Standard-of-care systemic therapy with or without stereotactic body radiotherapy in patients with oligoprogressive breast cancer or non-small-cell lung cancer (Consolidative Use of Radiotherapy to Block [CURB] oligoprogression): an open-label, randomised, controlled, phase 2 study.

BACKGROUND: Most patients with metastatic cancer eventually develop resistance to systemic therapy, with some having limited disease progression (ie, oligoprogression). We aimed to assess whether stereotactic body radiotherapy (SBRT) targeting oligoprogressive sites could improve patient outcomes. METHODS: We did a phase 2, open-label, randomised controlled trial of SBRT in patients with oligoprogressive metastatic breast cancer or non-small-cell lung cancer (NSCLC) after having received at least first-line systemic therapy, with oligoprogression defined as five or less progressive lesions on PET-CT or CT. Patients aged 18 years or older were enrolled from a tertiary cancer centre in New York, NY, USA, and six affiliated regional centres in the states of New York and New Jersey, with a 1:1 randomisation between standard of care (standard-of-care group) and SBRT plus standard of care (SBRT group). Randomisation was done with a computer-based algorithm with stratification by number of progressive sites of metastasis, receptor or driver genetic alteration status, primary site, and type of systemic therapy previously received. Patients and investigators were not masked to treatment allocation. The primary endpoint was progression-free survival, measured up to 12 months. We did a prespecified subgroup analysis of the primary endpoint by disease site. All analyses were done in the intention-to-treat population. The study is registered with ClinicalTrials.gov, NCT03808662, and is complete. FINDINGS: From Jan 1, 2019, to July 31, 2021, 106 patients were randomly assigned to standard of care (n=51; 23 patients with breast cancer and 28 patients with NSCLC) or SBRT plus standard of care (n=55; 24 patients with breast cancer and 31 patients with NSCLC). 16 (34%) of 47 patients with breast cancer had triple-negative disease, and 51 (86%) of 59 patients with NSCLC had no actionable driver mutation. The study was closed to accrual before reaching the targeted sample size, after the primary efficacy endpoint was met during a preplanned interim analysis. The median follow-up was 11·6 months for patients in the standard-of-care group and 12·1 months for patients in the SBRT group. The median progression-free survival was 3·2 months (95% CI 2·0-4·5) for patients in the standard-of-care group versus 7·2 months (4·5-10·0) for patients in the SBRT group (hazard ratio [HR] 0·53, 95% CI 0·35-0·81; p=0·0035). The median progression-free survival was higher for patients with NSCLC in the SBRT group than for those with NSCLC in the standard-of-care group (10·0 months [7·2-not reached] vs 2·2 months [95% CI 2·0-4·5]; HR 0·41, 95% CI 0·22-0·75; p=0·0039), but no difference was found for patients with breast cancer (4·4 months [2·5-8·7] vs 4·2 months [1·8-5·5]; 0·78, 0·43-1·43; p=0·43). Grade 2 or worse adverse events occurred in 21 (41%) patients in the standard-of-care group and 34 (62%) patients in the SBRT group. Nine (16%) patients in the SBRT group had grade 2 or worse toxicities related to SBRT, including gastrointestinal reflux disease, pain exacerbation, radiation pneumonitis, brachial plexopathy, and low blood counts. INTERPRETATION: The trial showed that progression-free survival was increased in the SBRT plus standard-of-care group compared with standard of care only. Oligoprogression in patients with metastatic NSCLC could be effectively treated with SBRT plus standard of care, leading to more than a four-times increase in progression-free survival compared with standard of care only. By contrast, no benefit was observed in patients with oligoprogressive breast cancer. Further studies to validate these findings and understand the differential benefits are warranted. FUNDING: National Cancer Institute.

Genetic diversity of the entomopathogenic fungus Metarhizium rileyi based on de novo microsatellite markers.

Epizootics of the entomopathogenic fungus Metarhizium rileyi regulate lepidopteran populations in soybean, cotton, and peanut agroecosystems to the point that insecticide applications could be unnecessary. However, the contribution and how different strains operate during the epizootic are unknown. Several unanswered questions remain: 1. How many genotypes of M. rileyi are present during an epizootic? 2. Which genotype is the most common among them? 3. Are the genotypes involved in annual epizootics at the same location the same? Therefore, the development of molecular markers to accurately identify these genotypes is very important to answer these questions. SSR primers were designed by prospecting in silico to discriminate genotypes and infer the genetic diversity of M. rileyi isolates from the collection kept at Embrapa Soybean. We tested 13 SSR markers on 136 isolates to identify 43 clones and 12 different genetic clusters, with genetic diversity ranging from Hs = 0.15 (cluster I) to Hs = 0.41 (cluster IV) and an average diversity of 0.24. No clusters were categorically distinguished based on hosts or geographical origin using Bayesian clustering analysis. Nonetheless, some clusters comprised most of the isolates with a common geographic origin; for example, cluster VIII was mainly composed of isolates from Central-western Brazil, cluster II from Southern Brazil, and cluster XII from Quincy, Northern Florida, in the United States. Underrepresented regions (few isolates) from Pacific Island nations of Japan, the Philippines, and Indonesia (specifically from Java) were placed into clusters IX and X. Although the analyzed isolates displayed evidence of clonal structure, the genetic diversity indices suggest a potential for the species to adapt to different environmental conditions.

