Plasma bile acids in healthy green iguanas and iguanas with chronic liver diseases.

The aim of the study was to establish reference values for plasma bile acid (BA) concentrations in a collection of healthy green iguanas and to compare the results with BA concentrations in iguana patients presented to the clinic with various types of chronic liver diseases, patients with other chronic diseases and healthy iguanas that were presented for routine or pre-surgical health check-up. The concentration of BA was determined using the enzymatic colorimetric method. Mean plasma bile acid concentration in 110 samples from healthy green iguanas fasted for 24 h was higher (15.89 ± 15.61 µmol/l) than plasma bile acid concentration in the same iguanas fasted for 48 h (9.56 ± 8.52 µmol/l) (P < 0.01). The 3α-hydroxy bile acid concentration was significantly altered in 9 patients suffering from chronic liver diseases (diagnosed by histology) (84.85 ± 22.29 µmol/l). BA concentration in one iguana with hepatocellular adenoma (13.0 µmol/l) was within the interval of BA in healthy iguanas. Mean plasma BA concentration in 10 green iguanas that were suffering from various types of chronic diseases, but without any hepatopathy was 7.85 ± 4.86 µmol/l. The mean plasma BA concentration in 18 samples from green iguanas presented to the clinic for routine health check-ups and 17 green iguana females with preovulatory follicle stasis (POFS) syndrome presented for ovariectomy was 11.95 ± 9.43 µmol/l and 12.97 ± 9.06 µmol/l, respectively. The data collected from this study suggest that plasma bile acids are significantly increased in green iguanas suffering from chronic liver diseases.

Molecular characterisation of a disseminated Cryptosporidium infection in a Koi carp (Cyprinus carpio).

Cryptosporidium is a protozoan parasite that infects a wide range of hosts, yet relatively little is known about the epidemiology of cryptosporidiosis in fish. Here we report a disseminated Cryptosporidium infection in a male Koi carp (Cyprinus carpio), with parasite stages identified deep within the epithelium of the intestine, kidneys, spleen, liver and gills causing severe granulomatous inflammatory lesions. Molecular characterization at two loci; 18S ribosomal RNA (rRNA) and actin, revealed this to be a novel Cryptosporidium genotype, most closely related to Cryptosporidium molnari.

VITAMIN B6 (PYRIDOXINE HYDROCHLORIDE) TOXICOSIS IN FALCONS.

This manuscript reports three independent accidental cases of vitamin (Vit) B6 toxicosis in gyrfalcons (Falco rusticolus) and peregrine falcons (Falco peregrinus) and a toxicology study that was conducted to characterize the clinical responses of gyrfalcons and gyrfalcon × peregrine falcons to a range of single intramuscular (IM) and oral (PO) doses of Vit B6. Both lethal and nonlethal doses were determined. Twelve female gyrfalcons died following IM injection of 1 ml of a vitamin B preparation. Within 30 min of injection, the birds passed pistachio green-colored urates and progressed to vomiting, anorexia, cessation of normal activity, ptosis, collapse, and death, occurring 24-36 hr post injections. Three individuals vomited frothy, partially digested blood and had clonic spasms and convulsions. Postmortem and histopathology revealed multifocal severe hepatic necrosis, splenic lymphoid tissue depletion and hemorrhages with arterial necrosis, and acute renal tubular necrosis. Following administration of a different, oral, mineral-vitamin supplement, a total of 21 peregrine falcons in two separate European facilities died suddenly. Histology of the liver showed diffuse congestion and multifocal coagulative necrosis with mild infiltration of heterophils. The particular nutritional supplement, used by both breeders, was analyzed and found to contain 5-9.7% Vit B6. Other randomly selected lots of the product contained 0.007-0.27% Vit B6. According to the product label, Vit B6 should have been present at 0.004%. To confirm the hypothesis that Vit B6 was responsible for the deaths of the falcons in Abu Dhabi, Vit B6 (British Pharmacopoeia [BP] grade) in powder form was diluted in water for injection and administered IM to four groups of falcons. Groups of four gyrfalcon × peregrine hybrid falcons or gyrfalcons (or both) were given a single IM dose of 5, 10, 15, or 20 mg/kg of Vit B6 or received an oral dose of 25, 50, or 75 mg of Vit B6. Only birds in the lowest-dose groups survived. The maximum nonlethal single doses of Vit B6 in falcons were 5 mg/kg i.m. and 25 mg/kg p.o.

Core and Shell Song Systems Unique to the Parrot Brain.

