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Lactococcus garvieae is the causative agent of lactococcosis in fish and an emerging zoonotic pathogen with high levels of antimicrobial resistance. We report a case of L. garvieae- associated septicemia in a central bearded dragon ( Pogona vitticeps ) confirmed via whole-blood PCR and direct sequencing. Following a 30-d course of ceftazidime (20 mg/kg IM q72h), the animal's clinical condition had not resolved; leukopenia persisted, with heterophil toxic change. Coelomic ultrasound findings were consistent with preovulatory follicular stasis, folliculitis, and coelomitis. Following surgical ovariectomy and an additional 30-d course of ceftazidime, the animal's behavior and appetite returned to normal, the animal tested negative via whole-blood PCR assay, and the CBC was unremarkable. To our knowledge, L. garvieae with L. garvieae- associated clinical disease has not been reported previously in a bearded dragon. We conclude that L. garvieae should be considered as a possible etiologic agent in cases of septicemia in bearded dragons, with the potential for zoonotic transmission warranting further investigation., Competing Interests: Declaration of conflicting interestsThe authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Although many studies have characterized catarrhine and platyrrhine primate herpesviruses, little is known about herpesviruses in prosimians. We aimed to identify and characterize herpesviruses in prosimians with proliferative lymphocytic disease. DNA was extracted from tissues of 9 gray mouse lemurs ( Microcebus murinus ) and 3 pygmy slow lorises ( Nycticebus pygmaeus ) with lymphoproliferative lesions, and we performed nested PCR and sequencing for detection of herpesviruses and polyomaviruses. We identified 3 novel herpesviruses and performed phylogenetic analyses to characterize their relationship with other herpesviruses. A gray mouse lemur herpesvirus clustered with other primate herpesviruses within the subfamily Betaherpesvirinae , just basal to the genus Cytomegalovirus . The other gray mouse lemur herpesvirus and the pygmy slow loris herpesvirus clustered within the subfamily Gammaherpesvirinae , although the relationships within the subfamily were less resolved. Quantitative PCR assays were developed for the 2 new gray mouse lemur viruses, providing specific, faster, less expensive, and quantitative detection tools. Further studies are needed to elucidate the relationship between the presence of these viruses and the severity or presence of lymphoproliferative lesions in prosimians.

Herpesviruses can be significant reptile pathogens. Herpesviral infection in a wild-caught, male spider tortoise ( Pyxis arachnoides ) under human care was detected during a routine wellness examination prior to transition between zoologic organizations. The tortoise had no clinical signs of illness. Oral swabs obtained during a physical examination as part of pre-shipment risk mitigation for infectious disease were submitted for consensus herpesvirus PCR assay and sequencing. Based on comparative sequence analysis, the novel herpesvirus identified is a member of the subfamily Alphaherpesvirinae . Studies of herpesviral phylogeny in chelonian species support branching patterns of turtle herpesviruses that closely mirror those of their hosts. The symmetry of these patterns is suggestive of close codivergence of turtle herpesviruses with their host species. The distribution of these viruses in both tortoises and emydids suggests a phylogenetic duplication event in the herpesviruses after host divergence of the Pleurodira and basal to the divergence of Americhelydia . Herpesviral infections have been documented to cause higher morbidity when introduced to aberrant host species, and significant consideration must be given to the presence of herpesviruses in the management of tortoise collections, particularly collections that include various species of testudines.

A deceased 9-wk-old male gray fox ( Urocyon cinereoargenteus ) with a history of decreased ambulation and diarrhea was submitted to the Texas A&M Veterinary Medical Diagnostic Laboratory. No significant gross findings were evident on postmortem examination. Histologically, the cerebrum and brainstem had mild necrotizing meningoencephalitis with protozoal schizonts and merozoites. Additionally, glial cells contained intracytoplasmic and intranuclear viral inclusion bodies. Sections of the cerebrum were positive for canine distemper virus (CDV) and negative for Sarcocystis neurona on immunohistochemistry. Bayesian analysis revealed that this Sarcocystis sp. clustered most closely with a clade of unnamed Sarcocystis sp. found in viperid snakes, with a posterior probability of 99%. CDV likely played a significant role in the expression of clinical sarcocystosis in this gray fox.