Insurance Denials and Patient Treatment in a Large Academic Radiation Oncology Center.

Importance: Insurance barriers to cancer care can cause significant patient and clinician burden. Objective: To investigate the association of insurance denial with changes in technique, dose, and time to delivery of radiation oncology treatment. Design, Setting, and Participants: In this single-institution cohort analysis, data were collected from patients with payer-denied authorization for radiation therapy (RT) from November 1, 2021, to December 8, 2022. Data were analyzed from December 15, 2022, to December 31, 2023. Exposure: Insurance denial for RT. Main Outcomes and Measures: Association of these denials with changes in RT technique, dose, and time to treatment delivery was assessed using χ2 tests. Results: A total of 206 cases (118 women [57.3%]; median age, 58 [range, 26-91] years) were identified. Most insurers (199 [96.6%]) were commercial payers, while 7 (3.4%) were Medicare or Medicare Advantage. One hundred sixty-one patients (78.2%) were younger than 65 years. Of 206 cases, 127 (61.7%) were ultimately authorized without any change to the requested RT technique or prescription dose; 56 (27.2%) were authorized after modification to RT technique and/or prescription dose required by the payer. Of 21 cases with required prescription dose change, the median decrease in dose was 24.0 (range, 2.3-51.0) Gy. Of 202 cases (98.1%) with RT delivered, 72 (34.9%) were delayed for a mean (SD) of 7.8 (9.1) days and median of 5 (range, 1-49) days. Four cases (1.9%) ultimately did not receive any authorization, with 3 (1.5%) not undergoing RT, and 1 (0.5%) seeking treatment at another institution. Conclusions and Relevance: In this cohort study of patients with payer-denied cases, most insurance denials in radiation oncology were ultimately approved on appeal; however, RT technique and/or effectiveness may be compromised by payer-mandated changes. Further investigation and action to recognize the time and financial burdens on clinicians and clinical effects on patients caused by insurance denials of RT is needed.

Disparities in Stage at Presentation Among Hispanic and Latinx Patients With Non-Small-Cell Lung Cancer in the United States.

PURPOSE: Hispanic and Latinx people in the United States are the fastest-growing ethnic group. However, previous studies in non-small-cell lung cancer (NSCLC) often analyze these diverse communities in aggregate. We aimed to identify differences in NSCLC stage at diagnosis in the US population, focusing on disaggregated Hispanic/Latinx individuals. METHODS: Data from the National Cancer Database from 2004 to 2018 identified patients with primary NSCLC. Individuals were disaggregated by racial and ethnic subgroup and Hispanic country of origin. Ordinal logistic regression adjusting for age, facility type, income, educational attainment, comorbidity index, insurance, and year of diagnosis was used to create adjusted odds ratios (aORs), with higher odds representing diagnosis at later-stage NSCLC. RESULTS: Of 1,565,159 patients with NSCLC, 46,616 were Hispanic/Latinx (3.0%). When analyzed in the setting of race and ethnicity, Hispanic patients were more likely to be diagnosed with metastatic disease compared with non-Hispanic White (NHW) patients: 47.0% for Hispanic Black, 46.0% Hispanic White, and 44.3% of Hispanic other patients versus 39.1% of non-Hispanic White patients (P < .001 for all). By country of origin, 51.4% of Mexican, 41.7% of Puerto Rican, 44.6% of Cuban, 50.8% of South or Central American, 48.4% of Dominican, and 45.6% of other Hispanic patients were diagnosed with metastatic disease, compared with 39.1% of NHWs. Conversely, 20.2% of Mexican, 26.9% of Puerto Rican, 24.2% of Cuban, 22.5% of South or Central American, 23.7% of Dominican, and 24.5% of other Hispanic patients were diagnosed with stage I disease, compared with 30.0% of NHWs. All Hispanic groups were more likely to present with later-stage NSCLC than NHW patients (greatest odds for Mexican patients, aOR, 1.44; P < .001). CONCLUSION: Hispanic/Latinx patients with non-small-cell lung cancer were more likely to be diagnosed with advanced disease compared with NHWs. Disparities persisted upon disaggregation by both race and country of origin, with over half of Mexican patients with metastatic disease at diagnosis. Disparities among Hispanic/Latinx groups by race and by country of origin highlight the shortcomings of treating these groups as a monolith and underscore the need for disaggregated research and targeted interventions.

An 18F-FDG PET/CT and Mean Lung Dose Model to Predict Early Radiation Pneumonitis in Stage III Non-Small Cell Lung Cancer Patients Treated with Chemoradiation and Immunotherapy.

