Project description:In 2014 we observed a noticeable increase in the number of sudden deaths among green tree pythons (Morelia viridis). Pathological examination revealed the accumulation of mucoid material within the airways and lungs in association with enlargement of the entire lung. We performed a full necropsy and histological examination on 12 affected green tree pythons from 7 different breeders to characterize the pathogenesis of this mucinous pneumonia. By histology we could show a marked hyperplasia of the airway epithelium and of faveolar type II pneumocytes. Since routine microbiological tests failed to identify a causative agent, we studied lung tissue samples from a few diseased snakes by next-generation sequencing (NGS). From the NGS data we could assemble a piece of RNA genome whose sequence was <85% identical to that of nidoviruses previously identified in ball pythons and Indian pythons. We then employed reverse transcription-PCR to demonstrate the presence of the novel nidovirus in all diseased snakes. To attempt virus isolation, we established primary cultures of Morelia viridis liver and brain cells, which we inoculated with homogenates of lung tissue from infected individuals. Ultrastructural examination of concentrated cell culture supernatants showed the presence of nidovirus particles, and subsequent NGS analysis yielded the full genome of the novel virus Morelia viridis nidovirus (MVNV). We then generated an antibody against MVNV nucleoprotein, which we used alongside RNA in situ hybridization to demonstrate viral antigen and RNA in the affected lungs. This suggests that in natural infection MVNV damages the respiratory tract epithelium, which then results in epithelial hyperplasia, most likely as an exaggerated regenerative attempt in association with increased epithelial turnover.IMPORTANCE Novel nidoviruses associated with severe respiratory disease were fairly recently identified in ball pythons and Indian pythons. Herein we report on the isolation and identification of a further nidovirus from green tree pythons (Morelia viridis) with fatal pneumonia. We thoroughly characterized the pathological changes in the infected individuals and show that nidovirus infection is associated with marked epithelial proliferation in the respiratory tract. We speculate that this and the associated excess mucus production can lead to the animals' death by inhibiting normal gas exchange in the lungs. The virus was predominantly detected in the respiratory tract, which renders transmission via the respiratory route likely. Nidoviruses cause sudden outbreaks with high rates of mortality in breeding collections, and most affected snakes die without prior clinical signs. These findings, together with those of other groups, indicate that nidoviruses are a likely cause of severe pneumonia in pythons.

Project description:Fireflies (Coleoptera, Lampyridae) are a globally threatened group of insects due to habitat loss and fragmentation, light pollution, climate change and pesticides. However, against all odds, some firefly populations persist in urbanized environments where all four of these factors are present simultaneously. In this work, we compiled several data sources to document the diversity of fireflies in the urbanized area of Morelia, characterize their current habitats, and determine the main stressors affecting these bioluminescent insects. We found seven genera and 26 species of fireflies (19 nocturnal, seven diurnal) associated with 32 urban, peri-urban and extra-urban areas; at least, 14 are new records for Michoacán, and the list for the state now includes nine genera and 41 species. Five additional sites were documented as extinction sites. We compared the characteristics of these five sites with those of the sites with extant populations. We found that in Morelia, fireflies are mainly associated with areas that have high to moderate proportions of vegetation cover, are near water bodies, have very gentle to moderate slopes, and are exposed to low levels of light pollution. In contrast, the extinction sites showed high proportions of artificial surfaces and high levels of light pollution. Because some fireflies are considered bioindicators of ecosystem integrity as they are associated to specific habitats, are highly diverse and due to their sensitivity to environmental changes, we consider that sites from Morelia's urban core and extinction sites show the highest levels of environmental degradation, threatening most fireflies and other insects living in the urban core with local extinction. At the same time, our results also suggest that implementing conservation strategies and sustainable planning for the urban development of Morelia in the short term could allow fireflies and other vital elements of the city's insect communities to persist for future generations. Restoration and conservation of green areas and nighttime environments are essential for biodiversity and human health, especially in intra-urban zones.

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Morelia spilota

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