Four Indian ringneck parakeets ( Psittacula krameri ; syn. ringneck parrots or rose-ringed parakeets) were submitted by 2 private owners for autopsy following a history of dyspnea and death. Gross findings were varied and included thickening of the left caudal thoracic air sac, white spots throughout the liver, mild dilation of the proventriculus, coelomic effusion, splenomegaly, and pulmonary congestion and edema. Microscopically, the submitted parakeets had significant lesions in the lower respiratory tract, including necrotizing bronchitis, parabronchitis, and interstitial pneumonia with numerous syncytia containing eosinophilic intranuclear inclusions. Electron microscopy of the lungs was compatible with a herpesviral infection and Psittacid alphaherpesvirus 5 (PsAHV5) was detected via PCR and sequencing. There has been inconsistent terminology used with Psittacid alphaherpesvirus 3 and PsAHV5; we attempt here to clarify the reported history of these viruses.

Herpesviruses are found in free-living and captive chelonian populations, often in association with morbidity and mortality. To date, all known chelonian herpesviruses fall within the subfamily Alphaherpesvirinae . We detected a novel herpesvirus in 3 species of chelonians: a captive leopard tortoise ( Stigmochelys pardalis ) in western TX, USA; a steppe tortoise ( Testudo [ Agrionemys ] horsfieldii ) found near Fort Irwin, CA, USA; and 2 free-living, three-toed box turtles ( Terrapene mexicana triunguis ) found in Forest Park, St. Louis, MO. The leopard tortoise was coinfected with the tortoise intranuclear coccidian and had clinical signs of upper respiratory tract disease. The steppe tortoise had mucopurulent nasal discharge and lethargy. One of the three-toed box turtles had no clinical signs; the other was found dead with signs of trauma after being observed with blepharedema, tympanic membrane swelling, cervical edema, and other clinical signs several weeks prior to death. Generally, the branching order of the turtle herpesviruses mirrors the divergence patterns of their hosts, consistent with codivergence. Based on phylogenetic analysis, this novel herpesvirus clusters with a clade of viruses that infect emydid hosts and is likely of box turtle origin. Therefore, we suggest the name terrapene alphaherpesvirus 3 (TerAHV3) for the novel virus. This virus also has the ability to host-jump to tortoises, and previously documented herpesviral morbidity tends to be more common in aberrant hosts. The relationship between clinical signs and infection with TerAHV3 in these animals is unclear, and further investigation is merited.

We identified a novel herpesvirus in a captive juvenile white stork ( Ciconia ciconia ) that experienced progressive weight loss followed by death. Histologic findings included severe splenic and hepatic necrosis, and intranuclear inclusion bodies in hepatocytes and unidentified splenic cells. The virus was amplified, sequenced, and subsequently accessioned as Ciconiid alphaherpesvirus 1. Phylogenetic analysis was performed and revealed that this virus is more closely related to mammalian herpesviruses than those within the genus Mardivirus . Additional sequence of viruses in this area may elucidate the ancestral virus that jumped from reptilian to mammalian hosts.

Avian coxiellosis is an emerging cause of morbidity and mortality among captive psittacines, and the utility of a rapid detection test using easily obtained samples is paramount in a clinical setting. New sequences were obtained from 3 genes: groEL, dnaK , and rpoB . We developed probe-hybridization quantitative PCR (qPCR) assays using groEL and dnaK genes. Samples, including splenic aspirates, liver aspirates, whole blood, and choanal, conjunctival, and cloacal swabs, were collected from 4 psittacine species including 3 blue-and-gold macaws ( Ara ararauna ), 2 scarlet-chested parrots ( Neophema splendida ), 1 Timneh African grey parrot ( Psittacus timneh ), and 1 yellow-naped Amazon parrot ( Amazona auropalliata ). Retrospective review of postmortem findings from 3 of these psittacines included splenomegaly, hepatitis, and/or transmission electron microscopy confirmation consistent with previous reports of avian coxiellosis. There was 100% agreement between these assays and consensus PCR with sequencing. A Wilcoxon rank-sum test found a strong correlation between groEL and dnaK cycle threshold values ( p < 0.001), validating these assays for detection of this avian Coxiella sp.

