Correction: A vesicular stomatitis virus-based prime-boost vaccination strategy induces potent and protective neutralizing antibodies against SARS-CoV-2.

[This corrects the article DOI: 10.1371/journal.ppat.1010092.].

A vesicular stomatitis virus-based prime-boost vaccination strategy induces potent and protective neutralizing antibodies against SARS-CoV-2.

The development of safe and effective vaccines to prevent SARS-CoV-2 infections remains an urgent priority worldwide. We have used a recombinant vesicular stomatitis virus (rVSV)-based prime-boost immunization strategy to develop an effective COVID-19 vaccine candidate. We have constructed VSV genomes carrying exogenous genes resulting in the production of avirulent rVSV carrying the full-length spike protein (SF), the S1 subunit, or the receptor-binding domain (RBD) plus envelope (E) protein of SARS-CoV-2. Adding the honeybee melittin signal peptide (msp) to the N-terminus enhanced the protein expression, and adding the VSV G protein transmembrane domain and the cytoplasmic tail (Gtc) enhanced protein incorporation into pseudotype VSV. All rVSVs expressed three different forms of SARS-CoV-2 spike proteins, but chimeras with VSV-Gtc demonstrated the highest rVSV-associated expression. In immunized mice, rVSV with chimeric S protein-Gtc derivatives induced the highest level of potent neutralizing antibodies and T cell responses, and rVSV harboring the full-length msp-SF-Gtc proved to be the superior immunogen. More importantly, rVSV-msp-SF-Gtc vaccinated animals were completely protected from a subsequent SARS-CoV-2 challenge. Overall, we have developed an efficient strategy to induce a protective response in SARS-CoV-2 challenged immunized mice. Vaccination with our rVSV-based vector may be an effective solution in the global fight against COVID-19.

The benefits of insect-swarm hunting to echolocating bats, and its influence on the evolution of bat echolocation signals.

Predation on swarms of prey, especially using visual information, has drawn much interest in studies of collective movement. Surprisingly, in the field of biosonar this aspect of prey detection, which is probably very common, has received little to no attention. Here, we combine computer simulations and actual echo measurements to accurately estimate the echo sound pressure of insect swarms of different size and density. We show that swarm echo sound pressure increases with 3dB for every doubling of insect number, irrespective of swarm density. Thus swarms will be much easier to detect than single insects. Many of the insects bats eat are so small that they are only detectable by echolocation at very short distances. By focusing on detection of swarms of insects, a bat may increase its operating range and diversify its diet. Interestingly, interference between the sound waves reflected from a swarm of insects can sometimes result in echoes that are much weaker than echoes from single insects. We show that bats can reduce this problem by increasing the bandwidth of their echolocation calls. Specifically, a bandwidth of 3-8 kHz would guarantee receiving loud echoes from any angle relative to the swarm. Indeed, many bat species, and specifically bats hunting in open spaces, where swarms are abundant, use echolocation signals with a bandwidth of several kHz. Our results might also explain how the first echolocating bats that probably had limited echolocation abilities, could detect insects through swarm hunting.

Host association influences variation at salivary protein genes in the bat ectoparasite Cimex adjunctus.

Parasite-host relationships create strong selection pressures that can lead to adaptation and increasing specialization of parasites to their hosts. Even in relatively loose host-parasite relationships, such as between generalist ectoparasites and their hosts, we may observe some degree of specialization of parasite populations to one of the multiple potential hosts. Salivary proteins are used by blood-feeding ectoparasites to prevent hemostasis in the host and maximize energy intake. We investigated the influence of association with specific host species on allele frequencies of salivary protein genes in Cimex adjunctus, a generalist blood-feeding ectoparasite of bats in North America. We analysed two salivary protein genes: an apyrase, which hydrolyses ATP at the feeding site and thus inhibits platelet aggregation, and a nitrophorin, which brings nitrous oxide to the feeding site, inhibiting platelet aggregation and vasoconstriction. We observed more variation at both salivary protein genes among parasite populations associated with different host species than among populations from different spatial locations associated with the same host species. The variation in salivary protein genes among populations on different host species was also greater than expected under a neutral scenario of genetic drift and gene flow. Finally, host species was an important predictor of allelic divergence in genotypes of individual C. adjunctus at both salivary protein genes. Our results suggest differing selection pressures on these two salivary protein genes in C. adjunctus depending on the host species.

