Maternal age, obesity and hyperglycaemia are associated with a delay in preimplantation embryo development in mouse.

In brief: Fertility has decreased due to advanced maternal age and the rising prevalence of the metabolic syndrome. Using quantitative image analysis methods, we show that these factors are associated with delayed preimplantation embryo development in a mouse model. Abstract: Delayed maternal age, obesity and diabetes are associated with reduced fertility. We investigated how age and obesity/metabolic syndrome impact fertility and hypothesized that its decrease is due to defects in preimplantation embryo development. Three groups of female C57Bl6 mice (12 weeks, 9 months and 1 year old) were fed either a high-fat diet for 8 weeks, to induce obesity and the metabolic syndrome, or a control chow diet. Body weight and composition, glucose tolerance and insulin resistance were assessed. Fecundity was evaluated by mating and pregnancy rates, as well as by the number of embryos. Embryo quality was assessed morphologically, and cell fate composition was analysed in preimplantation embryos by state-of-the-art single-cell quantitative confocal image analysis. The high-fat diet was associated with increased adiposity, glucose intolerance and insulin resistance, especially in the older mice. Fecundity was affected by age more than by the diet. Both age and high-fat diet were associated with reduced cell fate allocation, indicating a delay in the preimplantation embryo development, and with increased expression of GATA3, an inhibitor of placentation. These results support that age and the metabolic syndrome reduce fertility through mechanisms which are present at conception or very early in pregnancy.

Morphological Assessment of Concomitant Lesions Detected in Goat Herds Naturally Infected with Paratuberculosis (Johne's Disease).

Paratuberculosis (PTB), caused by Mycobacterium avium subspecies paratuberculosis (MAP), causes significant financial losses in the ruminant industry. The aim of this study is to describe the concomitant pathological findings as well as PTB-induced lesions in 39 naturally infected goats (15 vaccinated and 24 non-vaccinated). All animals exhibited MAP-induced microscopic lesions affecting target organs, although only 62% of those were detected grossly. Mainly concomitant inflammatory pathologies were recognized affecting the hemolymphatic, respiratory and gastrointestinal systems. Non-vaccinated animals exhibited both moderate and marked granulomatous enteritis in contrast with vaccinated ones which presented mild intestinal affection. Our results demonstrate that non-vaccinated animals presented pneumonia in all age groups studied (from 12 up to >48 months old). A significantly higher prevalence of ileocecal valve PTB lesions was detected in non-vaccinated animals with pneumonic lesions (p = 0.027). Furthermore, a reduction of gastrointestinal non-PTB processes was described in vaccinated goats. In conclusion, a PTB infected goat herd can be affected by a wide range of concomitant pathologies, mostly inflammatory in origin. Anatomic pathology is of crucial importance for correct herd diagnosis and histopathology is an indispensable tool for lesion detection. Additionally, anti-MAP vaccination could have a beneficial effect on the reduction of respiratory and gastrointestinal non-PTB diseases.

Veterinary Education and Training on Non-Traditional Companion Animals, Exotic, Zoo, and Wild Animals: Concepts Review and Challenging Perspective on Zoological Medicine.

The role of veterinarians is becoming more significant and necessary to support the welfare and health not only of non-traditional companion animals and wildlife animals, but also of humans and the environment. The importance of the One Health/One World concept and its social impact is increasing significantly, accompanied by the notoriety of new emerging and reemerging zoonoses. This paper aims to review and anchor the main concepts and professional applications of zoological medicine, which has been extensively discussed and adapted in recent decades. In addition, we analyse the main social demands, training, and educational needs and the perception of veterinary professionals relating to this specialised veterinary discipline. Our final goal is to reinforce the use of the term zoological medicine and contribute to highlight the need to foster and underpin specific educational policies and programs on this matter in the veterinary curricula. Zoological medicine should be the appropriate and agreed-upon term in the academic language concerning the veterinary medicine of pets, wild, or zoo species, excluding traditional domestic animals, and integrating the principles of ecology and conservation, applied to both natural and artificial environments. This discipline has suffered an intense evolution covering applications in private clinics, zoos, bioparks, and wildlife. All this implies current and future challenges for the veterinary profession that can only be addressed with greater and better attention from multiple perspectives, especially the education and training of professionals to improve and specialise in their professional scope of services.

Infection due to Buteo buteo herpesvirus in a common buzzard (Buteo buteo insularum).