Two male juvenile central bearded dragons ( Pogona vitticeps) were submitted for postmortem examination after dying at their respective homes. Dragon 1 had marked hemopericardium with restrictive epicarditis. The inner aspect of the distended pericardial sac was lined by a fibrinoheterophilic membrane. In addition, granulomas abutted the testes. Dragon 2 had acute hemopericardium and granulomatous arteritis of the great vessels exiting the heart. Histologically, both animals had granulomatous arteritis of the large arteries with intrahistiocytic gram-positive, slightly elongated, up to 2 μm long microorganisms that contained a vacuole. These microorganisms were also present in the paratesticular granulomas. On transmission electron microscopy, the microorganisms were identified as microsporidians given the presence of exospore, endospore, vacuole, nucleus, and a filament with 4-6 coils. The microsporidia were identified as Encephalitozoon pogonae based on sequencing of the internal transcribed spacer 1 of the ribosomal RNA genes. Microsporidia are agents of disease in bearded dragons. Intrapericardial arteritis of large arteries with hemopericardium or restrictive epicarditis is a fatal manifestation of this infection.

Astroviruses are viral pathogens that have been associated with enteric and neurologic disease in a variety of species. The domestic cat is a prominent host, with reports of astroviral infection being both highly prevalent and widely distributed in the feline population. Despite the potential for inducing significant disease, especially within shelter environments, there is currently only one reliable method of detection: standard reverse-transcription PCR using pan-astrovirus degenerate primers (consensus RT-PCR) with product sequencing. Unfortunately, this process is relatively slow and costly. Quantitative real-time PCR (qPCR) represents an efficient, economical alternative, with the added benefit of viral load quantification. We developed a RT-qPCR assay using probe hybridization technique to detect conserved regions of mamastrovirus 2 extracted from fecal samples of domestic cats. Known positive and negative samples were tested, and results were compared with gold standard consensus RT-PCR and sequencing. A standard curve was employed to determine limits of detection. In order to assess analytic specificity, we tested several additional samples that had been collected from non-felid species and were known to contain non-target astroviruses. Discrepant results between consensus RT-PCR and RT-qPCR testing were further analyzed with a validation RT-PCR assay, using mamastrovirus 2-specific primers. Our probe hybridization RT-qPCR assay is reliable and effective for the detection of mamastrovirus 2. This assay will allow rapid, affordable detection and facilitate further research on astroviral infection within domestic cats.

Astroviruses are small, nonenveloped RNA viruses that have been linked to numerous diseases in a variety of species, including enteric disease in humans and cheetahs. Species Mamastrovirus 2, previously known as feline astrovirus, has been isolated from the feces of domestic cats and cheetahs. A total of 122 cat fecal samples from Alachua County, FL Animal Services and the Veterinary Community Outreach Program at the University of Florida were analyzed, and 35 contained astroviral RNA that was amplified and identified using consensus RT-PCR and sequence analysis. Using phylogenetic analysis, 19 of the astroviral sequences were identified as Mamastrovirus 2, making it the most prevalent astrovirus in this population. Three samples were identified as an astrovirus similar to viruses previously identified in foxes in The Netherlands and a cat in California, and one was similar to a bat astrovirus. One astroviral sequence was identified as an Avastrovirus. Although a causative relationship between mamastroviruses and enteric disease in cats has yet to be established, it is clear that mamastroviruses are prevalent, and an understanding of prevalence of astroviral types may help direct future test development.

