An unprecedented cluster of Australian bat lyssavirus in Pteropus conspicillatus indicates pre‐flight flying fox pups are at risk of mass infection

In November 2017, two groups of P. conspicillatus pups from separate locations in Far North Queensland presented with neurological signs consistent with Australian bat lyssavirus (ABLV) infection. These pups (n = 11) died over an 11‐day period and were submitted to a government laboratory for testing where ABLV infection was confirmed. Over the next several weeks, additional ABLV cases in flying foxes in Queensland were also detected. Brain tissue from ABLV‐infected flying foxes during this period, as well as archived brain tissue, was selected for next‐generation sequencing. Phylogenetic analysis suggests that the two groups of pups were each infected from single sources. They were likely exposed while in crèche at night as their dams foraged. This study identifies crèche‐age pups at a potentially heightened risk for mass ABLV infection.     

Radiographic evaluation of cardiac size in flying fox species (Pteropus rodricensis, P. hypomelanus, and P. vampyrus).

Dilated cardiomyopathy is a relatively common pathology in captive flying foxes (Pteropus spp.). The goal of this study was to establish quantitative reference range measurements that could be used to support a diagnosis of cardiac disease in these animals. Lateral and ventrodorsal thoracic radiographs from apparently healthy flying foxes (n = 66) of three species (Rodriguez island flying fox, P. rodricensis, n = 18; small island flying fox, P. hypomelanus, n = 16; and Malaysian flying fox, P. vampyrus, n = 32) were evaluated objectively to describe the cardiac appearance. Absolute and relative cardiac dimensions also were measured. The same methods were used to evaluate radiographs from flying foxes (n = 9) with known dilated or acute cardiomyopathy. The following ratios were most appropriate for categorizing normal cardiac silhouette size. In the ventrodorsal projection, heart width to thoracic width and heart width to clavicle length were the preferred measurements. In the lateral projection, heart width compared with thoracic height was the preferred measurement. From radiographs of the bats with known dilated and acute cardiomyopathy, the apicobasilar heart length compared with thoracic height and heart width compared with thoracic height on lateral films were the most sensitive ratios for diagnosing cardiomegaly.     

Microvasculature of the medulla oblongata in the Lyle's flying fox (Pteropus lylei)

The microvasculature of the medulla oblongata in 15 adult Lyle's flying foxes (Pteropus lylei) was elucidated by using the vascular corrosion cast technique combined with scanning electron microscopy. The study showed that the medulla received the main arterial supply from branches of the vertebrobasilar system. The supplied areas were divided into three groups: ventral, lateral and dorsal groups. All vessel groups gave off circumferential and perforating branches; moreover, these branches anastomosed with one another in two fashions: end-to-end and side-by-side arrangements. In addition, the ramifications of the branches were L and Y types. The L type was more frequently found than the Y one. The density of capillaries in the nuclei was greater than that in the area of nerve fibres. Numerous arterial sphincters and smooth muscle cells were observed. Furthermore, capillaries in the medulla were of the continuous type, whereas those in the area postrema were fenestrated. The venous drainage system of the medulla was classified into caudal, middle and rostral parts. All of them emptied into both the sigmoid sinus and internal jugular vein. It was concluded that these vascular patterns provide sufficient blood supply to the medulla oblongata of P. lylei when abrupt changes in the position of this bat occurs.     

Fatal columbid herpesvirus-1 infections in three species of Australian birds of prey

We document columbid herpesvirus-1 (CoHV-1) infection in two barking owls (Ninox connivens), a powerful owl (Ninox strenua) and an Australian hobby (Falco longipennis). Antemortem signs of infection were non-specific and the birds either died soon after they were identified as ill or were found dead unexpectedly. Gross postmortem findings were also not specific. Microscopically, marked to massive splenic and hepatic necrosis with the presence of eosinophilic inclusion bodies in remaining splenocytes and hepatocytes was found in all birds. Herpesvirus virions were identified in liver sections from one of the boobook owls by electron microscopy. Using CoHV-1-specific primers and polymerase chain reaction, CoHV-1 DNA was amplified from tissue samples from all birds. A comparison of these sequences to previously reported sequences of CoHV-1 found them to be identical or to vary by a single base pair. These findings increase the number of known species of birds of prey that are susceptible to CoHV-1 infection and indicate that rock pigeons (Columbia livia) should not be included in the diet of captive Australian birds of prey.     

Migratory waterfowl as flying reservoirs of Yersinia species

Intestinal contents of tufted ducks and faecal samples from two other species of migratory birds were examined for Yersinia species of organisms. From 296 tufted ducks Yersinia species were isolated from 10 (3.4 per cent), from 223 swans Yersinia species were recovered from eight (3.6 per cent) but none were recovered from the faeces of 121 pintail ducks. Since the isolates differed from each other in their biochemical characteristics and, or, O serotypes, yersinia infection in these migratory waterfowl seemed to be sporadic.     

Cranial edema associated with a protein-losing nephropathy in a golden-mantled flying fox (Pteropus pumilus).