Radiation pneumonitis (RP) that develops early (i.e., within 3 mo) (RPEarly) after completion of concurrent chemoradiation (cCRT) leads to treatment discontinuation and poorer survival for patients with stage III non-small cell lung cancer. Since no RPEarly risk model exists, we explored whether published RP models and pretreatment 18F-FDG PET/CT-derived features predict RPEarly Methods: One hundred sixty patients with stage III non-small cell lung cancer treated with cCRT and consolidative immunotherapy were analyzed for RPEarly Three published RP models that included the mean lung dose (MLD) and patient characteristics were examined. Pretreatment 18F-FDG PET/CT normal-lung SUV featured included the following: 10th percentile of SUV (SUVP10), 90th percentile of SUV (SUVP90), SUVmax, SUVmean, minimum SUV, and SD. Associations between models/features and RPEarly were assessed using area under the receiver-operating characteristic curve (AUC), P values, and the Hosmer-Lemeshow test (pHL). The cohort was randomly split, with similar RPEarly rates, into a 70%/30% derivation/internal validation subset. Results: Twenty (13%) patients developed RPEarly Predictors for RPEarly were MLD alone (AUC, 0.72; P = 0.02; pHL, 0.87), SUVP10, SUVP90, and SUVmean (AUC, 0.70-0.74; P = 0.003-0.006; pHL, 0.67-0.70). The combined MLD and SUVP90 model generalized in the validation subset and was deemed the final RPEarly model (RPEarly risk = 1/[1+e(- x )]; x = -6.08 + [0.17 × MLD] + [1.63 × SUVP90]). The final model refitted in the 160 patients indicated improvement over the published MLD-alone model (AUC, 0.77 vs. 0.72; P = 0.0001 vs. 0.02; pHL, 0.65 vs. 0.87). Conclusion: Patients at risk for RPEarly can be detected with high certainty by combining the normal lung's MLD and pretreatment 18F-FDG PET/CT SUVP90 This refined model can be used to identify patients at an elevated risk for premature immunotherapy discontinuation due to RPEarly and could allow for interventions to improve treatment outcomes.

Biomarkers associated with pulmonary exacerbations in a randomized trial of nintedanib for radiation pneumonitis.

BACKGROUND AND PURPOSE: Radiation pneumonitis (RP) is a common side effect of thoracic radiotherapy and often has a long course characterized by acute exacerbations and progression to permanent lung fibrosis. There are no validated biomarkers of prognosis in patients diagnosed with RP. MATERIALS AND METHODS: We analyzed a time course of serum chemokines, cytokines, and other proteins from patients with grade 2+ RP in a randomized clinical trial of a steroid taper plus nintedanib, a multiple tyrosine kinase inhibitor, versus placebo plus a steroid taper for the treatment of RP. Weighted gene correlation network analysis (WGCNA) and univariable zero inflated Poisson models were used to identify groups of correlated analytes and their associations with clinical outcomes. RESULTS: Thirty enrolled patients had biomarker data available, and 17 patients had enough analytes tested for network analysis. WGNCA identified ten analytes, including transforming growth factor beta-1 (TGF-ß1), monocyte chemoattractant protein-1 (MCP-1), and platelet-derived growth factor (PDGF), that in aggregate were correlated with the occurrence of pulmonary exacerbations (p = 0.008), the total number of acute pulmonary exacerbations (p = 0.002), and treatment arm (p = 0.036). By univariable analysis, an increase in rate of change of two components of the RP module were associated with an increased incidence rate of pulmonary exacerbations: interleukin 5 (IL-5, incidence rate ratio (IRR) 1.02, 95% CI 1.01-1.04, p = 0.002), and tumor necrosis factor superfamily 12 (TNFSF12, IRR 1.06, CI 1-1.11, p = 0.036). An increased slope of epidermal growth factor (EGF) was associated with a decreased incidence rate of exacerbations (IRR 0.94, CI 0.89-1, p = 0.036). CONCLUSION: We identified a panel of serum biomarkers that showed association with nintedanib treatment and acute pulmonary exacerbations in patients with RP. A confirmatory study will be needed to validate this panel for use as a prognostic tool in patients with RP.

Clinical and Dosimetric Risk Factors Associated With Radiation-Induced Lung Toxicities After Multiple Courses of Lung Stereotactic Body Radiation Therapy.

Purpose: Data are limited on radiation-induced lung toxicities (RILT) after multiple courses of lung stereotactic body radiation therapy (SBRT). We herein analyze a large cohort of patients to explore the clinical and dosimetric risk factors associated with RILT in such settings. Methods and Materials: A single institutional database of patients treated with multiple courses of lung SBRT between January 2014 and December 2019 was analyzed. Grade 2 or higher (G2+) RILT after the last course of SBRT was the primary endpoint. Composite plans were generated with advanced algorithms including deformable registration and equivalent dose adjustment. Logistic regression analyses were performed to examine correlations between patient or treatment factors including dosimetry and G2+ RILT. Risk stratification of patients and lung constraints based on acceptable normal tissue complication probability were calculated based on risk factors identified. Results: Among 110 eligible patients (56 female and 54 male), there were 64 synchronous (58.2%; defined as 2 courses of SBRT delivered within 30 days) and 46 metachronous (41.8%) courses of SBRT. The composite median lung V20, lung V5, and mean lung dose were 9.9% (interquartile range [IQR], 7.3%-12.4%), 32.2% (IQR, 25.5%-40.1%), and 7.0 Gy (IQR, 5.5 Gy-8.6 Gy), respectively. With a median follow-up of 21.1 months, 30 patients (27.3%) experienced G2+ RILT. Five patients (4.5%) developed G3 RILT, and 1 patient (0.9%) developed G4 RILT, and no patients developed G5 RILT. On multivariable regression analysis, female sex (odds ratio [OR], 4.35; 95% CI, 1.49%-14.3%; P = .01), synchronous SBRT (OR, 8.78; 95% CI, 2.27%-47.8%; P = .004), prior G2+ RILT (OR, 29.8; 95% CI, 2.93%-437%; P = .007) and higher composite lung V20 (OR, 1.18; 95% CI, 1.02%-1.38%; P = .030) were associated with significantly higher likelihood of G2+ RILT. Conclusions: Our data suggest an acceptable incidence of G2+ RILT after multiple courses of lung SBRT. Female sex, synchronous SBRT, prior G2+ RILT, and higher composite lung V20 may be risk factors for G2+ RILT.

