Spiroplasmas as Biological Control Agents of Insect Pests

Author(s):  
Kevin Hackett ◽  
Shlomo Rottem ◽  
David L. Williamson ◽  
Meir Klein

Toward development of spiroplasmas as novel toxin-delivery systems for biocontrol of beetle pests in the United States (Leptinotarsa decemlineata) and Israel (Maladera matrida), media for cultivating beetle-associated spiroplasmas were improved and surveys of these spiroplasmas were conducted to provide transformable strains. Extensive surveys of spiroplasmas yielded promising extrachromosomal elements for vector constructs. One, plasmid pCT-1, was cloned, characterized, and used as a source of spiroplasma origin of replication in our shuttle vectors. The fibrillin gene was isolated and sequenced and its strong promoter was also used in the constructs. Means for transforming these vectors into spiroplasmas were developed and optimized, with electroporation found to be suitable for most applications. Development and optimization of means for using large unilamellar vesicles (LUVs) in spiroplasma transformation represents a breakthrough that should facilitate insertion of large clusters of virulence genes. With completion of the vector, we should thus be poised to genetically engineer spiroplasmas with genes that will express toxins lethal to our target beetles, thus providing an effective and inexpensive alternative to conventional means of beetle control.

2019 ◽  
Vol 82 (8) ◽  
pp. 1364-1368 ◽  
Author(s):  
RIZWANA TASMIN ◽  
PAUL A. GULIG ◽  
SALINA PARVEEN

ABSTRACT Salmonella enterica serovar Typhimurium is one of the leading causes of nontyphoidal gastroenteritis of humans in the United States. Commercially processed poultry carcasses are frequently contaminated with Salmonella serovar Kentucky in the United States. The aim of the study was to detect the Salmonella virulence plasmid containing the spv genes from Salmonella isolates recovered from commercially processed chicken carcasses. A total of 144 Salmonella isolates (Salmonella Typhimurium, n = 72 and Salmonella Kentucky, n = 72) were used for isolation of plasmids and detection of corresponding virulence genes (spvA, spvB, and spvC). Only four (5.5%) Salmonella Typhimurium isolates tested positive for all three virulence genes and hence were classified as possessing the virulence plasmid. All isolates of Salmonella Kentucky were negative for the virulence plasmid and genes. These results indicate that the virulence plasmid, which is very common among clinical isolates of Typhimurium and other Salmonella serovars (e.g., Enteritidis, Dublin, Choleraesuis, Gallinarum, Pullorum, and Abortusovis), may not be present in a significant portion of commercially processed chicken carcass isolates.


Helia ◽  
2016 ◽  
Vol 39 (64) ◽  
pp. 45-56 ◽  
Author(s):  
J. R. Prasifka ◽  
L. F. Marek ◽  
D. K. Lee ◽  
S. B. Thapa ◽  
V. Hahn ◽  
...  

AbstractDelayed planting is recommended to reduce damage from sunflower insect pests in the United States, including the sunflower moth, Homoeosoma electellum (Hulst) and banded sunflower moth, Cochylis hospes Walsingham. However, in some locations, planting earlier or growing later-maturing hybrids could improve yield or oil content of sunflowers which would partially offset any added costs from insect pests or their management. Because the abundance and distribution of some sunflower insects have changed since recommendations for delayed planting were developed, experimental plots were grown in 2012 and 2013 at sites in North Dakota, Nebraska, Iowa, and Illinois. Sunflowers were planted two to four weeks earlier than normal, including hybrids that flower two to three weeks later than elite commercial hybrids. The sum of seed damaged by sunflower moth, banded sunflower moth, and red sunflower seed weevil, Smicronyx fulvus LeConte, (i. e., total percentage) was influenced by location, but not the relative maturity of tested entries. However, when damage attributed solely to the red sunflower seed weevil was analyzed, more damaged seed were found for late-maturing entries in North Dakota and Nebraska. In addition to the trial data, current pest populations are lower than when delayed planting was first recommended and insecticide use during sunflower bloom is both common and effective. Together, these observations suggest factoring insect pests into planting time decisions may be unnecessary, except for areas with a history of problems with severe pests that cannot be managed using insecticides (e. g., sunflower midge, Contarinia schulzi Gagné).


2019 ◽  
Author(s):  
Justin Clements ◽  
Russell L. Groves ◽  
JoAnn Cava ◽  
Caroline Barry ◽  
Scott Chapman ◽  
...  