This study aimed to document the pathological findings observed in a common buzzard (Buteo buteo insularum) from Gran Canaria (Canary Islands, Atlantic Ocean), naturally infected with Buteo buteo herpesvirus (HV). Local authorities found the common buzzard alive, but it died after 10 days of specialized veterinary care. Postmortem investigation, including complete gross and histologic examination, immunohistochemistry, microbiology, and PCR, was performed. The animal presented necrotizing heterophilic and histiocytic bilateral conjunctivitis, stomatitis, pharyngitis, rhinitis, and sinusitis with secondary bacterial and fungal infections. Frequent eosinophilic intranuclear inclusion bodies were observed in the oral mucosa and esophagus epithelium. HV proteins and DNA were detected in tissues from this animal. The sequences obtained from the PCR product were identical to the reported sequences of Buteo buteo HV.

Toothed Whales Have Black Neurons in the Blue Spot.

Neuromelanin (NM) is a dark polymer pigment that is located mostly in the human substantia nigra, and in the locus ceruleus, referred to as "the blue spot". NM increases linearly with age, and has been described mainly in the human brain; however, it also occurs in the neurons of monkeys, horses, giraffes, cattle, sheep, goats, dogs, rats, and even in frogs. While in most of these mammals NM shows the histochemical and ultrastructural features typical of lipofuscins, human NM is confined within cytoplasmic organelles that are surrounded by a double membrane, suggesting an autophagic origin. In a study on the morphology of the locus ceruleus of the family Delphinidae, the presence of a variable quantity of NM in the interior of locus ceruleus neurons was observed for the first time; meanwhile, nothing is known about its ultrastructure and composition. Transmission electron microscopy demonstrated in two toothed whales-an Atlantic spotted dolphin (Stenella frontalis; family Delphinidae) and a Blainville's beaked whale (Mesoplodon densirostris; family Ziphiidae)-the presence of melanin granules associated with lipid droplets and membranes that were very similar to that of human NM. The relationship between NM and neuronal vulnerability must be studied in depth, and cetaceans may offer a new natural-spontaneous comparative model for the study of NM and its implication in neurodegenerative diseases.

Budd-Chiari-like pathology in dolphins.

Nearly two decades ago, pathologic examination results suggested that acoustic factors, such as mid-frequency active naval military sonar (MFAS) could be the cause of acute decompression-like sickness in stranded beaked whales. Acute systemic gas embolism in these whales was reported together with enigmatic cystic liver lesions (CLL), characterized by intrahepatic encapsulated gas-filled cysts, tentatively interpreted as "gas-bubble" lesions in various other cetacean species. Here we provide a pathologic reinterpretation of CLL in odontocetes. Among 1,200 cetaceans necropsied, CLL were only observed in four striped dolphins (Stenella coeruleoalba), with a low prevalence (2%, N = 179). Together, our data strongly suggest that CLL are the result of the combination of a pre-existing or concomitant hepatic vascular disorder superimposed and exacerbated by gas bubbles, and clearly differ from acute systemic gas embolism in stranded beaked whales that is linked to MFAS. Budd-Chiari-like syndrome in dolphins is hypothesized based on the present pathologic findings. Nonetheless, further researched is warranted to determine precise etiopathogenesis(es) and contributing factors for CLL in cetaceans.

Bacteremia and Aortic Valvular Endocarditis in a Eurasian Stone-Curlew (Burhinus oedicnemus distinctus) due to Streptococcus dysgalactiae.

Burhinus oedicnemus distinctus is an endemic subspecies of Eurasian Stone-curlew present in the Canary Islands. Their populations are rapidly declining, mainly because of anthropogenic impacts. This report describes valvular endocarditis and septicemia in a Eurasian Stone-Curlew with left foot loss and severe contralateral bumblefoot.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Assessment of Lung Disease in Finishing Pigs at Slaughter: Pulmonary Lesions and Implications on Productivity Parameters.

Swine respiratory disease is associated with productive losses. We evaluated the prevalence of lung lesions with an emphasis on Mycoplasma hyopneumoniae (Mh), porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), as well as the impact on productive parameters in 108 finishing pigs at slaughter. Pathologic, immunohistochemical (IHC) and serologic analyses were performed. Pneumonic processes were observed in 73.1% of the animals. They mainly consisted of cranioventral bronchopneumonia (CBP) (46.3%) and pleuritis (17.6%). Microscopically, bronchointerstitial pneumonia (67.4%) was common and was occasionally combined (27.9%) with interstitial pneumonia (IP). Mh and PCV2-antigens were detected in bronchointerstitial pneumonia (70.7%) and IP cases (33.3%). There were low titers against Mh (18%) and high titers against PRRSV (100%) and PCV2 (65%). Animals with CBP remained at the farm longer; those with >10% of lung parenchyma involvement were sent later (208.8 days old) and had a lower average carcass weight (74.1 kg) and a lower daily weight gain (500.8 gr/day) compared with animals without lesions (567.2 gr/day, 77.7 kg, 200.8 days old). We suggest that animals that do not reach the weight at slaughter should be sent to slaughter regardless to avoid further negative impacts of respiratory disease in productive parameters.