Clinical outcomes of stereotactic body radiation therapy for malignant pleural mesothelioma.

BACKGROUND: The objective of this study is to determine the outcomes and toxicities of patients with malignant pleural mesothelioma (MPM) treated with stereotactic body radiotherapy (SBRT). MATERIALS AND METHODS: Data were extracted from an institutional tumor registry for patients diagnosed with mesothelioma and treated with SBRT. Kaplan-Meier and Cox regression analyses were employed to determine local control (LC) and overall survival (OS). RESULTS: Forty-four patients with 59 total treated tumors from December 2006 to April 2022 were identified. Fifty-one (86.4 %) cases had oligoprogressive disease (five sites or less). The median prescription dose delivered was 3000 cGy in 5 fractions (range: 2700-6000 cGy in 3-8 fractions). Fifty-one (86.4 %) tumors were in the pleura, 4 (6.8 %) spine, 2 (3.4 %) bone, 1 (1.7 %) brain, and 1 (1.7 %) pancreas. The median follow-up from SBRT completion for those alive at last follow-up was 28 months (range: 14-52 months). The most common toxicities were fatigue (50.8 %), nausea (22.0 %), pain flare (15.3 %), esophagitis (6.8 %), dermatitis (6.8 %), and pneumonitis (5.1 %). There were no grade ≥ 3 acute or late toxicities. There were 2 (3.4 %) local failures, one of the pleura and another of the spine. One-year LC was 92.9 % (95 % CI: 74.6-98.2 %) for all lesions and 96.3 % (95 % CI: 76.5-99.5 %) for pleural tumors. One-year LC was 90.9 % (95 % CI: 68.1-97.6 %) for epithelioid tumors and 92.1 % (95 % CI: 72.1-98.0 %) for oligoprogressive tumors. One-year OS from time of SBRT completion was 36.4 % (95 % CI: 22.6-50.3 %). On multivariable analysis, KPS was the lone significant predictor for OS (p = 0.029). CONCLUSIONS: Our single-institutional experience on patients with MPM suggests that SBRT is safe with a low toxicity profile and potentially achieve good local control.

Stereotactic Body Radiation Therapy for Stage IIA to IIIA Inoperable Non-Small Cell Lung Cancer: A Phase 1 Dose-Escalation Trial.

PURPOSE: Larger tumors are underrepresented in most prospective trials on stereotactic body radiation therapy (SBRT) for inoperable non-small cell lung cancer (NSCLC). We performed this phase 1 trial to specifically study the maximum tolerated dose (MTD) of SBRT for NSCLC >3 cm. METHODS AND MATERIALS: A 3 + 3 dose-escalation design (cohort A) with an expansion cohort at the MTD (cohort B) was used. Patients with inoperable NSCLC >3 cm (T2-4) were eligible. Select ipsilateral hilar and single-station mediastinal nodes were permitted. The initial SBRT dose was 40 Gy in 5 fractions, with planned escalation to 50 and 60 Gy in 5 fractions. Adjuvant chemotherapy was mandatory for cohort A and optional for cohort B, but no patients in cohort B received chemotherapy. The primary endpoint was SBRT-related acute grade (G) 4+ or persistent G3 toxicities (Common Terminology Criteria for Adverse Events version 4.03). Secondary endpoints included local failure (LF), distant metastases, disease progression, and overall survival. RESULTS: The median age was 80 years; tumor size was >3 cm and ≤5 cm in 20 (59%) and >5 cm in 14 patients (41%). In cohort A (n = 9), 3 patients treated to 50 Gy experienced G3 radiation pneumonitis (RP), thus defining the MTD. In the larger dose-expansion cohort B (n = 25), no radiation therapy-related G4+ toxicities and no G3 RP occurred; only 2 patients experienced G2 RP. The 2-year cumulative incidence of LF was 20.2%, distant failure was 34.7%, and disease progression was 54.4%. Two-year overall survival was 53%. A biologically effective dose (BED) <100 Gy was associated with higher LF (P = .006); advanced stage and higher neutrophil/lymphocyte ratio were associated with greater disease progression (both P = .004). CONCLUSIONS: Fifty Gy in 5 fractions is the MTD for SBRT to tumors >3 cm. A higher BED is associated with fewer LFs even in larger tumors. Cohort B appears to have had less toxicity, possibly due to the omission of chemotherapy.

Optimal systemic treatment and real-world clinical application of ctDNA in patients with metastatic HER2-mutant lung cancer.

