Ornithine decarboxylase activity during the development of Anastrepha fraterculus (Diptera, Tephritidae)

Ornithine decarboxylase (ODC) (EC 4.1.1.17) is very important for polyamine biosynthesis, which is required for main biological events. In the present study, ODC activity was measured in samples of Anastrepha fraterculus's egg, larva, pupa body and abdomen, adult body, ovaries, and fat body of young females, and in ovaries of mature flies. The kinetic parameters (Km app and Vmax) for ODC activity were determined for pupa, larva, and young ovary. ODC activity showed fluctuations during A. fraterculus's life development. In its earlier stages, prior to emergence, the egg has high ODC-specific activity probably due to embryogenesis, which is characterized by a high rate of cell division. This enzyme activity is also significantly high in the ovary and fat body of young females possibly related to the increased oogenesis and vitellogenesis. The kinetic parameters (Km app and Vmax) had great variation. Our results using GTP showed that the great variation in kinetic parameters can be accounted for by post-translational modifications.     

Wolbachia endosymbiont in a species of the Anastrepha fraterculus complex (Diptera: Tephritidae)

Summary

In Anastrepha sp.2 aff. fraterculus, the egg-cell harbours a large population of endosymbionts. The bacteria were identified as belonging to genus Wolbachia by PCR assay using primers of the ftsZ gene followed by sequencing of the amplified band. Newly deposited eggs stained in toto by Hoechst show that the bacteria are unevenly dispersed throughout the egg-cell, with a higher accumulation at the posterior pole, and that the degree of infestation varies from egg to egg. Analysis by transmission electron microscopy shows that bacteria are present in the female germ line of embryonic and larval stages, as well as in the different cell types of the ovaries at the adult stage. Mature ova within the follicles harbour a large population of the symbionts. The results indicate the existence of a transovarian transmission of the endosymbionts in this fly.     

Competitive strategies between Anastrepha fraterculus and Anastrepha obliqua (Diptera: Tephritidae) regulating the use of host fruits

Interspecific competition between individuals of different species can result in reductions in their fecundity, growth or survival, reflecting differential exploitation of resources that become intensified due to spatial co-occurrence, ecological similarity and increased population densities. As two species cannot occupy the same niche, coexistence is only possible if the available resources are used in non-overlapping manners such as niche partitioning or the use of refuges. Among agricultural insect pests, such as fruit flies of the family Tephritidae, competitive interactions can result in competitive displacement, host changes, or the expansion or restriction of the numbers of hosts utilized that can have negative consequences for human agricultural activities. We evaluated the competitive interactions between two fruit fly species of the genus Anastrepha, Anastrepha obliqua (Macquart, 1835) and Anastrepha fraterculus (Wiedmann, 1830), on their respective preferred hosts (mangoes and guava). Experiments of larval competition and competition for ovipositioning sites by adult females were performed to compare the parameters of larval development time, numbers of pupae and emerged adults and numbers of ovipositions in the presence or absence of interspecific competition. We observed that the interactions between those species were asymmetrical and hierarchical, and our results suggest a competitive displacement of A. fraterculus by A. obliqua when those two species are present on the same fruit, whether mangoes or guavas.     

Intraspecific variation of cuticular hydrocarbon profiles in the Anastrepha fraterculus (Diptera: Tephritidae) species complex

The goal of this study was to define whether cuticular hydrocarbons (CHs) could be used for taxonomic determination of putative species hidden in the Anastrepha fraterculus cryptic species complex, widespread from Argentina to Mexico. Recently, increasing evidence of phenotypic and genetic variability has resulted in the characterization of eight morphotypes within this complex. The CH profiles of six A. fraterculus populations from Argentina, Brazil, Peru, Colombia and Mexico were analysed in this study by two‐dimensional gas chromatography coupled with mass spectrometry. In parallel, multiple factorial analyses were used to elucidate population structures. Vector populations segregated into four distinct groups. The analysis demonstrated that the studied populations from Peru, Argentina and southern Brazil (Vacaria) might be classified in accordance with the earlier division of the A. fraterculus complex into Peruvian, and Brazilian‐1 cryptic species, using the specific CH profiles. Population from south‐eastern Brazil (Piracicaba) formed separated group. Mexican and Andean (Colombian) putative species had similar CH signatures, when compared to each other.     

