunoprofile of all the tumors and suggested their ductal origin on the basis of cytokeratin 20 expression. Interestingly, cytokeratin 7 was detected in basal/myoepithelial cells. Further, all tumors were positive for type alpha estrogen and progesterone receptors, suggesting a role for steroid hormones in the development of these neoplasias in GP. This article describes the morphological and immunohistochemical features of the normal mammary gland and spontaneous mammary gland tumors in GP.

Correlating the immune response with the clinical-pathological course of persistent mastitis experimentally induced by Mycoplasma agalactiae in dairy goats.

To correlate the clinical course of mycoplasma mastitis with its immune response, right mammary glands of 15 lactating goats were inoculating with 10(10) colony-forming units (cfu) of Mycoplasma agalactiae (Ma). Before sacrificing the animals at 5, 15 or 45 days post-inoculation (dpi), blood Ma antibody titres and milk mycoplasma colony and somatic cell counts were monitored. Ma colonised the mammary gland and milk counts increased to over 10(12)cfu/ml within 5 dpi. During this period, an innate immune response involving neutrophils and macrophages was observed, and Ma antigen appeared in the degenerated acinar epithelium. From 7 dpi, a specific antibody response coincided with reduced viable mycoplasmas in milk. The humoral immune response was limited; by 37 dpi, all animals scored negative for anti-Ma antibodies, and around 10(8)cfu/ml were shed. Results indicate an early immune response to Ma inoculation unable to control mycoplasmal invasion. An ensuing humoral response, despite reducing the mycoplasma burden, leads to chronic, persistent infection.

Short communication: apoptosis regulates passive immune transfer in newborn kids.

Ten newborn kids were used to evaluate the relationship between apoptosis and passive immune transfer in neonatal enterocytes. Kids were slaughtered in groups of 2 at birth, 1, 2, 3, or 60 d postpartum, and samples of duodenal epithelium collected from each animal. Samples were fixed, dehydrated, and embedded in paraffin wax. Sections were assessed for apoptotic cells and immunostained for IgG. Our results suggest that IgG absorption is mediated by apoptotic enterocytes. Thus, delaying apoptosis may improve the success of passive immune transfer.

Mucor ramosissimus associated with feather loss in canaries (Serinus canarius).

Three canaries showing feather loss on legs, dorsum, neck, and head, and hyperkeratosis on the feet were sacrificed because of their poor corporal condition and submitted to the Unit of Histology and Anatomic Pathology at the Veterinary School of Las Palmas de Gran Canaria. Histologically, skin revealed pronounced epidermal and follicular infundibular hyperplasia associated with orthokeratotic hyperkeratosis. Numerous fungal spores were observed on the stratum corneum of the epidermis and within feather follicles, associated with destruction of the feathers. This fungus was identified as Mucor ramosissimus. To the best of authors' knowledge, this is the first report of dermatitis and feather loss associated with Mucor ramosissimus, not only in canaries but also in birds.

Malignant peripheral nerve sheath tumour (malignant schwannoma) in the diaphragm of a goat.

This report describes a malignant schwannoma in the diaphragm of a 2-year-old goat. The immunohistochemical and histological features indicated a diagnosis of malignant schwannoma rather than neurofibrosarcoma. The diaphragm represents an unusual location for neoplasia in both domestic animals and human beings. A possible anatomical origin from the phrenic nerve is discussed.

Rhabdomyolysis and myoglobinuric nephrosis (capture myopathy) in a striped dolphin.

This report describes delayed myoglobinuric capture myopathy in a striped dolphin (Stenella coeruleoalba) found stranded alive on the coast of Fuerteventura (Canary Islands, Spain). The animal was transported to Gran Canaria where it died 48 hr after stranding. The main lesions consisted of acute rhabdomyolysis affecting both cardiac and skeletal muscles, and myoglobinuric nephrosis. Using immunohistochemistry, degenerate myofibers with depletion of myoglobin, and an intracytoplasmatic immunoreaction for fibrinogen were observed. Orange-red pigmented casts in renal tubular lumens were strongly immunolabeled for myoglobin. To our knowledge, this is the first pathologic description of capture myopathy with myoglobinuric nephrosis in stranded cetaceans. Stress, exertion, trauma, and crush injury caused during the stranding, restraint, and transportation were the main causes of rhabdomyolysis in this case.

Benign Bilateral Adenomyoepithelioma of the Mammary Gland in a Ring-tailed Lemur (Lemur catta).