INTRODUCTION: No definitive answers currently exist regarding optimal first-line therapy for HER2-mutant NSCLC. Access to rapid tissue sequencing is a major barrier to precision drug development in the first-line setting. ctDNA analysis has the potential to overcome these obstacles and guide treatment. METHODS: We retrospectively analyzed patients with metastatic HER2-mutant NSCLC who underwent prospective clinical ctDNA sequencing and received systemic therapy at Memorial Sloan Kettering Cancer Center (MSK) from January 2016 to September 2022. HER2 mutations were identified by next-generation sequencing through MSK-IMPACT, MSK-ACCESS or Resolution ctDx LungTM assay. Primary endpoints were time to the next treatment (TTNT) and overall survival (OS). RESULTS: Sixty-three patients were included in the primary analysis. Chemoimmunotherapy (33/63, 52.4 %) was the predominant first-line treatment with a median TTNT of 5.1 months (95 %CI 4.1 - 6.1) whereas 55.0 % (22/40) of patients who received second-line T-DXd obtained a median TTNT of 9.2 m (95 % CI, 0-22.2). Plasma ctDNA was tested before first-line therapy in 40 patients with a median OS of 28.0 months (95 % CI 21-34), in whom 31 patients (78.0 %) had detectable ctDNA. HER2 mutations were detected on ctDNA with a median turnaround time of 13 days, occasionally co-occurred with EGFR and MET alterations and were tracked longitudinally correlating with treatment response. Patients with detectable baseline ctDNA had significantly shorter OS (hazard ratio (HR), 5.25; 95 % CI, 1.2-23.9; p = 0.019). CONCLUSION: Chemoimmunotherapy remains a major treatment option for metastatic HER2-mutant NSCLC. ctDNA can rapidly detect HER2 and co-mutations, and it has the potential to guide and monitor optimal first-line therapy. As a negative prognostic biomarker, detectable ctDNA at baseline would need to be taken into account for patient selection in future studies.

Genetic diversity of the sunflower caterpillar (Chlosyne lacinia saundersii Doubleday and Hewitson) (Lepidoptera: Nymphalidae) populations determined by molecular RAPD markers

Chlosyne lacinia saundersii is one of the most important pests of sunflower and it is the main target of insecticides applications. Larvae were collected in Londrina (PR), Santa Maria (RS), Dourados (MS), Ribeirão Preto (SP), Brasília (DF), Barreiras (BA), Uberaba (MG) and Vilhena (RO). Genomic DNA was extracted and amplified with ten-mer primers, which produced 101 loci. The size of the RAPD amplicons ranged from 180 to 2564 bp. Polymorphism among populations ranged from 31 percent to 67 percent, with the highest polymorphisms of 57 percent and 67 percent being detected in Uberaba and Vilhena populations, respectively. Populations with the highest similarity determined with Dice coefficient were from Ribeirão Preto and Barreiras, while insects from Londrina showed the highest similarity among them. Gene flow of C. lacinia saundersii 1.1 was lower than those previously observed for the noctuid Anticarsia gemmatalis Hübner, suggesting that C. lacinia saundersii populations are more isolated than the ones of this noctuid. Through the Analysis of Molecular Variance (AMOVA), RAPD variance was 33.64 percent among geographical populations and 66.36 percent within populations. These results suggest that populations of C. lacinia saundersii are genetically structured.

Chlosyne lacinia saundersii é uma das mais importantes pragas da cultura do girassol e o principal alvo das aplicações de inseticidas. As larvas foram coletadas em Londrina (PR), Santa Maria (RS), Dourados (MS), Ribeirão Preto (SP), Brasília (DF), Barreiras (BA), Uberaba (MG) e Vilhena (RO). O DNA genômico foi extraído e amplificado com dez primers, que produziram 101 locos. O tamanho das amplificações de RAPD variou de 180 a 2564 pb. O polimorfismo entre as populações variou de 31 por cento a 67 por cento, com maior polimorfismo 57 por cento e 67 por cento, detectado em populações de Uberaba e Vilhena, respectivamente. As populações com maior similaridade determinada com o coeficiente de Dice foram de Ribeirão Preto e Barreiras, enquanto os insetos coletados em Londrina apresentaram maior similaridade entre eles. O fluxo gênico de C. lacinia saundersii de 1,1 foi menor que o observado para a Anticarsia gemmatalis Hübner Noctuidae, sugerindo que as populações de C. lacinia saundersii estão mais isoladas do que estes noctuideos. Através da análise de variância molecular (AMOVA), RAPD a variação foi de 33,64 por cento entre as populações geográficas e 66,36 por cento dentro das populações. Estes resultados sugere que as populações de C. lacinia saundersii são geneticamente estruturadas.

Trypanosomatid prevalence in Nezara viridula (L), Euschistus heros (Fabricius) and Piezodorus guildinii (Westwood) (Heteroptera: Pentatomidae) populations in Northern Paraná, Brazil

The stink bug complex in soybeans has a great potential to cause damage, their control has been difficult due to high population densities at the end of the season. Studies related to entomopathogenic organisms in stink bugs are interesting for their possibilities to be used as biological control agents and because they could be an etiological agent in mass rearing. We determined the prevalence of trypanosomatids in the most important species of the stink bug complex in soybean, from Brazil, Nezara viridula (L.), Euschistus heros (Fabricius) and Piezodorus guildinii (Westwood). Insects were collected in soybean and on alternate hosts during the intercrop season. Hemolymph drops from each adult were mounted on slides and observed under microscope; stink bug numbers with and without flagellates were recorded. The flagellates found in P. guildinii or E. heros populations were smaller than that found in N. viridula. The higher prevalence of the trypanosomatids was observed on soybean reaching a peak of 30 percent of the sampled N. viridula specimens in January, 2001, and 44 percent prevalence in February, 2003. The maximum prevalences (16 percent) in P. guildinii were observed in January and February, 2003 and on July 27, 2003 with 20 percent of the specimens infected. Flagellates in the hemolymph of E. heros were less frequent (maximum prevalence of 8.7 percent), suggesting that this species is less susceptible than N. viridula and P. guildinii. Trypanosomatid prevalence was higher than that observed for entomopathogenic fungi, however they seem to be inefficient control agents in short term condition.