Trichechid herpesvirus 1 (TrHV-1) is currently the only known herpesvirus in any sirenian. We hypothesized that stress may lead to recrudescence of TrHV-1 in manatees, thus making TrHV-1 a potential biomarker of stress. We optimized and validated a TrHV-1 real-time quantitative probe hybridization PCR (qPCR) assay that was used to quantify TrHV-1 in manatee peripheral blood mononuclear cells (PBMCs). Average baseline TrHV-1 loads in a clinically healthy wild Florida manatee ( Trichechus manatus latirostris) population ( n = 42) were 40.9 ± SD 21.2 copies/100 ng DNA; 19 of 42 manatees were positive. TrHV-1 loads were significantly different between the 2 field seasons ( p < 0.025). This optimized and validated qPCR assay may be used as a tool for further research into TrHV-1 in Florida manatees.

California sea lion adenovirus 1 (CSLAdV-1) has been associated with hepatitis and enteritis in several wild and captive populations of diverse pinniped species. Currently available tests have been limited to pan-adenoviral polymerase chain reaction (PCR) followed by sequencing. We present the development of a quantitative probe-hybridization PCR (qPCR) assay for rapid, sensitive, and specific detection of this virus in California sea lions ( Zalophus californianus) and other pinnipeds. This assay did not amplify other mammalian adenoviruses and is able to detect consistently down to 10 viral copies per well. Compared with the gold standard conventional pan-adenovirus PCR/sequencing assay, diagnostic sensitivity and specificity of 100% and 88.2% were found, respectively. The lower diagnostic specificity of this qPCR assay may be the result of the lower limit of detection of this assay compared with the gold standard rather than the result of detection of true false-positives.

Mycobacteriosis is infrequently reported in free-ranging sea turtles. Nontuberculous Mycobacterium haemophilum was identified as the causative agent of disseminated mycobacteriosis in a juvenile leatherback turtle (Dermochelys coriacea) that was found stranded on the Atlantic coast of Florida. Disseminated granulomatous inflammation was identified histologically, most notably affecting the nervous system. Identification of mycobacterial infection was based on cytologic, molecular, histologic, and microbiologic methods. Among stranded sea turtles received for diagnostic evaluation from the Atlantic and Gulf of Mexico coasts of the United States between 2004 and 2015, the diagnosis of mycobacteriosis was overrepresented in stranded oceanic-phase juveniles compared with larger size classes, which suggests potential differences in susceptibility or exposure among different life phases in this region. We describe M. haemophilum in a sea turtle, which contributes to the knowledge of diseases of small juvenile sea turtles, an especially cryptic life phase of the leatherback turtle., (© 2016 The Author(s).)

A recently deceased juvenile male bottlenose dolphin (Tursiops truncatus) was found floating in the Gulf of Mexico, off Sand Key in Clearwater, Florida. At autopsy, we identified pneumonia and a focus of malacia in the right cerebrum. Cytologic evaluation of tissue imprints from the right cerebrum revealed fungal hyphae. Fungal cultures of the lung and brain yielded Aspergillus fumigatus, which was confirmed by amplification of a portion of the fungal nuclear ribosomal internal transcribed spacer 2 region sequence. Microscopic pulmonary lesions of bronchiolar epithelial cell syncytia with intracytoplasmic and intranuclear inclusions within bronchiolar epithelial cells were suggestive of Cetacean morbillivirus (CeMV) infection. The occurrence of CeMV infection was supported by positive immunohistochemical staining for morbillivirus antigen. CeMV detection was confirmed by amplification and sequencing a portion of the morbilliviral RNA-dependent RNA polymerase gene from lung tissue. This case provides CeMV sequence data available from the Gulf of Mexico and underscores the need for genomic sequencing across diverse host, temporospatial, and population (i.e., single animal vs. mass mortality events) scales to improve our understanding of these globally emerging pathogens., (© 2016 The Author(s).)

