Dataset Information

Development of a Rapid, Accurate, and On-Site Detection Protocol for Red Imported Fire Ants, Solenopsis invicta (Hymenoptera: Formicidae).

ABSTRACT: A rapid, accurate, and on-site molecular diagnostic protocol for red imported fire ants (Solenopsis invicta, Si) was developed using loop-mediated isothermal amplification (LAMP) assays. Si11977 (GenBank accession no. MK986826) was confirmed to be a Si-specific gene. Four-primer Si11977-LAMP (4pSi-LAMP) and six-primer Si11977-LAMP (6pSi-LAMP) assays specifically detected Si. The reaction time of 6pSi-LAMP assays was reduced by 5 min compared with 4pSi-LAMP assays. The optimal amount of polymerase and the detection limit for the 6pSi-LAMP assays were 0.1 unit/μL and 5 fg/μL, respectively. In addition, a method for extracting genomic DNA from ant tissues within 2 to 3 min and a protocol for performing on-site LAMP assays using a car heating mug and a LAMP observation box were described. The on-site Si detection protocol used in this study was possible within 30 min from DNA extraction to species identification.

SUBMITTER: Kim AY

PROVIDER: S-EPMC9495655 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Development of a Rapid, Accurate, and On-Site Detection Protocol for Red Imported Fire Ants, Solenopsis invicta (Hymenoptera: Formicidae).

Kim A-Young AY Koh Young Ho YH

Bioengineering (Basel, Switzerland) 20220902 9

A rapid, accurate, and on-site molecular diagnostic protocol for red imported fire ants (Solenopsis invicta, Si) was developed using loop-mediated isothermal amplification (LAMP) assays. Si11977 (GenBank accession no. MK986826) was confirmed to be a Si-specific gene. Four-primer Si11977-LAMP (4pSi-LAMP) and six-primer Si11977-LAMP (6pSi-LAMP) assays specifically detected Si. The reaction time of 6pSi-LAMP assays was reduced by ...[more]

PMID: 36134980

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Project description:BackgroundAdvances in sequencing and analysis tools have facilitated discovery of many new viruses from invertebrates, including ants. Solenopsis invicta is an invasive ant that has quickly spread worldwide causing significant ecological and economic impacts. Its virome has begun to be characterized pertaining to potential use of viruses as natural enemies. Although the S. invicta virome is the best characterized among ants, most studies have been performed in its native range, with less information from invaded areas.MethodsUsing a metatranscriptome approach, we further identified and molecularly characterized virus sequences associated with S. invicta, in two introduced areas, U.S and Taiwan. The data set used here was obtained from different stages (larvae, pupa, and adults) of S. invicta life cycle. Publicly available RNA sequences from GenBank's Sequence Read Archive were downloaded and de novo assembled using CLC Genomics Workbench 20.0.1. Contigs were compared against the non-redundant protein sequences and those showing similarity to viral sequences were further analyzed.ResultsWe characterized five putative new viruses associated with S. invicta transcriptomes. Sequence comparisons revealed extensive divergence across ORFs and genomic regions with most of them sharing less than 40% amino acid identity with those closest homologous sequences previously characterized. The first negative-sense single-stranded RNA virus genomic sequences included in the orders Bunyavirales and Mononegavirales are reported. In addition, two positive single-strand virus genome sequences and one single strand DNA virus genome sequence were also identified. While the presence of a putative tenuivirus associated with S. invicta was previously suggested to be a contamination, here we characterized and present strong evidence that Solenopsis invicta virus 14 (SINV-14) is a tenui-like virus that has a long-term association with the ant. Furthermore, based on virus sequence abundance compared to housekeeping genes, phylogenetic relationships, and completeness of viral coding sequences, our results suggest that four of five virus sequences reported, those being SINV-14, SINV-15, SINV-16 and SINV-17, may be associated to viruses actively replicating in the ant S. invicta.ConclusionsThe present study expands our knowledge about viral diversity associated with S. invicta in introduced areas with potential to be used as biological control agents, which will require further biological characterization.

Project description:The food-burying behavior has been reported in many mammals and birds, but was rarely observed in invertebrates. The red imported fire ants, Solenopsis invicta Buren, is an invasive pest in many areas of the world that usually performing food-burying during the foraging processes. However, the impacted factors and measureable patterns of this behavior is largely unknown. In the present study, food-burying vs food-transport behaviors of Solenopsis invicta were observed under laboratory and field conditions. When starved (no food was provided for 37 days) in the laboratory, food (sausage) was consumed by large numbers of ants, and few burying behaviors were observed. However, when food was provided until satiation of the colonies, food-transport was suppressed and significantly more soil particles were relocated on the food and graph paper square (where the food was placed) when compared with these colonies exposed to starved conditions. Videotapes showed that soil particles (1.47 ± 0.09 mm2) were preferentially placed adjacent to (in contact with) the food items at the beginning; and after the edges were covered, ants transported significantly smaller soil particles (1.13 ± 0.06 mm2) to cover the food. Meanwhile, larger particles (1.96 ± 0.08 mm2) were pulled/dragged around (but not in contact with) the food. Interestingly, only a small number of ants, mainly the small workers, were involved in food-burying, and the ants tended to repeatedly transport soil particles. A total of 12 patterns of particle transport were identified, and soil particles were most frequently picked from the foraging arena and subsequently placed adjacent to the food. In the field, almost all released food was actively transported by Solenopsis invicta workers, and no burying behavior was observed. Our results show that the food-burying behavior of Solenopsis invicta may be associated with the suppressed foraging activity, and the burying task may be carried out by certain groups of workers.

Dataset Information

Development of a Rapid, Accurate, and On-Site Detection Protocol for Red Imported Fire Ants, Solenopsis invicta (Hymenoptera: Formicidae).

Publications

Development of a Rapid, Accurate, and On-Site Detection Protocol for Red Imported Fire Ants, <i>Solenopsis invicta</i> (Hymenoptera: Formicidae).

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