Periodic pattern formation during embryonic development.

During embryonic development many organs and structures require the formation of series of repeating elements known as periodic patterns. Ranging from the digits of the limb to the feathers of the avian skin, the correct formation of these embryonic patterns is essential for the future form and function of these tissues. However, the mechanisms that produce these patterns are not fully understood due to the existence of several modes of pattern generation which often differ between organs and species. Here, we review the current state of the field and provide a perspective on future approaches to studying this fundamental process of embryonic development.

A Symptom-Based Natural Language Processing Surveillance Pipeline for Post-COVID-19 Patients.

Post-acute sequelae of SARS CoV-2 (PASC) are a group of conditions in which patients previously infected with COVID-19 experience symptoms weeks/months post-infection. PASC has substantial societal burden, including increased healthcare costs and disabilities. This study presents a natural language processing (NLP) based pipeline for identification of PASC symptoms and demonstrates its ability to estimate the proportion of suspected PASC cases. A manual case review to obtain this estimate indicated our sample incidence of PASC (13%) was representative of the estimated population proportion (95% CI: 19±6.22%). However, the high number of cases classified as indeterminate demonstrates the challenges in classifying PASC even among experienced clinicians. Lastly, this study developed a dashboard to display views of aggregated PASC symptoms and measured its utility using the System Usability Scale. Overall comments related to the dashboard's potential were positive. This pipeline is crucial for monitoring post-COVID-19 patients with potential for use in clinical settings.

Novel Aspects in Pattern Formation Arise from Coupling Turing Reaction-Diffusion and Chemotaxis.

Recent experimental studies on primary hair follicle formation and feather bud morphogenesis indicate a coupling between Turing-type diffusion driven instability and chemotactic patterning. Inspired by these findings we develop and analyse a mathematical model that couples chemotaxis to a reaction-diffusion system exhibiting diffusion-driven (Turing) instability. While both systems, reaction-diffusion systems and chemotaxis, can independently generate spatial patterns, we were interested in how the coupling impacts the stability of the system, parameter region for patterning, pattern geometry, as well as the dynamics of pattern formation. We conduct a classical linear stability analysis for different model structures, and confirm our results by numerical analysis of the system. Our results show that the coupling generally increases the robustness of the patterning process by enlarging the pattern region in the parameter space. Concerning time scale and pattern regularity, we find that an increase in the chemosensitivity can speed up the patterning process for parameters inside and outside of the Turing space, but generally reduces spatial regularity of the pattern. Interestingly, our analysis indicates that pattern formation can also occur when neither the Turing nor the chemotaxis system can independently generate pattern. On the other hand, for some parameter settings, the coupling of the two processes can extinguish the pattern formation, rather than reinforce it. These theoretical findings can be used to corroborate the biological findings on morphogenesis and guide future experimental studies. From a mathematical point of view, this work sheds a light on coupling classical pattern formation systems from the parameter space perspective.

Early Season Monitoring of Tarnished Plant Bug, Lygus lineolaris, in Wild Hosts Using Pheromone Traps.

The tarnished plant bug, Lygus lineolaris (Hemiptera: Miridae), has a wide host range of over 700 plant species, including 130 crops of economic importance. During early spring, managing the field edges with weeds and other wild hosts is important in preventing early-season infestations of L. lineolaris in cotton to prevent damage to the squares and other fruiting structures. Scouting fields for L. lineolaris is time- and labor-intensive, and end-user variability associated with field sampling can lead to inaccuracies. Insect traps that combine visual cues and pheromones are more accurate, sustainable, and economically feasible in contrast to traditional insect detection methods. In this study, we investigated the application of red or white sticky cards baited with the female-produced sex pheromone to monitor overwintering L. lineolaris populations in early spring. Field experiments demonstrated that the red sticky cards baited with a pheromone blend containing hexyl butyrate, (E)-2-hexenyl butyrate, and (E)-4-oxo-2-hexenal in 4:10:7 ratio are highly effective in trapping L. lineolaris adults in early spring before the row crops are planted, and in monitoring their movement into a cotton crop. The monitoring of L. lineolaris should help growers to make judicious decisions on insecticide applications to control early pest infestations, thereby reducing economic damage to cotton.

