Non-destructive insect metabarcoding for surveillance and biosecurity in citrus orchards: recording the good, the bad and the psyllids
Background The Australian citrus industry remains one of the few in the world to be unaffected by the African and the Asian citrus psyllids, Trioza erytreae Del Guercio and Diaphorina citri Kuwayama, respectively, and the diseases their vectored bacteria can cause. Surveillance, early detection, and strict quarantine measures are therefore fundamental to safeguard Australian citrus. However, long-term targeted surveillance for exotic citrus pests can be a time-consuming and expensive activity, often relying on manually screening large numbers of trap samples and morphological identification of specimens, which requires a high level of taxonomic knowledge. Methods Here we evaluated the use of non-destructive insect metabarcoding for exotic pest surveillance in citrus orchards. We conducted an 11-week field trial, between the months of December and February, at a horticultural research farm (SuniTAFE Smart Farm) in the Northwest of Victoria, Australia, and processed more than 250 samples collected from three types of invertebrate traps across four sites. Results The whole-community metabarcoding data enabled comparisons between different trapping methods, demonstrated the spatial variation of insect diversity across the same orchard, and highlighted how comprehensive assessment of insect biodiversity requires use of multiple complimentary trapping methods. In addition to revealing the diversity of native psyllid species in citrus orchards, the non-targeted metabarcoding approach identified a diversity of other pest and beneficial insects and arachnids within the trap bycatch, and recorded the presence of the triozid Casuarinicola cf warrigalensis for the first time in Victoria. Ultimately, this work highlights how a non-targeted surveillance approach for insect monitoring coupled with non-destructive DNA metabarcoding can provide accurate and high-throughput species identification for biosecurity and biodiversity monitoring.
Martoni et al. (2023) Non-destructive insect metabarcoding for surveillance and biosecurity in citrus orchards: recording the good, the bad and the psyllids. PeerJ 11:e15831 https://doi.org/10.7717/peerj.15831
Evaluation of a biosecurity survey approach for contamination by Chlamydia pecorum in koala rehabilitation, field capture, and captive settings
Transmission of Chlamydia pecorum between koalas is a potential risk in field capture or rehabilitation settings, where koalas are held in proximity to each other, or equipment is shared between animals. Given the impact of C. pecorum on koala welfare and population viability it is surprising that quarantine and disinfection protocols in a koala rehabilitation facility or capture settings have not previously been evaluated. This study aimed to evaluate an approach, based on the detection of chlamydial DNA and cell viability, to determine the degree of environmental contamination within a koala care facility. Various fomite sites associated with koala care at a koala rehabilitation facility in New South Wales, Australia were identified as potential sources of chlamydial contamination, following exposure to koalas known to be infected with C. pecorum. Fomite sites were swabbed following exposure, and again after decontamination procedures were carried out. Samples were tested for the presence of chlamydial DNA using qPCR and viability using both RT-qPCR and cell culture. From a total of 239 sampling events, 30 tested qPCR positive for chlamydial DNA, with 19 and 11 samples corresponding to pre-decontamination and post-decontamination events respectively. Detection of chlamydial DNA appeared to be most common in the examination room, especially on fomite sites in direct contact with koalas. Physical removal of chlamydial DNA, or its degradation by the elements, appeared to be more common on outdoor enclosures, clothing, and hands. Based on the cell culture assay, of the pre-decontamination samples with chlamydial DNA, eight had viable chlamydial cells, two of these at low levels. Of the post-decontamination samples with chlamydial DNA, one had a moderate number, and one had a very low number of viable chlamydial cells. RT-qPCR was unsuccessful in determining cell viability due to low yields of RNA and high levels of contaminants from the environmental samples. The outcomes of this study provide a knowledge base for the design of future biosecurity evaluation guidelines in captive and koala rehabilitation facilities. The higher incidence of chlamydial DNA detection by qPCR than viable organism highlights the need to use viability assays in similar studies. However, further investment is still needed to optimise these methods and improve sensitivity for complex environmental samples.