An adult golden-mantled flying fox (Pteropus pumilus) was diagnosed with nephrotic syndrome on the basis of the findings of proteinuria, hypoalbuminemia, hypercholesterolemia, and cranial edema. Membranoproliferative glomerulitis and interstitial nephritis were confirmed antemortem by renal biopsy. The bat had received seven injections of oxytocin in the period immediately prior to presentation. The possible role of oxytocin in the development of the nephropathy is discussed. Supportive care and treatment with a single plasma transfusion, furosemide, and prednisone led to a gradual but complete resolution of the nephrotic syndrome in this animal.     

Viral-derived DNA invasion and individual variation in an Indonesian population of large flying fox Pteropus vampyrus

Here, we performed next-generation sequencing (NGS) on six large flying foxes (Pteropus vampyrus) collected in Indonesia. Seventy-five virus species in the liver tissue of each specimen were listed. Viral homologous sequences in the bat genome were identified from the listed viruses. This finding provides collateral evidence of viral endogenization into the host genome. We found that two of the six specimens bore partial sequences that were homologous to the plant pathogens Geminiviridae and Luteoviridae. These sequences were absent in the P. vampyrus chromosomal sequences. Hence, plant viral homologous sequences were localized to the hepatocytes as extrachromosomal DNA fragments. Therefore, this suggests that the bat is a potential carrier or vector of plant viruses. The present investigation on wild animals offered novel perspectives on viral invasion, variation, and host interaction.     

Chemical characterization of milk oligosaccharides of the island flying fox (Pteropus hypomelanus) (Chiroptera: Pteropodidae)

Although a considerable amount of information has accumulated about oligosaccharides in the milk and colostrum of representatives of various mammalian orders, nothing is so far known concerning these sugars in the milk of any bat species (order Chiroptera). In this study, we determined that the following oligosaccharides occur in milk of the island flying fox, Pteropus hypomelanus (Chiroptera: Pteropidae): Gal(α1–3)Gal(β1–4)Glc (isoglobotriose), Gal(β1–4)GlcNAc(β1–3)Gal(β1–4)Glc (lacto‐N‐neotetraose), Gal(β1–4)GlcNAc(β1–3)[Gal(β1–4)GlcNAc(β1–6)]Gal(β1–4)Glc (lacto‐N‐neohexaose) and Neu5Gc(α2–3)Gal(β1–4)Glc (3′‐NGc‐SL). However, lactose was found to be the dominant saccharide in this milk, as in most eutherian mammals. The biologic importance of oligosaccharides in Chiropteran milks warrants further study.     

No Evidence of Hendra Virus Infection in the Australian Flying‐fox Ectoparasite Genus Cyclopodia

Hendra virus (HeV) causes potentially fatal respiratory and/or neurological disease in both horses and humans. Although Australian flying‐foxes of the genus Pteropus have been identified as reservoir hosts, the precise mechanism of HeV transmission has yet to be elucidated. To date, there has been limited investigation into the role of haematophagous insects as vectors of HeV. This mode of transmission is particularly relevant because Australian flying‐foxes host the bat‐specific blood‐feeding ectoparasites of the genus Cyclopodia (Diptera: Nycteribiidae), also known as bat flies. Using molecular detection methods, we screened for HeV RNA in 183 bat flies collected from flying‐foxes inhabiting a roost in Boonah, Queensland, Australia. It was subsequently demonstrated that during the study period, Pteropus alecto in this roost had a HeV RNA prevalence between 2 and 15% (95% CI [1, 6] to [8, 26], respectively). We found no evidence of HeV in any bat flies tested, including 10 bat flies collected from P. alecto in which we detected HeV RNA. Our negative findings are consistent with previous findings and provide additional evidence that bat flies do not play a primary role in HeV transmission.     

Pathogenesis studies with Australian bat lyssavirus in grey-headed flying foxes (Pteropus poliocephalus)

OBJECTIVE: To examine the susceptibility of the grey-headed flying fox (Pteropus poliocephalus) to Australian bat lyssavirus (ABL), and to provide preliminary observations on the pathogenesis of the disease in flying foxes. PROCEDURE: Ten flying foxes were inoculated intramuscularly with ABL, and four with a bat-associated rabies virus. Inoculated animals were observed daily, and clinical samples collected every 9 to 14 days. Animals with abnormal clinical signs were euthanased, and samples collected for histological, serological, virological and immunohistological examinations. At 3 months post inoculation (PI), all survivors were euthanased, and each submitted to a similar examination. RESULTS: Three ABL-inoculated flying foxes, and two rabies-inoculated animals developed abnormal clinical signs between 15 and 24 days PI. All three ABL-inoculated animals had histological lesions consistent with a lyssavirus infection, and lyssaviral antigen was identified in the central nervous system (CNS) of each. Virus was isolated from the brain of two affected animals. Of the rabies-inoculated flying foxes, both had histological lesions and viral antigen in the CNS. Virus was recovered from the brain of only one. None of the five affected flying foxes developed anti-lyssavirus antibodies, but, by 3 months PI, five of the seven ABL-inoculated survivors, and one of the two rabies virus-inoculated survivors, had seroconverted. The dynamics of the immune responses were quite variable. CONCLUSIONS: The response of flying foxes to ABL, administered by a peripheral route of inoculation, was similar to that of bats inoculated peripherally with bat-derived rabies viruses.     

