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1.  CTLA4 overexpressing adipose tissue-derived mesenchymal stem cell therapy in a dog with steroid-refractory pemphigus foliaceus 
Canine pemphigus foliaceus is an autoimmune antibody-mediated skin disease characterized by acantholysis. The objective of this case report is to present the successful management of steroid refractory pemphigus foliaceus with cytotoxic T-lymphocyte antigen 4 (CTLA4)-overexpressing adipose tissue mesenchymal stem cells (ATMSCs).
Case presentation
A 10-year-old, 12.3-kg, castrated male Shih Tzu presented with severe pruritus and anorexia. The diagnosis of pemphigus foliaceus was made based on its history, physical examination, and histopathology results of a skin biopsy. Treatment with prednisolone and combination therapy of other immunosuppressive drugs had failed; therefore, immunosuppressive gene, CTLA4 overexpressing ATMSCs (CTLA4-ATMSCs) and/or naive ATMSCs administration was performed with the consent of the owner. ATMSCs were administered 21 times over a period of 20 months with intervals of 2 to 8 week. Prednisolone was gradually tapered concurrently and no relapse of the clinical signs was observed. After the termination of CTLA4-ATMSCs and/or naive ATMSCs treatment, the skin lesions had improved and could be managed with a low dose of prednisolone for 12 months.
CTLA4-ATMSCs or naive ATMSCs transplantation may be beneficial as adjunctive therapy to initiate and maintain the remission of skin lesions caused by pemphigus foliaceus in veterinary medicine.
PMCID: PMC4417222  PMID: 25889154
Autoimmune disease; CTLA4; Dog; Mesenchymal stem cell; Pemphigus foliaceus
2.  Outbreak of Marek’s disease in a vaccinated broiler breeding flock during its peak egg-laying period in China 
BMC Veterinary Research  2015;11:157.
Outbreaks of Marek’s disease (MD), caused by Marek’s disease virus (MDV), primarily occur in 10–12-week-old hens.
Case presentation
We report a case of MD in a breeding flock of 24–30-week-old vaccinated broilers in China. The clinical signs in the affected chickens appeared at 24 weeks, and the incidence of tumours peaked at 30 weeks. The morbidity and mortality of the hens were 5 % and 80 %, respectively. Hematoxylin–eosin staining of the tissues showed the typical characteristics of MD. MDV infection was confirmed in the hens with an agar gel diffusion precipitation assay for the MD antigen in the feather follicle epithelium. An MDV strain, designated AH1410, was isolated from the blood lymphocytes. Sequence analyses of the pp38, meq, and gB genes revealed that strain AH1410 had molecular features consistent with a virulent, previously identified MDV.
Our data provide evidence that not only is MDV becoming more virulent, but that the period of its onset in chickens is expanding. These findings provide the basis the molecular surveillance and further study of virulent MDV mutants and control strategies for MD in China.
PMCID: PMC4511970  PMID: 26202662
Marek’s disease virus; Isolation; Gene analysis; Broiler breeding chicken; peak egg-laying period
3.  Intentional fatal metallic phosphide poisoning in a dog—a case report 
BMC Veterinary Research  2015;11:158.
Metallic phosphides are extremely toxic pesticides that are regulated in their usage. Information concerning the impact of metallic phosphides on human health is abundant. Data regarding the clinical pathology of phosphide poisoning in humans or domestic and wild animals is largely incomplete with only a few cases of metallic phosphide poisoning being reported every year, especially in humans. For the majority of cases reported in dogs the data are vague or incomplete. Here we report a complete and detailed description of pathological changes in a case of intentional metallic phosphide poisoning in a dog including an exhaustive examination of the brain.
Case presentation
A 1 year old, male, Belgian Shepherd crossbreed dog with a clean medical history and no observed clinical signs prior to death, was submitted for post mortem examination. The dog was found dead by the owner. Near the body a suspect mix of bread, fat and a blackish powder was found. The owner announced the authorities and submitted the animal and the possible bait for forensic examination. At necropsy, multisystemic necrotic and degenerative lesions were observed. Histological exam confirmed the presence of necrotic and degenerative lesions of variable severity in all of the examined organs. The toxicological forensic examination revealed the presence of the phosphine gas in the gastric content and the bait.
Metallic phosphide poisoning is a rarely reported entity, since the diagnosis of intentional poisoning with these compounds is a great challenge for forensic pathologists and toxicologists. To our knowledge, this is the first study describing the lesions completely in veterinary forensic toxicology. We assume that the toxic shows systemic endotheliotropism and damage of the endothelial cells responsible for the hemorrhagic lesions and for the secondary ischemic necrosis in various organs. This report will contribute to a better understanding of the pathogenesis in cases of acute metallic phosphide exposure in animals.
PMCID: PMC4512158  PMID: 26202780
Canine; Phosphine; Poisoning; Veterinary forensic medicine; Lesions
4.  First report of Edwardsiellosis in cage-cultured sharpsnout sea bream, Diplodus puntazzo from the Mediterranean 
BMC Veterinary Research  2015;11:155.
Edwardsiella tarda, is a serious bacterial pathogen affecting a broad range of aquaculture fish species. The bacterium has also been reported as a human pathogen, however recent studies have dissociated the fish pathogenic Edwardsiella from those isolated from humans by placing them in a new species, E. piscicida. Here we report the first case of Edwardsiellosis in cultured sharpsnout sea breams, Diplodus puntazzo in Greece.