Naturally occurring mammary tumours are uncommon in prosimians. A 20-year-old female ring-tailed lemur (Lemur catta) developed bilateral enlargement of the mammary glands. Surgical removal revealed that both masses were comprised of multiple nodules and cystic areas that entirely replaced the normal glands. Histologically, a benign neoplastic biphasic cellular proliferation, composed of luminal-epithelial and basal-myoepithelial components, was identified. Immunohistochemical analysis for expression of cytokeratin (CK) AE1/AE3, CK7, CK5 + 8, CK14, vimentin, p63 and 14-3-3σ highlighted the biphasic nature of the neoplasm. A low mitotic count, low Ki67 labelling index, expression of oestrogen receptor-α, lack of expression of human epidermal growth factor receptor and a 3-year disease-free period without recurrence supported the benign nature of the tumour. Macroscopically, histologically and immunohistochemically this neoplasm resembled benign adenomyoepithelioma of the breast in women. This is the first complete report of a naturally occurring mammary tumour in a ring-tailed lemur.

Fibrino-necrotic typhlitis caused by Escherichia fergusonii in ostriches (Struthio camelus).

Two adult ostriches developed anorexia, prostration, and severe hemorrhagic diarrhea, dying 24 hr after the onset of clinical signs. On postmortem examination, the cecal mucosa showed locally extensive areas of hemorrhages and fibrino-necrotic typhlitis with a white-yellowish material covering the mucosal surface. Multiple serosal petequial hemorrhages and fibrinous peritonitis were present. Histologic examination revealed an intense mononuclear infiltration in the lamina propria and submucosa of the cecum and extensive superficial necrosis associated with fibrin and serocellular deposits. Several gram-negative bacterial colonies were observed within the necrotic areas. Samples from intestinal lesions were collected, and pure growth of Escherichia fergusonii was obtained. Escherichia fergusonii is a member of Enterobacteriaceae, closely related to Escherichia coli and Shigella sp., established as a new species of the genus Escherichia in 1985. In veterinary medicine, E. fergusonii has been reported in calves and sheep from dinical cases suggestive of salmonellosis. To our knowledge, this report represents the first description of E. fergusonii associated with enteritis in ostrich.

Hepatosplenic large cell immunoblastic lymphoma in a bottlenose dolphin (Tursiops truncatus) with high levels of polychlorinated biphenyl congeners.

This report describes a large cell immunoblastic lymphoma in a bottlenose dolphin found stranded alive in Gran Canaria, Spain. Diffuse infiltration of round neoplastic cells was observed in the splenic cords and sinuses and in hepatic sinusoids, resulting in moderate organ enlargement. The tumour cells (immunophenotype IgG+ and CD3-) showed scant, lightly eosinophilic or basophilic cytoplasm, distinct cell boundaries and hyperchromatic nuclei, each with one or more nucleoli. Mitoses were common. On the basis of histopathological, immunohistochemical and ultrastructural features, the tumour was classified as an immunoblastic lymphoma. Eleven polychlorinated biphenyl (PCB) congeners, 23 organochlorine pesticides and 16 polycyclic aromatic hydrocarbons (PAHs) were detected in the blubber and liver. High concentrations of PCBs 153, 180, 138 and 187 found in the liver may have been associated with the hepatosplenic lymphoma.

Decay of sperm obtained from epididymes of wild ruminants depending on postmortem time.

We have carried out a study on the effect of postmortem time (PT) in some characteristics of epididymal sperm salvaged from hunted Iberian red deer and roe deer. Testis were collected, identified, refrigerated down to 5 degrees C, and sent to our laboratory by the wardens of the hunting reserves. This way, samples were delivered at different times postmortem. Sperm were extracted from the cauda epididymis by means of cuts. Analyzed parameters were: osmolality, pH, motility-both subjectively and with CASA, HOS test reactivity, acrosomal status and viability (assessed with propidium iodide). Osmolality and pH rose with prolonged postmortem time, possibly due to tissue decomposition. Most sperm quality parameters negatively correlated with PT. Besides, when comparing PT classes (groups of 24 h for red deer and 30 h for roe deer), we could appreciate that motility was more affected by PT than other quality variables. Progressive motility was especially impaired. We also classified the samples in high, medium and low quality for each PT group (considering progressive motility, intact acrosomes and reactivity to the HOS test), and it was clear that after 2 days the number of high quality samples was testimonial, and after several days, we almost found only low quality samples. In conclusion, epididymal sperm from Iberian red deer and roe deer undergo a decrease of quality with PT, but it could stay acceptable within many hours postmortem. There are implications for wildlife conservation programs, as epididymal sperm is a good source of germplasm. If valuable animals die and it is not possible to process their sperm immediately, it may still be possible to obtain viable spermatozoa many hours later.