Rapid mechanosensitive migration and dispersal of newly divided mesenchymal cells aid their recruitment into dermal condensates.

Embryonic mesenchymal cells are dispersed within an extracellular matrix but can coalesce to form condensates with key developmental roles. Cells within condensates undergo fate and morphological changes and induce cell fate changes in nearby epithelia to produce structures including hair follicles, feathers, or intestinal villi. Here, by imaging mouse and chicken embryonic skin, we find that mesenchymal cells undergo much of their dispersal in early interphase, in a stereotyped process of displacement driven by 3 hours of rapid and persistent migration followed by a long period of low motility. The cell division plane and the elevated migration speed and persistence of newly born mesenchymal cells are mechanosensitive, aligning with tissue tension, and are reliant on active WNT secretion. This behaviour disperses mesenchymal cells and allows daughters of recent divisions to travel long distances to enter dermal condensates, demonstrating an unanticipated effect of cell cycle subphase on core mesenchymal behaviour.

UMAD1 contributes to ESCRT-III dynamic subunit turnover during cytokinetic abscission.

Abscission is the final stage of cytokinesis whereby the midbody, a thin intercellular bridge, is resolved to separate the daughter cells. Cytokinetic abscission is mediated by the endosomal sorting complex required for transport (ESCRT), a conserved membrane remodelling machinery. The midbody organiser CEP55 recruits early acting ESCRT factors such as ESCRT-I and ALIX (also known as PDCD6IP), which subsequently initiate the formation of ESCRT-III polymers that sever the midbody. We now identify UMAD1 as an ESCRT-I subunit that facilitates abscission. UMAD1 selectively associates with VPS37C and VPS37B, supporting the formation of cytokinesis-specific ESCRT-I assemblies. TSG101 recruits UMAD1 to the site of midbody abscission, to stabilise the CEP55-ESCRT-I interaction. We further demonstrate that the UMAD1-ESCRT-I interaction facilitates the final step of cytokinesis. Paradoxically, UMAD1 and ALIX co-depletion has synergistic effects on abscission, whereas ESCRT-III recruitment to the midbody is not inhibited. Importantly, we find that both UMAD1 and ALIX are required for the dynamic exchange of ESCRT-III subunits at the midbody. Therefore, UMAD1 reveals a key functional connection between ESCRT-I and ESCRT-III that is required for cytokinesis.

Entomopathogenicity of Ascomycete Fungus Cordyceps militaris on the Cotton Bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae).

This study investigated the exposure of the cotton bollworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) to a novel pathogenic fungal agent historically associated with human medicinal value, a commercial strain of Cordyceps militaris ((L.) Fr.) Vuill. (Hypocreales). A series of comparative studies were conducted to evaluate the efficacy of two different exposure methods using four concentrations (n × 109, n × 108, n × 107, n × 106) of C. militaris, where n × 109 provided a concentration of approximately 420 ± 37 spores per mm2 with 398 ± 28 viable spores. Survival of cotton bollworms of all stages was not affected by C. militaris at any concentration 1 d post-exposure. The greatest reduction in survival and highest sporulation rates were observed primarily on or after 7 d post-exposure for early instars (first and second). Significant declines in the survival of early instars were observed for all concentrations at 7 d, and 95% mortality by 10 d, with the exception of the fifth instars that experienced a less severe reduction in survival (35%) when exposed to any concentrations used in the study. Survival of late instars (third to fifth) ranged from 44% to 68% on day 10, while adult survival was near 99% across the duration of the experiment. The relatively narrow range observed for both the lethal concentration and sporulation of second, third, and fifth instar cotton bollworms exposed to the C. militaris strain may demonstrate potential field application for control of larval populations of cotton bollworms.

A chromosome scale assembly of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), genome.

OBJECTIVE: The tarnished plant bug (TPB), Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), is a pest damaging many cultivated crops in North America. Although partial transcriptome data are available for this pest, a genome assembly was not available for this species. This assembly of a high-quality chromosome-length genome of TPB is aimed to develop the genetic resources that can provide the foundation required for advancing research on this species. RESULTS: The initial genome of TPB assembled with paired-end nucleotide sequences generated with Illumina technology was scaffolded with Illumina HiseqX reads generated from a proximity ligated (HiC) library to obtain a high-quality genome assembly. The final assembly contained 3963 scaffolds longer than 1 kbp to yield a genome of 599.96 Mbp. The N50 of the TPB genome assembly was 35.64 Mbp and 98.68% of the genome was assembled into 17 scaffolds larger than 1 Mbp. This megabase scaffold number is the same as the number of chromosomes observed in karyotyping of this insect. The TPB genome is known to have high repetitive DNA content, and the reduced assembled genome size compared to flowcytometric estimates of approximately 860 Mbp may be due to the collapsed assembly of highly similar regions.