The ability to imitate complex sounds is rare, and among birds has been found only in parrots, songbirds, and hummingbirds. Parrots exhibit the most advanced vocal mimicry among non-human animals. A few studies have noted differences in connectivity, brain position and shape in the vocal learning systems of parrots relative to songbirds and hummingbirds. However, only one parrot species, the budgerigar, has been examined and no differences in the presence of song system structures were found with other avian vocal learners. Motivated by questions of whether there are important differences in the vocal systems of parrots relative to other vocal learners, we used specialized constitutive gene expression, singing-driven gene expression, and neural connectivity tracing experiments to further characterize the song system of budgerigars and/or other parrots. We found that the parrot brain uniquely contains a song system within a song system. The parrot "core" song system is similar to the song systems of songbirds and hummingbirds, whereas the "shell" song system is unique to parrots. The core with only rudimentary shell regions were found in the New Zealand kea, representing one of the only living species at a basal divergence with all other parrots, implying that parrots evolved vocal learning systems at least 29 million years ago. Relative size differences in the core and shell regions occur among species, which we suggest could be related to species differences in vocal and cognitive abilities.

Marek's disease virus associated ocular lymphoma in Roulroul partridges (Rollulus rouloul).

Two 1-year old Roulroul partridges (Rollulus rouloul), one male and one female, were presented because of eye problems and anorexia. Twenty of the 30 Roulroul partridges in the owner's collection had already died. The affected birds stopped eating, became thinner, and eventually died. Antibiotic treatment, which started because of the suspicion of a septicaemic process, was unsuccessful. At clinical examination of the two partridges it was found that in both birds, one eye ball was filled with a whitish yellow amorphous material and the other eye ball of the female showed a distinct corneal opacity. Both presented birds were euthanized. Necropsy revealed no significant abnormalities in addition to the eye lesions. Histology and immunohistochemistry of the female's eye revealed an infiltrate of T-lymphocytes corresponding to ocular lymphoma. Herpesvirus genus-specific PCR, followed by Sanger sequencing confirmed the presumptive diagnosis of Marek's disease in both birds. To our knowledge, this is the first confirmed case of infection with Gallid Herpesvirus 2 (Marek's disease virus) in partridges and the first case in this specific species.

Worldwide phylogenetic relationship of avian poxviruses.

Poxvirus infections have been found in 230 species of wild and domestic birds worldwide in both terrestrial and marine environments. This ubiquity raises the question of how infection has been transmitted and globally dispersed. We present a comprehensive global phylogeny of 111 novel poxvirus isolates in addition to all available sequences from GenBank. Phylogenetic analysis of the Avipoxvirus genus has traditionally relied on one gene region (4b core protein). In this study we expanded the analyses to include a second locus (DNA polymerase gene), allowing for a more robust phylogenetic framework, finer genetic resolution within specific groups, and the detection of potential recombination. Our phylogenetic results reveal several major features of avipoxvirus evolution and ecology and propose an updated avipoxvirus taxonomy, including three novel subclades. The characterization of poxviruses from 57 species of birds in this study extends the current knowledge of their host range and provides the first evidence of the phylogenetic effect of genetic recombination of avipoxviruses. The repeated occurrence of avian family or order-specific grouping within certain clades (e.g., starling poxvirus, falcon poxvirus, raptor poxvirus, etc.) indicates a marked role of host adaptation, while the sharing of poxvirus species within prey-predator systems emphasizes the capacity for cross-species infection and limited host adaptation. Our study provides a broad and comprehensive phylogenetic analysis of the Avipoxvirus genus, an ecologically and environmentally important viral group, to formulate a genome sequencing strategy that will clarify avipoxvirus taxonomy.

Filarial tenosynovitis caused by Pelecitus species (Spirurida, Filarioidea, Onchocercidae) in the legs of a channel-billed toucan (Ramphastos vitellinus).

An adult channel-billed toucan (Ramphastos vitellinus), imported from Surinam, South America, 18 months earlier, was found dead in its cage. At necropsy, the hock joints were swollen bilaterally and had a reduced range of motion. Small curled worms were present in subcutaneous tissue around the joints, with edema and fibrin formation. These worms were classified as the filarioid nematode, Pelecitus species. Blood-borne microfilariae were seen in cytologic impression smears of the lung and the leg lesions.

Multiple papillomas in a diamond python, Morelia spilota spilota.

A 4-yr-old male diamond python (Morelia spilota spilota) was evaluated for multiple black papillated exophytic skin proliferations and signs of pneumonia. The histopathologic structure of the skin biopsy specimens led to the diagnosis of a benign papilloma-like neoplasia. In this case, papillomavirus DNA could be amplified from a biopsy sample with a broad range polymerase chain reaction. Nested pan-herpes polymerase chain reaction was negative, and herpesvirus inclusion bodies were not found. Because of the histologically benign nature of the papilloma, the skin proliferations were left untreated. Ten mo after the first presentation, the skin lesions had regressed almost completely; 34 mo later, only scars from the biopsies were left.

Microsatellite typing of avian clinical and environmental isolates of Aspergillus fumigatus.

