Meningoencephalitis in a Boelen's python (Morelia boeleni) associated with paramyxovirus infection.

An adult male Boelen's python, Morelia boeleni, presented with acute neurologic disease and was euthanatized. Histologic examination revealed nonsuppurative meningoencephalitis. Occasional eosinophilic intracytoplasmic inclusions were noted in glial cells. On the basis of clinical signs and histopathology, inclusion body disease of boid snakes was suspected, but inclusions were not seen in other organs commonly affected with the disease. Moreover, electron microscopy revealed that the inclusions contained stacks of filaments 13-14 nm wide. With the use of a generic paramyxovirus cDNA probe, sections of brain and esophageal ganglion demonstrated hybridization. The findings indicate that paramyxovirus was the likely cause of the encephalomyelitis in this python, and this virus should be included in the differential diagnosis of pythons exhibiting central nervous system disease.     

Predation by carpet pythons (Morelia spilota) is an important cause of mortality in a free‐living koala (Phascolarctos cinereus) population in South East Queensland

Koalas (Phascolarctos cinereus) are experiencing significant declines across the northern part of their range. However, unbiased, population‐level estimates of mortality are rarely reported, as it's difficult to quantify causes of mortality robustly in this cryptic species. We aimed to determine the relative importance of carpet python (Morelia spilota) predation in a free‐living koala population and describe the characteristic pathological findings during necropsy. In total, 503 koalas were captured, underwent veterinary examination and telemetric tagging, and were monitored after release over a four‐year period. Mortalities were detected when activity data reported by K‐Tracker® biotelemetry collars indicated low or zero activity, or during routine field monitoring events. Experienced koala veterinarians performed thorough, standardised necropsy examinations on retrieved carcasses to determine causes of death. The three, sometimes subtle, cardinal signs used to definitively diagnose carpet python‐caused deaths of koalas were a U‐shaped primary bite site, slicking of the fur by python saliva (particularly around the face), and diffuse, uniform pulmonary congestion. We found that carpet pythons were important predators of koalas, second only to wild dogs (dingoes and dingo hybrids (Canis familiaris dingo)), accounting for 11.6% of predation deaths and 7.2% of total deaths. Less than half (38%) of the koalas killed by carpet pythons were ingested. Though carpet pythons are known predators of koalas, their relative importance as a cause of mortality hasn't previously been recognised. Population viability analyses and conservation management plans benefit from robust cause‐of‐death data collected during longitudinal monitoring studies, requiring telemetry methods that facilitate rapid detection of mortalities.     

Computed tomography for the diagnosis and treatment monitoring of bacterial pneumonia in Indian pythons (Python molurus)

Eight Indian pythons (Python molurus) with clinical and microbiological evidence of pneumonia were examined by computed tomography (ct) before and after treatment. The results were assessed subjectively and measurements were taken following a standard protocol. Changes in the lung tissue of all the pythons were diagnosed, and the extent of the disease could be assessed. ct examinations after treatment showed an improvement in the six pythons whose clinical condition had improved, but in the other two pythons they demonstrated the severity of the disease. The subjective assessments were superior to the evaluation of measurements of attenuation in regions of interest. However, the average and the maximum attenuation provided additional information on the extent of the disease. Except for one python with only mild clinical signs, the attenuation after successful treatment was still higher than in healthy pythons.     

Evidence for the vertical transmission of Sunshine virus

Sunshine virus is a paramyxovirus of pythons associated with neurorespiratory disease and mortalities. This report provides evidence for its vertical transmission. In a collection of over 200 Australian pythons, a dam and a sire, both carpet pythons (Morelia spilota), were PCR-positive for Sunshine virus at a time when the dam was likely to have been gravid. A clutch of 21 eggs was laid and three non-viable eggs were tested for the presence of Sunshine virus by PCR. One egg had been incubating for 34 days while the other two had been incubating for 49 days. The surface of all three eggs was negative for Sunshine virus but swabs of the allantois and amnion were positive in all three eggs. Embryo tissue samples were tested from the two 49 day old eggs. From one embryo, a sample of brain and a pooled sample of lung, liver, kidney and intestine were positive, while for the other embryo, a pooled sample of lung, liver, kidney, intestine and brain was positive. Fourteen of the 21 eggs hatched and all hatchlings were tested by PCR at least once between the ages of 53 and 229 days old. All hatchlings were PCR-negative for Sunshine virus.     