The developmental basis of fingerprint pattern formation and variation.

Fingerprints are complex and individually unique patterns in the skin. Established prenatally, the molecular and cellular mechanisms that guide fingerprint ridge formation and their intricate arrangements are unknown. Here we show that fingerprint ridges are epithelial structures that undergo a truncated hair follicle developmental program and fail to recruit a mesenchymal condensate. Their spatial pattern is established by a Turing reaction-diffusion system, based on signaling between EDAR, WNT, and antagonistic BMP pathways. These signals resolve epithelial growth into bands of focalized proliferation under a precociously differentiated suprabasal layer. Ridge formation occurs as a set of waves spreading from variable initiation sites defined by the local signaling environments and anatomical intricacies of the digit, with the propagation and meeting of these waves determining the type of pattern that forms. Relying on a dynamic patterning system triggered at spatially distinct sites generates the characteristic types and unending variation of human fingerprint patterns.

Lethal Concentration and Sporulation by Contact and Direct Spray of the Entomopathogenic Fungus Beauveria bassiana on Different Stages of Nezara viridula (Heteroptera: Pentatomidae).

The southern green stink bug, Nezara viridula (L.) (Heteroptera: Pentatomidae) is the most significant pest of soybean worldwide. The present study was conducted to compare the effectiveness of a Delta native strain NI8 of Beauveria bassiana by contact and direct spray on nymphs (2nd to 5th instar) and adults of N. viridula. Water control and four concentrations of B. bassiana were used to evaluate the survival, mortality, and molting percentage and to estimate median lethal concentration (LC50), median lethal sporulation (LS50), and resistance ratio (RR50). Direct spray at all concentrations observed the greatest reduction in survival on all life stages. Mortality and sporulation were positively correlated by concentration, while molting was highly variable with a significantly lower negative correlation on insects that were directly sprayed. Pathogenicity exhibited reduction as young stages developed and emerged to adult. The LC50 (Contact: 612 spores/mm2; Direct spray: 179 spores/mm2) and LS50 (Contact: 1960 spores/mm2 Spray: 3.3 × 106) values showed that adults of N. viridula were highly resistant than any other life stage when exposed to either contact or direct spray. Fourth instar was the most susceptible (LC50: Contact: 18 spores/mm2; Direct spray: 23 spores/mm2) (LS50: Contact: 53 spores/mm2; Direct spray: 26 spores/mm2) followed by second, third, and fifth instars.

Susceptibility of Different Life Stages of Kudzu Bug Megacopta cribraria (F.) (Hemiptera: Plataspidae) to Two Different Native Strains of Beauveria bassiana.

This is the first study that examined and compared the survival, LC50, and RR50 estimates of Megacopta cribraria F. (Hemiptera: Plataspidae) nymphs and adults that were exposed to two native Beauveria bassiana isolates (Previously codified as NI8 and KUDSC strains) at four concentrations. The greatest reduction in survival and mortality was observed primarily on or after 10 d post-exposure to B. bassiana isolates. Survival of early instars (2nd, 3rd) were not affected by either strains or concentration at 3 d and 5 d post-exposure. Survival of later instars (5th) and adults was significantly reduced when exposed to the KUDSC strain at all concentrations. Comparison of dose−mortality values (LC50) using resistance ratios (RR50) were significantly different between life stages of the kudzu bug for both strains of B. bassiana. The LC50 values showed that kudzu bug adults are more susceptible than any other life stage when exposed to either strain. The KUDSC strain was more pathogenic than NI8 10 d after exposure, but NI8 exhibited significantly higher pathogenicity than KUDSC 20 d after exposure. Our results suggest potential field application of B. bassiana for kudzu bug control and their integration into pest management strategies to suppress them before they cause economic damage to soybean crops.