Case presentation
The disease has affected cultured sharpsnout sea breams of a commercial fish farm in a single location in East Greece. Two populations of sharpsnout sea breams stocked in two consecutive years in floating cages presented signs of disease which included nodules and abscesses in spleen and kidney, morbidity and cumulative mortality reaching 5.3 %. Using microbiological, biochemical and molecular tools we have identified Edwardsiella sp. as the main aetiological factor of the disease. Following phylogenetic analysis the bacterial isolates are grouped with the newly described Edwardsiella piscicida species.
This is the first report of Edwardsiellosis in this species but most importantly in sea cage-cultured fish in the Mediterranean which may pose a serious threat for aquaculture fish species in this region.
PMCID: PMC4508803  PMID: 26193880
Edwardsiella; Bacterial disease; Aquaculture; Fish
5.  Surgical resection of peripheral odontogenic fibromas in African pygmy hedgehog (Atelerix albiventris): a case study 
BMC Veterinary Research  2015;11:145.
Neoplastic lesions of the mammary gland, lymph nodes, or oral cavity in African pygmy hedgehogs (Atelerix albiventris) are common in captive animals. Chemotherapy and radiotherapy protocols have not yet been established for the African pygmy hedgehog. Thus, surgical resection is the current treatment of choice in this species.
Case presentation
A 5-year-old male African pygmy hedgehog showed multiple erythematous, round small tumors located in the oral cavity, on both sides of maxilla. The treatment of choice was surgical resection of tumors using a surgical knife under general anesthesia. Excised neoplastic lesions were diagnosed as peripheral odontogenic fibroma by histopathology. Six months after surgery relapse of tumors in the oral cavity was not observed.
The treatment adopted in this case report is safe for the patient and provides the best solution for mild proliferative lesions of the oral cavity. To our knowledge this is the first report of surgical resection of oral tumors (peripheral odontogenic fibroma) in the African pygmy hedgehog.
PMCID: PMC4490659  PMID: 26141314
African pygmy hedgehog; Peripheral odontogenic fibroma; Oral disorders; Oral tumors and histopathology
6.  Emergence of porcine epidemic diarrhea virus in southern Germany 
BMC Veterinary Research  2015;11:142.
Over the last years, porcine epidemic diarrhea virus (PEDV) has caused devastating enteric diseases in the US and several countries in Asia, while outbreaks in Europe have only been reported sporadically since the 1980s. At present, only insufficient information is available on currently circulating PEDV strains in Europe and their impact on the European swine industry. In this case report, we present epidemic outbreaks of porcine epidemic diarrhea in three farms in South-Western Germany.
Case presentation
Epidemic outbreaks of diarrhea affecting pigs of all age groups were reported in three farms, one fattening farm and two piglet producing farms, in South-Western Germany between May and November 2014. In the fattening farm yellowish, watery diarrhea without evidence of mucus or blood was associated with a massive reduction of feed consumption. Severity of clinical signs and mortality in young suckling pigs varied significantly between the two affected sow farms. While mortality in suckling piglets reached almost 70 % in one sow herd, no increase in suckling piglet mortality was observed in the second sow farm. In all three cases, PEDV was confirmed in feces and small intestines by RT-qPCR. Phylogenetic analyses based on full-length PEDV genomes revealed high identity among strains from all three herds. Moreover, the German strains showed very high nucleotide identity (99.4 %) with a variant of PEDV (OH851) that was isolated in the United States in January 2014. This strain with insertions and deletions in the S-gene (so called INDEL strains) was reported to show lower virulence. Slightly lower identities were found with other strains from the US and Asia.
Phylogenetic information on the distribution of PEDV strains in Europe is severely lacking. In this case report we demonstrate that acute outbreaks of PEDV occurred in southern Germany in 2014. Current strains were clearly different from isolates found in the 1980s and were closely related to a PEDV variant found in the US in 2014. Moreover, the present case report indicates that variant strains of PEDV, containing insertions and deletions in the S gene, which were reported to be of lower virulence, might be able to cause high mortality in suckling piglets.
PMCID: PMC4487554  PMID: 26135732
Porcine epidemic diarrhea virus; Southern Germany; Phylogenetic analysis; S INDEL PEDV
7.  Hepatopathy following consumption of a commercially available blue-green algae dietary supplement in a dog 
BMC Veterinary Research  2015;11:136.
Dietary supplement use in both human and animals to augment overall health continues to increase and represents a potential health risk due to the lack of safety regulations imposed on the manufacturers. Because there are no requirements for demonstrating safety and efficacy prior to marketing, dietary supplements may contain potentially toxic contaminants such as hepatotoxic microcystins produced by several species of blue-green algae.
Case presentation
An 11-year-old female spayed 8.95 kg Pug dog was initially presented for poor appetite, lethargy polyuria, polydipsia, and an inability to get comfortable. Markedly increased liver enzyme activities were detected with no corresponding abnormalities evident on abdominal ultrasound. A few days later the liver enzyme activities were persistently increased and the dog was coagulopathic indicating substantial liver dysfunction. The dog was hospitalized for further care consisting of oral S-adenosylmethionine, silybin, vitamin K, and ursodeoxycholic acid, as well as intravenous ampicillin sodium/sulbactam sodium, dolasetron, N-acetylcysteine, metoclopramide, and intravenous fluids. Improvement of the hepatopathy and the dog’s clinical status was noted over the next three days. Assessment of the dog’s diet revealed the use of a commercially available blue-green algae dietary supplement for three-and-a-half weeks prior to hospitalization. The supplement was submitted for toxicology testing and revealed the presence of hepatotoxic microcystins (MCs), MC-LR and MC-LA. Use of the supplement was discontinued and follow-up evaluation over the next few weeks revealed a complete resolution of the hepatopathy.