Cardiopulmonary effects of three different anaesthesia protocols in cats.

To develop an alternative anaesthetic regimen for cats with cardiomyopathy, the cardiopulmonary effects of three different premedication-induction protocols, followed by one hour maintenance with isoflurane in oxygen: air were evaluated in six cats. Group I: acepromazine (10 microg/kg) + buprenorphine (10 microg/kg) IM, etomidate (1-2 mg/kg) IV induction. Group II: midazolam (1 mg/kg) + ketamine (10 mg/kg) IM induction. Group III: medetomidine (1.5 mg/m2 body surface) IM, propofol (1-2 mg/kg) IV induction. Heart rate, arterial blood pressure, arterial blood gases, respiration rate, and temperature were recorded for the duration of the experiment. In group I the sedative effect after premedication was limited. In the other groups the level of sedation was sufficient. In all groups premedication resulted in a reduced blood pressure which decreased further immediately following induction. The reduction in mean arterial pressure (MAP) reached statistical significance in group I (142+/-22 to 81+/-14 mmHg) and group II (153+/-28 to 98+/-20 mmHg) but not in group III (165+/-24 to 134+/-29 mmHg). Despite the decrease in blood pressure, MAP was judged to have remained within an acceptable range in all groups. During maintenance of anaesthesia, heart rate decreased significantly in group III (from 165+/-24 to 125+/-10 b.p.m. at t=80 min). During anaesthesia the PCO2 and PO2 values increased significantly in all groups. On the basis of the results, the combination acepromazine-buprenorphine is preferred because heart rate, MAP, and respiration are acceptable, it has a limited sedative effect but recovery is smooth.     

Anaesthesia in two species of large Australian skink

The use of ketamine hydrochloride and sodium pentobarbitone in the anaesthesia of two species of Australian skink was examined. The effects of ketamine at ambient temperatures of 15 degrees C and 30 degrees C were studied. Ketamine produced consistent responses up to and including anaesthesia at dose rates of 170 to 230 mg/kg at 30 degrees C. The effect of temperature on the anaesthetic dose, respiratory and cardiac rates, muscle relaxation, analgesia and the onset and duration of anaesthesia was examined. Respiration in both species was depressed but heart rate was increased in Bobtail skinks (Tiliqua rugosa) and depressed in King's skinks (Egernia kingii). Muscle relaxation was good when anaesthetic doses were given. Generally, the onset and duration of anaesthesia were extended at 15 degrees C while the dose rates required for this effect were reduced. Although there was individual variation in the response to ketamine, it was found to be a useful and practical agent for the anaesthesia of large skinks. Pentobarbitone was found to be unsuitable as an anaesthetic agent because it produced inconsistent results and several fatalities.     

Field anaesthesia of leatherback sea turtles (Dermochelys coriacea)

Ten nesting leatherback sea turtles on Trinidad were anaesthetised for electroretinogram (ERG) measurements, using ketamine and medetomidine, reversed with atipamezole. They weighed 242 to 324 kg and were given initial doses of 3 to 8 mg/kg ketamine and 30 to 80 microg/kg medetomidine administered into an external jugular vein; six of the turtles received supplementary doses of 2.6 to 3.9 mg/kg ketamine combined with 0 to 39 microg/kg medetomidine. The lower doses were used initially to ensure against overdosage and reduce the chances of residual effects after the turtles returned to the water, but successful ergs called for step-wise dose increases to the required level of anaesthesia. Respiratory rate, heart rate, electrocardiogram, cloacal temperature, and venous blood gases were monitored, and blood was collected for plasma biochemistry. At the end of the erg procedure, atipamezole was administered at 150 to 420 microg/kg (five times the dose of medetomidine), half intramuscularly and half intravascularly. The turtles were monitored and prevented from re-entering the water until their behaviour was normal. No apparent mortalities or serious anaesthetic complications occurred. The observed within-season return nesting rate of the anaesthetised turtles was comparable with that of unanaesthetised turtles.     

兰狐和银黑狐免疫球蛋白的提纯与鉴定

采用硫酸铵盐析和Agarose-Protein G亲和层析相结合的方法，从兰狐和银黑狐血清中提取和纯化了免疫球蛋白（IgG），应用SDS-PAGE分析表明：提纯的免疫球蛋白纯度高，重链和轻链的分子量分别为45ku和21ku左右；以犬瘟热病毒（CDV）为包被抗原的间接ELISA试验结果表明：提纯的蛋白免疫小鼠分别制备的抗兰狐和银黑狐免疫球蛋白抗体作为二抗检测CDV阳性血清具有良好的生物活性。本研究制备的高纯度的兰狐和银黑狐免疫球蛋白，对今后狐源CDV等疫病血清流行病学的调查、诊断试剂盒的开发及疫苗免疫效果的评价奠定了基础。     

Dexmedetomidine chemical restraint of two raptor species undergoing inhalation anaesthesia

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