A novel herpesvirus was detected in a 17-day-old Chilean flamingo (Phoenicopterus chilensis) with pneumonia, hepatopathy, and severe anemia that was housed in California. Postmortem examination identified a pale, enlarged liver, mildly increased fluid in the lungs, and red foci in the spleen. Histologic examination revealed marked hepatic necrosis with syncytia, splenic necrosis, and interstitial pneumonia with eosinophilic intranuclear inclusions within hepatocytes and in unidentified cells of the lung. Transmission electron microscopy identified virions consistent with a herpesvirus in the nucleus and cytoplasm of degenerative hepatocytes. Nested consensus PCR, sequencing, and phylogenetic analysis identified a novel herpesvirus within the genus Iltovirus in the subfamily Alphaherpesvirinae., (© 2016 The Author(s).)

Herpesviruses are important pathogens of chelonians, and include Chelonid herpesvirus 5, which is associated with fibropapillomatosis in sea turtles. Spirorchid trematodes are blood flukes that reside within the cardiovascular system of marine turtles and may be associated with severe disease. An eastern box turtle (Terrapene carolina) at the South Florida Wildlife Care Center (Fort Lauderdale, Florida) was presented to the facility with papillomatous growths behind both rear legs. Surgical removal resulted in remission for 8 months; however, lesions recurred, prompting a second surgery and acyclovir therapy. Surgical biopsies revealed subacute superficial inflammation associated with the supporting stroma of the cutaneous papillomas and granulomas within the superficial dermis containing fragmented and collapsed brown trematode eggs surrounded by multinucleated giant cells and epithelioid macrophages. Pan-herpesviral and pan-trematode consensus polymerase chain reaction and sequencing were run on tissue samples. Comparative sequence analysis revealed a novel alphaherpesvirus and a novel trematode in the genus Spirorchis. The animal became anorexic and was euthanized due to poor quality of life. While we do not yet have a complete understanding of the effects of herpesvirus and trematode infections in eastern box turtles, the findings thus presented provide initial insights into the disease relationships among these chelonians., (© 2015 The Author(s).)

Agamid adenovirus 1 (AgAdv-1) is a significant cause of disease in bearded dragons (Pogona sp.). Clinical manifestations of AgAdv-1 infection are variable and often nonspecific; the manifestations range from lethargy, weight loss, and inappetence, to severe enteritis, hepatitis, and sudden death. Currently, diagnosis of AgAdv-1 infection is achieved through a single published method: standard nested polymerase chain reaction (nPCR) and sequencing. Standard nPCR with sequencing provides reliable sensitivity, specificity, and validation of PCR products. However, this process is comparatively expensive, laborious, and slow. Probe hybridization, as used in a TaqMan assay, represents the best option for validating PCR products aside from the time-consuming process of sequencing. This study developed a real-time PCR (qPCR) assay using a TaqMan probe-based assay, targeting a highly conserved region of the AgAdv-1 genome. Standard curves were generated, detection results were compared with the gold standard conventional PCR and sequencing assay, and limits of detection were determined. Additionally, the qPCR assay was run on samples known to be positive for AgAdv-1 and samples known to be positive for other adenoviruses. Based on the results of these evaluations, this assay allows for a less expensive, rapid, quantitative detection of AgAdv-1 in bearded dragons., (© 2015 The Author(s).)

Three adult central bearded dragons (Pogona vitticeps) originating from a commercial breeding facility presented with clinical signs, including anorexia, dehydration, white multifocal lesions on the dorsal aspect of the tongue, blepharospasm, and weight loss. In 1 of 3 lizards, a marked regenerative anemia was noted, and all 3 bearded dragons had erythrocytic intracytoplasmic inclusion bodies. Nine bearded dragons housed in contact also had identical, but fewer intraerythrocytic inclusions. Inclusion bodies examined by electron microscopy had particles consistent with iridoviruses. Attempts to culture the virus were unsuccessful; however, amplification and sequencing of regions of the viral DNA polymerase by polymerase chain reaction confirmed the presence of an iridovirus. One of the bearded dragons died, while the 2 others showing clinical signs were euthanized. The remaining 9 infected bearded dragons of the teaching colony were also euthanized. Postmortem examination revealed a moderate, multifocal, lymphoplasmacytic or mononuclear adenitis of the tongue in the 3 bearded dragons, and a lymphohistiocytic hepatitis with bacterial granulomas in 2 lizards., (© 2014 The Author(s).)