To the authors’ knowledge, this is the first case report of microcystin intoxication in a dog after using a commercially available blue-green algae dietary supplement. Veterinarians should recognize the potential harm that these supplements may cause and know that with intervention, recovery is possible. In addition, more prudent oversight of dietary supplement use is recommended for our companion animals to prevent adverse events/intoxications.
PMCID: PMC4472392  PMID: 26087767
Dog; Microcystin; Blue-green algae; Hepatopathy; Toxicosis; Dietary supplement
8.  Alexander disease in a dog: case presentation of electrodiagnostic, magnetic resonance imaging and histopathologic findings with review of literature 
BMC Veterinary Research  2015;11:115.
Alexander disease is a rare neurodegenerative disorder that has not often been described in dogs. None of the existing descriptions include electrodiagnostic or magnetic resonance imaging workup. This is the first presentation of the results of an electrodiagnostic evaluation including electromyography, motor nerve conduction velocity, F-wave, the brainstem auditory evoked response and magnetic resonance imaging of a dog with Alexander disease.
Case presentation
A six month old male entire Bernese mountain dog was presented with central nervous system symptoms of generalized tremor, general stiffness, decreased proprioceptive positioning, a reduced menace response, decreased physiological nystagmus, myotonic spasms and increased spinal reflexes which progressed to lateral recumbency. The electromyography revealed normal muscle activity and a decreased motor nerve conduction velocity, temporal dispersion of the compound muscle action potential, prolonged F-wave minimal latency, lowered F-ratio, decreased latency, and lowered amplitude of the brainstem auditory evoked potentials. The magnetic resonance imaging examination revealed ventriculomegaly and linear hyperintensity on the border of the cortical grey and white matter. The histopathological examination confirmed the presence of diffuse degenerative changes of the white matter throughout the neuraxis. A proliferation of abnormal astrocytes was found at the border between the white matter and cortex. There was also a massive accumulation of eosinophilic Rosenthal fibers as well as diffuse proliferation of abnormally large astrocytes and unaffected neurons.
This is the first histopathologically confirmed case of Alexander disease in a dog with a full neurological workup. The results of the electrodiagnostic and magnetic resonance imaging examinations allow for a high-probability antemortem diagnosis of this neurodegenerative disorder in dogs.
PMCID: PMC4448536  PMID: 25985984
Rosenthal fibers; Leucodystrophy; Electrodiagnostic; Magnetic resonance imaging; Dog
9.  Another potential carp killer?: Carp Edema Virus disease in Germany 
BMC Veterinary Research  2015;11:114.
Infections with carp edema virus, a pox virus, are known from Japanese koi populations since 1974. A characteristic clinical sign associated with this infection is lethargy and therefore the disease is called “koi sleepy disease”. Diseased koi also show swollen gills, enophthalmus, and skin lesions. Mortality rates up to 80 % are described. For a long period of time, disease outbreaks seemed to be restricted to Japan. However, during the last years clinical outbreaks of koi sleepy disease also occurred in the UK and in the Netherlands.
Case presentation
In spring 2014 koi from different ponds showing lethargic behavior, skin ulcers, inflammation of the anus, enophthalmus, and gill necrosis were presented to the laboratory for diagnosis. In all cases, new koi had been purchased earlier that spring from the same retailer and introduced into existing populations. Eleven koi from six ponds were examined for ectoparasites and for bacterial and viral infections (cyprinid herpesviruses in general and especially koi herpesvirus (KHV) known formally as Cyprinid herpesvirus 3 (CyHV–3); and Carp Edema Virus). In most of the cases parasites were not detected from skin and gills. Only opportunistic freshwater bacteria were isolated from skin ulcers. In cell cultures no cytopathic effect was observed, and none of the samples gave positive results in PCR tests for cyprinid herpesviruses. By analyzing gill tissues for CEV in seven out of eleven samples by a nested PCR, PCR products of 547 bp and 180 bp (by using nested primers) could be amplified. An outbreak of Koi Sleepy Disease was confirmed by sequencing of the PCR products. These results confirm the presence of CEV in German koi populations.
A clinical outbreak of “koi sleepy disease” due to an infection with Carp Edema Virus was confirmed for the first time in Germany. To avoid transmission of CEV to common carp testing of CEV should become part of fish disease surveillance programs.
Electronic supplementary material
The online version of this article (doi:10.1186/s12917-015-0424-7) contains supplementary material, which is available to authorized users.
PMCID: PMC4431602  PMID: 25976542
Carp Edema Virus; Koi sleepy disease; Cyprinus carpio
10.  Two cases of meningocele and meningoencephalocele in Jeju native pigs 
Meningocele and meningoencephalocele of the skull are congenital deformities. Various species, such as pigs, dogs, and cats, are susceptible to congenital meningocele and meningoencephalocele and the incidence is higher in large white and landrace pigs.