AbstractThe Colorado Potato Beetle, Leptinotarsa decemlineata, is a major agricultural pest of solanaceous crops in the United States. Historically, a multitude of insecticides have been used to control problematic populations. Due to increasing resistance to insecticides, novel compounds and methodologies are warranted for the control of beetle populations. Mixed-isomer conjugated linoleic acid has been studied in-depth for its beneficial properties to mammalian systems. At the same time, studies have demonstrated that conjugated linoleic acid can manipulate fatty acid composition in non-mammalian systems, resulting in embryo mortality. Consequently, experiments were conducted to assess the effects of foliar-applied conjugated linoleic acid on larval growth, embryogenesis, and feeding preference in Colorado potato beetle. Both maternal and deterrent effects of dietary conjugated linoleic acid were assessed. Conjugated linoleic acid demonstrated desirable insecticidal properties, including increased larval mortality, slowed larval development, antifeedant effects, and decreased egg viability after maternal ingestion.


Insects ◽  
2019 ◽  
Vol 10 (9) ◽  
pp. 259 ◽  
Author(s):  
Mikel Domínguez-Arrizabalaga ◽  
Maite Villanueva ◽  
Ana Beatriz Fernandez ◽  
Primitivo Caballero

The genome of the Bacillus thuringiensis BM311.1 strain was sequenced and assembled in 359 contigs containing a total of 6,390,221 bp. The plasmidic ORF of a putative cry gene from this strain was identified as a potential novel Cry protein of 1138 amino acid residues with a 98% identity compared to Cry7Aa1 and a predicted molecular mass of 129.4 kDa. The primary structure of Cry7Aa2, which had eight conserved blocks and the classical structure of three domains, differed in 28 amino acid residues from that of Cry7Aa1. The cry7Aa2 gene was amplified by PCR and then expressed in the acrystalliferous strain BMB171. SDS-PAGE analysis confirmed the predicted molecular mass for the Cry7Aa2 protein and revealed that after in vitro trypsin incubation, the protein was degraded to a toxin of 62 kDa. However, when treated with digestive fluids from Leptinotarsa decemlineata larvae, one major proteinase-resistant fragment of slightly smaller size was produced. The spore and crystal mixture produced by the wild-type BM311.1 strain against L. decemlineata neonate larvae resulted in a LC50 value of 18.8 μg/mL, which was statistically similar to the estimated LC50 of 20.8 μg/mL for the recombinant BMB17-Cry7Aa2 strain. In addition, when this novel toxin was activated in vitro with commercial trypsin, the LC50 value was reduced 3.8-fold to LC50 = 4.9 μg/mL. The potential advantages of Cry7Aa2 protoxin compared to Cry7Aa1 protoxin when used in the control of insect pests are discussed.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Maria I. Zucchi ◽  
Erick M. G. Cordeiro ◽  
Clint Allen ◽  
Mariana Novello ◽  
João Paulo Gomes Viana ◽  
...  

Abstract Unravelling the details of range expansion and ecological dominance shifts of insect pests has been challenging due to the lack of basic knowledge about population structure, gene flow, and most importantly, how natural selection is affecting the adaptive process. Piezodous guildinii is an emerging pest of soybean in the southern region of the United States, and increasingly important in Brazil in recent years. However, the reasons P. guildinii is gradually becoming more of a problem are questions still mostly unanswered. Here, we have genotyped P. guildinii samples and discovered 1,337 loci containing 4,083 variant sites SNPs that were used to estimate genetic structure and to identify gene candidates under natural selection. Our results revealed the existence of a significant genetic structure separating populations according to their broad geographic origin, i.e., U.S. and Brazil, supported by AMOVA (FGT = 0.26), STRUCTURE, PCA, and FST analyses. High levels of gene flow or coancestry within groups (i.e., within countries) can be inferred from the data, and no spatial pattern was apparent at the finer scale in Brazil. Samples from different seasons show more heterogeneous compositions suggesting mixed ancestry and a more complex dynamic. Lastly, we were able to detect and successfully annotated 123 GBS loci (10.5%) under positive selection. The gene ontology (GO) analysis implicated candidate genes under selection with genome reorganization, neuropeptides, and energy mobilization. We discuss how these findings could be related to recent outbreaks and suggest how new efforts directed to better understand P. guildinii population dynamics.


2017 ◽  
Vol 83 (6) ◽  
Author(s):  
James R. Johnson ◽  
Stephen B. Porter ◽  
Brian Johnston ◽  
Paul Thuras ◽  
Sarah Clock ◽  
...  