A Study on the Pathological Effects of Trypanorhyncha Cestodes in Dusky Groupers Epinephelus marginatus from the Canary Islands.

Trypanorhyncha are cestodes commonly infecting marine fish. Numerous studies have detailed the biology of Trypanorhyncha species, but information on the pathological changes produced by these parasites is limited. Dusky groupers are keystone species necessary for the preservation of several marine ecosystems. Considering their vulnerable state of conservation and the efforts being made to culture them, identification of the effects caused by Trypanorhyncha is vital. Here, we aimed to determine the prevalence and pathological changes produced by Trypanorhyncha in dusky groupers from the Canary Islands. The prevalence of trypanorhynch plerocerci was 96%. Grossly, in the abdominal cavity, there were numerous larvae-filled cysts and nodules. These were embedded in abundant fibrosis, producing visceral adhesions. Histologically, affecting the peritoneum, stomach, and intestine there were numerous degenerated encysted plerocerci and extensive deposition of mature connective tissue. These findings indicate that Trypanorhyncha is highly prevalent in adult dusky groupers from the Canary Islands, producing a progressive and chronic response. Furthermore, fish immune system appears to attempt to eliminate the parasites through fibrous encapsulation. Nonetheless, extensive fibrosis may have a detrimental impact on fish health when adjacent cells or tissues are compressed and their functions impaired.

Studying the heterogeneous pathogenesis of canine diabetes: Observational characterization of an island population.

BACKGROUND: Canine diabetes mellitus has mostly been studied in northern European, Australian and American populations, whereas other regions have received less attention. OBJECTIVES: We evaluated the epidemiological, clinical and histopathological features of diabetic dogs in Gran Canaria, Spain. METHODS: Prevalence and incidence were estimated. Clinical features were analysed, and serum and genomic DNA were obtained. Dogs with presumed idiopathic or immune-mediated diabetes, were DLA-typed and antibodies against GAD65 and IA-2 were assessed. Pancreases from ten diabetic dogs were examined and compared with pancreases from non-diabetic dogs. RESULTS AND CONCLUSIONS: Twenty-nine diabetic dogs were identified in a population of 5,213 (prevalence: 0.56%; incidence: 0.37%). Most were female (79%) and sexually intact (87% of females, 83% of males). Diabetes secondary to dioestrus (55.2%) and insulin-deficient diabetes (20.7%) were the most frequent types. Antibodies against GAD65 and IA-2 were identified in two out of five cases and DLA-genotyping revealed novel haplotypes. Breed distribution differed between diabetic and non-diabetic dogs. Reduced number of pancreatic islets and ß-cell mass were observed, with vacuolation of islet cells and ductal epithelium. In this population, where neutering is not standard practice, diabetes secondary to dioestrus is the most frequent diabetes subtype. Genetic susceptibility also differed from previous studies. These results support the heterogeneous pathogenesis of canine diabetes.

Comparative Immunopathology of Cetacean morbillivirus Infection in Free-Ranging Dolphins From Western Mediterranean, Northeast-Central, and Southwestern Atlantic.