O complexo de percevejos da soja possui um grande potencial de dano, seu controle é difícil ao final da safra por suas elevadas densidades populacionais. Trabalhos relacionados com organismos entomopatogênicos em percevejos são interessantes devido a suas possibilidades como agentes de controle e porque eles podem ser agentes causais de doenças em criação massal. A prevalência de tripanossomatídeos foi determinada nas espécies mais importantes da cultura da soja, no Brasil, Nezara viridula (L.), Euschistus heros (Fabricius) e Piezodorus guildinii (Westwood). Os insetos foram coletados na soja e hospedeiros alternativos durante a entressafra. Gotas de hemolinfa de cada adulto foram montadas em lâminas e observadas ao microscópio; o número de percevejos com e sem flagelados foi registrado. Os flagelados encontrados nas populações de P. guildinii e E. heros foram menores que os observados em N. viridula. A maior prevalência dos tripanossomatídeos foi observada nos espécimes de N. viridula coletados na soja alcançando 30 por cento de infecção em janeiro, 2001, e 44 por cento prevalência em fevereiro de 2003. Em P. guildinii as máximas prevalências foram observadas em janeiro e fevereiro de 2003 (16 por cento) e em 27 de julho de 2003 com 20 por cento dos espécimes infectados. Os flagelados na hemolinfa de E. heros foram menos freqüentes (máxima prevalência de 8,7 por cento), sugerindo que esta espécie é menos suscetível que N. viridula e P. guildinii. Embora os níveis de prevalência sejam mais elevados que os observados para fungos entomopatogênicos, os tripanossomatídeos não parecem ser eficientes reguladores das populações de percevejos de percevejos em curto prazo.

Intraspecific variation and population structure of the Velvetbean Caterpillar, Anticarsia gemmatalis Hübner, 1818 (Insecta: Lepidoptera: Noctuidae)

The velvetbean caterpillar (VBC), Anticarsia gemmatalis Hübner, 1818 (Insecta: Lepidoptera: Noctuidae), is one of the most important New World soybean agro-ecosystems pests, occurring from 40° N in the USA to 39° S in Argentina. Information on the migration patterns of the VBC moth may be important for managing the resistance of VBC populations to insecticides or plants carrying the Bacillus thuringiensis insecticide genes, especially since sedentary populations have a higher potential to became resistant than migratory populations. We studied intraspecific variations of geographically distinct VBC populations in order to determine the genetic distance between them and to assess the variability of VBC populations from near the city of Londrina (Paraná (PR) state, Brazil). Samples of the VBC were obtained from sites near the following towns or cities: Marianna and Quincy (Florida, USA); La Virginia (Tucumán province, Argentina); Londrina (PR), Passo Fundo (Rio Grande do Sul, Brazil) and Planaltina (Goiás, Brazil). The VBC samples were used to construct a genetic similarity matrix based on random amplified polymorphic DNA (RAPD) allele frequencies, the cotton leafworm, Alabama argillacea, Hübner 1823 (Insecta: Lepidoptera: Noctuidae), being used as an outgroup. Interestingly, despite the great distance (about 6,500 km) between Planaltina and Quincy some of the specimens from the Quincy population clustered in a group genetically close to the Planaltina populations. Larvae collected on peanuts in Marianna and on soybean in Quincy, 70 km apart, appeared genetically similar. The population from Planaltina was the most heterogeneous (polymorphism = 85.6 percent; heterozygosity = 0.1505). The Argentinean VBC population was entirely different from the Brazilian populations. The genetic similarities found between individuals from geographically distant populations and effective migration rate values (2.0566 > Nm < 15.2618) indicate that migration occurs.

Insecticide resistance to endosulfan, monocrotophos and metamidophos in the neotropical brown stink bug, Euschistus heros (F. ) / Resistência do percevejo marrom Euschistus heros (F. ) aos inseticidas endosulfan, monocrotofós e metamidofós

As populações do percevejo marrom, Euschistus heros (F.), são normalmente controladas com inseticidas fosforados e ciclodienos (endossulfam). A ocorrência de casos de insucesso de E. heros, o percevejo mais comum nas regiões produtoras de soja do Paraná, São Paulo e da Região Central do Brasil, tem levado à investigação da resistência a esses inseticidas. Com essa finalidade foi avaliada a suscetibilidade de adultos provenientes de campos de soja, aos inseticidas endossulfam, monocrotofós e metamidofós. A população mais suscetível aos três inseticidas foi coletada em Londrina. A população mais resistente ao endossulfam (TR= 8,7) e ao metamidofós (TR= 2,7) foi proveniente de Pedrinhas Paulista (SP); para monocrotofós a população mais resistente foi coletada em Centenário do Sul (PR) (TR= 3,1). Percevejos E. heros provenientes de Toledo (PR) apresentaram níveis de suscetibilidade ao metamidofós comparáveis aos de Londrina (PR).