Range-wide genetic structure and demographic history in the bat ectoparasite Cimex adjunctus.

BACKGROUND: Evolutionary histories of parasite and host populations are intimately linked such that their spatial genetic structures may be correlated. While these processes have been relatively well studied in specialist parasites and their hosts, less is known about the ecological and evolutionary consequences of relationships between generalist ectoparasites and their hosts. The aim of this study was to investigate the genetic structure and demographic history of a bat ectoparasite, Cimex adjunctus, whose host affinity is weak but the biology of the potential hosts have been well studied. This ectoparasite has been hypothesized to rely on its hosts for dispersal due to its low inherent dispersal potential. Here we describe genetic diversity and demographic history in C. adjunctus through most of its range in North America. We investigated variation at the cytochrome c oxidase 1 mitochondrial gene and nine microsatellite markers, and tested the prediction that genetic diversity in C. adjunctus is spatially structured. We also tested the prediction that demographic history in C. adjunctus is characterized by range and demographic expansion as a consequence of post-Pleistocene climate warming. RESULTS: We found stronger spatial structuring of genetic diversity in C. adjunctus than has been quantified in two of its hosts, but contrast in amount of variation explained by host association with different genetic markers (i.e., nuclear vs mitochondrial DNA). Also, C. adjunctus' history is not primarily characterized by demographic and range expansion, as is the case with two of its key hosts. CONCLUSIONS: Our study shows different patterns of genetic structure and demographic history in C. adjunctus than have been detected in two of its key hosts. Our results suggest an effect of a loose parasite-host relationship and anti-parasitism strategies on genetic structure and post-Pleistocene recovery of population size.

Opening the black box of bird-window collisions: passive video recordings in a residential backyard.

Collisions with windows on buildings are a major source of bird mortality. The current understanding of daytime collisions is limited by a lack of empirical data on how collisions occur in the real world because most data are collected by recording evidence of mortality rather than pre-collision behaviour. Based on published literature suggesting a causal relationship between bird collision risk and the appearance of reflections on glass, the fact that reflections vary in appearance depending on viewing angle, and general principles of object collision kinematics, we hypothesized that the risk and lethality of window collisions may be related to the angle and velocity of birds' flight. We deployed a home security camera system to passively record interactions between common North American bird species and residential windows in a backyard setting over spring, summer and fall seasons over 2 years. We captured 38 events including 29 collisions and nine near-misses in which birds approached the glass but avoided impact. Only two of the collisions resulted in immediate fatality, while 23 birds flew away immediately following impact. Birds approached the glass at variable flight speeds and from a wide range of angles, suggesting that the dynamic appearance of reflections on glass at different times of day may play a causal role in collision risk. Birds that approached the window at higher velocity were more likely to be immediately killed or stunned. Most collisions were not detected by the building occupants and, given that most birds flew away immediately, carcass surveys would only document a small fraction of window collisions. We discuss the implications of characterizing pre-collision behaviour for designing effective collision prevention methods.

Chemical characterisation of potential pheromones from the shoulder gland of the Northern yellow-shouldered-bat, Sturnira parvidens (Phyllostomidae: Stenodermatinae).

