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Biology subjects

Cook, S. C.

Publications and source records attributed to Cook, S. C..

7 recordsLinked to original sources

Comparing morphological and molecular methods for stream macroinvertebrate biodiversity assessment across seven watersheds in the southern United States

Freshwater biodiversity is declining at alarming rates globally, yet our capacity to monitor it depends heavily on the methods used for biodiversity assessment. Stream benthic macroinvertebrates are widely used as bioindicators, but traditional morphological identification is labor-intensive and limited by taxonomic expertise, while DNA metabarcoding offers a promising but incompletely evaluated alternative. Here, we compared biodiversity patterns detected by four macroinvertebrate sampling and identification methods (morphological identification of benthic samples, morphological identification of combined benthic and stream-edge samples, eDNA metabarcoding of bulk benthic samples, and eDNA metabarcoding of streamwater) across 62 sites in seven watersheds spanning the southern continental United States. We compared methods across three facets of biodiversity (gamma diversity, alpha diversity, and taxonomic composition) and three taxonomic levels (family, genus, and species). We found that which method detected more taxa shifted with taxonomic resolution: morphological methods detected more families and genera, while metabarcoding methods detected more species. Bulk benthic metabarcoding detected more families and genera than water eDNA metabarcoding, consistent with the "watered-down biodiversity" effect of DNA dilution in water samples, but the two metabarcoding methods did not differ significantly in species richness. All four methods detected largely distinct suites of taxa, with metabarcoding methods yielding more unique taxa than morphological methods. Notably, taxa uniquely detected by water eDNA were dominated by soft-bodied organisms (oligochaetes, leeches, earthworms) and water-column-associated taxa, while benthic eDNA uniquely detected hard-bodied EPT insects. We attribute this pattern to fundamental differences in eDNA shedding, transport, and deposition dynamics across organism types. These results demonstrate that no single method captures a complete picture of stream macroinvertebrate biodiversity, and that method choice should be guided by the taxonomic resolution and community components most relevant to the goals of the bioassessment program.

ecology↗

Evaluation of efficacy of formic acid and thermal remediation for management of Tropilaelaps and Varroa mites in central Thailand

The western honey bee, Apis mellifera, faces a new threat from the spread of parasitic Tropilaelaps (Tropi) mites, specifically T. mercedesae, which adds additional complexity to an apicultural landscape heavily impacted by Varroa destructor. In this study conducted in central Thailand, we investigated the efficacy of two methods of applying formic acid and a thermal remediation technique in controlling Tropi mites and Varroa, focusing our attention on the reproductive stage of the mites which is restricted to capped brood cells. Results revealed that both formic acid treatments (Formic Pro and liquid formic acid) demonstrated an immediate and substantial reduction in live Tropi and Varroa populations, maintaining near-zero levels for the 3- week duration of the study. In contrast, thermal remediation, employing heating pads, exhibited a more gradual decline, achieving an 85.42% reduction in Tropi mites and a 92.33% reduction in Varroa mites by week three. Notably, heat-treated colonies experienced an unexpected resurgence in mite populations during week two. The findings contribute valuable insights into potential strategies for mitigating the threat of Tropi mites and highlight the urgency of further research to safeguard global honey bee populations.

zoology↗

Kleptocytosis: A Novel Parasitic Strategy for Accelerated Reproduction via Host Protein Stealing in Varroa destructor

Varroa destructor must produce mature offspring within the inflexible nine-day window framing the pupal development of their honey bee host. Missing this deadline renders the foundress mites fitness zero, establishing evolutionary pressure to accelerate reproduction and development. Through micro-computed tomography and modeling Varroas energy budget, we found each egg grows to constitute [~]18% of the foundresss body volume in 30 hours, yet accounts for less than 10% of her energy usage. We hypothesized that this small energy investment is a successful strategy because of Varroas long-ignored ability to traffic intact host proteins. Through gel electrophoresis, concomitant immunodetection, and MS/MS, we detected several intact, bee-derived proteins in mite eggs, including metamorphic proteins (hexamerins) and egg-yolk precursors (vitellogenin and apolipophorin) which likely reduce the parasites direct reproductive investment. We then expressed recombinant Halotag labelled Apis vitellogenin to determine the route conveying exogenous proteins into the mites oocyte. We detected fluorescent vitellogenin in the lyrate organ and observed a tube-like extension of the lyrate organ connecting to the ovum, likely providing an avenue for intact host proteins. Finally, we tested the hypothesis that exogenous proteins maintain functionality in the parasite. Varroa metamorphose despite their inability to produce hexamerins, critical metamorphic proteins. Through label-free quantification of proteins in metamorphosing mites, we observed a hexamerin depletion pattern consistent with usage as a metamorphic amino acid reservoir. We describe this process as "kleptocytosis," denoting movement of stolen intact macromolecules from host to a parasite cell. Given their fixed developmental timeline, this pathway presents a promising target for novel Varroa management strategies. Significance StatementGlobal honey bee health is threatened by the parasitic mite Varroa destructor; its success due in part to its rapid reproduction and accelerated development. By combining biological modeling, micro-computed tomography, fluorescence imaging, and quantitative proteomics, we show that Varroa siphon intact, ostensibly functional host proteins conveying them directly to their oocyte. This avoids the energetically inefficient process of digesting and reconstructing ingested proteins and affords the capacity to utilize proteins that it does not produce de novo facilitating rapid reproduction and accelerated development. We call this process "kleptocytosis," and identify anatomical adaptations which apparently facilitate protein movement. This work exposes a new target in Varroas physiology, providing a promising direction for more effective management strategies.

zoology↗

The vectoring competence of the mite Varroa destructor for Deformed wing virus of honey bees is highly dynamic and affects survival of the mite.