A total of 5 psittacine birds in an enclosed zoological exhibit, including 2 princess parrots and 3 cockatoos of 2 different species, developed severe central nervous system clinical signs over a 2-3-month period and died or were euthanized. Histologically, all birds had a lymphoplasmacytic and histiocytic encephalitis with intralesional protozoa consistent with a Sarcocystis species in addition to intramuscular tissue sarcocysts. By immunohistochemical staining, merozoites in brain and tissue cysts in muscle did not react with polyclonal antisera against Sarcocystis falcatula, Sarcocystis neurona, Toxoplasma gondii, and Neospora caninum, or with a monoclonal antibody to S. neurona. Transmission electron microscopy on sarcocyst tissue cyst walls from 2 birds was morphologically consistent with Sarcocystis calchasi. Polymerase chain reaction (PCR) amplification and sequencing of partial 18S ribosomal RNA from muscle tissue cysts and brain schizonts from 3 birds was consistent with a clade containing S. calchasi and Sarcocystis columbae but could not distinguish these closely related Sarcocystis species. However, PCR amplification and sequencing of the internal transcribed spacer 1 RNA segment in the brain from 2 birds and muscle from 2 birds specifically identified the isolates as S. calchasi. The current report documents that multiple psittacine species are susceptible intermediate hosts of S. calchasi, and that infection can cause encephalitis resulting in significant morbidity and mortality in psittacine aviaries.

A novel gammaherpesvirus was identified in a large flying fox (Pteropus vampyrus) with conjunctivitis, blepharitis, and meibomianitis by nested polymerase chain reaction and sequencing. Polymerase chain reaction amplification and sequencing of 472 base pairs of the DNA-dependent DNA polymerase gene were used to identify a novel herpesvirus. Bayesian and maximum likelihood phylogenetic analyses indicated that the virus is a member of the genus Percavirus in the subfamily Gammaherpesvirinae. Additional research is needed regarding the association of this virus with conjunctivitis and other ocular pathology. This virus may be useful as a biomarker of stress and may be a useful model of virus recrudescence in Pteropus spp.

A novel herpesvirus was detected in sun bears (Helarctos malayanus) with oral squamous cell carcinoma. Five captive sun bears from 4 institutions in the United States presented with oral lesions ranging from erythema and mild erosions to nodular, ulcerated masses. All 5 were diagnosed with squamous cell carcinoma. The tumors were treated with surgical resection but recurrence, local extension, or appearance of new lesions was noted in all cases. Intralesional chemotherapy was administered in 2 cases, and the nonsteroidal anti-inflammatory drug piroxicam was administered in 3 cases. Virus was detected in 4 of the 5 bears' tissue samples using a consensus herpesvirus polymerase chain reaction. Nucleotide sequencing and phylogenetic analysis showed that this herpesvirus is in the subfamily Gammaherpesvirinae and distinct from other known herpesviruses. The association between the herpesvirus and squamous cell carcinoma is unknown. The current study presents a novel gammaherpesvirus within the order Ursidae, with the name Ursid herpesvirus 1 proposed.

A novel adenovirus was identified in a cotton-top tamarin (Saguinus oedipus) with diarrhea by negative-staining electron microscopy of feces, consensus polymerase chain reaction, and sequencing. Partial sequences were obtained from the DNA-dependent DNA polymerase, the p52k gene, and the hexon. Bayesian and maximum likelihood phylogenetic analyses indicated that the virus is a member of the genus Mastadenovirus, and is herein termed Saguinus siadenovirus 1. The phylogeny of the mastadenoviruses is similar to that of their hosts, supporting coevolution. Support for this was strongest in the analysis of the predicted hexon protein. The obtained sequences were GC-rich, which may suggest a lack of recent host jumps. The diversity and evolution of the adenoviruses of platyrrhine primates merits further investigation. Additional study of the association of this virus with diarrhea is indicated.

