Recovery of ranavirus dsDNA from formalin-fixed archival material.
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The extraction and amplification of nucleic acid from formalin-fixed and paraffin-embedded tissues has become an important exercise in the collection of retrospective epidemiological data. A protocol is described that enables the extraction and amplification of dsDNA from fixed tissues within paraffin blocks and from specimens stored in 10% (aq) formalin. The procedure can be used for the examination of ranavirus DNA within archival tissues thereby providing valuable data for identifying the origin and tracing the spread of ranaviruses. (+info)
Salmon: a new autosomal mutation demonstrating incomplete dominance in the boine snake Boa constrictor.
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An unusual and attractive pigmentation pattern mutation termed "salmon" has been identified in the United States in several captive colonies of the common neotropical boine snake boa constrictor [Boa constrictor (Boidae)]. Boa constrictors expressing the Sa pigmentation pattern appear to be restricted to regions of Panama. Animals with the Sa phenotype exhibit a sharp decrease in melanophore pigments (e.g., melanin) and an increase of xanthophore pigments (e.g., pteridines and carotenoids) throughout the body, including ventral and caudal regions. Moreover, the dorsal saddles (blotches) and lateral diamond patterns are greatly reduced and/or absent. Our study was initiated using a female B. constrictor born in captivity and expressed the Sa pigmentation pattern. Results from breeding experiments indicated an inheritance mode of autosomal incomplete dominance for the Sa and Wt alleles. (+info)
Ventilatory and cardiovascular responses of a python (Python molurus) to exercise and digestion.
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To investigate the potential limiting steps of peak metabolic rates, we examined gas exchange rates ( vdot (O2), vdot (CO2)), respiratory exchange ratio (RER), breathing frequency, tidal volume, minute ventilation volume (V.e) as well as the heart rate, systemic blood flow and stroke volume of Burmese pythons (Python molurus) while fasting at rest, exercising, digesting and exercising while digesting. All measured variables increased significantly during exercise (crawling at 0.4 km h(-)(1) and at vdot (O2max)), highlighted by a 17-fold increase in vdot (CO2) and a 24-fold increase in V.e. During the digestion of a meal equivalent to 25 % of the snake's body mass, pythons responded with increases in vdot (O2) and heart rate similar to those experienced during exercise, along with a 4.5-fold increase in systemic blood flow. Interestingly, pythons hyperventilated while exercising, whereas they hypoventilated during digestion. The combined demands of exercise and digestion resulted in significantly higher vdot (O2), vdot (CO2), breathing frequency and heart rate than during either exercise or digestion alone. Evidently, the capacities of the ventilatory and cardiovascular systems to transport oxygen to locomotor muscles are not a limiting factor in the attainment of peak metabolic rates during exercise in pythons (+info)
Isolation and characterization of an antigenically distinct 68-kd protein from nonviral intracytoplasmic inclusions in Boa constrictors chronically infected with the inclusion body disease virus (IBDV: Retroviridae).
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The relationship between a retroviral infection and the development of nonviral intracytoplasmic inclusion bodies was studied in a Boa constrictor model. Twelve juvenile age- and size-matched inclusion body disease (IBD)-negative boas were randomly divided into three groups. Each group was inoculated intraperitoneally with 1 ml of an IBD virus (IBDV)-infected liver homogenate or 1 ml of normal boa liver homogenate (sham-inoculated control) or was left untreated. All boas were monitored for development of IBD by daily examination and serial liver biopsy over 1 year. The 4 IBDV-inoculated boas became IBDV and inclusion positive by 10 weeks postinoculation. The average size and density of inclusion bodies increased with the duration of infection. Ultrastructurally, inclusion bodies <2 microm in diameter consisted of intracytoplasmic aggregates of granular electron-dense material that were not membrane limited. Larger inclusions (3-6 microm in diameter) were characterized as membrane-bound aggregates of amorphous to granular electron-dense material admixed with membranelike fragments. The sham-inoculated and untreated control snakes did not become inclusion or IBDV positive. Direct comparison of the protein electrophoretograms of IBDV-infected and normal boa tissues demonstrated a prominent 68-kd protein band unique to infected inclusion-positive tissues. Monoclonal antibodies directed against the 68-kd protein band specifically labeled inclusion bodies. The results of this study demonstrate that IBD inclusions represent an intracytoplasmic accumulation of an antigenically distinct IBDV-associated protein. (+info)
Luminal and systemic signals trigger intestinal adaptation in the juvenile python.
