Population genetics of Leptoglossus clypealis (Hemiptera: Coreidae) in the Western United States.

The leaffooted bug, Leptoglossus clypealis Heidemann, is a seed-feeding economic pest of crops including almonds and pistachios. The historical distribution of L. clypealis has been considered to be West of the Mississippi in the United States. L. clypealis was recently found in sorghum in the Coastal Bend of Texas, representing a new host record and new collection locality. This study investigated the genetic diversity of L. clypealis museum voucher samples from the Western United States (i.e., Texas, California, and Idaho) collected from 1994 to 2019, including the L. clypealis samples from the Coastal Bend. Eleven new sequences were obtained. Sample sequences were compared with public sequences of L. clypealis from the Western United States. The mitochondrial DNA cytochrome oxidase 1 (mtDNA COI) barcode gene region revealed differences among and within the collection regions. Texas, Idaho, and California all had samples with unique genotypes, and the combined dataset had a haplotype diversity of 1.0. The Texas specimens recently collected in the Coastal Bend did not match genotypes from California or Idaho, and it is unlikely they were recently introduced. Overall, L. clypealis from Texas, Idaho, and California have a high level of genetic diversity, and the 3 regions appear to be within the native range of the species.

Innervation of the southern green stink bug [Nezara viridula (L.) (Hemiptera: Pentatomidae)] stylet bundle.

The southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae), continues to threaten high-value cash crops, including cotton. Earlier reports confirmed ingestion and transmission of disease-causing pathogens of cotton, including elucidation of the dimensions for the food and salivary canals of the southern green stink bug stylet bundle. During this earlier work, innervation of the stylet bundle was observed. Here, we present the first imagery and descriptions of the innervations (i.e., dendrites) within the southern green stink bug stylets. Two types of dendrites innervate each mandibular stylet, and the number of dendrites differed depending on location. Within the head, six dendrites (3 within a thick-walled and 3 within a thin-walled dendrite sheath) are present in each mandibular stylet; only 3 dendrites within a thin-walled sheath are present at the most distal labial segment. Transmission electron microscopy (TEM) suggests innervation of the maxillary stylets, and the presence of stained tissue within the dendritic canal of the maxillary stylets was observed via light microscopy, thereby supporting the TEM analyses. These new observations regarding types and spatial differences in numbers of dendrites within the mandibular stylets - and the new revelation of innervation within maxillary stylets - improve the current knowledge base regarding internal stylet morphology and feeding mechanics.

Acquisition and Transmission of Fusarium oxysporum f. sp. vasinfectum VCG 0114 (Race 4) by Stink Bugs.

Fusarium oxysporum f. sp. vasinfectum VCG 0114 (race 4; i.e., FOV4) is an emerging pathogen that causes severe root rot and wilt of cotton. FOV4 is seed-borne, but the mode of seed invasion is uncertain. In an initial study, seeds in bolls that were puncture inoculated with FOV4 conidia when they were 25- or 30-days old became infected but remained viable. Because stink bugs can ingest and introduce bacterial and yeast pathogens into cotton bolls, we hypothesized that stink bugs may ingest and transmit FOV4. Southern green stink bugs and brown stink bugs were exposed to potato dextrose agar cultures of FOV4 and subsequently caged with cotton bolls to assess transmission potential. Both species fed on the cultures and acquired FOV4, and brown stink bugs transmitted FOV4 to cotton bolls. Thus, management of FOV4 may require management of stink bugs to mitigate the spread of the disease in cotton.

Retention of Pantoea agglomerans Sc1R across stadia of the southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae).

Southern green stink bug [Nezara viridula (L.)] adults and other pentatomid pests can transmit pathogens (e.g., the bacterium Pantoea agglomerans) that cause disease in cotton (Gossypium hirsutum L.) and other high-value cash crops worldwide. First instars of N. viridula were recently shown to ingest P. agglomerans strain Sc1R with rifampicin-resistance, and to retain the pathogen to the 2nd instar. The objective of this study was to determine the acquisition of P. agglomerans Sc1R by early instars of N. viridula and determine persistence of P. agglomerans Sc1R across subsequent stadia. In three trials, early instars (1st and 2nd) were exposed to P. agglomerans Sc1R and subsequently maintained to adulthood; cohorts were sampled at 3rd and 5th instars, as well as adults. In every trial, P. agglomerans Sc1R was detected in all stadia, including adults, but significantly higher frequencies of infection than expected were observed at the initial stage of infection (either 1st or 2nd instar). Higher densities of P. agglomerans Sc1R were detected in 1st and 2nd instars, and lower densities were observed in subsequent stadia. Densities of innate microbiota were generally lower when the initial stage of exposure was at 1st instar than when the initial stage of exposure was at the 2nd instar. Overall, half of the adults possessed P. agglomerans Sc1R. These findings demonstrated that N. viridula nymphs can acquire P. agglomerans Sc1R and retain the pathogen to adulthood. Potential avenues of research to further elucidate the implications of nymphs harboring pathogens to adulthood are discussed.