Case presentation
In this study, swelling was observed in the fontanel areas of the median planes of the skull cap in two female piglets of the same litter. Gross clinical examination, neurological examination, computed tomography (CT), and magnetic resonance imaging (MRI) were conducted on the symptomatic piglets. The gross clinical and neurological examinations revealed no specific findings, except for the swellings. According to the CT results, the length of the defect on the sagittal section of the skull was 4.7 mm in case 1 and 20.62 mm in case 2. Connected flow between the skull swellings and the cerebrospinal fluid (CSF) of the lateral ventricles was observed, and partial herniation was identified in case 2. On MRI, CSF with high T2 signals was identified in the arachnoid spaces between the cerebrum and the cerebellum in the two cases, which is consistent with intracranial hypertension. The size of the swelling formed in the parietal bones was 1.6 × 1.1 × 1.8 cm3 (case 1) and 1.2 × 1.38 × 1.7 cm3 (case 2). The increase in intracranial pressure was more obvious in case 2 than in case 1, and was accompanied by posterior displacements of the mesencephalon and cerebellum.
Case 1 was diagnosed as meningocele resulting from meningeal herniation and case 2 was diagnosed as meningoencephalocele caused by brain tissue herniation.
PMCID: PMC4399108  PMID: 25889174
11.  Fatal disease associated with Swine Hepatitis E virus and Porcine circovirus 2 co-infection in four weaned pigs in China 
In recent decades, Porcine circovirus 2 (PCV2) infection has been recognized as the causative agent of postweaning multisystemic wasting syndrome, and has become a threat to the swine industry. Hepatitis E virus (HEV) is another high prevalent pathogen in swine in many regions of the world. PCV2 and HEV are both highly prevalent in pig farms in China.
Case presentation
In this study, we characterized the HEV and PCV2 co-infection in 2–3 month-old piglets, based on pathogen identification and the pathological changes observed, in Hebei Province, China. The pathological changes were severe, and general hyperemia, hemorrhage, inflammatory cell infiltration, and necrosis were evident in the tissues of dead swine. PCR was used to identify the pathogen and we tested for eight viruses (HEV, Porcine reproductive and respiratory syndrome virus, PCV2, Classical swine fever virus, Porcine epidemic diarrhea virus, Transmissible gastroenteritis coronavirus, Porcine parvovirus and Pseudorabies virus) that are prevalent in Chinese pig farms. The livers, kidneys, spleens, and other organs of the necropsied swine were positive for HEV and/or PCV2. Immunohistochemical staining showed HEV- and PCV2-antigen-positive signals in the livers, kidneys, lungs, lymph nodes, and intestine.
HEV and PCV2 co-infection in piglets was detected in four out of seven dead pigs from two pig farms in Hebei, China, producing severe pathological changes. The natural co-infection of HEV and PCV2 in pigs in China has rarely been reported. We speculate that co-infection with PCV2 and HEV may bring some negative effect on pig production and recommend that more attention should be paid to this phenomenon.
PMCID: PMC4379595  PMID: 25889526
Weaned pigs; Hepatitis E virus; Porcine circovirus 2; Co-infection; High mortality
12.  Isolation of Leptospira interrogans serovar Hardjoprajitno from a calf with clinical leptospirosis in Chile 
Although Leptospira isolation has been reported in Chilean cattle, only serological evidence of serovar Hardjo bovis infection has been routinely reported. The present report provides characterization of the pathological presentation and etiology of a clinical case of leptospirosis in a calf from the Los Rios Region in Chile.
Case presentation
In a dairy herd in southern Chile, 11 of 130 calves died after presenting signs such as depression and red-tinged urine. One of these calves, a female of eight months, was necropsied, and all the pathological findings were consistent with Leptospira infection. A urine sample was submitted to conventional bacteriological analysis together with highly specific molecular biology typing tools, in order to unravel the specific Leptospira specie and serovar associated with this clinical case.
A significant finding of this study was that the obtained isolate was confirmed by PCR as L. interrogans, its VNTR profile properly matching with L. interrogans Hardjoprajitno as well as its specific genomic identity revealed by secY gen.
Leptospira interrogans serovar Hardjoprajitno was associated with the investigated calf clinical case.
This information adds to the value of serologic results commonly reported, which encourage vaccination improvements to match circulating strains. In addition, this finding represents the first case report of this serovar in Chilean cattle.
PMCID: PMC4374366  PMID: 25888965
Leptospira; Cattle; Clinical; Hardjoprajitno
13.  Use of a novel silk mesh for ventral midline hernioplasty in a mare 
Ventral midline hernia formation following abdominal surgery in horses is an uncommon complication; however, it can have serious consequences leading to increased morbidity and mortality. Currently, mesh hernioplasty is the treatment of choice for large ventral midline hernias in horses to allow potential return to normal function. Complications following mesh hernioplasty using polypropylene or polyester mesh in horses can be serious and similar to complications seen in human patients, including persistent incisional drainage, mesh infection, hernia recurrence, intra-abdominal adhesions, mesh or body wall failure, recurrent abdominal pain (colic), and peritonitis. This report describes the use of a novel bioresorbable silk mesh for repair of a large ventral midline incisional hernia in a mature, 600-kg horse. To our knowledge, this is the first report of its kind in the literature.