The Neotropical brown stink bug, Euschistus heros (F.), is usually controlled by organophosphate (OP) and cyclodiene (endosulfan) insecticides. Some unsuccessful control cases lead us to search for resistance to these insecticides in E. heros, the most common stink bug in the soybean growing regions of Paraná State, São Paulo State and Central Region of Brazil. Bioassays were performed with endosulfan, monocrotophos and metamidophos against adults. The most susceptible population was found in Londrina. Populations from Pedrinhas Paulista (SP) showed the highest resistance ratio (RR) to endosulfan (RR= 8.7) and metamidophos (RR=2.7); resistance to monocrotophos was detected on bugs from Centenário do Sul (PR) (RR= 3.1). Stink bugs collected from Toledo (PR), showed susceptibility to metamidophos comparable to those from Londrina.

Natural Occurrence of the Entomopathogenic Fungi Metarhizium, Beauveria and Paecilomyces in Soybean Under Till and No-till Cultivation Systems / Ocorrência Natural de Fungos Entomopatogênicos Metarhizium, Beauveria e Paecilomyces na Cultura da Soja sob Plantio Direto e Convencional

A ocorrência de fungos entomopatogênicos Metarhizium, Beauveria e Paecilomyces foi estudada em condições de semeadura direta e convencional da soja. Foi determinada a densidade de unidades formadoras de colônia por g de solo e por cm² de folíolos de soja. Verificou-se que no solo sob semeadura direta ocorreu maior incidência dos entomopatógenos, mas sobre os folíolos essa diferença não ocorreu, proporcionando as mesmas possibilidades de infecção nos insetos da parte aérea suscetíveis que ocorrem nas duas condições de cultivo.

The occurrence of the entomopathogenic fungi Metarhizium, Beauveria and Paecilomyces in soybean under till and no-till cultivation systems was studied. The number of colony forming units per g of soil and per cm² of leaf area was determined. Soil under no-till system showed a higher incidence of entomopathogenic fungi, but no difference in fungal density was observed on leaves. Thus the probability of infection for susceptible leaf inhabiting insects should be similar in both systems.

Genetic differentiation among Brazilian populations of Euschistus heros (Fabricius) (Heteroptera: Pentatomidae) based on RAPD analysis / Diferenciação genética entre populações de Euschistus heros (Fabricius) (Heteroptera: Pentatomidae) do Brasil mediante marcadores de RAPD

The Neotropical Brown Stink Bug (NBSB), Euschistus heros (Fabricius), has a wide distribution in Brazilian soybean fields, being more important in the central region of the country. The species is the main target of insecticide applications for stink bug pest control. We determined the variability among and within populations of the NBSB by random amplification of polymorphic DNA (RAPD) analysis. Samples of NBSB were collected in soybean fields from Ubiratã (PR), Londrina (PR), Centenário do Sul (PR), Cândido Mota (SP), Ponta Porã (MS) and Sapezal (MT). Genomic DNA was extracted from the head to minimize DNA contamination by endoparasites. DNA was amplified with 10 mer primers. Fifteen primers produced 246 bands. The genetic similarity matrix was obtained, based on RAPD allele frequencies, using Nei's 1972 genetic distance. NSBS collected from the same geographical region clustered together. The populations from Londrina and Centenário do Sul were genetically closer than the others and stink bugs collected in Cândido Mota were closer to Ponta Porã population. The Sapezal geographical population was the most divergent from the others. Females and males clustered separately inside each geographical population, implying that RAPD permits gender discrimination. We did not observe individuals from one region clustering together with stink bugs from another region. The number of polymorphic loci from the different populations ranged between 40.6 percent and 52.1 percent. The gene flow indexes (overall Nm = 0.8307) were lower than that observed for Anticarsia gemmatalis Hübner and Helicoverpa armigera (Hübner) suggesting that in stink bug populations gene flow is lower than in the noctuid moths

O percevejo marrom da soja, Euschistus heros (Fabricius), possui ampla distribuição na região sojicola do Brasil, sendo mais importante na região central do País. Tem sido o alvo principal das aplicações de inseticidas dirigidas contra o complexo de pentatomídeos praga. A variabilidade nos RAPD entre e dentro das populações do percevejo marrom foi determinada. Amostras da espécie foram coletadas nos campos de soja de Ubiratã (PR), Londrina (PR), Centenário do Sul (PR), Cândido Mota (SP), Ponta Porã (MS) e Sapezal (MT). O DNA genômico foi extraído da cabeça para minimizar a contaminação do DNA proveniente dos endoparasitóides e parasitas que ocorrem na hemocele. O DNA foi amplificado com iniciadores de 10 nucleotídeos. Quinze iniciadores produziram 246 bandas. A similaridade genética foi obtida com base na freqüência alélica dos RAPD utilizando-se a distância de Nei 1972. Os percevejos coletados da mesma região geográfica apresentaram a maior similaridade genética. As populações de Londrina e Centenário do Sul foram geneticamente mais próximas que as restantes. Também, percevejos coletados em Cândido Mota foram próximos das populações de Ponta Porã. A população geográfica de Sapezal foi a mais divergente das outras. As fêmeas e os machos agruparam-se em grupos diferentes dentro de cada população geográfica, o que significa que os RAPD possibilitam a diferenciação dos sexos. Não foram observados indivíduos de uma população agrupando-se com indivíduos de outra região. O número de loci polimórficos das diferentes populações variou entre 40,6 por cento e 52,1 por cento. Os índices de fluxo gênico de E. heros (Nm = 0,8307) foram menores que os observados anteriormente para Anticarsia gemmatalis Hübner e Helicoverpa armigera (Hübner) sugerindo que as populações do pentatomídeo apresentam maior isolamento geográfico que nos referidos noctuídeos, por exemplo