Diapause Response of the Boll Weevil (Coleoptera: Curculionidae) to Feeding Period Duration and Cotton Square Size.

Distribution of the boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), in the United States has been greatly reduced by eradication efforts. Still, it remains a key pest of cotton (Gossypium spp., [Malvales: Malvaceae]) in the New World, and has proven difficult to eliminate from Mexico and from southern Texas. In those regions, improved knowledge of boll weevil overwintering ecology may benefit efforts by eradication and management programs. Adult diapause in the boll weevil is well documented, but influences of the feeding period duration between adult eclosion and assessment of diapause remain unstudied. We examined diapause incidence and associated survival for weevils fed for 7, 14, or 21 d after adult eclosion. Diapause incidence of females was less influenced by feeding duration compared with males. For males, highest diapause incidence occurred after 14 d of feeding compared with 7 or 21 d. Host-free survival tended to be higher after 14 d of feeding compared with 7 or 21 d, although many weevils were long-lived (≥80 d) after each feeding period duration. Males exhibited higher survival compared with females, and survival was higher for weevils fed large flower buds (squares) compared with smaller squares. Survival was most influenced by temperature; longevity increased with decreasing temperature except at the lowest temperature (12.8°C). These results suggest an optimal feeding period for induction of diapause and maximized host-free longevity. These findings may permit improved timing of late-season insecticide treatments aimed at reducing overwintering populations, and thereby improve effectiveness of eradication and management programs.

Poultry litter and the environment: Physiochemical properties of litter and soil during successive flock rotations and after remote site deposition.

The U.S. broiler meat market has grown over the past 16 years and destinations for U.S. broiler meat exports expanded to over 150 countries. This market opportunity has spurred a corresponding increase in industrialized poultry production, which due to the confined space in which high numbers of animals are housed, risks accumulating nutrients and pollutants. The purpose of this research was to determine the level of pollutants within poultry litter and the underlying soil within a production facility; and to explore the impact of spent litter deposition into the environment. The study follows a production facility for the first 2.5 years of production. It monitors the effects of successive flocks and management practices on 15 physiochemical parameters: Ca, Cu, electrical conductivity, Fe, K, Mg, Mn, moisture, Na, NO3(-)/N, organic matter, P, pH, S, and Zn. Litter samples were collected in-house, after clean-outs and during stockpiling. The soil before house placement, after the clean-outs and following litter stockpiling was monitored. Management practices markedly altered the physiochemical profiles of the litter in-house. A canonical discriminant analysis was used to describe the relationship between the parameters and sampling times. The litter profiles grouped into five clusters corresponding to time and management practices. The soil in-house exhibited mean increases in all physiochemical parameters (2-297 fold) except Fe, Mg, %M, and pH. The spent litter was followed after deposition onto a field for use as fertilizer. After 20 weeks, the soil beneath the litter exhibited increases in EC, Cu, K, Na, NO3(-)/N, %OM, P, S and Zn; while %M decreased. Understanding the impacts of industrialized poultry farms on the environment is vital as the cumulative ecological impact of this land usage could be substantial if not properly managed to reduce the risk of potential pollutant infiltration into the environment.

Relationship Between Piercing-Sucking Insect Control and Internal Lint and Seed Rot in Southeastern Cotton (Gossypium hirsutum L.).

In 1999, crop consultants scouting for stink bugs (Hemiptera spp.) in South Carolina discovered a formerly unobserved seed rot of cotton that caused yield losses ranging from 10 to 15% in certain fields. The disease has subsequently been reported in fields throughout the southeastern Cotton Belt. Externally, diseased bolls appeared undamaged; internally, green fruit contain pink to dark brown, damp, deformed lint, and necrotic seeds. In greenhouse experiments, we demonstrated transmission of the opportunistic bacterium Pantoea agglomerans by the southern green stink bug, Nezara viridula (L.). Here, green bolls were sampled from stink bug management plots (insecticide protected or nontreated) from four South Atlantic coast states (North Carolina, South Carolina, Georgia, and Florida) to determine disease incidence in the field and its association with piercing-sucking insects feeding. A logistic regression analysis of the boll damage data revealed that disease was 24 times more likely to occur (P = 0.004) in bolls collected from plots in Florida, where evidence of pest pressure was highest, than in bolls harvested in NC with the lowest detected insect pressure. Fruit from plots treated with insecticide, a treatment which reduced transmission agent numbers, were 4 times less likely to be diseased than bolls from unprotected sites (P = 0.002). Overall, punctured bolls were 125 times more likely to also have disease symptoms than nonpunctured bolls, irrespective of whether or not plots were protected with insecticides (P = 0.0001). Much of the damage to cotton bolls that is commonly attributed to stink bug feeding is likely the resulting effect of vectored pathogens.