Case presentation
A 9-year-old, 600-kg Warmblood mare presented with a ventral midline hernia following emergency exploratory celiotomy 20 months prior. The mare was anesthetized and a hernioplasty was performed using a novel bioresorbable silk mesh (SERI® Surgical Scaffold; Allergan Medical, Boston, MA). No complications were encountered either intra- or postoperatively. The mare was discharged from the hospital at 3 days postoperatively in an abdominal support bandage. At 8 and 20 weeks postoperatively, ultrasonographic assessment showed evidence of tissue ingrowth within and around the mesh. The mare was able to be bred 2 years in a row, carrying both foals to full gestation with no complications. Following both foalings, the abdomen has maintained a normal contour with no evidence of hernia recurrence.
Ventral abdominal hernias can be repaired in horses using a bioresorbable silk mesh, which provides adequate biomechanical strength while allowing for fibrous tissue ingrowth. The use of a bioresorbable silk mesh for the repair of ventral hernias can be considered as a realistic option as it potentially provides significant benefits over traditional non-resorbable mesh.
PMCID: PMC4426545  PMID: 25879822
Abdominal; Hernia; Hernioplasty; Horse; Mesh; Silk
14.  First report of junctional epidermolysis bullosa (JEB) in the Italian draft horse 
Epitheliogenesis imperfecta in horses was first recognized at the beginning of the 20th century when it was proposed that the disease could have a genetic cause and an autosomal recessive inheritance pattern. Electron microscopy studies confirmed that the lesions were characterized by a defect in the lamina propria and the disease was therefore reclassified as epidermolysis bullosa. Molecular studies targeted two mutations affecting genes involved in dermal–epidermal junction: an insertion in LAMC2 in Belgians and other draft breeds and one large deletion in LAMA3 in American Saddlebred.
Case presentation
A mechanobullous disease was suspected in a newborn, Italian draft horse foal, which presented with multifocal to coalescing erosions and ulceration on the distal extremities. Histological examination of skin biopsies revealed a subepidermal cleft formation and transmission electron microscopy demonstrated that the lamina densa of the basement membrane remained attached to the dermis. According to clinical, histological and ultrastructural findings, a diagnosis of junctional epidermolysis bullosa (JEB) was made. Genetic tests confirmed the presence of 1368insC in LAMC2 in the foal and its relatives.
This is the first report of JEB in Italy. The disease was characterized by typical macroscopic, histologic and ultrastructural findings. Genetic tests confirmed the presence of the 1368insC in LAMC2 in this case: further investigations are required to assess if the mutation could be present at a low frequency in the Italian draft horse population. Atypical breeding practices are responsible in this case and played a role as odds enhancer for unfavourable alleles. Identification of carriers is fundamental in order to prevent economic loss for the horse industry.
PMCID: PMC4372232  PMID: 25889423
Junctional epidermolysis bullosa; Horse; Mechanobullous disease; Electron microscopy; Lamina densa; LAMC2; Italian draft horse; Inbreeding
15.  Novel adenovirus detected in captive bottlenose dolphins (Tursiops truncatus) suffering from self-limiting gastroenteritis 
Adenoviruses are common pathogens in vertebrates, including humans. In marine mammals, adenovirus has been associated with fatal hepatitis in sea lions. However, only in rare cases have adenoviruses been detected in cetaceans, where no clear correlation was found between presence of the virus and disease status.
Case presentation
A novel adenovirus was identified in four captive bottlenose dolphins with self-limiting gastroenteritis. Viral detection and identification were achieved by: PCR-amplification from fecal samples; sequencing of partial adenovirus polymerase (pol) and hexon genes; producing the virus in HeLa cells, with PCR and immunofluorescence detection, and with sequencing of the amplified pol and hexon gene fragments. A causative role of this adenovirus for gastroenteritis was suggested by: 1) we failed to identify other potential etiological agents; 2) the exclusive detection of this novel adenovirus and of seropositivity for canine adenoviruses 1 and 2 in the four sick dolphins, but not in 10 healthy individuals of the same captive population; and 3) the virus disappeared from feces after clinical signs receded. The partial sequences of the amplified fragments of the pol and hexon genes were closest to those of adenoviruses identified in sea lions with fatal adenoviral hepatitis, and to a Genbank-deposited sequence obtained from a harbour porpoise.
These data suggest that adenovirus can cause self-limiting gastroenteritis in dolphins. This adenoviral infection can be detected by serology and by PCR detection in fecal material. Lack of signs of hepatitis in sick dolphins may reflect restricted tissue tropism or virulence of this adenovirus compared to those of the adenovirus identified in sea lions. Gene sequence-based phylogenetic analysis supports a common origin of adenoviruses that affect sea mammals. Our findings suggest the need for vigilance against adenoviruses in captive and wild dolphin populations.
PMCID: PMC4352565  PMID: 25888777
Adenovirus; Cetacean; Bottlenose dolphin; Tursiops truncatus
16.  Quadricuspid aortic valve and a ventricular septal defect in a horse 
BMC Veterinary Research  2014;10:142.
Quadricuspid aortic valve (QAV) and ventricular septal defect (VSD) are congenital heart defects and have been described in both human and veterinary medical literature.
Case presentation
A 5-year-old half-bred bay stallion was referred for surgical castration. Cardiac murmurs were heard on the presurgical clinical examination and the cardiac examination revealed subcutaneous oedema, tachycardia with a precodrial thrill and a grade 5/6 pansystolic murmur, which was heard on auscultation of the right and left side of the chest. Examination of the B-mode echocardiograms revealed the presence of a QAV (one small cusp, two equal-sized cusps, and one large cusp) and VSD in the membranous portion of the intraventricular septum. These two congenital cardiac defects were accompanied by mild aortic valve regurgitation and severe tricuspid regurgitation. Despite the presence of these cardiac defects, the horse underwent surgical castration under general anesthesia. Surgery, anaesthesia and recovery from anaesthesia were uneventful. The gelding was euthanasied after 17 months because of a progressive loss of body weight, weakness and recumbency.
