Staging classification of omicron variant SARS-CoV-2 infection based on dual-spectrometer LIBS (DS-LIBS) combined with machine learning.

Effective differentiation of the infection stages of omicron can provide significant assistance in transmission control and treatment strategies. The combination of LIBS serum detection and machine learning methods, as a novel disease auxiliary diagnostic approach, has a high potential for rapid and accurate staging classification of Omicron infection. However, conventional single-spectrometer LIBS serum detection methods focus on detecting the spectra of major elements, while trace elements are more closely related to the progression of COVID-19. Here, we proposed a rapid analytical method with dual-spectrometer LIBS (DS-LIBS) assisted with machine learning to classify different infection stages of omicron. The DS-LIBS, including a broadband spectrometer and a narrowband spectrometer, enables synchronous collection of major and trace elemental spectra in serum, respectively. By employing the RF machine learning models, the classification accuracy using the spectra data collected from DS-LIBS can reach 0.92, compared to 0.84 and 0.73 when using spectra data collected from single-spectrometer LIBS. This significant improvement in classification accuracy highlights the efficacy of the DS-LIBS approach. Then, the performance of four different models, SVM, RF, IGBT, and ETree, is compared. ETree demonstrates the best, with cross-validation and test set accuracies of 0.94 and 0.93, respectively. Additionally, it achieves classification accuracies of 1.00, 0.92, 0.92, and 0.89 for the four stages B1-acute, B1-post, B2, and B3. Overall, the results demonstrate that DS-LIBS combined with the ETree machine learning model enables effective staging classification of omicron infection.

Females prefer males producing a high-rate song with shorter timbal-stridulatory sound intervals in a cicada species.

Uncovering mate choice and factors that lead to the choice are very important to understanding sexual selection in evolutionary change. Cicadas are known for their loud sounds produced by males using the timbals. However, males in certain cicada species emit 2 kinds of sounds using respectively timbals and stridulatory organs, and females may produce their own sounds to respond to males. What has never been considered is the mate choice in such cicada species. Here, we investigate the sexual selection and potential impact of predation pressure on mate choice in the cicada Subpsaltria yangi Chen. It possesses stridulatory sound-producing organs in both sexes in addition to the timbals in males. Results show that males producing calling songs with shorter timbal-stridulatory sound intervals and a higher call rate achieved greater mating success. No morphological traits were found to be correlated with mating success in both sexes, suggesting neither males nor females display mate preference for the opposite sex based on morphological traits. Males do not discriminate among responding females during mate searching, which may be due to the high energy costs associated with their unusual mate-seeking activity and the male-biased predation pressure. Females generally mate once but a minority of them re-mated after oviposition which, combined with the desirable acoustic traits of males, suggest females may maximize their reproductive success by choosing a high-quality male in the first place. This study contributes to our understanding mechanisms of sexual selection in cicadas and other insects suffering selective pressure from predators.

The best of both worlds: cicada males change costly signals to achieve mates while females choose a mate based on both calling and courtship songs.

Cicadas usually sing and mate in the higher parts of trees. Studies addressing the effects of different acoustic signals on mate choice in Cicadidae are very limited. We investigated the effects of both acoustical features and morphological traits on mate choice in an East Asian cicada Platypleura kaempferi. Males produce high-rate calling songs that attract females, then produce low-rate courtship songs to secure mating when a female is attracted. Higher calling song rate (CR), shorter single-pulse duration, and shorter pulse period of the calling song, together with lower courtship song rate and longer echeme period of the courtship song, are the most desirable traits used by females to choose a mate. These traits indicate that the more a male can raise the rate of song production, the higher the probability he is sexually selected by the female. No correlation was found between morphological traits and mating success. After mating, a minority of males started emitting calling songs again, but the CR was significantly lower than before mating and none of them attracted a new mate later. This promotes females mating with unmated males. We hypothesize that P. kaempferi may have the best of both worlds due to the unique song modulation and the mechanism of female mate choice: males change energetically, costly acoustic signals to achieve mates, while females choose a mate based on males' acoustic properties. Our results contribute to better understanding the diversity of mating preference and enrich the mechanism of mate choice in acoustic insects.

Gut Bacteria Associated With Monochamus saltuarius (Coleoptera: Cerambycidae) and Their Possible Roles in Host Plant Adaptations.