The EDA-deficient mouse has Zymbal's gland hypoplasia and acute otitis externa.

In mice, rats, dogs and humans, the growth and function of sebaceous glands and eyelid Meibomian glands depend on the ectodysplasin signalling pathway. Mutation of genes encoding the ligand EDA, its transmembrane receptor EDAR and the intracellular signal transducer EDARADD leads to hypohidrotic ectodermal dysplasia, characterised by impaired development of teeth and hair, as well as cutaneous glands. The rodent ear canal has a large auditory sebaceous gland, the Zymbal's gland, the function of which in the health of the ear canal has not been determined. We report that EDA-deficient mice, EDAR-deficient mice and EDARADD-deficient rats have Zymbal's gland hypoplasia. EdaTa mice have 25% prevalence of otitis externa at postnatal day 21 and treatment with agonist anti-EDAR antibodies rescues Zymbal's glands. The aetiopathogenesis of otitis externa involves infection with Gram-positive cocci, and dosing pregnant and lactating EdaTa females and pups with enrofloxacin reduces the prevalence of otitis externa. We infer that the deficit of sebum is the principal factor in predisposition to bacterial infection, and the EdaTa mouse is a potentially useful microbial challenge model for human acute otitis externa.

Limb development genes underlie variation in human fingerprint patterns.

Fingerprints are of long-standing practical and cultural interest, but little is known about the mechanisms that underlie their variation. Using genome-wide scans in Han Chinese cohorts, we identified 18 loci associated with fingerprint type across the digits, including a genetic basis for the long-recognized "pattern-block" correlations among the middle three digits. In particular, we identified a variant near EVI1 that alters regulatory activity and established a role for EVI1 in dermatoglyph patterning in mice. Dynamic EVI1 expression during human development supports its role in shaping the limbs and digits, rather than influencing skin patterning directly. Trans-ethnic meta-analysis identified 43 fingerprint-associated loci, with nearby genes being strongly enriched for general limb development pathways. We also found that fingerprint patterns were genetically correlated with hand proportions. Taken together, these findings support the key role of limb development genes in influencing the outcome of fingerprint patterning.

Biology, Ecology, and Pest Management of the Tarnished Plant Bug, Lygus lineolaris (Palisot de Beauvois) in Southern Row Crops.

The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), (Hemiptera: Miridae) is considered the most damaging pest of cotton (Gossypium hirsutum L.) in the mid-southern United States, although it is established throughout the United States, southern Canada, and northern Mexico. The introduction of transgenic crops for the control of moths in the Heliothine complex and eradication of the boll weevil, Anthonomus grandis, from much of the United States led to greatly reduced pesticide use in cotton fields, which allowed L. lineolaris to emerge as a new primary pest of cotton in the mid-southern United States. Since the publication of a review by Layton (2000) on damage caused by Lygus lineolaris, many new studies have been published on the changes in host range, population dynamics, sampling methods and thresholds, cultural practices, sex pheromones and attractant blends, novel pesticides and insecticide resistance mechanisms, olfactory and feeding behaviors, introduction of biological control agents, host-plant resistance mechanisms, and new molecular and genetic tools for integrated pest management of Lygus species in cotton and other important crops. Here, we review and discuss the latest developments in L. lineolaris research in the last two decades.

Intrawound vancomycin application after spinal surgery: a propensity score-matched cohort analysis.