The impact of fungicides on Nomuraea rileyi (Farlow) Samson epizootics and on populations of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), on soybean / Impacto de fungicidas sobre epizootias de Nomuraea rileyi (Farlow) Samson e sobre populações de Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae), em soja

The fungus Nomuraea rileyi (Farlow) Samson is one of the most important natural enemies of soybean caterpillars, mainly under humid weather conditions. Outbreaks of the fungus Microsphaera diffusa Cooke & Peck have demanded fungicide applications on soybeans, which could result in outbreaks of noctuid populations by reduction of the natural inocula of N. rileyi. The recommended fungicides have shown to be detrimental to beneficial fungi, reducing infection, delaying epizootics, and resulting in increased host population densities. In laboratory assays, benomyl, difenoconazole, sulphur and carbendazim affected conidial germination of N. rileyi, being the latter less deleterious. To assess the impact of fungicides used to control M. diffusa, on N. rileyi, two tests were carried on, spraying difenoconazole (75 g a.i./ha) and benomyl (262.5 g a.i./ha) on soybean plots. In the 1997/98 trial, fungicide was sprayed once on soybean plants at R1-R2 developmental stages. In the 1998/99 test, two applications were made, when plants were at V5 and V7 developmental stages, respectively. The number of VBC larvae was significantly higher in the fungicide treated plots than in the control plots. In the 1997/98 test, benomyl treated plots resulted in higher populations of VBC than in the control or in the difenoconazole plots. In the 1998/99 test, VBC population was higher from 7 to 12 days after the first application, and remained high until 19 days after the second application. In general, fungicide treatments delayed the begining of N. rileyi epizootics from 2 to 14 days.

O fungo Nomuraea rileyi (Farlow) Samson é um dos inimigos naturais mais importantes de lagartas da soja, principalmente em condições de tempo úmido. Surtos de oídio têm demandado a aplicação de fungicidas na soja, especialmente nas etapas iniciais do ciclo agrícola, época na qual a interferência sobre N. rileyi pode favorecer a ocorrência de lagarta por redução do inóculo desse fungo. Ensaios de laboratório mostraram que benomil, difenoconazole, enxofre e carbendazim reduziram a germinação de N. rileyi sendo o último menos deletério. Para avaliar o impacto dos fungicidas utilizados no controle do oídio sobre N. rileyi, foram realizadas aplicações com difenoconazole (75 g i.a./ha) ou benomil (262,5 g i.a./ha) em parcelas de soja. No ensaio realizado em 1997/1998 foi realizada apenas uma aplicação de fungicida, estando as plantas no estágio R1-R2. No ensaio realizado em 1998/99 realizaram-se duas aplicações, espaçadas de 19 dias, quando as plantas estavam nos estágios V5 e V7, respectivamente. O número de lagartas foi significativamente maior nas parcelas tratadas com os fungicidas que nas parcelas testemunhas. Em 1997/98, a parcela tratada com benomil apresentou população mais elevada da lagarta-da-soja que nas parcelas testemunhas ou na tratada com difenoconazole. Em 1998/99, a população da lagarta-da-soja foi maior entre o sétimo e décimo segundo dia após a primeira aplicação, e continuou alta até o décimo nono dia após a segunda aplicação. Os fungicidas causaram um atraso de 2 a 14 dias na iniciação da epizootia de N. rileyi.

Produção de anticorpos policlonais para lectina de hemolinfa de Anticarsia gemmatalis / Production of polyclonal antibodies for lectin from Anticarsia gemmatalis hemolymph

A lagarta de Anticarsia gemmatalis promove extensos danos na cultura da soja e seu controle é geralmente baseado na aplicação de inseticidas químicos. Devido aos riscos à saúde humana, animal e ao meio ambiente, métodos alternativos de controle tem sido desenvolvidos como o bioinseticida Baculovirus anticarsia. Há relatos de desenvolvimento de resistência em populações de A. gemmatalis submetidas, em laboratório, ao tratamento com baculovirus durante várias gerações. Os insetos apresentam mecanismos elaborados de proteção contra agentes infecciosos, como as lectinas, que atuam como moléculas de reconhecimento. Assim, o objetivo deste estudo foi desenvolver anticorpos policlonais para lectina de A. gemmatalis. A atividade de lectina de hemolinfa de lagartas de A. gemmatalis foi avaliada frente a hemácias humanas, de coelho, camundongo, carneiro e boi de ensaio de hemaglutinação. Apenas as hemácias de bovino nåo foram aglutinadas pela lectina. As hemácias de coelho apresentaram maior reatividade com a lectina (1:512) e portanto os anticorpos policlonais foram produzidos em coelho imunizado com hemácias autólogas sensibilizadas com lectina. O anticorpo anti-lectina apresentou título de 1:8 em reação de precipitação em gel. Assim neste estudo foi possível produzir anticorpos para lectina de A. gemmatalis sem necessidade de emprego de técnicas dispendiosas de purificação.