Cetacean morbillivirus (CeMV; Paramyxoviridae) causes epizootic and interepizootic fatalities in odontocetes and mysticetes worldwide. Studies suggest there is different species-specific susceptibility to CeMV infection, with striped dolphins (Stenella coeruleoalba), bottlenose dolphins (Tursiops truncatus), and Guiana dolphins (Sotalia guianensis) ranking among the most susceptible cetacean hosts. The pathogenesis of CeMV infection is not fully resolved. Since no previous studies have evaluated the organ-specific immunopathogenetic features of CeMV infection in tissues from infected dolphins, this study was aimed at characterizing and comparing immunophenotypic profiles of local immune responses in lymphoid organs (lymph nodes, spleen), lung and CNS in CeMV-molecularly (RT-PCR)-positive cetaceans from Western Mediterranean, Northeast-Central, and Southwestern Atlantic. Immunohistochemical (IHC) analyses targeted molecules of immunologic interest: caspase 3, CD3, CD20, CD57, CD68, FoxP3, MHCII, Iba1, IFNγ, IgG, IL4, IL10, lysozyme, TGFß, and PAX5. We detected consistent CeMV-associated inflammatory response patterns. Within CNS, inflammation was dominated by CD3+ (T cells), and CD20+ and PAX5+ (B cells) lymphocytes, accompanied by fewer Iba1+, CD68+, and lysozyme+ histiocytes, mainly in striped dolphins and bottlenose dolphins. Multicentric lymphoid depletion was characterized by reduced numbers of T cells and B cells, more pronounced in Guiana dolphins. Striped dolphins and bottlenose dolphins often had hyperplastic (regenerative) phenomena involving the aforementioned cell populations, particularly chronically infected animals. In the lung, there was mild to moderate increase in T cells, B cells, and histiocytes. Additionally, there was a generalized increased expression of caspase 3 in lymphoid, lung, and CNS tissues. Apoptosis, therefore, is believed to play a major role in generalized lymphoid depletion and likely overt immunosuppression during CeMV infection. No differences were detected regarding cytokine immunoreactivity in lymph nodes, spleen, and lung from infected and non-infected dolphins by semiquantitative analysis; however, there was striking immunoreactivity for IFNγ in the CNS of infected dolphins. These novel results set the basis for tissue-specific immunophenotypic responses during CeMV infection in three highly susceptible delphinid species. They also suggest a complex interplay between viral and host's immune factors, thereby contributing to gain valuable insights into similarities, and differences of CeMV infection's immunopathogenesis in relation to body tissues, CeMV strains, and cetacean hosts.

Comparative histopathologic and viral immunohistochemical studies on CeMV infection among Western Mediterranean, Northeast-Central, and Southwestern Atlantic cetaceans.

Cetacean morbillivirus (CeMV) is a major natural cause of morbidity and mortality in cetaceans worldwide and results in epidemic and endemic fatalities. The pathogenesis of CeMV has not been fully elucidated, and questions remain regarding tissue tropism and the mechanisms of immunosuppression. We compared the histopathologic and viral immunohistochemical features in molecularly confirmed CeMV-infected Guiana dolphins (Sotalia guianensis) from the Southwestern Atlantic (Brazil) and striped dolphins (Stenella coeruleoalba) and bottlenose dolphins (Tursiops truncatus) from the Northeast-Central Atlantic (Canary Islands, Spain) and the Western Mediterranean Sea (Italy). Major emphasis was placed on the central nervous system (CNS), including neuroanatomical distribution of lesions, and the lymphoid system and lung were also examined. Eleven Guiana dolphins, 13 striped dolphins, and 3 bottlenose dolphins were selected by defined criteria. CeMV infections showed a remarkable neurotropism in striped dolphins and bottlenose dolphins, while this was a rare feature in CeMV-infected Guiana dolphins. Neuroanatomical distribution of lesions in dolphins stranded in the Canary Islands revealed a consistent involvement of the cerebrum, thalamus, and cerebellum, followed by caudal brainstem and spinal cord. In most cases, Guiana dolphins had more severe lung lesions. The lymphoid system was involved in all three species, with consistent lymphoid depletion. Multinucleate giant cells/syncytia and characteristic viral inclusion bodies were variably observed in these organs. Overall, there was widespread lymphohistiocytic, epithelial, and neuronal/neuroglial viral antigen immunolabeling with some individual, host species, and CeMV strain differences. Preexisting and opportunistic infections were common, particularly endoparasitism, followed by bacterial, fungal, and viral infections. These results contribute to understanding CeMV infections in susceptible cetacean hosts in relation to factors such as CeMV strains and geographic locations, thereby establishing the basis for future neuro- and immunopathological comparative investigations.

Pathologic findings and causes of death of stranded cetaceans in the Canary Islands (2006-2012).

This study describes the pathologic findings and most probable causes of death (CD) of 224 cetaceans stranded along the coastline of the Canary Islands (Spain) over a 7-year period, 2006-2012. Most probable CD, grouped as pathologic categories (PCs), was identified in 208/224 (92.8%) examined animals. Within natural PCs, those associated with good nutritional status represented 70/208 (33.6%), whereas, those associated with significant loss of nutritional status represented 49/208 (23.5%). Fatal intra- and interspecific traumatic interactions were 37/208 (17.8%). Vessel collisions included 24/208 (11.5%). Neonatal/perinatal pathology involved 13/208 (6.2%). Fatal interaction with fishing activities comprised 10/208 (4.8%). Within anthropogenic PCs, foreign body-associated pathology represented 5/208 (2.4%). A CD could not be determined in 16/208 (7.7%) cases. Natural PCs were dominated by infectious and parasitic disease processes. Herein, our results suggest that between 2006 and 2012, in the Canary Islands, direct human activity appeared responsible for 19% of cetaceans deaths, while natural pathologies accounted for 81%. These results, integrating novel findings and published reports, aid in delineating baseline knowledge on cetacean pathology and may be of value to rehabilitators, caregivers, diagnosticians and future conservation policies.