Casteriano et al. (2023) Evaluation of a biosecurity survey approach for contamination by Chlamydia pecorum in koala rehabilitation, field capture, and captive settings. PeerJ 11:e15842 https://doi.org/10.7717/peerj.15842
Morphological changes in female reproductive organs in the African monarch butterfly, host to a male-killing Spiroplasma
Background Sexual selection and conflicts within and between sexes promote morphological diversity of reproductive traits within species. Variation in the morphology of diagnostic reproductive characters within species offer an excellent opportunity to study these evolutionary processes as drivers of species diversification. The African monarch, Danaus chrysippus (Linnaeus, 1758), is widespread across Africa. The species is polytypic, with the respective geographical ranges of the four colour morphs only overlapping in East Africa. Furthermore, some of the populations host an endosymbiotic bacterium, Spiroplasma, which induces son-killing and distorts the local host population sex-ratio, creating sexual conflicts between the females seeking to optimize their fecundity and the limited mating capacity of the rare males. Methods We dissected females from Kenya, Rwanda and South Africa, where Spiroplasma vary in presence and prevalence (high, variable and absent, respectively), and conducted microscopy imaging of their reproductive organs. We then characterized the effect of population, female body size, and female mating status, on the size and shape of different genitalia characters of the D. chrysippus female butterflies. Results We showed that although the general morphology of the organs is conserved in D. chrysippus, female genitalia vary in size and shape between and within populations. The virgin females have smaller organs, while the same organs were expanded in mated females. Females from highly female-biased populations, where the male-killing Spiroplasma is prevalent, also have a larger area of their corpus bursae covered with signa structures. However, this pattern occurs because a larger proportion of the females remains virgin in the female-biased populations rather than because of male depletion due to the symbiont, as males from sex-ratio distorted populations did not produce significantly smaller nutritious spermatophores.
Malmberg et al. (2023) Morphological changes in female reproductive organs in the African monarch butterfly, host to a male-killing Spiroplasma. PeerJ 11:e15853 https://doi.org/10.7717/peerj.15853
Repetition velocity as a measure of loading intensity in the free weight and Smith machine Bulgarian split squat
This study investigated the grouped and individualized load-velocity profile (GLVP vs. ILVP) in Bulgarian split squat using Smith machine and free weight. Seventy five recreational male lifters completed two incremental loading tests of Bulgarian split squat. Mean velocity was measured by a linear-position transducer (GymAware). Linear regression equation was applied to construct the GLVP and ILVP. The agreement of predicted %1RM and measured %1RM was assessed by a combination of intraclass correlation coefficient (ICC), coefficient of variation (CV), standard error of measurement (SEM) and Bland-Altman analysis. Acceptable validity was defined as ICC > 0.75, CV ≤ 10% and p ≥ 0.05 (a paired Wilcoxon signed-rank test). A very high level of inverse load-velocity relationships were demonstrated in Bulgarian split squat (r = − 0.92) with free weights and a Smith machine. ILVP (ICC ≥ 0.98, CV ≤ 8.73%, p ≥ 0.56) was valid enough to predict the %1RM, but GLVP of both limbs revealed large CVs in free weights (CV: 15.4%,15.63%) and a Smith machine (CV: 11.24%, 12.25%). Cross-validation between the actual %1RM and predicted %1RM using free weights and a Smith machine ILVP was not acceptable (p ≤ 0.03, CV ≥ 14.07%). A very high level of inverse relationship were observed between %1RM and MV in Bulgarian split squat using free weights and a Smith machine, indicating individualized load velocity properties, and the ILVP showed high between-devices variability in both scenarios. Using velocity as a measure of loading intensity in Bulgarian split squat needs to consider the individualized load velocity properties, and difference between free weights and a Smith machine.
Liao et al. (2023) Repetition velocity as a measure of loading intensity in the free weight and Smith machine Bulgarian split squat. PeerJ 11:e15863 https://doi.org/10.7717/peerj.15863
Parent perceptions regarding virtual pediatric dental clinics during COVID-19 pandemic: a cross-sectional study
Objective To assess the attitudes and responses of parents of pediatric patients towards virtual dental clinics during COVID-19. Material and Method A total of 102 parents of pediatric patients who were scheduled for virtual (video or telephonic) clinic appointments for new patient consultations and follow-up clinics were included in the study. Parents and patients could attend the virtual clinic from a personal computer, tablet, or smartphone. An electronic self-administered questionnaire was sent to the parents through email after consultation (video or telephonic) which consisted of demographic data and a ten-item, five-point Likert-scale assessing: (i) parent satisfaction; (ii) ease of use; (iii) the effectiveness including increasing access to clinical services; (iv) reliability of the teledentistry system and (v) usefulness for patients. Statistical analyses used were t-test, one-way ANOVA test, Shapiro–Wilk and histogram. Result Out of 102 parents, 52 attended video clinics and 50 attended the telephonic clinic. The majority of the parents were between 30–39 years of age, and about 73% had no previous experience with either virtual or telephonic consultation. Ninety-four percent of parents were satisfied with vide clinic consultation, and most of them agreed that accessing clinical services through a video clinic was easy to understand, comfortable, and time-saving. The majority of the parents (94%) agreed and strongly agreed that they will use video clinics again in the future for consultation. Conclusion Parents’ response to the use of virtual clinics for pediatric dentistry during COVID was positive. The majority of the parents stated that they would consider using teledentistry for future consultation.