Demonstration of rabies viral antigen in paraffin tissue sections: comparison of the immunofluorescence technique with the unlabeled antibody enzyme method

The immunofluorescence technique and the peroxidase-antiperoxidase method were used to demonstrate rabies antigen in a retrospective study on formalin-fixed, paraffin-embedded brain tissues from 34 naturally infected wild and domestic animals. Rabies was confirmed with immunofluorescent staining on fresh brain tissue at the time of necropsy of the animals. There was a perfect correlation (serial sections from a given brain area were always positive by both methods), but the peroxidase-antiperoxidase technique was preferred, since no trypsin digestion was required. Twenty six of the 34 animals were immunohistochemically positive and had encephalitis, and in 21 of these 26, the hematoxylin and eosin-stained sections contained detectable intracytoplasmic inclusion bodies in at least 1 brain area. Of the remaining 8 animals (with no inflammatory lesions), 7 were positive for rabies antigen and 2 had no inclusion bodies. Rabies antigen was apparent in 62% of the brain areas in which inclusion bodies were not found in the corresponding hematoxylin and eosin stained sections. Thus, together with the inclusion body positive areas, which were all immunohistochemically positive, it was possible to diagnose rabies in a total 84% of the areas examined. Both techniques greatly facilitate the diagnosis of rabies and may be a reliable help to the diagnostic pathologist when only formalin-fixed tissues are available. However, the methods should not be considered substitutes for the immunofluorescence technique and the mouse inoculation test with fresh brain tissue.     

速眠新和异氟烷对蟒蛇的全麻效果研究

本试验通过使用速眠新和异氟烷两种常用全身麻醉剂对缅甸蟒的麻醉效果进行了研究。对15条蟒蛇肌肉注射和腹腔注射(0.1、0.2、0.4 mL/kg)速眠新麻醉剂和对6条蟒蛇使用异氟烷吸入性麻醉后,进行麻醉效果的评估。试验结果表明,常规动物2~4倍的速眠新Ⅱ注射剂对蟒蛇的麻醉效果不明显;4%的异氟烷吸入性麻醉剂可用于蟒蛇的诱导麻醉,2.5%的异氟烷可用于蟒蛇的维持麻醉,其麻醉效果显著,具有诱导麻醉迅速、维持麻醉稳定、肌松作用好、安全性高、可控性强、苏醒快、副作用小等优点。结果显示,异氟烷吸入性麻醉剂可运用于蟒蛇的临床麻醉保定中。     

Environmental temperatures, physiology and behavior limit the range expansion of invasive Burmese pythons in southeastern USA

A well-established population of Burmese pythons resides in the Everglades of southern Florida. Prompted in part by a report that identified much of southern USA as suitable habitat for expansion or establishment of the Burmese python, we examined the plausibility of this snake to survive winters at sites north of the Everglades. We integrated daily low and high temperatures recorded from October to February from 2005-2011 at Homestead, Orlando and Gainesville, Florida; and Aiken, South Carolina, with minimum temperatures projected for python digestion (16 °C), activity (5 °C) and survival (0 °C). Mean low and high temperatures decreased northward from Homestead to Aiken and the number of days of freezing temperatures increased northward. Digestion was impaired or inhibited for 2 months in the Everglades and up to at least 5 months in Aiken, and activity was increasingly limited northward during these months. Reports of overwinter survivorship document that a single bout of low and freezing temperatures results in python death. The capacity for Burmese pythons to successfully overwinter in more temperate regions of the USA is seemingly prohibited because they lack the behaviors to seek refuge from, and the physiology to tolerate, cold temperatures. As tropical Southeast Asia is the source of the Everglades Burmese pythons, we predict it is unlikely that they will be able to successfully expand to or colonize more temperate areas of Florida and adjoining states due to their lack of behavioral and physiological traits to seek refuge from cold temperatures.     

An outbreak of adenoviral infection in inland bearded dragons (Pogona vitticeps) coinfected with dependovirus and coccidial protozoa (Isospora sp.).

Thirty of 200 (15%) hatchling inland bearded dragons were found dead after a short period (48 hours) of weakness and lethargy. The most common clinical signs were head tilt and circling. Six bearded dragons with neurological signs were euthanized, and postmortem examination revealed no gross abnormalities. Microscopically, severe, randomly distributed hepatocellular necrosis with large basophilic intranuclear inclusion bodies in numerous hepatocytes was noted. Small-intestinal enterocytes contained intracytoplasmic coccidial protozoa (Isospora sp.) and occasional enterocytes had basophilic intranuclear inclusion bodies. Transmission electron microscopy revealed both 80- and 20-nm-diameter viral particles, which were consistent with adenoviruses and dependoviruses, respectively. Adenoviral outbreaks in groups of animals are uncommon. An adverse synergistic effect of the coccidiosis with the adenoviral infection may have played a critical role in the high morbidity and mortality in this case.     

