Laura Varone

Fire Ants (Solenopsis spp.) and Their Natural Enemies in Southern South America

We review the fire ant research conducted by the ARS-South American Biological Control Laboratory (SABCL) since 1987 to find a complex of natural enemies in southern South America and evaluate their specificity and suitability for field release as self-sustaining biological control agents. We also include those studies conducted by the ARS-Center for Medical, Agriculture, and Veterinary Entomology in the United States with the SABCL collaboration. Ecological and biological information is reported on local fire ants and their microsporidia, nematodes, viruses, phorid flies, eucharitid wasps, strepsiptera, and parasitic ants. Their biology, abundance, distribution, detrimental effect, field persistence, specificity, and phenology are discussed. We conclude that the objectives of the ARS program in South America are being achieved and that the pioneering studies have served to encourage further investigations in the United States and other countries and advanced the implementation of biological control programs to decrease imported fire ant densities and damage. Still, several promising organisms should be further investigated for eventual field release in the near future.

Egg limitation and the functional response of the parasitoid Campoletis grioti (Hym: Ichneumonidae)

Abstract The hypothesis that mature egg availability during a parasitoids lifetime affects the functional response, as well as a parasitoids handling time and searching efficiency, was tested. Campoletis grioti Blanchard (Hym: Ichneumonidae) female parasitoids and Spodoptera frugiperda Smith (Lep: Noctuidae) host larvae were used as an example. Each female of C. grioti was confined with different host densities (6, 10, 30, 50, 80, 120 and 160 larvae/day) of first and second S. frugiperda instars until the female died. We found that C. grioti showed a type II functional response, with a reduction in the number of parasitized hosts in relation to female ageing. Handling time and searching efficiency increased over time. When we standardized the data simulating unlimited egg supply, the functional response, handling time and searching efficiency became constant throughout the parasitoids life. We discuss the relationship between egg availability and functional response and its associated parameters, and its consequence on biological control programs.

DNA barcoding and morphological identification of Argentine species of Apanteles (Hymenoptera : Braconidae), parasitoids of cactus-feeding moths (Lepidoptera : Pyralidae : Phycitinae), with description of a new species

Abstract. The gregarious species of Apanteles that attack cactus-feeding phycitine moths (Lepidoptera : Pyralidae) from Argentina are identified using DNA barcodes and morphological data. Sequences of specimens from 10 different populations were generated. Corrected genetic divergences showed two clusters of specimens separated by COI divergences higher than 6%. Our morphological examinations were congruent with the molecular evidence and therefore two species were confidently identified: Apanteles alexanderi Brèthes and a new species, Apanteles opuntiarum Martínez & Berta, sp. nov., which we describe and illustrate. Despite these two taxa being considered to represent a single polyphagous species for at least 50 years, they can be easily distinguished by the length of their ovipositor sheaths. A key to the species of microgastrine parasitoids of cactus-feeding Lepidoptera in South America is provided. The recognition of the new species reveals a narrower host range for the species involved in this system, making it necessary to re-evaluate their potential as biocontrol agents of the prickly pear moth.

Complete genome sequence of an Argentinean isolate of Solenopsis invicta virus 3

Solenopsis invicta virus 3 (SINV-3) is a recently described positive-strand RNA virus that infects the red imported fire ant, S. invicta. The genome of an Argentinean isolate of Solenopsis invicta virus 3 (SINV-3ArgSF) obtained from the Santa Fe region of Argentina was sequenced in entirety. Assembly of nine overlapping fragments yielded a consensus genome sequence 10,386 nucleotides long, excluding the poly(A) tail present on the 3′ end (Genbank accession number GU017972). With the exception of the poly(A) tail, the genome length of SINV-3ArgSF was identical to the North American isolate (SINV-3USDM). The SINV-3ArgSF genome possessed three major open reading frames (ORFs) (comprised of ≥100 codons) in the sense orientation; SINV-3USDM possessed only two. ORFs 1 and 2 had identical start and stop genome positions for both isolates. Blastp analysis of the translated ORF 1 of SINV-3ArgSF recognized conserved domains for helicase, protease, and RNA-dependent RNA polymerase. These domains and their corresponding positions were identical to those reported for SINV-3USDM. ORF 2a, unique to the SINV-3ArgSF genome, was also found in frame 2 and had a canonical start codon located at nucleotide position 8,351 and a stop codon ending at position 8,827. Blastp analysis of the translated amino acid sequence of ORF 2a revealed no significant similarity in the Genbank database. The two SINV-3 isolates exhibited 96.2% nucleotide sequence identity across the entire genome. The amino acid sequences of ORFs 1 and 2 exhibited higher identities (99.0 and 98.2%, respectively) than the corresponding nucleotide regions within the genome. These data indicated that the nucleotide differences between the SINV-3 isolates were largely synonymous. This observation was corroborated by codon substitution rate analysis. Thus, the majority of the SINV-3 codon changes were silent in the two polyproteins, indicating purifying selection pressure on the viral genome.