OBJECTIVE: Surgical site infection (SSI) following spine surgery is associated with increased morbidity and healthcare costs. In an effort to reduce SSI rates, the application of intrawound vancomycin powder has gained popularity. However, there is limited high-quality evidence to support the safety and efficacy of this practice. The authors sought to determine if intrawound application of vancomycin powder improves 90-day overall SSI rates. METHODS: The authors performed a retrospective, vancomycin exposure-matched cohort study at a single tertiary care hospital over 21 months. They included all patients undergoing elective spinal surgery and stratified the patients into two groups: those who received intrawound vancomycin powder application and those who received no application of vancomycin powder. The primary outcome of interest was the 90-day overall SSI rate. Secondary outcomes included rates of superficial SSI, deep SSI, wound disruption, and a post hoc analysis of the microbiology and minimum inhibitory concentrations. Baseline patient demographics, clinical presentation, comorbidities, perioperative factors, and 90-day postoperative outcomes were manually abstracted from patient charts. To mitigate bias, we performed 1:1 matching after calculating propensity scores and identified 1 patient from the no-vancomycin cohort for each patient in the vancomycin cohort. RESULTS: A total of 997 patients met our inclusion criteria (473 patients receiving vancomycin and 524 patients not receiving vancomycin). Propensity score matching produced 221 matched pairs. Risk-adjusted analysis demonstrated similar overall SSI rates between the groups (OR 1.9, p = 0.329). On unadjusted analysis, the overall 90-day SSI rate was greater in the vancomycin group (n = 10 [4.5%]) than in the no-vancomycin group (n = 5 [2.3%]) (p < 0.001), as were the superficial SSI rate (7 [3.2%] vs 4 [1.8%], p < 0.001), deep SSI rate (3 [1.4%] vs 1 [0.5%], p < 0.001), and wound disruption rate (5 [2.3%] vs 1 [0.5%], p < 0.001). No cultured isolate demonstrated vancomycin resistance. CONCLUSIONS: The authors observed no difference in SSI rates after the intrawound application of vancomycin powder during spine surgery. Vancomycin use did not contribute to antimicrobial resistance; however, it may select out gram-negative bacteria and increase rates of wound disruption.

Complete Genome Sequence of Serratia sp. Strain CC119, Associated with Inner Cotton Boll Rot via Insect Vector Transmission.

Serratia species are Gram-negative bacteria that can infect both animals and plants. The annotated genome presented is the first for a Serratia sp. strain (called CC119) that is a cotton boll pathogen. The opportunistic strain is associated with the boll-piercing-sucking insect Creontiades signatus.

Effect of Oxytetracycline-Hydrochloride on Phytoplasma Titer and Symptom Progression of the 16SrIV-D Phytoplasma in Cabbage Palms from Florida.

Lethal bronzing (LB) is a phytoplasma disease of palms in Florida, U.S.A. Historically, the use of the antibiotic oxytetracycline-hydrochloride (OTC-HCl) was optimized to treat for LY, and currently label rates for OTC-HCl developed for LY are being used to treat palms preventatively against LB. Because of the economic impact of LB, assessing OTC-HCl against LB is essential for developing and optimizing management options for this disease. In this study, Sabal palmetto palms declining from LB were injected with OTC-HCl to assess efficacy on LB. Four groups of palms were selected that represented healthy palms, early symptomatic palms, palms with moderate symptoms, and palms with late-stage symptoms. Within each group, treatment palms injected with OTC-HCl and a control group with no antibiotics were tested weekly by quantitative PCR for 1 year. For asymptomatic palms, treated palms never developed symptoms or tested positive, whereas one of the control palms did. The early symptomatic palms that were treated had similar levels of phytoplasma to the control group but had much slower symptom development. Palms with moderate symptoms had no difference in titer between the treatment and the control group, but treated palms had much slower symptom development. Palms with late-stage symptoms showed no difference in phytoplasma titer or symptom progression between treated and control palms. These results suggest that label rates of OTC-HCl appear useful as a preventative against LB, but once symptoms develop, label rates cannot cure palms. In the meantime, removal of symptomatic palms is recommended.

Predictors of elderly mortality after trauma: A novel outcome score.

