Optimization of Flavonoid Extraction from Salix babylonica L. Buds, and the Antioxidant and Antibacterial Activities of the Extract.

The present study was designed to evaluate the chemical extraction, chemical composition, and antioxidant and antibacterial properties of the total flavonoids in Willow Buds (TFW). We investigated the optimal extraction of TFW using response surface methodology (RSM). Chemical compounds were analyzed using Q-Orbitrap LC-MS/MS. The DPPH radical scavenging capacity, hydroxy radical inhibitory ability, and superoxide anion radical inhibitory ability were explored to determine the antioxidant properties of flavonoid extractions. The antibacterial effect was assessed via minimal inhibitory concentration. The results demonstrated that the optimal extraction conditions were an ethanol concentration of 50%, a time of 35 min, and a liquid/material ratio of 70:1 mL/g. Under these conditions, the yield of TFW was 7.57%. Eight flavonoids, a phenolic glycoside, and an alkaloid were enriched in the Willow Buds. The TFW exhibited significant antioxidant activity, with IC50 values of 0.18-0.24 mg/mL and antimicrobial activity against Escherichia coli, Salmonella enterica, Staphylococcus aureus, and Streptococcus pneumoniae. TFW may be explored as potential and natural compounds in food and pharmacological applications.

Assessing the health risks of reintroduction: The example of the Amur leopard, Panthera pardus orientalis.

Translocation of wildlife as a means of reintroducing or reinforcing threatened populations is an important conservation tool but carries health risks for the translocated animals and their progeny, as well as wildlife, domestic animals and humans in the release area. Disease risk analyses (DRA) are used to identify, prioritize and design mitigation strategies to address these threats. Here, we use a DRA undertaken for Amur leopards (Panthera pardus orientalis) to illustrate how specific methodology can optimize mitigation strategy design. A literature review identified a total of 98 infectious hazards and 28 non-infectious hazards. Separate analyses were undertaken for disease risks in leopards from hazards of source origin (captive zoo collections and the transit pathway to the Russian Far East), or of destination origin (in breeding enclosures and wider release areas); and for disease risks in other wildlife, domesticated species or humans, similarly from hazards of source or destination origin. Hazards were assessed and ranked as priority 1, priority 2, priority 3 or low priority in each of the defined scenarios. In addition, we undertook a generic assessment of stress on individual leopards. We use three examples to illustrate the process: Chlamydophila felis, canine distemper virus (CDV) and feline immunodeficiency virus (FIV). We found that many potentially expensive screening procedures could be performed prior to export of leopards, putting the onus of responsibility onto the zoo sector, for which access to diagnostic testing facilities is likely to be optimal. We discuss how our methods highlighted significant data gaps relating to pathogen prevalence in the Russian Far East and likely future unpredictability, in particular with respect to CDV. There was emphasis at all stages on record keeping, meticulous planning, design, staff training and enclosure management, which are relatively financially inexpensive. Actions to minimize stress featured at all time points in the strategy and also focussed on planning, design and management.

Canine distemper virus: an emerging disease in wild endangered Amur tigers (Panthera tigris altaica).

UNLABELLED: Fewer than 500 Amur tigers (Panthera tigris altaica) remain in the wild. Due to low numbers and their solitary and reclusive nature, tiger sightings across their range in the Russian Far East and China are rare; sightings of sick tigers are rarer still. Serious neurologic disease observed in several wild tigers since 2001 suggested disease emergence in this endangered species. To investigate this possibility, histology, immunohistochemistry (IHC), in situ hybridization (ISH), and reverse transcription-PCR (RT-PCR) were performed on tissues from 5 affected tigers that died or were destroyed in 2001, 2004, or 2010. Our results reveal canine distemper virus (CDV) infection as the cause of neurologic disease in two tigers and definitively establish infection in a third. Nonsuppurative encephalitis with demyelination, eosinophilic nuclear viral inclusions, and positive immunolabeling for CDV by IHC and ISH were present in the two tigers with available brain tissue. CDV phosphoprotein (P) and hemagglutinin (H) gene products were obtained from brains of these two tigers by RT-PCR, and a short fragment of CDV P gene sequence was detected in lymph node tissue of a third tiger. Phylogenetically, Amur tiger CDV groups with an Arctic-like strain in Baikal seals (Phoca siberica). Our results, which include mapping the location of positive tigers and recognition of a cluster of cases in 2010, coupled with a lack of reported CDV antibodies in Amur tigers prior to 2000 suggest wide geographic distribution of CDV across the tiger range and recent emergence of CDV as a significant infectious disease threat to endangered Amur tigers in the Russian Far East. IMPORTANCE: Recognition of disease emergence in wildlife is a rare occurrence. Here, for the first time, we identify and characterize a canine distemper virus (CDV), the second most common cause of infectious disease death in domestic dogs and a viral disease of global importance in common and endangered carnivores, as the etiology of neurologic disease and fatal encephalitis in wild, endangered Amur tigers. We establish that in 2010 CDV directly or indirectly killed -1% of Amur tigers. Location of positive cases over an expansive geographic area suggests that CDV is widely distributed across the tiger range. Interspecies interactions are increasing as human populations grow and expand into wildlife habitats. Identifying animal reservoirs for CDV and identifying the CDV strains that are transmissible to and among wildlife species, including Amur tigers and sympatric critically endangered Amur leopards (Panthera pardus orientalis), is essential for guiding conservation and mitigation efforts.