Aspergillosis is one of the most common causes of death in captive birds. Aspergillosis in birds is mainly caused by Aspergillus fumigatus, a ubiquitous and opportunistic saprophyte. Currently it is not known whether there is a link between the environmental isolates and/or human isolates of A. fumigatus and those responsible for aspergillosis in birds. Microsatellite typing was used to analyse 65 clinical avian isolates and 23 environmental isolates of A. fumigatus. The 78 genotypes that were obtained were compared with a database containing genotypes of 2514 isolates from human clinical samples and from the environment. There appeared to be no specific association between the observed genotypes and the origin of the isolates (environment, human or bird). Eight genotypes obtained from isolates of diseased birds were also found in human clinical samples. These results indicate that avian isolates of A. fumigatus may cause infection in humans.

Whole-genome characterization of a novel polyomavirus detected in fatally diseased canary birds.

Polyomaviruses of birds are aetiological agents of acute inflammatory diseases in non-immunocompromised hosts, which is in contrast to mammalian polyomaviruses. VP4, an additional structural protein encoded by the viral genomes of the known avian polyomaviruses, has been suggested to contribute to pathogenicity through loss of cells following induction of apoptosis. Four distinct bird polyomaviruses have been identified so far, which infect crows, finches, geese and parrots. Using broad-spectrum PCR, a novel polyomavirus, tentatively designated canary polyomavirus (CaPyV), was detected in diseased canary birds (Serinus canaria) that died at an age of about 40 days. Intranuclear inclusion bodies were found in the liver, spleen and kidneys. The entire viral genome was amplified from a tissue sample using rolling-circle amplification. Phylogenetic analysis of the genome sequence indicated a close relationship between CaPyV and other avian polyomaviruses. Remarkably, an ORF encoding VP4 could not be identified in the CaPyV genome. Therefore, the mechanism of pathogenicity of CaPyV may be different from that of the other avian polyomaviruses.

Fatal systemic cladosporiosis in a merino sheep flock.

Systemic cladosporiosis is described in 25 merino sheep from a flock consisting 250 animals. The fungal pneumonia appeared after an intensive antibiotic treatment, because of a respiratory system disorder. The pen of the flock was humid and crowded, and animals had signs of respiratory distress, coughing, fever and anorexia. All of the ill animals died, and necropsy was performed on 10 sheep. The lesions were characterized by a multifocal pyogranulomatous pneumonia and an abomasitis. Severe hemorrhages were observed in the lungs. At the histopathological examination, severe vasculitis with thrombosis was observed in various organs, especially in the lungs and abomasums, suggestive for a hematogenous dissemination of the infection in these organs. Numerous PAS-positive fungal elements were seen in the pyogranulomatous foci. Dark green fungal colonies were seen in the blood agar and Sabouraud dextrose agar that were identified as Cladosporium cladosporioides. This report highlights that phaeohyphomycosis can cause a severe systemic and fatal disease in merino sheep under insufficient management conditions.

Pathology of aging psittacines.

Aging processes leading to specific organ problems are not obvious in aging psittacines. In general, birds live long and age slowly despite their high metabolic rates and very high total lifetime energy expenditures. Most pathologic processes seen in older parrots are generally not specific for aging because they are seen in young birds as well. Pathologic processes that have a tendency to occur more in older psittacines are atherosclerosis and repeated injury processes, such as chronic pulmonary interstitial fibrosis, pneumoconiosis, liver fibrosis, and lens cataracts. Also, some neoplasms are more often seen at an older age.

Characterization of atherosclerosis by histochemical and immunohistochemical methods in African grey parrots (Psittacus erithacus) and Amazon parrots (Amazona spp.).

The aim of the study was to characterize atherosclerotic changes in African grey parrots (Psittacus erithacus) and Amazon parrots (Amazona spp.) by histochemical and immunohistochemical methods. Samples of the aorta ascendens and trunci brachiocephalici from 62 African grey parrots and 35 Amazon parrots were stained by hematoxylin and eosin and Elastica van Gieson for grading of atherosclerosis in these birds. Four different stages were differentiated. The incidence of atherosclerosis in the examined parrots was 91.9% in African grey parrots and 91.4% in Amazon parrots. To evaluate the pathogenesis in birds, immunohistochemical methods were performed to demonstrate lymphocytes, macrophages, smooth muscle cells, and chondroitin sulfate. According to the missing lymphocytes and macrophages and the absence of invasion and proliferation of smooth muscle cells in each atherosclerotic stage, "response-to-injury hypothesis" seems inapplicable in parrots. Additionally, we found alterations of vitally important organs (heart, lungs) significantly correlated with atherosclerosis of the aorta ascendens.

Bacterial and parasitic diseases of passerines.