Temporal effects of 3 commonly used anticoagulants on hematologic and biochemical variables in blood samples from macaws and Burmese pythons

BACKGROUND: Few studies have been done to evaluate anticoagulants for use with blood samples from birds and reptiles. Heparin currently is the most commonly used anticoagulant in practice, but may adversely affect blood cell staining and quantitation. OBJECTIVE: The purpose of this study was to evaluate the effects of lithium heparin, K3-EDTA, and sodium citrate, with and without the addition of albumin, on hematologic variables in macaw (Ara sp) and python (Python molurus bivittatus) blood samples. METHODS: Blood samples from 10 macaws and 10 Burmese pythons were collected in heparin-coated syringes and placed into tubes containing either lithium heparin, K3-EDTA, or sodium citrate with and without the addition of 0.25 mL of a 22% bovine serum albumin solution. Cell lysis was determined by counting the number of lysed cells/200 WBCs in Wright's-Giemsa-stained blood smears and by qualitative evaluation of pink plasma in microhematocrit tubes. A CBC was done after 3, 12, and 24 hours of storage at 4 degrees C in anticoagulant-containing tubes and results were compared with those obtained at 0 hour for the heparin-coated syringe sample. A biochemical panel also was done at each time point in similarly stored lithium-heparin samples. RESULTS: Hemolysis was significantly increased in citrated samples from both macaws and pythons beginning at 12 hours. At 24 hours, 19 of 30 (63%) macaw samples in all anticoagulants had >100 lysed cells/200 WBCs. There were no significant differences in hematologic values in samples from pythons collected in heparin or EDTA at any time point. No significant differences were found in the number of lysed cells or in other hematologic data in samples with albumin. Glucose concentration decreased and potassium concentration increased significantly over time in heparinized blood samples. CONCLUSIONS: Based on the results of this study, whole blood samples anticoagulated with lithium heparin or EDTA should be evaluated within 12 hours (macaws) or 24 hours (pythons) of collection and stored at 4 degrees C for best results. Citrate should be avoided as it may result in increased cell lysis. The addition of albumin does not prevent cell lysis.     

Pathologic changes in pigs with prednisolone-induced recrudescence of herpesvirus infection

In pigs inoculated with Aujeszky's disease virus (ADV), recrudescence of herpesvirus infection was induced by daily administration of 1,000 mg of prednisolone for 5 days at 2 (group A) or 5 (group B) months after the primary infection. At necropsy in group A pigs, ADV was recovered from nasal secretions 3 to 9 days after prednisolone treatment initiation and from the brain cortex 10 days after treatment initiation; ADV was not recovered from group B pigs. In pigs of both groups killed 10 days after treatment initiation, 2 types of characteristic lesions were found. One type was a nonsuppurative encephalitis that consisted of neuronal necrosis, neuronophagia, and formation of eosinophilic intranuclear inclusion bodies. The 2nd type had basophilic intranuclear inclusion bodies in the enlarged endothelial cells of the kidneys, liver, lungs, adrenal glands, and lymph node sinusoids. Cells containing intranuclear inclusion bodies had immature and mature herpesvirus particles. Therefore, the brain lesions containing the eosinophilic inclusion bodies were considered to be due to ADV. Basophilic inclusion bodies in the endothelial cells were due to porcine cytomegalovirus. These observations indicated that prednisolone treatment resulted in recrudescence of ADV and porcine cytomegalovirus infections.     