A QAV in combination with VSD in a horse is an interesting finding, because to the best of our knowledge, this has not been previously described in equine literature.
PMCID: PMC4134333  PMID: 24981768
Horse; Quadricuspid aortic valve; Ventricular septal defect; Congenital heart defect; Echocardiography
17.  Methamphetamine intoxication in a dog: case report 
BMC Veterinary Research  2014;10:139.
Methamphetamine abuse has undergone a dramatic worldwide increase, and represents a significant and global issue for public health. Incidents of methamphetamine intoxication and death in humans are relatively commonplace. Because of its increasing illicit availability, together with legitimate use in human medicine, accidental or intentional exposure to methamphetamine in dogs is becoming a more likely scenario.
Case presentation
A 3-year-old, 3.7 kg intact female Miniature Poodle who had been intentionally fed an unknown amount of a crystalline-like substance developed extreme agitation, seizures, tachycardia, hyperthermia, hypertension, disseminated intravascular coagulation (DIC), bloody diarrhea, and dilated pupils. Blood work revealed leukocytosis, erythropenia, lymphocytosis, thrombocytopenia, coagulation abnormalities, but all to a mild extent, together with mild elevation in both alanine aminotranferease (ALT) and alkaline phosphatase (ALKP), and a mild decreased in glucose. Radiologic diagnosis revealed generalized, severe distension of the stomach and small intestinal tract with air. Immunochromatographic screening tests and gas chromatography mass spectrometry analysis confirmed methamphetamine intoxication and revealed concentrations of methamphetamine in blood and urine of 0.32 μg/mL and 2.35 μg/mL respectively. The dog demonstrated progressive improvement after supportive care, with the high fever resolved over the initial 24 hours of hospitalization, and agitation was successfully controlled beyond 48 hours after initial hospitalization. Hemostatic abnormalities were progressive improved after heparin therapy and supportive care. By the sixth day of hospitalization the dog was clinically well, and all laboratory data had returned to normal with the exception of a mild elevateion of ALKP.
To the authors’ knowledge, this is the second case report of methamphetamine intoxication in dogs presented in veterinary practice in open literature so far. Although rare, methamphetamine intoxication should be considered as a differential diagnosis in dogs with a toxic substance ingestion history and with typical nervous and cardiovascular system symptoms. In such cases rapid diagnosis and aggressive intervention is important for prognosis. Blood methamphetamine concentration may be a helpful value for assessment of the severity of intoxication and prediction of clinical outcomes.
PMCID: PMC4078017  PMID: 24962469
Methamphetamine; Intoxication; Dog; Agitation; Tachycardia; Hyperthermia; Disseminated intravascular coagulation; Urine screen test; Gas chromatography mass spectrometry
18.  Molecular and pathological insights into Chlamydia pecorum-associated sporadic bovine encephalomyelitis (SBE) in Western Australia 
BMC Veterinary Research  2014;10:121.
Despite its global recognition as a ruminant pathogen, cases of Chlamydia pecorum infection in Australian livestock are poorly documented. In this report, a C. pecorum specific Multi Locus Sequence Analysis scheme was used to characterise the C. pecorum strains implicated in two cases of sporadic bovine encephalomyelitis confirmed by necropsy, histopathology and immunohistochemistry. This report provides the first molecular evidence for the presence of mixed infections of C. pecorum strains in Australian cattle.
Case presentation
Affected animals were two markedly depressed, dehydrated and blind calves, 12 and 16 weeks old. The calves were euthanized and necropsied. In one calf, a severe fibrinous polyserositis was noted with excess joint fluid in all joints whereas in the other, no significant lesions were seen. No gross abnormalities were noted in the brain of either calf. Histopathological lesions seen in both calves included: multifocal, severe, subacute meningoencephalitis with vasculitis, fibrinocellular thrombosis and malacia; diffuse, mild, acute interstitial pneumonia; and diffuse, subacute epicarditis, severe in the calf with gross serositis. Immunohistochemical labelling of chlamydial antigen in brain, spleen and lung from the two affected calves and brain from two archived cases, localised the antigen to the cytoplasm of endothelium, mesothelium and macrophages. C. pecorum specific qPCR, showed dissemination of the pathogen to multiple organs. Phylogenetic comparisons with other C. pecorum bovine strains from Australia, Europe and the USA revealed the presence of two genetically distinct sequence types (ST). The predominant ST detected in the brain, heart, lung and liver of both calves was identical to the C. pecorum ST previously described in cases of SBE. A second ST detected in an ileal tissue sample from one of the calves, clustered with previously typed faecal bovine isolates.
This report provides the first data to suggest that identical C. pecorum STs may be associated with SBE in geographically separated countries and that these may be distinct from those found in the gastrointestinal tract. This report provides a platform for further investigations into SBE and for understanding the genetic relationships that exist between C. pecorum strains detected in association with other infectious diseases in livestock.