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Juvenile pythons undergo large rapid upregulation of intestinal mass and intestinal transporter activities upon feeding. Because it is also easy to do surgery on pythons and to maintain them in the laboratory, we used a python model to examine signals and agents for intestinal adaptation. We surgically isolated the middle third of the small intestine from enteric continuity, leaving its mesenteric nerve and vascular supply intact. Intestinal continuity was restored by an end-to-end anastomosis between the proximal and distal thirds. Within 24 h of the snake's feeding, the reanastomosed proximal and distal segments (receiving luminal nutrients) had upregulated amino acid and glucose uptakes by up to 15-fold, had doubled intestinal mass, and thereby soon achieved total nutrient uptake capacities equal to those of the normal fed full-length intestine. At this time, however, the isolated middle segment, receiving no luminal nutrients, experienced no changes from the fasted state in either nutrient uptakes or in morphology. By 3 days postfeeding, the isolated middle segment had upregulated nutrient uptakes to the same levels as the reanastomosed proximal and distal segments, but it still lacked any appreciable morphological response. These contrasting results for the reanastomosed intestine and for the isolated middle segment suggest that luminal nutrients and/or pancreatic biliary secretions are the agents triggering rapid upregulation of transporters and of intestinal mass and that systemic nerve or hormonal signals later trigger transporter regulation but no trophic response. (+info)
Adenovirus hepatitis in a boa constrictor (Boa constrictor).
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A boa constrictor was submitted for postmortem evaluation. At necropsy, there were no substantial lesions except in the liver. Light microscopy revealed severe multifocal to coalescing coagulative necrotic hepatitis, with basophilic and eosinophilic intranuclear inclusions in hepatocytes within the necrotic foci. The histopathological findings suggested a viral hepatitis. An adenoviral infection was diagnosed by means of transmission electronic microscopy and in situ hybridization techniques. (+info)
Structural flexibility of the intestine of Burmese python in response to feeding.
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The small intestine of Burmese pythons, Python molurus bivittatus, undergoes a remarkable size increase shortly after feeding. We studied the dynamics, reversibility and repeatability of organ size changes using noninvasive imaging techniques. We employed light and electron microscopy, flow cytometry and immunohistology to study the cytological mechanisms that drive the size changes of the small intestine. Within 2 days of feeding, the size of the small intestine increased to up to three times the fasting value. The size changes were fully reversible and could be elicited repeatedly by feeding. These enormous size changes were possible because the mucosal epithelium of the small intestine is a transitional epithelium that allows for considerable size changes without cell proliferation. Histological evidence suggested that a fluid pressure-pump system (lymphatic, blood pressure) was the driving force that inflated the intestinal villi. The rates of cell proliferation were not elevated immediately after feeding but peaked 1 week later when small intestine size was already declining. In contrast to the current paradigm, we suggest that the small intestine is not part of the previously proposed 'pay-before-pumping' model. Instead, the size of the python's small intestine may be upregulated without major metabolic investment. It can occur even if the individual is energetically exhausted. An evolutionary perspective of the transitional epithelium mechanism suggests superior functionality compared with the pay-before-pumping model because it allows for long periods of fasting and depletion of energy reserves, while still enabling the snake to digest prey and absorb nutrients. (+info)
Retroviral particles in neoplasms of Burmese pythons (Python molurus bivittatus).
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Neoplastic diseases associated with retroviruses were diagnosed in four Burmese pythons (Python molurus bivattatus) from a single collection. Snake No. 1 was a 7-year-old female with recurrent undifferentiated mesenchymal round cell tumor (lymphosarcoma) of the oral cavity. At necropsy, similar neoplastic masses were evident in the uterus and ovary, and there was diffuse involvement of the spleen. Snake No. 2 was a 4.5-year-old female that was euthanatized because of complications following resection of a segmental colonic adenocarcinoma. Snake No. 3 was a 5-year-old female that was euthanatized because of a large transitional cell carcinoma of the right kidney. Snake No. 4 was a 19-year-old female that was euthanatized following recurrence of an intermandibular fibrosarcoma. Ultrastructural examination revealed few to numerous extracellular and intracellular (intravacuolar) type C-like retroviral particles in all tumors. Tumors were about 90-95 nm in diameter, with an electron-dense core and bilaminar external membrane. The relationship of the intraneoplastic viral particles to the etiology of the tumors is uncertain. (+info)