Exploitation of a high genomic mutation rate in Solenopsis invicta virus 1 to infer demographic information about its host, Solenopsis invicta.

The RNA-dependent RNA polymerase (RdRp) region of Solenopsis invicta virus 1 (SINV-1) was sequenced from 47 infected colonies of S. invicta, S. richteri, S. geminata, and S. invicta/richteri hybrids collected from across the USA, northern Argentina, and northern Taiwan in an attempt to infer demographic information about the recent S. invicta introduction into Taiwan by phylogenetic analysis. Nucleotide sequences were calculated to exhibit an overall identity of >90% between geographically-separated samples. A total of 171 nucleotide variable sites (representing 22.4% of the region amplified) were mapped across the SINV-1 RdRp alignment and no insertions or deletions were detected. Phylogenetic analysis at the nucleotide level revealed clustering of Argentinean sequences, distinct from the USA sequences. Moreover, the SINV-1 RdRp sequences derived from recently introduced populations of S. invicta from northern Taiwan resided within the multiple USA groupings implicating the USA as the source for the recent introduction of S. invicta into Taiwan. Examination of the amino acid alignment for the RdRp revealed sequence identity >98% with only nine amino acid changes observed. Seven of these changes occurred in less than 4.3% of samples, while 2 (at positions 1266 and 1285) were featured prominently. Changes at positions 1266 and 1285 accounted for 36.2% and 34.0% of the samples, respectively. Two distinct groups were observed based on the amino acid residue at position 1266, Threonine or Serine. In cases where this amino acid was a Threonine, 90% of these sequences possessed a corresponding Valine at position 1285; only 10% of the Threonine(1266)-containing sequences possessed an Isoleucine at the 1285 position. Among the Serine(1266) group, 76% possessed an Isoleucine at position 1285, while only 24% possessed a Valine. Thus, it appears that the Threonine(1266)/Valine(1285) and Serine(1266)/Isoleucine(1285) combinations are predominant phenotypes.

Field Host Range of Apanteles opuntiarum (Hymenoptera: Braconidae) in Argentina, a Potential Biocontrol Agent of Cactoblastis cactorum (Lepidoptera: Pyralidae) in North America

Summary Field exploration in Argentina for cactophagous lepidopteran hosts parasitized by the recently described braconid parasitoid Apanteles opuntiarum Martínez & Berta (Hymenoptera: Braconidae) revealed a host range restricted to Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae) and C. doddi. Field collections of cactophagous larvae parasitized by the congeneric and sympatric parasitoid A. alexanderi included the host species C. bucyrus, Sigelgaita nr. chilensis, Tucumania sp., Tucumania tapiacola and Salambona sp. If a narrow host range for A. opuntiarum is confirmed in quarantine with North American cactus-feeding species, then this parasitoid could be released as a biological control agent for C. cactorum with little or no risk to non-target species.