Monochamus saltuarius (Coleoptera: Cerambycidae) is an important native pest in the pine forests of northeast China and a dispersing vector of an invasive species Bursaphelenchus xylophilus. To investigate the bacterial gut diversity of M. saltuarius larvae in different host species, and infer the role of symbiotic bacteria in host adaptation, we used 16S rRNA gene Illumina sequencing and liquid chromatography-mass spectrometry metabolomics processing to obtain and compare the composition of the bacterial community and metabolites in the midguts of larvae feeding on three host tree species: Pinus koraiensis, Pinus sylvestris var. mongolica, and Pinus tabuliformis. Metabolomics in xylem samples from the three aforementioned hosts were also performed. Proteobacteria and Firmicutes were the predominant bacterial phyla in the larval gut. At the genus level, Klebsiella, unclassified_f__Enterobacteriaceae, Lactococcus, and Burkholderia-Caballeronia-Paraburkholderia were most dominant in P. koraiensis and P. sylvestris var. mongolica feeders, while Burkholderia-Caballeronia-Paraburkholderia, Dyella, Pseudoxanthomonas, and Mycobacterium were most dominant in P. tabuliformis feeders. Bacterial communities were similar in diversity in P. koraiensis and P. sylvestris var. mongolica feeders, while communities were highly diverse in P. tabuliformis feeders. Compared with the other two tree species, P. tabuliformis xylems had more diverse and abundant secondary metabolites, while larvae feeding on these trees had a stronger metabolic capacity for secondary metabolites than the other two host feeders. Correlation analysis of the association of microorganisms with metabolic features showed that dominant bacterial genera in P. tabuliformis feeders were more negatively correlated with plant secondary metabolites than those of other host tree feeders.

Comparative transcriptome analysis of the newly discovered insect vector of the pine wood nematode in China, revealing putative genes related to host plant adaptation.

BACKGROUND: In many insect species, the larvae/nymphs are unable to disperse far from the oviposition site selected by adults. The Sakhalin pine sawyer Monochamus saltuarius (Gebler) is the newly discovered insect vector of the pine wood nematode (Bursaphelenchus xylophilus) in China. Adult M. saltuarius prefers to oviposit on the host plant Pinus koraiensis, rather than P. tabuliformis. However, the genetic basis of adaptation of the larvae of M. saltuarius with weaken dispersal ability to host environments selected by the adult is not well understood. RESULTS: In this study, the free amino and fatty acid composition and content of the host plants of M. saltuarius larvae, i.e., P. koraiensis and P. tabuliformis were investigated. Compared with P. koraiensis, P. tabuliformis had a substantially higher content of various free amino acids, while the opposite trend was detected for fatty acid content. The transcriptional profiles of larval populations feeding on P. koraiensis and P. tabuliformis were compared using PacBio Sequel II sequencing combined with Illumina sequencing. The results showed that genes relating to digestion, fatty acid synthesis, detoxification, oxidation-reduction, and stress response, as well as nutrients and energy sensing ability, were differentially expressed, possibly reflecting adaptive changes of M. saltuarius in response to different host diets. Additionally, genes coding for cuticle structure were differentially expressed, indicating that cuticle may be a potential target for plant defense. Differential regulation of genes related to the antibacterial and immune response were also observed, suggesting that larvae of M. saltuarius may have evolved adaptations to cope with bacterial challenges in their host environments. CONCLUSIONS: The present study provides comprehensive transcriptome resource of M. saltuarius relating to host plant adaptation. Results from this study help to illustrate the fundamental relationship between transcriptional plasticity and adaptation mechanisms of insect herbivores to host plants.

De novo comparative transcriptome analysis of a rare cicada, with identification of candidate genes related to adaptation to a novel host plant and drier habitats.

BACKGROUND: Although the importance of host plant chemistry in plant-insect interactions is widely recognized, our understanding about the genetic basis underlying the relationship between changes in midgut proteins and adaptation of plant-feeding insects to novel host plants and habitats is very limited. To address this knowledge gap, the transcriptional profiles of midguts among three populations of the cicada Subpsaltria yangi Chen were compared. Among which, the Hancheng (HC) and Fengxiang (FX) populations occurring in the Loess Plateau feed on Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chow, while the population occurring in a much drier habitat in the Helan (HL) Mountains is locally specialized on a chemically divergent plant, Ephedra lepidosperma C. Y. Cheng. RESULTS: Based on comparative analysis, 1826 (HL vs HC) differentially expressed genes (DEGs) and 723 DEGs (HL vs FX) were identified between the populations utilizing different host plants, including 20, 36, 2, 5 and 2 genes related to digestion, detoxification, oxidation-reduction, stress response and water-deprivation response, respectively, and 35 genes presumably associated with osmoregulation. However, only 183 DEGs were identified between the HC and FX populations, including two genes related to detoxification, two genes related to stress response, and one gene presumably associated with osmoregulation. These results suggest that the weakest expression differences were between the populations utilizing the same host plant and occurring in the closest habitats, which may help explain the metabolic mechanism of adaptation in S. yangi populations to novel host plants and new niches. CONCLUSIONS: The observed differences in gene expression among S. yangi populations are consistent with the hypothesis that the host plant shift and habitat adaptation in the HL population was facilitated by differential regulation of genes related to digestion, detoxification, oxidation-reduction, stress response, water-deprivation response and osmoregulation. The results may inform future studies on the molecular mechanisms underlying the relationship between changes in midgut proteins and adaptation of herbivorous insects to novel host plants and new niches.