Alghamdi (2023) Parent perceptions regarding virtual pediatric dental clinics during COVID-19 pandemic: a cross-sectional study. PeerJ 11:e15289 https://doi.org/10.7717/peerj.15289
Chemical diversity and antifouling activity of geniculate calcareous algae (Corallinales, Rhodophyta) from Brazil
Marine biofouling is a natural process by which many organisms colonize and grow in submerged structures, causing serious economic consequences for the maritime industry. Geniculate calcareous algae (GCA; Corallinales, Rhodophyta) produce bioactive secondary metabolites and are a promise for new antifouling compounds. Here, we investigated the antifouling activity of four GCA species—Amphiroa beauvoisii, Jania sagittata (formerly Cheilosporum sagittatum), Jania crassa, and Jania prolifera (formerly Amphiroa flabellata)—from the Brazilian coast against macro- and microorganisms. Simultaneously, metabolomic tools were applied to assess the chemical profiles of these seaweeds using gas chromatography coupled to mass spectrometry (GC-MS). Data analysis by principal component and molecular networking analyses used the global natural products social molecular networking platform (GNPS). Our results showed that all extracts were active against different strains of marine bacteria and that the J. sagittata (JsSI) extract showed the highest percentage of bacterial inhibition. The J. sagittata (JsSI) extract was the most active against the mussel Perna perna, showing 100% byssus inhibition. Regarding toxicity, only the J. crassa (JcP) extract showed a 20% mortality rate. The chemical profiles of the evaluated GCA extracts differed qualitatively and quantitatively. Yet, the steroid (3β)-cholest-5-en-3-ol was the major compound commonly identified in all extracts, with the exception of J. sagittata (JsSI). Moreover, we observed intra- and interspecific chemical variabilities among GCA extracts for the different populations, which could explain their antifouling activity variability. This study contributed new information about the chemical compounds produced by this group of seaweeds and showed its antifouling potential. These GCA species may be the subject of future studies to obtain new bioactive compounds with biotechnological potential in maritime areas.
Oliveira et al. (2023) Chemical diversity and antifouling activity of geniculate calcareous algae (Corallinales, Rhodophyta) from Brazil. PeerJ 11:e15731 https://doi.org/10.7717/peerj.15731
Green inspired synthesis of zinc oxide nanoparticles using Silybum marianum (milk thistle) extract and evaluation of their potential pesticidal and phytopathogens activities
Background The green approaches for the synthesis of nanoparticles are gaining significant importance because of their high productivity, purity, low cost, biocompatibility, and environmental friendliness. Methods The aim of the current study is the green synthesis of zinc oxide nanoparticles (ZnO-NPs) using seed extracts of Silybum marianum, which acts as a reducing and stabilizing agent. central composite design (CCD) of response surface methodology (RSM) optimized synthesis parameters (temperature, pH, reaction time, plant extract, and salt concentration) for controlled size, stability, and maximum yields of ZnO-NPs. Green synthesized ZnO-NPs was characterized using UV-visible spectroscopy and Zetasizer analyses. Results The Zetasizer confirmed that green synthesized ZnO-NPs were 51.80 nm in size and monodispersed in nature. The UV-visible results revealed a large band gap energy in the visible region at 360.5 nm wavelength. The bioactivities of green synthesized ZnO-NPs, including antifungal, antibacterial, and pesticidal, were also evaluated. Data analysis confirmed that these activities were concentration dependent. Bio-synthesized ZnO-NPs showed higher mortality towards Tribolium castaneum of about 78 ± 0.57% after 72 h observation as compared to Sitophilus oryzae, which only displayed 74 ± 0.57% at the same concentration and time intervals. Plant-mediated ZnO-NPs also showed high potential against pathogenic gram-positive bacteria (Clavibacter michiganensis), gram-negative bacteria (Pseudomonas syringae), and two fungal strains such as Fusarium oxysporum, and Aspergillums niger with inhibition zones of 18 ± 0.4, 25 ± 0.4, 21 ± 0.57, and 19 ± 0.4 mm, respectively. Conclusion The results of this study showed that Silybum marianum-based ZnO-NPs are cost-effective and efficient against crop pests.