Pyogranulomatous obliterative laryngotracheitis by Rhizopus arrhizus (syn. R. oryzae) in a free-ranging Atlantic spotted dolphin Stenella frontalis.

We report the gross and microscopic findings and molecular identification of fungal hyphate infection in a juvenile female Atlantic spotted dolphin Stenella frontalis found dead off Arguineguin, Gran Canaria (Canary Islands, Spain). On necropsy examination, the animal had a large cranial intrathoracic mass and multiple variably-sized nodules throughout the larynx and trachea that obliterated the lumen. Microscopically, the masses were composed of abundant pyogranulomatous inflammation with numerous fungal hyphae. These were pauciseptate (coenocytic) and had non-parallel walls, non-dichotomous irregular to right angle branching, and bulbous dilations. PCR analysis from these inflammatory foci yielded Rhizopus arrhizus (syn. R. oryzae). This fungal pathogen is often ascribed to opportunistic infections in immunosuppressed humans and animals. In the present case, a potential cause for immunosuppression was not identified; PCR analysis for cetacean morbillivirus was negative. Herein, we report the first confirmed case of R. arrhizus infection in a free-living Atlantic cetacean. These findings add to the body of knowledge on fungal disease in cetaceans in general and, in particular, in odontocetes, where respiratory involvement is common.

Spontaneous multicentric soft tissue sarcoma in a captive African pygmy hedgehog (Atelerix albiventris): case report and literature review.

This report describes the clinical, macroscopic, histopathological and immunohistochemical features of a spontaneous multicentric extraskeletal sarcoma in an adult male African hedgehog (Atelerix albiventris). It also provides a succinct up-to-date review on neoplasia in this species. On autopsy examination, main gross findings included a moderately demarcated cranial mass and a multilobulated, caudal intra-abdominal mass. The cranial mass had perforated the underlying temporal and occipital bones and had extended into the cranial vault and was compressing the surface of the cerebellum and cerebrum. Histologic, histochemical and immunohistochemical analyses supported a diagnosis of multicentric poorly differentiated spindle cell sarcoma with fibrosarcomatous, storiform and myxoid foci. The high incidence of neoplasia and cross similarities renders the African hedgehog a suitable species for comparative pathology studies.

Disseminated avian mycobacteriosis in a free-living grey heron (Ardea cinerea).

Wild birds share with humans the capacity for moving fast over large distances. During migratory movements, birds carry pathogens that can be transmitted between species. One of these concerning pathogens is Mycobacterium spp. A necropsy was performed in a grey heron (Ardea cinerea) that had been medically treated for a polyarthritic process. Grossly, firm white-yellowish nodules of various size, resembling granulomas, were observed in right carpal joint, both patellar joints, neck musculature, palate, pharynx, larynx, nasal sinuses, pericardial sac, air sacs, proventriculus and intestinal serosa, pancreas, kidneys, adrenal glands, and oviduct. Microscopically, these lesions were composed of multinucleated giant cells, large macrophages, and lymphocytes, with a central zone of necrosis and in some cases with peripheral fibrosis. Acid-fast bacilli were detected within these lesions. Lesions were cultured and Mycobacterium avium subsp. avium was identified. To the best of our knowledge this is the first description of mycobacteriosis in grey herons. Although the grey heron is currently considered a least-concern species, avian mycobacteriosis remains a threat on the conservation management of endangered avian species, and constitutes a public health concern as well.

Pulmonary angiomatosis and hemangioma in common dolphins (Delphinus delphis) stranded in Canary Islands.

Vascular tumors and disorders, like angiomatosis, are rarely described in cetacean species. A retrospective histological study was carried out on lung samples from 35 common dolphins (Delphinus delphis) stranded in the Canary Islands coasts looking for morphological vascular changes and likely related causes. Twenty-five out of thirty-five (71%) common dolphins showed focal or multifocal angiomatosis-like lesions. A high association between this type of vascular proliferation and parasitic infestation was observed. In addition, a single pulmonary cavernous hemangioma not previously reported in common dolphins is presented.