Bats of the genus Sturnira (Family Phyllostomidae) are characterised by shoulder glands that are more developed in reproductively mature adult males. The glands produce a waxy secretion that accumulates on the fur around the gland, dyeing the fur a dark colour and giving off a pungent odour. These shoulder glands are thought to play a role in their reproductive behaviour. Using gas chromatography-mass spectrometry, we analysed solvent extracts of fur surrounding the shoulder gland in the northern-shouldered bat, Sturnira parvidens to (i) characterise the chemical composition of shoulder gland secretions for the first time, and (ii) look for differences in chemical composition among and between adult males, sub-adult/juvenile males and adult females. Fur solvent extracts were analysed as liquids and also further extracted using headspace solid-phase microextraction to identify volatile components in the odour itself. Odour fingerprint analysis using non-metric multidimensional scaling plots and multivariate analysis revealed clear and significant differences (P < 0.001) between adult males vs both juvenile males and adult females. The chemical components of the shoulder gland secretion included terpenes and phenolics, together with alcohols and esters, most likely derived from the frugivorous diet of the bat. Many of the compounds identified were found exclusively or in elevated quantities among adult (reproductive) males compared with adult females and non-reproductive (juvenile) males. This strongly suggests a specific role in male-female attraction although a function in male-male competition and/or species recognition is also possible.

Comparative analysis of landscape effects on spatial genetic structure of the big brown bat and one of its cimicid ectoparasites.

Identification of landscape features that correlate with genetic structure permits understanding of factors that may influence gene flow in a species. Comparing effects of the landscape on a parasite and host provides potential insights into parasite-host ecology. We compared fine-scale spatial genetic structure between big brown bats (Eptesicus fuscus) and their cimicid ectoparasite (Cimex adjunctus; class Insecta) in the lower Great Lakes region of the United States, in an area of about 160,000 km2. We genotyped 142 big brown bat and 55 C. adjunctus samples at eight and seven microsatellite loci, respectively, and inferred effects of various types of land cover on the genetic structure of each species. We found significant associations between several land cover types and genetic distance in both species, although different land cover types were influential in each. Our results suggest that even in a parasite that is almost entirely reliant on its hosts for dispersal, land cover can affect gene flow differently than in the hosts, depending on key ecological aspects of both species.

Host association and selection on salivary protein genes in bed bugs and related blood-feeding ectoparasites.

Reciprocal selective pressures can drive coevolutionary changes in parasites and hosts, and result in parasites that are highly specialized to their hosts. Selection and host co-adaptation are better understood in endoparasites than in ectoparasites, whose life cycles may be more loosely linked to that of their hosts. Blood-feeding ectoparasites use salivary proteins to prevent haemostasis in the host, and maximize energy intake. Here we looked for signals of selection in salivary protein genes of ectoparasite species from a single genus (Cimex) that associate with a range of hosts including mammals (bats and humans) and birds (swallows). We analysed two genes that code for salivary proteins that inhibit platelet aggregation and vasoconstriction and may directly affect the efficiency of blood feeding in these species. Significant positive selection was detected at five codons in one gene in all bat-associated species groups. Our results suggest association with bats, versus humans or swallows, has posed a selective pressure on the salivary apyrase gene in species of Cimex.

Vision impairs the abilities of bats to avoid colliding with stationary obstacles.

BACKGROUND: Free-flying insectivorous bats occasionally collide with stationary objects they should easily detect by echolocation and avoid. Collisions often occur with lighted objects, suggesting ambient light may deleteriously affect obstacle avoidance capabilities. We tested the hypothesis that free-flying bats may orient by vision when they collide with some obstacles. We additionally tested whether acoustic distractions, such as "distress calls" of other bats, contributed to probabilities of collision. METHODOLOGY/PRINCIPAL FINDINGS: To investigate the role of visual cues in the collisions of free-flying little brown bats (Myotis lucifugus) with stationary objects, we set up obstacles in an area of high bat traffic during swarming. We used combinations of light intensities and visually dissimilar obstacles to verify that bats orient by vision. In early August, bats collided more often in the light than the dark, and probabilities of collision varied with the visibility of obstacles. However, the probabilities of collisions altered in mid to late August, coincident with the start of behavioural, hormonal, and physiological changes occurring during swarming and mating. Distress calls did not distract bats and increase the incidence of collisions. CONCLUSIONS/SIGNIFICANCE: Our findings indicate that visual cues are more important for free-flying bats than previously recognized, suggesting integration of multi-sensory modalities during orientation. Furthermore, our study highlights differences between responses of captive and wild bats, indicating a need for more field experiments.