自发性急性犬瘟热的原发性脱髓性脑病

为了进一步观察犬瘟热病毒引起的原发性脑损伤和包涵体形成的特点，调查脑组织的损伤与神经症状的关系，对10只急性犬瘟热病犬的脑组织进行了详细的病理学研究。为了仔细地观察病变，本试验按照解剖学关系将脑组织分成3个大部分和11个切面，即大脑(4个切面)，脑干(5个切面)和小脑(2个切面)。组织切片经HE、LFB和免疫组织化学染色后进行检查，结果表明：在大脑，脱髓呈弥漫性发生，程度较轻；脑干的周围或靠近第三脑室的白质脱髓较重；小脑在轻度或中度脱髓的基础上常出现严重的多发性脱髓灶。脱髓部呈空泡或海绵状，有少量胶质细胞存在，但无炎性反应。脱髓性病损是非时称性发生，对神经束没有特殊的亲和力。在脑室的室管膜细胞内发现有较多的嗜酸性胞浆或核内包涵体。用抗犬瘟热病毒抗体染色，带有包涵体的室管膜细胞呈现强阳性反应。部分锥体细胞，神经核细胞和漓氏细胞变性、溶解或胞浆深染。胞核浓缩。这种变化以小锥体神经细胞表现得最为明显。根据此研究结果，作者认为由犬瘟热病毒引起的原发性脑组织损伤是一种脱髓性脑病，而不是脑炎变化；位于室管膜细胞内的包涵体对于脑组织犬瘟热的确诊具有重要的作用；由于犬瘟热病毒引起神经细胞的损伤是非特异性的，对脑组织的侵害是非对称性的。对神经束的作用无特殊的亲和力，所以患犬瘟热的犬在临床上可出现不同的神经症状。     

Structure of inclusion bodies and electron microscopic virus detection in naturally occurring inclusion body hepatitis in chickens]

Electron microscopy was used to examine the liver of chickens with spontaneous inclusion body hepatitis. Eosinophilic inclusion bodies only were established from two flocks, mainly amphophilic from one flock, and primarily basophilic from another two flocks. Eosinophilic inclusion bodies were predominant in broiler chickens with dystrophic fatty degeneration of the liver, while basophile inclusion bodies were recorded primarily from parental or laying-hen chickens with reduced metabolic stress of the liver and more focal necrosis. The eosinophilic inclusion bodies consitsed of a filamentous matrix, with virus particles not safely detectable. The amphophilic inclusion bodies contained parvovirus particles, most likely adenoassociated virus, while the basophilic inclusion bodies inclused parvoviruses or adenoviruses (in flock NO. IV) or adenoviruses only (in flock No. V) in an amorphous chromatin matrix. The presence of parvoviruses in field material was taken to suggest a possible role of those pathogens in inclusion body hepatitis.     

缅甸蟒沙门氏菌的分离与鉴定

为探究缅甸蟒肺炎病因,本研究对海南文昌缅甸蟒养殖场的4条发病缅甸蟒进行了肺和心血组织的病原菌分离纯化、菌落及染色形态观察、生化、和16S rRNA分子鉴定,最后对各分离菌株进行了体外药敏实验。结果显示,全部分离株菌落及细胞形态特征一致,均为革兰氏阴性杆菌;糖醇发酵等生理生化特性与肠道沙门氏菌一致;根据16S rRNA基因的同源性比对,发现分离菌株与肠道沙门氏菌Salmonella enterica subsp.houtenae DSM 9221菌株同源性达到99%,可确定分离株全部为肠道沙门氏菌。通过23种抗菌药物的药物敏感性显示,所分离菌株对氟罗沙星等12种抗生素完全敏感。本研究结果表明海南文昌缅甸蟒肺炎病因为沙门氏菌感染,体外药敏实验可为该病的抗生素治疗提供使用参考。     

Atrioventricular valvular insufficiency and congestive heart failure in a carpet python

Right atrioventricular valve insufficiency and bilateral congestive heart failure were identified in a carpet python (Morelia spilota variegata) with the aid of colour Doppler echocardiography, electrocardiography and radiography. The snake failed to respond to diuretic therapy and was euthanased. Based on this case, it appears that bilateral congestive failure is feasible in univentricular animals with lesions restricted to one side of the heart. Loop diuretic therapy may be inappropriate in non-crocodilian reptiles because reptiles lack a loop of Henle.     

Computed tomography of the lungs of Indian pythons (Python molurus)

OBJECTIVE: To evaluate the use of computed tomography (CT) for detection of pneumonia in snakes. ANIMALS: 8 clinically normal Indian pythons (Python molurus) and 5 pythons with evidence of respiratory tract disease. PROCEDURES: Preliminary examinations (clinical examination, conventional radiography, and microbiologic examination of a transtracheal wash sample) were performed. The lungs of each snake were then examined by use of CT performed in accordance with a standardized protocol. Structures of the lungs were assessed, and thickness and attenuation of the parenchyma were determined. RESULTS: It was possible to assess lung parenchyma in all pythons. Mean +/- SD attenuation in healthy pythons was -744.4 +/- 47.1 Hounsfield units. Significant differences were not evident between the right and left lungs or among measurement areas within a lung. In all Indian pythons with clinical signs of dyspnea and microbiologic detection of pathogens, hyperattenuation of the alveolar tissue was evident. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of the results revealed the benefit of CT for use in the diagnosis of pneumonia in snakes. A standardized protocol and reference values were established as a basis for CT assessment of the lungs of snakes.     

