Physiologic and biochemical measurements and response to noxious stimulation at various concentrations of MS‐222 in Koi (Cyprinus carpio)

ObjectiveTo evaluate the physiological effect and response to noxious stimulation at five concentrations of MS-222 in koi (Cyprinus carpio).Study designProspective experimental study.AnimalsTwenty-one healthy adult unknown sex koi fish weighing mean 450 ± SD 120 g.MethodsEach fish was exposed to five different concentrations of MS-222 (50, 70, 110, 150 and 190 mg L^?1) in a random sequence during the same anaesthetic event. For each concentration of MS-222, vital functions such as heart rate (HR) (via Doppler) and opercular rate (OpR) were recorded after a standardized induction period. Response to two noxious stimuli in the form of haemostat clamp pressure applied on the tail and the lip was evaluated, and blood was drawn to measure biochemical and blood gas values.ResultsDecrease in response to noxious stimulation with an increase of MS-222 concentration both for the lip (p = 0.0027) and the tail (p < 0.0001) stimulus was observed. Biochemical values were unaffected by the concentration of MS-222 with the exception of lactate concentration which was weakly correlated with the duration of anaesthesia (r = 0.31, p < 0.001) and the number of times the fish was clamped or bled prior to sampling (r = 0.23, p < 0.001). Opercular rate decreased with the increase in anaesthetic concentration, and HR was not affected.Conclusions and clinical relevanceOur results indicated a decrease in response to stimulus and a decrease in OpR that were associated with increased concentrations of MS-222. This may assist in establishing anaesthetic protocols using MS-222 in fish and supports the use of supramaximal pressure stimuli to teleost fish under variable MS-222 concentrations as a model for future studies.     

Cloacal papillomatosis in the absence of herpesvirus and papillomavirus in a sulphur-crested cockatoo (Cacatua galerita)

CASE HISTORY A 21-year-old male sulphur-crested cockatoo (Cacatua galerita) was presented following the sudden appearance of blood associated with the passage of faeces and urates.

CLINICAL AND PATHOLOGICAL FINDINGS: There was fresh blood-staining of the feathers around the vent. The dorsal mucosal wall of the proctodeum was erythematous and roughened in appearance. An endoscopic biopsy was performed, and histological examination revealed multiple fronds of epithelium; the mucosa varied from simple to pseudostratified columnar epithelium, with diffuse hyperplasia of goblet cells. The underlying connective tissue stroma was well vascularised and was infiltrated with mixed inflammatory cells, comprising granulocytic cells and macrophages. PCR testing for both herpesvirus and papillomavirus, using consensus primers, was negative.

DIAGNOSIS: Cloacal papillomatosis.

CLINICAL RELEVANCE: This case manifested typical clinical signs and histological lesions of cloacal papillomatosis in the absence of demonstrable herpesvirus or papillomavirus. Veterinarians need to consider this disease in the differential diagnosis of blood in the droppings of parrots and cockatoos even in countries where psittacine herpesviruses are exotic diseases.     

Detection of bovine herpesvirus type 4 antibodies and bovine lymphotropic herpesvirus in New Zealand dairy cows

AIM: To detect the presence of bovine herpesvirus (BoHV) type 4 in New Zealand dairy cows with clinical metritis.

METHODS: Serum samples taken from 92 dairy cows with clinical metritis, each from a different farm, were tested for the presence of antibodies against BoHV-4 using a commercially available, indirect ELISA. Peripheral blood mononuclear cells (PBMC) were collected from 10 BoHV-4 seropositive cows, and PBMC were examined by a pan-herpesvirus nested PCR to detect herpesvirus. PCR products were sequenced directly and a proportion of the PCR products were cloned and sequenced to identify the virus present.

RESULTS: Antibodies to BoHV-4 were detected in 23/92 (25%) serum samples. The pan-herpesvirus PCR was positive in 8/10 PBMC samples. Cloning and sequencing identified that all of the eight PCR-positive PBMC contained bovine lymphotropic herpesvirus (BLHV); no BoHV-4 DNA was detected.

CONCLUSIONS: This study reports the finding of the presence of apparent antibodies to BoHV-4, and BLHV DNA in New Zealand dairy cows affected by metritis.

CLINICAL RELEVANCE: Bovine herpesvirus type 4 and BLHV are reported to have the potential to cause reproduction failure in cows. This is the first report of apparent BoHV-4 antibodies, and BLHV in New Zealand. The importance and epidemiology of these viruses in cattle in New Zealand requires further investigation.     