Jahan et al. (2023) Green inspired synthesis of zinc oxide nanoparticles using Silybum marianum (milk thistle) extract and evaluation of their potential pesticidal and phytopathogens activities. PeerJ 11:e15743 https://doi.org/10.7717/peerj.15743
Lethal and sublethal effects of flubendiamide and spirotetramat against the leaf worm, Spodoptera litura(Fabricius) under laboratory conditions
The leaf worm, Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) is a notorious insect pest of many economically important cultivated crops like cotton, maize, tobocco and vegetables causing severe economic losses from 50–100%. In most crops, damage arises due to voracious feeding by the larvae and leads to the skeletonizing of leaves. Toxicological studies were performed to estimate lethal and sublethal levels of flubendiamide and spirotetramat against S. litura. Effects of these estimated values were assessed on different biological traits of S. litura including life duration, survival and next generation potential. Both flubendiamide and spirotetramat showed toxic responses against second instar larvae of S. litura under laboratory conditions. Larval duration and survival rate of S. litura to were significantly different. Exposure to test insecticides resulted in negative effect on the demography of S. litura as longer life cycle and decreased fecundity. Changes in net reproductive rate and intrinsic rate of increase also helped to decide the fate of these insecticides. Low reproductive potential and low hatching percentage due to exposure to test insecticides can help to manage next generation of target pest. These two new chemistry insecticides can be recommended for their effective and long-term utilization against this important leaf feeder which may help its management and decrease economic losses faced by the growers. Their impact on larval duration and low survival rate at lethal levels guides about their potential in pest control.
un-Nisa et al. (2023) Lethal and sublethal effects of flubendiamide and spirotetramat against the leaf worm, Spodoptera litura(Fabricius) under laboratory conditions. PeerJ 11:e15745 https://doi.org/10.7717/peerj.15745
Response surface methodology for the mixed fungal fermentation of Codonopsis pilosula straw using Trichoderma reesei and Coprinus comatus
The objective of this study was to investigate the cellulose degradation rate (CDR) and lignin degradation rate (LDR) of Codonopsis pilosula straw (CPS) and the optimal fermentation parameters for mixed fungal fermentation. Single-factor tests were used to study the effects of the fungal ratio (Trichoderma reesei: Coprinus comatus), fungal inoculum, corn flour content, and fermentation time on the degradation rate of cellulose and lignin. Based on the results of this experiment, the optimal fermentation factors were identified, and the effects of various factors and their interactions on the degradation rates of cellulose and lignin were further evaluated using the response surface method. The quadratic polynomial mathematical model of degradation rates of the cellulose and lignin in CPS by mixed fungus fermentation was established using Design Expert software v8.0.6. Under the optimal parameters for fungal fermentation of CPS straw (fungal ratio 4:6, fungal inoculum 8%, corn flour content 10%, fermentation time of 15 d), the CDR and LDR reached 13.65% and 10.73%, respectively. Collectively, the mixed fungal fermentation of CPS resulted in decreased lignin and cellulose content, better retention of nutrients, and enhanced fermentation quality. The results of this study indicate that fermentation using Trichoderma reesei and Coprinus comatus is a productive method for straw degradation, providing a theoretical basis for the development of CPS as feed.
Wei et al. (2023) Response surface methodology for the mixed fungal fermentation of Codonopsis pilosula straw using Trichoderma reesei and Coprinus comatus. PeerJ 11:e15757 https://doi.org/10.7717/peerj.15757
Tailored modulation of S100A1 and RASSF8 expression by butanediamide augments healing of rotator cuff tears
Objectives This investigation sought to elucidate promising treatment modalities for rotator cuff tears (RCTs) by delving into the molecular machinations instigating the affliction. The focus was on differentially expressed genes (DEGs) linked to RCTs, and the exploration of their roles and operative pathways. Methods DEGs were discerned from GEO datasets, followed by the establishment of a protein-protein interaction (PPI) network. Subsequently, the network’s core genes were determined employing a Venn diagram. Enrichment analysis facilitated the unveiling of the biological roles and signal transduction pathways of these pivotal genes, thus shedding light on molecular strategies for RCT-targeted treatment. The Discovery Studio 2019 software was employed to sift through FDA-sanctioned drugs targeting these essential proteins. Moreover, the efficaciousness of these FDA-endorsed drugs vis-à-vis RCTs was corroborated by the construction of an in vivo animal model of the injury and the in vitro cultivation of tendon-derived stem cells. Results Bioinformatics outcomes revealed a significant overexpression of S100A1 and RASSF8 in RCT patients. The FDA drug repository indicated that Butanediamide has a selective affinity for S100A1 and RASSF8. Subsequent in vivo and in vitro experimentation demonstrated that Butanediamide could suppress S100A1 expression and bolster TDSC proliferation, thereby facilitating RCT healing. Conclusions S100A1 and RASSF8 are pivotal genes implicated in RCTs, and their roles have been elucidated. The FDA-approved compound, Butanediamide, may represent a prospective therapeutic agent for RCTs by targeting S100A1 and RASSF8, respectively.
Lin et al. (2023) Tailored modulation of S100A1 and RASSF8 expression by butanediamide augments healing of rotator cuff tears. PeerJ 11:e15791 https://doi.org/10.7717/peerj.15791