Influence of construction and demolition waste on fitness and community structure of cicada nymphs: New bioindicators of soil pollution.

Construction and demolition (C&D) waste is a novel, widespread environmental stress that negatively affects environment, organisms and ecosystem. Yet effects of cumulative consequences of C&D waste on the fitness and community structure of most underground insects remain unclear. With relatively sessile life underground lasting several years, cicadas can be an important aspect in forest nutrient cycles. Whether cicada nymphs are easily affected by C&D waste, and whether they have evolved any adaptations to cope with the adverse environments merit exploration. Here, we investigated the biodiversity, community structure, population dynamics and morphology of cicada nymphs in both uncontaminated and contaminated habitats by C&D waste in Guanzhong Plain, China since 2011. In total, 1,573 cicada nymphs were collected from 2011 to 2015, including 62 malformed nymphs. The malformed nymphs can be divided into three types: physically damaged individuals (still alive) (3.2%), fungus-infected individuals (dead) (64.5%), and bacterium-infected individuals (dead) (32.3%). The proportion of malformed nymphs increased year by year in the habitats contaminated by C&D waste. In the uncontaminated habitats, although no significant differences of population density among the investigated years were observed, yet there was a distinct increasing trend of population of Meimuna mongolica, whereas populations of both Cryptotympana atrata and Platypleura kaempferi distinctly decreased. This indicates that M. mongolica is possibly evolving into the most dominant species in the ecological niche when it competes with other sympatric species, but more researches are needed to establish whether there is a shift in the species composition of cicadas. In the habitats contaminated by C&D waste, a higher ratio of malformed individuals and a decline of population of both M. mongolica and C. atrata were revealed; P. kaempferi was not found in the contaminated habitats, indicating a weaker resistance of this species against C&D waste. The negative responses of cicada nymphs to C&D waste have significant implications for the habitat destruction. Cicada nymphs may be suitable bioindicators for underground-habitat-quality monitoring, as merits further research to reveal the association between the magnitude of C&D waste contamination with the fitness and population dynamics of cicada nymphs.

Sexual pair-formation in a cicada mediated by acoustic behaviour of females and positive phonotaxis of males.

The functions of female song found in a few cicadas have rarely been studied. In the cicada Subpsaltria yangi we investigated the acoustic behaviour and signal structure of songs produced by females, the phonotaxis of males, and mate choice, as well as the selective pressure imposed on this species by predators. Pair-formation in S. yangi occurs when males signal, females respond, then males move to signaling females, which is opposite to that in most other cicadas where females move to calling males. Females only mate once and are sexually unreceptive after copulation. Most males mate once, but ~25% mate multiply. Females display little direct evidence of mate preference or choice of males, and all mate encounters led to a successful mating. Only males are attacked by a robber fly, Philonicus albiceps, while flying to females. This imposes strong selection on males - only males who can evade predators mate. Males are also attracted to human simulations of female calls. This behaviour exposes the mating system to impacts from anthropogenic noise systems which could disrupt mating activity of this species. Our results improve the understanding of mate choice/competition in cicadas, and are valuable for future studies of the evolution of sound communication in the Cicadoidea.

Morphology and identification of the final instar nymphs of three cicadas (Hemiptera, Cicadidae) in Guanzhong Plain, China based on comparative morphometrics.

The present investigation provides comparative morphometrics of the final instar nymphs of three dominant cicada species, i.e., Cryptotympana atrata (Fabricius), Meimuna mongolica (Distant) and Platypleura kaempferi (Fabricius), in Guanzhong Plain, China. Particularly, characters on the antennae, legs, and apex of abdomen of both males and females of these three species were investigated and analyzed. In addition, the numbers of hind tibial spines of the final instar nymphs of 21 representatives of Cicadoidea were compared. The results provide useful characteristics for nymph identification of related species and for further taxonomic and phylogenetic analysis of Cicadoidea.