牛疱疹病毒I型gB基因的原核表达

根据GenBank中BHV-1 Colorado 1毒株的全基因组序列,针对gB基因的主要B细胞免疫区域设计一对引物。提取BHV-1 Colorado 1毒株的基因组DNA,PCR扩增大小为585 bp的gB片段,将其克隆至pET-22b（+）原核表达载体并测序分析,同时用Nde I和Not I酶切鉴定。SDS-PAGE和Western blot试验结果证明,获得的可溶性gB重组蛋白能与牛传染性鼻气管炎病毒标准阳性WEI血清发生特异性反应,为ELISA等免疫诊断方法的建立奠定了物质基础。     

A novel herpesvirus associated with chronic superficial keratitis and proliferative conjunctivitis in a great horned owl (Bubo virginianus)

An adult great‐horned owl (Bubo virginianus; GHOW) presented with a history of recurrent corneal ulceration of the right eye (OD). Findings included ulcerative superficial keratitis, proliferative conjunctivitis, and iris pigmentary changes. The ulcer was initially nonresponsive to medical therapy, but showed rapid and appropriate healing following diamond burr debridement. Proliferative conjunctivitis markedly improved following topical antiviral therapy with cidofovir 1%, interferon alpha 2B ophthalmic solutions, and oral l ‐lysine. Histopathologic evaluation of a conjunctival biopsy revealed epithelial features suspicious for viral cytopathic changes and intranuclear structures suspicious for viral inclusions, suggestive of a possible viral‐induced papillomatous conjunctivitis. A novel alphaherpesvirus, referred to as Strigid Herpesvirus 1 (StrHV1), was identified using PCR and gene sequencing. This case represents a new clinical manifestation of a previously unreported herpesvirus in the GHOW. Identification of the herpes virus was critical to administration of appropriate therapy and resolution of the conjunctivitis, and corneal epithelial debridement promoted resolution of the chronic corneal epithelial defect.     

Ultrastructural analysis of sequential cyprinid herpesvirus 3 morphogenesis in vitro

Cyprinid herpesvirus 3 (CyHV‐3) is an alloherpesvirus, and it is the aetiological agent of koi herpesvirus disease. Although the complex morphogenic stages of the replication cycle of CyHV‐3 were shown to resemble that of other members of the Herpesvirales, detailed analysis of the sequence and timing of these events was not definitively determined. This study describes these features through a time course using cyprinid cell cultures (KF‐1 and CCB) infected with CyHV‐3 (KHV isolate, H361) and analysed by transmission electron microscopy. Rapid viral entry was noted, with high levels of intracellular virus within 1–4 h post‐infection (hpi). Intranuclear capsid assembly, paracrystalline array formation and primary envelopment of capsids occurred within 4 hpi. Between 1 and 3 days post‐infection (dpi), intracytoplasmic secondary envelopment occurred, as well as budding of infectious virions at the plasma membrane. At 5–7 dpi, the cytoplasm contained cytopathic vacuoles, enveloped virions within vesicles, and abundant non‐enveloped capsids; also there was frequent nuclear deformation. Several morphological features are suggestive of inefficient viral assembly, with production of non‐infectious particles, particularly in KF‐1 cells. The timing of this alloherpesvirus morphogenesis is similar to other members of the Herpesvirales, but there may be possible implications of using different cell lines for CyHV‐3 propagation.     

Cyprinid herpesvirus 3 as a potential biological control agent for carp (Cyprinus carpio) in Australia: susceptibility of non‐target species

Carp (Cyprinus carpio L.) is a pest species in Australian waterways, and cyprinid herpesvirus 3 (CyHV‐3) is being considered as a potential biological control (biocontrol) agent. An important consideration for any such agent is its target specificity. In this study, the susceptibility to CyHV‐3 of a range of non‐target species (NTS) was tested. The NTS were as follows: 13 native Australian, and one introduced, fish species; a lamprey species; a crustacean; two native amphibian species (tadpole and mature stages); two native reptilian species; chickens; and laboratory mice. Animals were exposed to 100–1000 times the approximate minimum amount of CyHV‐3 required to cause disease in carp by intraperitoneal and/or bath challenge, and then examined clinically each day over the course of 28 days post‐challenge. There were no clinical signs, mortalities or histological evidence consistent with a viral infection in a wide taxonomic range of NTS. Furthermore, there was no molecular evidence of infection with CyHV‐3, and, in particular, all RT‐PCRs for viral mRNA were negative. As a consequence, the results encourage further investigation of CyHV‐3 as a potential biocontrol agent that is specific for carp.