PMCID: PMC4064815  PMID: 24884687
Sporadic bovine encephalomyelitis; Chlamydia pecorum; Cattle; MLSA; Histopathology; Immunohistochemistry
19.  Acute gastroesophageal intussusception in a juvenile Australian shepherd dog: endoscopic treatment and long-term follow-up 
BMC Veterinary Research  2014;10:109.
Canine gastroesophageal intussusception (GEI) is a rare and potentially fatal disease usually affecting puppies or young dogs < 3 months of age and of medium to large breeds. Surgical intervention has been advocated as the therapy of choice by most authors. Endoscopic treatment may offer an advantageous or alternative method of treatment.
Case presentation
GEI was diagnosed in a nine-week-old Australian Shepherd dog with an acute onset of vomiting and regurgitation and compatible radiographic findings on thoracic radiography. Treatment consisted of endoscopic gastric repositioning and placement of a percutaneous endoscopic gastrostomy (PEG) tube to prevent repeated dislocation of the stomach, and to allow for nutritional supplementation During a follow- up period of eight months, thoracic radiographs were obtained showing persistent esophageal dilatation in the absence of compatible clinical signs.
Endoscopic intervention is an effective, alternative in selected canine GEI- cases, allowing for rapid, minimally invasive confirmation of diagnosis and therapy. After initial treatment, radiographic long-term follow-up seems prudent even in asymptomatic patients.
PMCID: PMC4024095  PMID: 24885648
Esophageal obstruction; Canine; Eso2phagoscopy; PEG tube; Gastroesophageal intussusception
20.  Cryptococcal meningitis in a goat – a case report 
Cryptococcus spp. are saprophytic and opportunistic fungal pathogens that are known to cause severe disease in immunocompromised animals. In goats there are reports of clinical cryptococcal pneumonia and mastitis but not of meningitis.
Case presentation
The following report describes a case of a five year old buck showing severe neurological signs, including paraplegia and strong pain reaction to touch of the hindquarters region. Treatment with antibiotics was unsuccessful and the animal was euthanized for humanitarian reasons. Postmortem examination revealed lumbar meningitis, lung nodules and caseous lymphadenitis lesions. Encapsulated Cryptococcus neoformans were identified from the lungs and meninges, showing that cryptococcal meningitis should be included in the differential diagnosis of goats showing paresis and hyperesthesia. The possibility of concurrent immunosuppression due to Corynebacterium pseudotuberculosis infection is raised.
Cryptoccocal meningitis should be included in the differential diagnosis list of goat diseases with ataxia and hyperesthesia.
PMCID: PMC4020613  PMID: 24708822
Cryptococcus; Meningitis; Goat; Allodynia; Ataxia; Caseous lymphadenitis
21.  Japanese encephalitis in a 114-month-old cow: pathological investigation of the affected cow and genetic characterization of Japanese encephalitis virus isolate 
Japanese encephalitis virus (JEV) is classified into the genus Flavivirus in the family Flaviviridae. JEV can cause febrile illness and encephalitis mainly in humans and horses, and occasionally in cattle.
Case presentation
In late September 2010, a 114-month-old cow showed neurological symptoms similar to the symptoms observed in previous bovine cases of Japanese encephalitis (JE); therefore, we conducted virological and pathological tests on the cow. As a result, JEV was isolated from the cerebrum of the affected cow. We determined the complete genome sequence of the JEV isolate, which we named JEV/Bo/Aichi/1/2010, including the envelope (E) gene region and 3’ untranslated region (3’UTR). Our phylogenetic analyses of the E region and complete genome showed that the isolate belongs to JEV genotype 1 (G1). The isolate, JEV/Bo/Aichi/1/2010, was most closely related to several JEV G1 isolates in Toyama Prefecture, Japan in 2007–2009 by the phylogenetic analysis of the E region. In addition, the nucleotide alignment revealed that the deletion in the 3’UTR was the same between JEV/Bo/Aichi/1/2010 and several other JEV G1 isolates identified in Toyama Prefecture in 2008–2009. A hemagglutination inhibition (HI) test was conducted for the detection of anti-JEV antibodies in the affected cow, and the test detected 2-mercaptoethanol (2-ME)-sensitive HI antibodies against JEV in the serum of the affected cow. The histopathological investigation revealed nonsuppurative encephalomyelitis in the affected cow, and the immunohistochemical assay detected JEV antigen in the cerebrum.
We diagnosed the case as JE of a cow based on the findings of nonsuppurative encephalomyelitis observed in the central nervous system, JEV antigen detected in the cerebrum, JEV isolated from the cerebrum, and 2-ME-sensitive HI antibodies against JEV detected in the serum. This is the first reported case of JE in a cow over 24 months old.
PMCID: PMC4007642  PMID: 24618225
Japanese encephalitis; Cow; Arthropod-borne virus; Viral encephalomyelitis
22.  Spinal dermoid sinus in a Dachshund with vertebral and thoracic limb malformations 
Dermoid sinus is an uncommon epithelial-lined fistula that may be associated with vertebral malformations. In humans, Klippel-Feil syndrome (KFS) is a rare condition characterized by congenital cervical vertebral fusion and may be associated with other developmental defects, including dermoid sinus. The present case report describes an adult Dachshund with cervical and cranial thoracic vertebral malformations as well as thoracic limb malformations resembling KFS with a concurrent type IV dermoid sinus.