Many veterinarians are relatively unfamiliar with the passerines. The aviculture, diagnostic procedures, and common diseases, and their treatment have been discussed in several recent publications. Owners of passerines (songbirds) are using veterinary care in increasing numbers as aviculturists recognize the advances in avian medical and surgical treatment of these patients. This article discusses the bacterial and parasitic diseases of passerines.

Mortality due to polyomavirus infection in two nightjars (Caprimulgus europaeus).

Two nightjars (Caprimulgus europaeus) from a bird park in the Netherlands died suddenly, with no clinical signs, within 1 month of each other. The main pathologic findings at necropsy were splenomegaly and hepatic necrosis. On histologic examination, intranuclear viral inclusion bodies consistent with avian polyomavirus were observed in the liver, spleen, and kidneys. Polymerase chain reaction testing of samples from the liver, spleen, and kidneys detected avian polyomaviral DNA, and sequence analysis showed that the virus had a sequence homology of 99% to psittacine avian polyomavirus strains. To our knowledge, this is the first report of avian polyomavirus infection in the order Caprimulgiformes. Lovebirds (Agapornis species), which were housed near the nightjars, were considered as the possible source of infection.

Presence of avian bornavirus RNA and anti-avian bornavirus antibodies in apparently healthy macaws.

Recently a novel avian bornavirus has been described that has been suggested to be the possible etiological agent for proventricular dilatation disease or macaw wasting disease. This article describes two macaws that shed avian bornaviral RNA sequences and demonstrated anti-avian bornavirus antibodies as revealed by reverse transcriptase polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and Western blot, yet are free of outward clinical signs of the disease.

Reovirus infections associated with high mortality in psittaciformes in The Netherlands.

In The Netherlands between January 2002 and December 2004, numerous psittaciformes died showing severe splenomegaly and hepatomegaly with multifocal acute necrosis. At the start of the outbreaks mostly parakeets were affected, but later larger parrots were also involved. Seventy-eight birds showed the same features and six were examined completely, including a virological examination. Tests for polyomavirus, Pacheco's disease (herpesvirus) and circovirus psittacine beak and feather disease (PBFD) viruses and Chlamydophila psittaci were carried out. All results were negative, except for two cases of circovirus infection. Many concurrent bacterial and parasitic infections were seen. Immunohistochemistry revealed reovirus antigen in intralesional mononuclear cells, and reovirus-like particles could be observed by negative contrast electron microscopy. A reovirus was grown and the isolates reacted with polyclonal reovirus antiserum but did not react with monoclonal antibodies against chicken reovirus. The virus was therefore considered a psittacine reovirus. Because reoviruses were seen consistently, they seemed to be the most probable cause of the outbreaks. Climate, the introduction of new birds and the transportation of birds might be other factors involved in the disease seen in The Netherlands. No regional influence could be seen; therefore, we suggested that the virus might be widespread and carriers could be a source of re-introduction.

Zoo and wildlife medical education: a European perspective.

Europe has a long and distinguished history in veterinary science and education, and it was here that the first professional investigations of pathological conditions in zoo animals took place. However, despite an increasing number of veterinarians working with wildlife, education in zoological and wildlife medicine has only recently become part of formal veterinary training at the undergraduate level. Consequently, current educational opportunities in zoological and wildlife medicine vary widely throughout Europe, both in availability and in composition. The need to establish agreed standards in education across Europe and to foster the mobility of students and teaching staff are reflected by international agreements such as the Bologna Treaty and the ERASMUS-SOCRATES program. Europe is also home to a number of voluntary professional organizations, such as the European Wildlife Disease Association and the European Association of Zoo and Wildlife Veterinarians, that actively promote the inclusion of zoological and wildlife medicine in veterinary education. Zoo and wildlife medicine is currently a veterinary specialization in Europe, but educational opportunities are likely to increase in availability in the future.

Hydrometra in a ferret-case report.

A desexed female ferret was presented with bilateral alopecic changes. Clinical examination revealed generalized alopecia and abdominal distension. A polycystic mass was found behind the right kidney, and the whole abdomen was filled with a large turgid mass. Radiography and ultrasonography confirmed the presumptive diagnosis of a hydrometra. Hematology and serum biochemistry showed regenerative anemia with light azotemia. Laparotomy showed the presence of a neoplastic mass at the location of the right ovary, a massive enlargement of the uterus filled with a clear fluid, and a subcapsular cyst on the left kidney. After surgery, histopathologic examination of the tissues diagnosed a leiomyoma of the right ovary with hyperplasia of the uterine wall.

Cowpox virus transmission from rats to monkeys, the Netherlands.

We report an outbreak of cowpox virus among monkeys at a sanctuary for exotic animals. Serologic analysis and polymerase chain reaction were performed on blood and swab samples from different rodent species trapped at the sanctuary during the outbreak. Sequence comparison and serologic results showed that brown rats (Rattus norvegicus) transmitted the virus to monkeys.