Bacteria of the genus Helicobacter are associated with disease in humans and animals. Reports of infection in reptiles are very limited. In the present study, pathological findings and molecular characterization are reported for a Helicobacter species associated with septicemia in a pancake tortoise (Malacochersus tornieri). Disseminated infection resulted in regional cellulitis and edema of the head and neck, and pericarditis. Spiral bacteria were identified in cytological preparations and Warthin-Starry-stained sections of pathological lesions. Amplification of partial sequence of the 16S ribosomal gene using polymerase chain reaction identified the organism as Helicobacter and suggest that it is a novel species.

An adult female California sea lion (Zalophus californianus) that stranded in central California was found to have a small glossal polypoid mass on gross necropsy. Histologically, the mass was consistent with a fibropapilloma, and intranuclear inclusions were found within endothelial cells lining small arterioles within the mass. Electron microscopy revealed 40-nm virions within endothelial intranuclear inclusions. Rolling circle amplification was used to obtain a partial viral genomic sequence. Sequence analysis identified the virus as a novel polyomavirus, tentatively named California sea lion polyomavirus 1. In addition, the sea lion had a severely thickened small intestine and swollen pale kidneys on gross examination. Severe renal amyloidosis with chronic interstitial nephritis was diagnosed histologically as well as T-cell intestinal lymphoma, which was confirmed via immunophenotyping and molecular clonality. The relationship, if any, between polyomavirus infection and the other disease processes in this sea lion is not known, but it is considered unlikely that the polyomavirus induced the lymphoma.

Reoviruses are nonenveloped, segmented, double-stranded RNA viruses capable of infecting a wide range of invertebrate, vertebrate, fungus, and plant hosts. Though sporadic infection has been reported in a variety of reptilian species, infection of rough green snakes (Opheodrys aestivus) has not been previously described. Five wild-caught, adult rough green snakes were obtained by a zoological institution. Clinical deterioration was first noted in all snakes after 3 weeks in quarantine. Despite treatment, clinical decline progressed, and all 5 snakes died or were euthanized by 48 days post-arrival. Moderate, multifocal, acute, necrotizing hepatitis with hepatocellular syncytia was diagnosed in 1 snake. Two additional snakes had severe, diffuse, subacute to chronic pancreatitis. All 5 snakes had gastroenteric cryptosporidiosis. Electron microscopic examination of liver from the snake with hepatic lesions revealed scattered hepatocytes containing 1 or more intranuclear clusters of approximately 90 nm in diameter viral particles arranged in loose arrays. Polymerase chain reaction (PCR) amplification of a segment of the reovirus RNA-dependent RNA polymerase gene was performed on RNA extracted from tissues of all 5 snakes. PCR amplification of samples extracted from the snake with hepatic lesions resulted in a 109-base pair (bp) product. Phylogenetic analyses indicated the virus was a novel strain distinct from other reoviruses at a level consistent with species difference. The source of infection was unknown. PCR amplification of samples extracted from the other 4 snakes was negative.