Case presentation
A 1.5 year-old Dachshund with congenital thoracic limbs deformities and cervical-thoracic vertebral malformations presented with cervical hyperesthesia, rigidity of the cervical musculature and tetraparesis. Neurologic, radiographic, and computed tomography (CT) (2D, 3D, CT fistulography) examinations revealed skeletal anomalies, a dermoid sinus in the cranial thoracic region and epidural gas within the vertebral canal. Surgical resection and histopathological evaluation of the sinus tract were performed and confirmed a type IV dermoid sinus. The clinical signs progressively recovered postoperatively, and no recurrent signs were observed after 6 months of follow-up.
Cervical vertebral malformations associated with limbs anomalies have not been reported in dogs and may represent a condition similar to KFS in humans. KFS can occur concurrently with other congenital conditions including dermoid sinus and should be included among the complex congenital anomalies described in dogs.
PMCID: PMC3975874  PMID: 24593884
Dermoid sinus; Klippel-Feil syndrome; Limbs malformations; Dog
23.  Management of a congenital tracheoesophageal fistula in a young Spanish water dog 
Tracheoesophageal fistula (TEF) in dogs is a rare disease with only few reports in the literature. This report aims to contribute to the current literature on suitable diagnostic methods for TEF and to provide follow-up information after successful surgical treatment.
Case presentation
A seven-month-old intact female Spanish Water Dog was presented for further investigation of recurrent respiratory symptom. Bronchoscopy revealed a small hole-like defect in the tracheal wall at the bifurcation. The finding of the contrast material swallow study under fluoroscopy was indicative of a TEF. To further evaluate the connection between the trachea and esophagus, a computed tomography scan was performed. The TEF was surgically approached by thoracotomy through the right lateral sixth intercostal space. The fistula was identified, double ligated and divided. Histopathology confirmed the process to originate from the esophagus and to be patent. The dog was re-examined two weeks and ten months after surgery, with no evidence of recurring clinical signs.
Contrast material swallow study using fluoroscopy was the most reliable diagnostic method. Bronchoscopy may allow the fistula to be visualized, but due to a small fistular opening it can lead to a false negative result. Surgical correction by ligation and dividing of the fistula suggests a good prognosis for early diagnosed and operated TEF.
PMCID: PMC3895692  PMID: 24423070
Congenital tracheoesophageal fistula; Dog; Bronchoscopy; Fluoroscopy; Computed tomography; Histopathology
24.  Outbreak of Listeria monocytogenes in an urban poultry flock 
Listeria monocytogenes infection is most commonly recognized in ruminants, including cattle, sheep, and goats; but it is rarely diagnosed in poultry. This report describes an outbreak of L. monocytogenes in a backyard poultry flock. Also, it points out the importance of collaboration between veterinarians and public health departments and the possible implications of zoonotic diseases.
Case presentation
Depression, lack of appetite, labored breathing, and increased mortality were noted for 5 months in several affected birds within the flock. The pathologic changes in the internal organs of infected birds included severe myocarditis, pericarditis, pneumonia, hepatitis, and splenitis. No lesions were noted in the brain. Gram-positive organisms were seen in histologic sections of the heart and spleen. Listeria monocytogenes was detected by real time PCR from formalin fixed heart and spleen, and was isolated from fresh lung, spleen, and liver. This isolate was identified as L. monocytogenes serotype 4b by 16S rDNA sequencing and by PCR-based serotyping assay.
This is the first report describing outbreak of L. monocytogenes in backyard poultry flock in Washington State and use of molecular methods to confirm L. monocytogenes infection from formalin fixed tissues.
PMCID: PMC3852838  PMID: 24119838
25.  Unusual presentation of alveolar echinococcosis as prostatic and paraprostatic cysts in a dog 
Alveolar echinococcosis (AE) is caused by the larval stage (metacestode) of Echinococcus multilocularis. The domestic dog can act as a definitive host and harbor adult cestodes in its small intestine or become an aberrant intermediate host carrying larval stages that may cause severe lesions in the liver, lungs and other organs with clinical signs similar to AE in humans.
Case presentation
A case of canine AE, affecting the liver and prostate with development of multilocular hydatid paraprostatic cysts and possible lung involvement is described in an 8–year-old neutered male Labrador retriever dog.
The dog presented with progressive weight loss, acute constipation, stranguria and a suspected soft tissue mass in the sublumbar region. Further evaluation included computed tomography of the thorax and abdomen, which revealed cystic changes in the prostate, a paraprostatic cyst, as well as lesions in the liver and lungs. Cytological examination of fine-needle aspirates of the liver, prostate and paraprostatic cyst revealed parasitic hyaline membranes typical of an Echinococcus infection and the presence of E. multilocularis-DNA was confirmed by PCR.
The dog was treated with albendazole and debulking surgery was considered in case there was a good response to antiparasitic treatment. Constipation and stranguria resolved completely. Six months after the definitive diagnosis, the dog was euthanized due to treatment-resistant ascites and acute anorexia and lethargy.
To the authors’ knowledge, this is the first publication of an E. multilocularis infection in a dog causing prostatic and paraprostatic cysts. Although rare, E. multilocularis infection should be considered as an extended differential diagnosis in dogs presenting with prostatic and paraprostatic disease, especially in areas where E. multilocularis is endemic.
PMCID: PMC3765175  PMID: 23938212
Echinococcus multilocularis; Prostatic cyst; Paraprostatic cysts; Cytology; Radiology; PCR; Canine alveolar echinococcosis; Dog

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