The ectoparasitic mite, Varroa destructor and the viruses it vectors, including types A and B of Deformed wing virus (DWV), pose a major threat to honey bees, Apis mellifera. Analysis of 256 mites collected from the same set of colonies on five occasions from May to October 2021 showed that less than a half of them, 39.8% (95% confidence interval (CI): 34.0 - 46.0%), were able to induce an overt-level DWV infection with more than 109 viral genomes per bee in the pupa after 6 days of feeding, with both DWV-A and DWV-B being vectored at similar rates. To investigate the effect of the phoretic stage on the mites ability to vector DWV, the mites from two field collection events were divided into two groups, one of which was tested immediately for their infectiveness, and the other was kept with adult worker bees in cages for 12 days prior to testing their infectiveness. We found that while 39.2 % (95% CI: 30.0 - 49.1%) of the immediately tested mites induced overt-level DWV infections, 12-day phoretic passage significantly increased the infectiousness of mites to 89.8% (95% CI: 79.2 - 95.6%). It is likely that Varroa mites that survive brood interruptions in field colonies are increasingly infectious. We found that mite lifespan was significantly affected by the type of DWV it transmitted to pupae. The mites, which induced overt DWV-B but not DWV-A infection had an average lifespan of 15.5 days (95% CI: 11.8 - 19.2 days), which was significantly shorter than those of the mites which induced overt DWV-A but not DWV-B infection, with an average lifespan of 24.3 days (95% CI: 20.2 - 28.5), or the mites which did not induce high levels of DWV-A or DWV-B, with an average survival of 21.2 days (95% CI: 19.0 - 23.5 days). The mites which transmitted high levels of both DWV-A and DWV-B had an intermediate average survival of 20.5 days (95% CI: 15.1 - 25.9 days). The negative impact of DWV-B on mite survival could be a consequence of the ability of DWV-B, but not DWV-A to replicate in Varroa mites.

microbiology↗

Resistance to amitraz in the parasitic honey bee mite Varroa destructor is associated with mutations in the β-adrenergic-like octopamine receptor

Varroa destructor is considered a major reason for high loss rate of Western honey bee (Apis mellifera) colonies. To prevent colony losses caused by V. destructor it is necessary to actively manage the mite population. Beekeepers, particularly commercial beekeepers, have few alternative treatments other than synthetic acaricides to control the parasite, resulting in intensive treatment regimens that led to the evolution of resistance in mite populations. To investigate the mechanism of the resistance to amitraz detected in V. destructor mites from French and U.S. apiaries, we identified and characterized octopamine and tyramine receptors (the known targets of amitraz) in this species. The comparison of sequences obtained from mites collected from different apiaries with different treatment regimens, showed that the amino acid substitutions N87S or Y215H in the Oct{beta}R were associated with treatment failures reported in French or U.S. apiaries, respectively. Based on our findings, we have developed and tested two high throughput diagnostic assays based on TaqMan(R) able to accurately detect mites carrying the mutations in this receptor. This valuable information may be of help for beekeepers when selecting the most suitable acaricide to manage V. destructor.

genetics↗

A novel system for maintaining Varroa destructor mites on artificial diets and its application for studying mites as a vector for honey bee viruses.

The mite Varroa destructor is one of the most destructive parasites of the honey bee (Apis mellifera) and the primary cause of colony collapse in most regions of the world. These mites cause serious injury to their hosts, especially during the larval and pupal stages, and serve as the vector for several viruses, which affect honey bee health causing colony death. Attempts by beekeepers to control these mites have yielded limited success. The inability to rear populations of mites in vitro that excludes contact with their honey bee hosts has stymied research of Varroa biology. Previous attempts to rear and/or maintain Varroa mites in vitro by feeding them on artificial diets have had limited success. Several methods were plagued by mechanical failures including leaking membranes and, thus far, none have been widely adopted. Here we report a robust system for maintaining Varroa mites that includes an artificial diet, which does not contain honey bee tissue-derived components, thus making it particularly valuable in studying mite vectoring of honey bee viruses. With our system we demonstrated for the first time that Varroa mites maintained on an artificial diet supplemented with the particles of honey bee viruses, cDNA clone-derived genetically tagged Varroa destructor virus-1 and wild-type Deformed wing virus, can acquire and later transmit these viruses to recipient honey bee pupae. Along with providing an opportunity to study parasites and pathogens in the absence of honey bee hosts, this in vitro system for Varroa mite maintenance is both scalable and consistent. These features can be used to better understand mite nutritional needs, metabolic activity, responses to chemicals and other biological functions.

zoology↗

Deformed wing virus type A, a major honey bee pathogen, is vectored by the mite Varroa destructor in a non-propagative manner.

Honey bees, the primary managed insect pollinator, suffer considerable losses due to Deformed wing virus (DWV), an RNA virus vectored by the mite Varroa destructor. Mite vectoring has resulted in the emergence of virulent DWV variants. The basis for such changes in DWV is poorly understood. Most importantly, it remains unclear whether replication of DWV occurs in the mite. In this study, we exposed Varroa mites to DWV type A via feeding on artificially infected honey bees. A significant, 357-fold increase in DWV load was observed in these mites after 2 days. However, after 8 additional days of passage on honey bee pupae with low viral loads, the DWV load dropped by 29-fold. This decrease significantly reduced the mites ability to transmit DWV to honey bees. Notably, negative-strand DWV RNA, which could indicate viral replication, was detected only in mites collected from pupae with high DWV levels but not in the passaged mites. We also found that Varroa mites contain honey bee mRNAs, consistent with the acquisition of honey bee cells which would additionally contain DWV replication complexes with negative-strand DWV RNA. We propose that transmission of DWV type A by Varroa mites occurs in a non-propagative manner.

microbiology↗