Geographical Range and Laboratory Studies on Apanteles opuntiarum (Hymenoptera: Braconidae) in Argentina, a Candidate for Biological Control of Cactoblastis cactorum (Lepidoptera: Pyralidae) in North America

Abstract The cactus moth, Cactoblastis cactorum (Berg) (Lepidoptera: Pyralidae), is a pest that threatens native Opuntia spp. in North America. Control tactics developed and implemented against this invasive pest successfully eradicated the moth in Mexico and on barrier islands in the United States. However, with the cancellation of the regional management program in the United States, no control tactics are being implemented to mitigate the expansion of the moths geographical range. Hence, an integrated approach including biological control is proposed to regulate the population of C. cactorum in North America. Field surveys of the recently described parasitoid, Apanteles opuntiarum Martínez & Berta, were carried out within the C. cactorum native range in Argentina, and laboratory studies were conducted to develop a parasitoid rearing protocol. Apanteles opuntiarum was the most common parasitoid of C. cactorum and their field distributions were similar. In the laboratory, the parasitoids reproductive success was maximized when one or two female wasps were exposed to 30 host larvae within a 500 ml container. Laboratory reared females were less successful at parasitizing hosts than field collected females. In spite of the success achieved with laboratory rearing, male bias was observed throughout the experiments. Because this bias might be related to the presence of the reproductive parasite Wolbachia, both laboratory colony and field collected individuals were screened and Wolbachia was detected. This study provides useful field and laboratory information on (1) laboratory rearing techniques for A. opuntiarum; (2) developing host specificity test protocols for studies under quarantine conditions; and (3) selecting parasitoid populations that best match the climatic conditions present in the C. cactorum invaded areas of North America.

Larval morphology and host use confirms ecotypic variation in Cactoblastis cactorum (Berg)

Despite their recognized importance in the literature, the contribution of native-range species interactions to invasion success has been inadequately studied. Previous authors have suggested that biases in the sampling of propagules from the native range might influence invasion success, but most contemporary invasion hypotheses focus on the development of novel interactions or a release from native consumers and competitors. When ecotypic variation exists in native host-consumer associations, the specific pattern of sampling across ecotypes could determine invasion success, especially when the genetic diversity among exotic propagules is low. The South American cactus moth, Cactoblastis cactorum (Berg), is an oligophagous consumer whose larvae feed on prickly pear cacti (subfamily Opuntioideae). The moth was collected from a small geographic area along the Argentina-Uruguay border in 1925 and was introduced to multiple continents as a biological control species, which has subsequently invaded North America. Here we show that groups defined by genetic structure in this species’ native range are concordant with distinct patterns of host association and larval morphology. Furthermore, in Florida populations, morphological traits have diverged from those found in the native range, and patterns of host association suggest that strong biases in host preference also occur in invasive populations. The documented history of C. cactorum introductions confirms that multiple attempts were made to export the moth, but that only a single ecotype was exported successfully. Additional work will be necessary to determine whether the observed host biases in North America reflect a rapid adaptation to naïve hosts or a conservation of traits related to specific aspects of the host-consumer association.

Geographic distribution suggests that Solenopsis invicta is the host of predilection for Solenopsis invicta virus 1

Solenopsis invicta virus 1 (SINV-1) was found regularly and prevalently in S. invicta. In sampled locations where S. invicta and S. geminata are sympatric (specifically, Gainesville, FL and Travis, TX), SINV-1 was detected in S. geminata. Conversely, in areas in which S. geminata and S. invicta are allopatric, SINV-1 was not detected in S. geminata; these locations included north Australia (n=12), southern Mexico (n=107), Hawaii (n=48), Taiwan (n=12), and the Johnston Atoll (n=6). A similar relationship was observed for S. richteri. In areas in which S. invicta and S. richteri were sympatric, SINV-1 was detected in the S. richteri population, but in areas in which S. invicta and S. richteri were allopatric, SINV-1 was not detected. These occurrences suggest that S. invicta is the host of predilection, or preferred host for SINV-1, and that the congenerics, S. geminata and S. richteri serve as either accidental, reservoir, or transfer hosts. The minus genome strand of SINV-1 was detected in S. geminata and S. richteri indicating that these species may serve as functional hosts capable of supporting SINV-1 replication. SINV-1 was not detected in S. xyloni regardless of its proximity to S. invicta. These results suggest that SINV-1 may be an example of pathogen spillover or pollution.