Epizootic outbreaks of gizzard erosion associated with adenovirus infection in chickens

Two outbreaks of gizzard erosion in slaughtered broiler chickens in Japan were examined pathologically and microbiologically. The prevalences of such lesions were 9%-11% and 4%-50% in the affected flocks. Affected chickens had no clinical signs. Group I fowl adenovirus (FAV) serotype 1 was isolated from gizzard lesions. Histologically, gizzard mucosa were necrotic. Intranuclear inclusion bodies were seen in the enlarged nuclei of degenerating epithelial cells of the gizzard. The keratinoid layer in the erosion was edematous and desquamated and contained degenerative cells. Moderate to marked inflammatory cell infiltration was observed in the lamina propria and perivascular connective tissue in the submucosa and muscle layer. Immunohistochemical staining showed evidence of FAV antigens in the intranuclear inclusion bodies within degenerating epithelial cells. Ultrastructurally, numerous viral particles were demonstrated in the inclusions.     

Ecological correlates of invasion impact for Burmese pythons in Florida

An invasive population of Burmese pythons (Python molurus bivittatus) is established across several thousand square kilometers of southern Florida and appears to have caused precipitous population declines among several species of native mammals. Why has this giant snake had such great success as an invasive species when many established reptiles have failed to spread? We scored the Burmese python for each of 15 literature-based attributes relative to predefined comparison groups from a diverse range of taxa and provide a review of the natural history and ecology of Burmese pythons relevant to each attribute. We focused on attributes linked to spread and magnitude of impacts rather than establishment success. Our results suggest that attributes related to body size and generalism appeared to be particularly applicable to the Burmese python's success in Florida. The attributes with the highest scores were: high reproductive potential, low vulnerability to predation, large adult body size, large offspring size and high dietary breadth. However, attributes of ectotherms in general and pythons in particular (including predatory mode, energetic efficiency and social interactions) might have also contributed to invasion success. Although establishment risk assessments are an important initial step in prevention of new establishments, evaluating species in terms of their potential for spreading widely and negatively impacting ecosystems might become part of the means by which resource managers prioritize control efforts in environments with large numbers of introduced species.     

Pharmacokinetics of a long-acting ceftiofur formulation (ceftiofur crystalline free acid) in the ball python (Python regius)

The objective of this study was to determine the pharmacokinetics of a long-acting formulation of ceftiofur crystalline-free acid (CCFA) following intramuscular injection in ball pythons (Python regius). Six adult ball pythons received an injection of CCFA (15 mg/kg) in the epaxial muscles. Blood samples were collected by cardiocentesis immediately prior to and at 0.5, 1, 2, 4, 8, 12, 18, 24, 48, 72, 96, 144, 192, 240, 288, 384, 480, 576, 720, and 864 hr after CCFA administration. Plasma ceftiofur concentrations were determined by high-performance liquid chromatography. A noncompartmental pharmacokinetic analysis was applied to the data. Maximum plasma concentration (Cmax) was 7.096 +/- 1.95 microg/ml and occurred at (Tmax) 2.17 +/- 0.98 hr. The area under the curve (0 to infinity) for ceftiofur was 74.59 +/- 13.05 microg x h/ml and the elimination half-life associated with the terminal slope of the concentration-time curve was 64.31 +/- 14.2 hr. Mean residence time (0 to infinity) was 46.85 +/- 13.53 hr. CCFA at 15 mg/kg was well tolerated in all the pythons. Minimum inhibitory concentration (MIC) data for bacterial isolates from snakes are not well established. For MIC values of < or =0.1 microg/ml, a single dose of CCFA (15 mg/kg) provides adequate plasma concentrations for at least 5 days in the ball python. For MICs > or =0.5 microg/ml, more frequent dosing or a higher dosage may be required.     

Reptilian viruses: adenovirus-like agent isolated from royal python (Python regius).

An adenovirus-like agent was isolated from a moribund royal python (Python regius). The DNA containing virus replicated in IgH2-cells at 30 degrees C forming eosinophilic intranuclear inclusion bodies. The virus proved to be stabile to treatment with chloroform, pH3 und pH 12 but it was labile to heat (56 degrees C). Infected IgH2 cells revealed symmetric hexagonal virus particles measuring 67-79 nm in the nucleus. The isolate shared characteristics with the viruses of the family Adenoviridae.