A novel siadenovirus was identified in the Sulawesi tortoise (Indotestudo forsteni). A group of 105 Sulawesi tortoises was obtained by the Turtle Survival Alliance. Many of the tortoises were in poor health. Clinical signs included anorexia, lethargy, mucosal ulcerations and palatine erosions of the oral cavity, nasal and ocular discharge, and diarrhea. Initial diagnostic tests included fecal testing for parasites, complete blood count and plasma biochemical analysis, mycoplasma serology, and polymerase chain reaction (PCR) testing for intranuclear coccidia and chelonian herpesvirus. Treatment included administration of antibiotics, antiparasitic medications, parenteral fluids, and nutritional support. Tissue samples from animals that died were submitted for histopathologic evaluation. Histopathologic examination revealed systemic inflammation and necrosis associated with intranuclear inclusions consistent with a systemic viral infection in 35 tortoises out of 50 examined. Fecal testing results and histopathologic findings revealed intestinal and hepatic amoebiasis and nematodiasis in 31 animals. Two of 5 tortoises tested by PCR were positive for Chlamydophila sp. Aeromonas hydrophila and Escherichia coli were cultured from multiple organs of 2 animals. The mycoplasma serology and PCR results for intranuclear coccidia and chelonian herpesvirus were negative. Polymerase chain reaction testing of tissues, plasma, and choanal/cloacal samples from 41 out of 42 tortoises tested were positive for an adenovirus, which was characterized by sequence analysis and molecular phylogenetic inference as a novel adenovirus of the genus Siadenovirus. The present report details the clinical and anatomic pathologic findings associated with systemic infection of Sulawesi tortoises by this novel Siadenovirus, which extends the known reptilian adenoviruses to the chelonians and extends the known genera of reptilian Adenoviridae beyond Atadenovirus to include the genus Siadenovirus.

Intranuclear coccidia and Mycoplasma spp. were identified from the nasal cavity of 5 Sulawesi tortoises (Indotestudo forsteni) affected by chronic rhinosinusitis and oronasal fistulae. This study provides the first antemortem diagnosis of intranuclear coccidiosis in tortoises and the first cytomorphologic descriptions of this disease. Histopathologic and ultrastructural morphology of the intranuclear coccidia were identical to those previously described in tortoises. Nucleic acid sequence data of a 1715 base-pair fragment of the 18S small subunit rRNA gene identify this coccidian as a novel species.

Multifocal hyperemic nodules and plaques associated with the cloacal mucosa of juvenile alligators (Alligator mississippiensis) at a public aquarium were investigated. Grossly, pale pink to dark red multifocal, circular lesions of varying degrees of severity were identified on the cloacal and, in males, phallus mucosa. Cloacal mucosa biopsies were obtained from 2 of the alligators. These samples were examined histologically and by polymerase chain reaction (PCR) using consensus primers targeting a conserved region of the herpesvirus polymerase gene. Microscopically, the lesions were characterized as submucosal lymphoid follicles with hyperemia and hemorrhage. No inclusion bodies were observed. Minimal to no anisokaryosis was present, and no etiologic agents were identified. Through PCR, a band consistent in size with herpesvirus was observed. Tissues showing similar clinical, histopathologic, and PCR findings were collected from animals at an alligator farm several months later. Sequencing of the PCR amplicon resulted in a 180-base pair sequence that shared 85% sequence identity with tortoise herpesvirus-1.

This report documents a case of spontaneous, fatal, and likely recrudescent human herpesvirus type 1 (HHV-1) infection in a captive white-handed gibbon (Hylobates lar) confirmed by polymerase chain reaction (PCR). An approximately 44-year-old, captive, female, white-handed gibbon with a history of recurrent conjunctivitis and occasional seizures became acutely weak, disoriented, and ataxic. A postictal state was suspected by caretakers and veterinary staff, and euthanasia was ultimately elected because of lack of clinical improvement with supportive care. No significant abnormalities were detected at necropsy. Histologically, sections of cerebrum and midbrain contained minimal to mild, multifocal lymphoplasmacytic meningoencephalitis with numerous intranuclear viral inclusions within astrocytes and some neurons. The presumptive diagnosis of HHV-1-induced encephalitis was strengthened by nested PCR amplification of a segment of the herpesvirus DNA polymerase gene. Sequences from this region have been found to be unique to each herpesvirus species, thus identifying HHV-1 as the likely etiologic agent in this case. Positive HHV-1 serology from several years before the terminal episode suggested that the disease was most likely due to recrudescence of latent HHV-1 infection.