ABSTRACT Chicken meat products are hypothesized to be vehicles for transmitting antimicrobial-resistant and extraintestinal pathogenic Escherichia coli (ExPEC) to consumers. To reassess this hypothesis in the current era of heightened concerns about antimicrobial use in food animals, we analyzed 175 chicken-source E. coli isolates from a 2013 Consumer Reports national survey. Isolates were screened by PCR for ExPEC-defining virulence genes. The 25 ExPEC isolates (12% of 175) and a 2:1 randomly selected set of 50 non-ExPEC isolates were assessed for their phylogenetic/clonal backgrounds and virulence genotypes for comparison with their resistance profiles and the claims on the retail packaging label (“organic,” “no antibiotics,” and “natural”). Compared with the findings for non-ExPEC isolates, the group of ExPEC isolates had a higher prevalence of phylogroup B2 isolates (44% versus 4%; P < 0.001) and a lower prevalence of phylogroup A isolates (4% versus 30%; P = 0.001), a higher prevalence of multiple individual virulence genes, higher virulence scores (median, 11 [range, 4 to 16] versus 8 [range, 1 to 14]; P = 0.001), and higher resistance scores (median, 4 [range, 0 to 8] versus 3 [range, 0 to 10]; P < 0.001). All five isolates of sequence type 131 (ST131) were ExPEC (P = 0.003), were as extensively resistant as the other isolates tested, and had higher virulence scores than the other isolates tested (median, 12 [range, 11 to 13] versus 8 [range, 1 to 16]; P = 0.005). Organic labeling predicted lower resistance scores (median, 2 [range, 0 to 3] versus 4 [range, 0 to 10]; P = 0.008) but no difference in ExPEC status or virulence scores. These findings document a persisting reservoir of extensively antimicrobial-resistant ExPEC isolates, including isolates from ST131, in retail chicken products in the United States, suggesting a potential public health threat. IMPORTANCE We found that among Escherichia coli isolates from retail chicken meat products purchased across the United States in 2013 (many of these isolates being extensively antibiotic resistant), a minority had genetic profiles suggesting an ability to cause extraintestinal infections in humans, such as urinary tract infection, implying a risk of foodborne disease. Although isolates from products labeled “organic” were less extensively antibiotic resistant than other isolates, they did not appear to be less virulent. These findings suggest that retail chicken products in the United States, even if they are labeled “organic,” pose a potential health threat to consumers because they are contaminated with extensively antibiotic-resistant and, presumably, virulent E. coli isolates.


1983 ◽  
Vol 115 (11) ◽  
pp. 1539-1541 ◽  
Author(s):  
M. A. Latheef ◽  
J. H. Ortiz

Okra, Abelmoschus esculentus (L.) Moench, is a member of the Malvaceae. It is an important crop in the southern United States where vegetable production becomes precarious during the summer owing to extreme conditions of temperature and drought. Okra is grown in most southern home gardens and is consumed as a main dish vegetable or used in soups and gumbos.Because of its popularity and environmental adaptability, okra may have the potential for attracting fresh market premium prices for limited resource farm operations. However, little has been published on insect pests of okra in the United States. In an investigation at Petersburg, Virginia, during 1979 and 1980, we found that the corn earworm, Heliothis zea (Boddie), was an important pest of okra.


1979 ◽  
Vol 111 (7) ◽  
pp. 863-864 ◽  
Author(s):  
M.A. Latheef ◽  
R.D. Irwin

Companionate planting is one of many insect control strategies propounded by organic growers. A variety of herbs are purported to possess repellent properties to many insect pests of vegetables (Rodale 1975; Yepsen 1976; Hill 1976; Anonymous 1977a). According to Atsatt and O'Dowd (1976), repellent plants are members of plant defense guilds which prevent herbivores from locating their hosts. Perrin and Phillips (1978) reviewed the elements of natural pest control characteristic of mixed cropping systems and outlined suggestions for transferring these elements to “agronomically acceptable” monocultures. However, very little work has been done to substantiate these claims and field evidence of the protective properties of companionate plants is, therefore, warranted. This becomes all the more important because of the dramatic increase in home gardening recently in North America. It is estimated that 33 million home and community gardens were grown in the United States during 1976 and 1977 (Anonymous 1977b). Here we describe the results of field experiments in which we sought to quantify the effect of companionate plants against major lepidopteran pests of cabbage in southside Virginia.


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