Temporal analysis of cotton boll symptoms resulting from southern green stink bug feeding and transmission of a bacterial pathogen.

The southern green stink bug, Nezara viridula (L.), is a significant pest of cotton, Gossypium hirsutum L., and is becoming an increasing challenge due to the decrease in use of broad-spectrum insecticides on the crop. The southern green stink bug can vector an opportunistic Pantoea agglomerans strain (designated Sc 1-R) into cotton bolls, resulting in infection. The appearance of stink bug damage varies, and pest managers cannot readily identify its source. This research reports a systematic depiction of green, immature boll responses at various stages of maturity (1, 2, and 3 wk post-anthesis [WPA]) to stink bug injury and to infection by the vectored cotton pathogen by demonstrating the progression of effects 1, 2, and 3 wk after exposure (WAE). When laboratory-reared adult southern green stink bug not harboring Sc 1-R deposited bacteria into greenhouse-grown bolls at 1, 2, or 3 WPA during feeding/probing, bacteria reached concentrations of 10(9), 10(9), and 10(3) colony-forming units (CFUs)/g tissue, respectively, at 3 WAE, yet caused minimal seed and lint damage regardless of the age of the bolls that were penetrated. Bolls at a maturity of 1 or 2 WPA showed similar susceptibility when exposed to stink bugs that vectored Sc 1-R. After a week of infection, seeds were salmon-pink with normal white lint and up to 10(4) CFUs/g tissue when Sc 1-R was detected. Necrosis of the entire inoculated locule(s) with a maximum Sc 1-R concentration detected at 10(8) CFUs/g tissue occurred in samples harvested 2 or 3 WAE. Conversely, seed and lint deterioration due to the transmitted opportunist into bolls exposed 3 WPA was confined to the puncture site. In summary, after a week of development, bolls were tolerant to southern green stink bug feeding/ probing damage and to nonpathogenic bacteria, but they were severely damaged when the opportunistic pathogen Sc 1-R was transmitted. At 3 WPA, the fruit was immune to the spread of the pathogen with infections confined to the puncture site.

The acquisition and internalization of Salmonella by the lesser mealworm, Alphitobius diaperinus (Coleoptera: Tenebrionidae).

In poultry broiler production facilities, it is important to understand the sources and contribution of reservoir populations of pathogens. The lesser mealworm beetle, Alphitobius diaperinus (Panzer), is a common pest in poultry litter that is reported to carry pathogens affecting both human and animal health. This study investigates whether the carriage of a bacterial pathogen occurs by the harboring of bacteria internally by these insects. Beetles were exposed to a marker bacterium, Salmonella enterica serovar Typhimurium-green fluorescent protein (ST-GFP), at concentrations up to 10(7) colony-forming units (cfu)/mL for 0.5 to 12 h, and then subsequently surface disinfected and dissected. The head, gastrointestinal tract and hemolymph were cultured for the presence of ST-GFP. This study definitively demonstrates the internal carriage of Salmonella by this insect and found that the beetles rapidly acquired bacteria from external sources and harbored the bacteria within their alimentary canal after exposure for 30 min at 10(4) cfu/mL and within the hemolymph after exposure for 2 h at 10(6) cfu/mL. Beetles internalized an average of 9.5 × 10(1) and 3.2 × 10(3) after a 2-h exposure to 2 × 10(4) and 2 × 10(6) cfu/mL, respectively. The lesser mealworm is a serious pest within the poultry brooder and laying industry and because of their mobility, voracious feeding habits, and prey potential may represent an active source facilitating the dissemination of Salmonella.

Identification of the first neuropeptides from the CNS of Hemiptera: CAPA peptides of the southern green stinkbug Nezara viridula (L.).

A direct mass spectrometric investigation of nerve homologs of the abdominal perisympathetic organs was employed to reveal the first and complete sequences of CAPA peptides from a hemipteran species, the southern green stinkbug Nezara viridula. Side-chain fragmentations allowed the assignment of internal Leu/Ile; on-plate acetylation was used to distinguish between the mass-related Lys and Gln. The following sequences were obtained: DQLFPFPRV-NH(2) (CAPA-PVK-1), EQLIPFPRV-NH(2) (CAPA-PVK-2), and NGSAGNGGLWFGPRL/I-NH(2) (CAPA-PK). CAPA-PVKs are associated with the regulation of diuresis in insects, and identification of those native to a hemipteran will provide the experimental basis to better understand regulation of water balance in this family of insects.