INTRODUCTION: Elderly trauma patients are at high risk for mortality, even when presenting with minor injuries. Previous prognostic models are poorly used because of their reliance on elements unavailable during the index hospitalization. The purpose of this study was to develop a predictive algorithm to accurately estimate in-hospital mortality using easily available metrics. METHODS: The National Trauma Databank was used to identify patients 65 years and older. Data were split into derivation (2007-2013) and validation (2014-2015) data sets. There was no overlap between data sets. Factors included age, comorbidities, physiologic parameters, and injury types. A two-tiered scoring system to predict in-hospital mortality was developed: a quick elderly mortality after trauma (qEMAT) score for use at initial patient presentation and a full EMAT (fEMAT) score for use after radiologic evaluation. The final model (stepwise forward selection, p < 0.05) was chosen based on calibration and discrimination analysis. Calibration (Brier score) and discrimination (area under the receiving operating characteristic curve [AuROC]) were evaluated. Because National Trauma Databank did not include blood product transfusion, an element of the Geriatric Trauma Outcome Score (GTOS), a regional trauma registry was used to compare qEMAT versus GTOS. A mobile-based application is currently available for cost-free utilization. RESULTS: A total of 840,294 patients were included in the derivation data set and 427,358 patients in the validation data set. The fEMAT score (median, 91; S.D., 82-102) included 26 factors, and the qEMAT score included eight factors. The AuROC was 0.86 for fEMAT (Brier, 0.04) and 0.84 for qEMAT. The fEMAT outperformed other trauma mortality prediction models (e.g., Trauma and Injury Severity Score-Penetrating and Trauma and Injury Severity Score-Blunt, age + Injury Severity Score). The qEMAT outperformed the GTOS (AuROC, 0.87 vs. 0.83). CONCLUSION: The qEMAT and fEMAT accurately estimate the probability of in-hospital mortality and can be easily calculated on admission. This information could aid in deciding transfer to tertiary referral center, patient/family counseling, and palliative care utilization. LEVEL OF EVIDENCE: Epidemiological Study, level IV.

Transmission of Cotton Seed and Boll Rotting Bacteria by the Verde Plant Bug (Hemiptera: Miridae).

Field experiments and supporting laboratory work were conducted to characterize the ability of the verde plant bug, Creontiades signatus (Distant), a boll-feeding sucking bug, to transmit a cotton seed and boll rot bacterial pathogen, Serratia marcescens (Bizio) (Enterobacteriales: Enterobacteriaceae). Serratia marcescens was originally isolated from bolls infested with verde plant bug in south Texas, and a Rifampicin resistant S. marcescens strain was used in transmission and retention experiments. Serratia-exposed and nonexposed adult verde plant bugs from a laboratory colony were placed individually on 5-, 6-, 7-, and 8-d-old bolls (postanthesis). The bacterial acquisition process did not apparently affect insect vigor based on similar average boll injury ratings observed across both exposed and nonexposed bugs. Cotton bolls caged with Serratia-exposed verde plant bugs had significantly greater presence of S. marcescens and cotton boll rot symptoms than bolls caged without bugs (no-insect controls) or nonexposed bugs. Transmission of the disease agent by verde plant bug was confirmed across all boll ages assayed. Incidence of diseased locules on 5- and 6-d-old bolls was the same or greater than on 7- and 8-d-old bolls. Verde plant bug was able to harbor the disease agent from 24- to 96-h postinfection, and transmission efficiency rates ranged from 54 to 62% during initial transmission and retention (transmission across two bolls fed upon consecutively) studies. Along with photographic evidence, the experimental data supported that boll damage associated with verde plant bug infestations was magnified when insects transmitted the cotton pathogen S. marcescens as demonstrated in this 2-yr field experiment.

Photoperiod-Specific Within-Plant Distribution of the Green Stink Bug (Hemiptera: Pentatomidae) on Cotton.

Sampling methods for detecting stink bugs are intensive, time-consuming, and yield variable results. In a 2-yr mark-release-observe experiment, over 500 adult green stink bugs, Chinavia hilaris (Say) (Hemiptera: Pentatomidae), were used to test for variation in nocturnal and diurnal insect distribution patterns on cotton. Field-collected stink bugs were marked or unmarked with nontoxic fluorescent sharpie markers, released, and monitored in cotton fields at peak bloom. Stink bugs were monitored visually during day and night, aided by a handheld blacklight for nighttime observations. Within-cotton distribution insect observations were categorized by plant section (i.e., bottom, middle, and top branches), by fruiting positions and leaf surface, and by concealed or exposed orientation on floral bracts and leaf surfaces. Green stink bugs were primarily distributed on the middle and top branches irrespective of photoperiod, and on bolls in first position from the main stem. Differences in stink bugs observed concealed or exposed on fruiting structures were detected. During daytime, stink bugs were primarily observed inside the bract of bolls, and when detected on leaves concealed on the lower surface. In contrast, stink bugs were primarily outside the bract of bolls at night, and when detected on leaves were exposed on an upper surface. These results support focus on assessing internal boll injury for evaluating stink bug injury to avoid the challenges in stink bug detection observed here, and point to additional study to refine stink bug density estimation when needed.