Comparative histology of muscle in free ranging cetaceans: shallow versus deep diving species.

Different marine mammal species exhibit a wide range of diving behaviour based on their breath-hold diving capabilities. They are classically categorized as long duration, deep-diving and short duration, shallow-diving species. These abilities are likely to be related to the muscle characteristics of each species. Despite the increasing number of publications on muscle profile in different cetacean species, very little information is currently available concerning the characteristics of other muscle components in these species. In this study, we examined skeletal muscle fiber type, fiber size (cross sectional area and lesser diameter), intramuscular substrates, and perimysium-related structures, by retrospective study in 146 stranded cetaceans involving 15 different species. Additionally, we investigated diving profile-specific histological features. Our results suggest that deep diving species have higher amount of intramyocyte lipid droplets, and evidence higher percentage of intramuscular adipose tissue, and larger fibre sizes in this group of animals.

Heart regeneration after myocardial infarction using synthetic biomaterials.

Myocardial infarction causes almost 7.3 million deaths each year worldwide. However, current treatments are more palliative than curative. Presently, cell and protein therapies are considered the most promising alternative treatments. Clinical trials performed until now have demonstrated that these therapies are limited by protein short half-life and by low transplanted cell survival rate, prompting the development of novel cell and protein delivery systems able to overcome such limitations. In this review we discuss the advances made in the last 10years in the emerging field of cardiac repair using biomaterial-based delivery systems with focus on the progress made on preclinical in vivo studies. Then, we focus in cardiac tissue engineering approaches, and how the incorporation of both cells and proteins together into biomaterials has opened new horizons in the myocardial infarction treatment. Finally, the ongoing challenges and the perspectives for future work in cardiac tissue engineering will also be discussed.

Ovine cutaneous squamous cell carcinoma: immunohistochemical expression of CD3, CD4, CD8 and MHC class II antigens in the associated inflammatory infiltrate.

The immunohistochemical expression of CD3, CD4, CD8 and MHC class II antigens in the cellular inflammatory infiltrate associated with early and advanced ovine squamous cell carcinomas (OSCC), as well as actinic keratosis was analyzed. The majority of the peritumoral and intratumoral lymphocytes reacted with the anti-human CD3 polyclonal antibody. The number of CD8+ T lymphocytes increased in advanced OSCC compared with that of actinic keratosis and early OSCC, whereas the number of CD4+ lymphocytes was similar in early and advanced OSCC. Tumor cells were unreactive with the anti-MHC class II antibody, but the majority of the mononuclear cellular infiltrate expressed this antigen in early and advanced tumors.

Cross-reactivity of human and bovine antibodies in striped dolphin paraffin wax-embedded tissues.

This study evaluates the cross-reactivity of seven anti-human and one anti-bovine antibodies in formalin-fixed, paraffin-embedded tissue samples of liver and mesenteric lymph nodes of 13 striped dolphins (Stenella coeruleoalba). Four antibodies (CD3, IgG, lysozyme and S100 protein) reacted with striped dolphin lymph nodes in a similar pattern to that observed in the species of origin. The anti-human MHC class II mAb reacted strongly with macrophages and dendritic-like cells of striped dolphins, whereas a small number of lymphocytes were labelled with this antibody. These antibodies were used to study the immunophenotype of the inflammatory infiltrated in non-specific chronic reactive hepatitis (eight cases) and chronic parasite cholangitis (two cases) and normal liver (three cases) of striped dolphins. Non-specific chronic reactive hepatitis was composed of inflammatory infiltration of CD3+ T lymphocytes and IgG+ plasma cells in portal spaces and hepatic sinusoids. Lymphonodular aggregates observed in chronic parasitic cholangitis showed a cellular distribution similar to that found in lymph node cortex, including the presence of S100+ and MHC class II+ dendritic-like cells in lymphoid follicles and interfollicular areas. This result suggests that those inflammatory infiltrates are highly organised to enhance antigen presentation to B and T cells.