Morphometry and distribution of sensilla on the antennae of Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae)

Antennal sensilla of Anastrepha fraterculus (Wied.) were examined using scanning electron microscopy. In the flagellum, there are trichoid, basiconic, clavate type I and II, and styloconic sensilla and microtrichia. Only microtrichiae and chaetica sensilla were observed in the scape and pedicel. The number of sensilla in the flagellum was similar between sexes. At the apex there was a higher density of trichoid and an absence of clavate sensilla, while basiconic sensilla were more abundant in the proximal region.     

Comparative demography of three neotropical larval-prepupal parasitoid species associated with Anastrepha fraterculus (Diptera: Tephritidae)

The South American fruit fly, Anastrepha fraterculus (Wiedemann), is one of two fruit fly species of economic importance in Argentina, which along with the exotic Mediterranean fruit fly, Ceratitis capitata (Wiedemann) produce annual losses of nearly US$ 90 million for fruit production in this country.Biological control may contribute to integrated approaches to manage both pestiferous species.Information on survival, reproduction and population growth parameters, critical for successful augmentation of natural enemies, is provided for three fruit fly neotropical parasitoids, Aganaspis pelleranoi (Hymenoptera: Figitidae), Opius bellus and Doryctobracon crawfordi (Hymenoptera: Braconidae). A. pelleranoi had the highest intrinsic reproductive rate (r), survival time and reproductive time, followed by O. bellus. Meanwhile D. crawfordi showed the lowest values for reproductive and population growth parameters.     

Multispectral imaging for quality control of laboratory‐reared Anastrepha fraterculus (Diptera: Tephritidae) pupae

The sterile insect technique (SIT) has been widely used to suppress several fruit fly species. In southern Brazil, millions of sterile flies of the South American fruit fly, Anastrepha fraterculus Wiedemann (Dipetra: Tephritidae), will be produced in a mass‐rearing facility called MOSCASUL to suppress wild populations from commercial apple orchards. In spite of standard rearing conditions, the quality of pupal batches can be inconsistent due to various factors. The quantification of poor quality material (e.g. empty pupae, dead pupae or larvae) is necessary to track down rearing issues, and pupal samples must be taken randomly and evaluated individually. To speed up the inspection of pupal samples by replacing the manual testing with the mechanized one, this study assessed a multispectral imaging (MSI) system to distinguish the variations in quality of A. fraterculus pupae and to quantify the variations based on reflectance patterns. Image acquisition and analyses were performed by the VideometerLab4 system on 7‐d‐old pupae by using 19 wavelengths ranging from 375 to 970 nm. The image representing the near infrared wavelength of 880 nm clearly distinguished among high‐quality pupae and the other four classes (i.e. low‐quality pupae, empty pupae, dead pupae and larvae). The blind validation test indicated that the MSI system can classify the fruit fly pupae with high accuracy. Therefore, MSI‐based classification of A. fraterculus pupae can be used for future pupal quality assessments of fruit flies in mass‐rearing facilities.     

Measurement of the aculeus for the characterization of five Anastrepha species of the fraterculus group (Diptera: Tephritidae)

Species identification of the genus Anastrepha Schiner is based mostly on the shape of the aculeus apex. In some species groups, such as fraterculus, species are separated by subtle differences in the aculeus apex, namely Anastrepha fraterculus (Wied.), A. obliqua (Macquart), A. sororcula Zucchi, A. zenildae Zucchi and A. turpiniae Stone. In order to help the identification, the aculei of these five species from 25 localities of 17 Brazilian states were measured. The aculeus and apex lengths of these species vary along geographical distribution and even from specimens reared from same host. For this reason and due to superimposition, these Anastrepha species cannot be separated based on the two measures exclusively.     

Population genetics of Anastrepha fraterculus (Diptera,Tephritidae) in different hosts: Genetic differentiation and heterozygosity

An allozymic survey of 11 populations of Anastrepha fraterculus Wied. (Diptera, Tephritidae) in 8 different hosts from the same orchard revealed very small levels of genetic distance (D = 0.002±0.001) between hosts. The data thus provide no evidence for host-dependent genetic differentiation. Low values of heterozygosity were observed (H = 0.050±0.28) and 6 loci out of 11 were monomorphic. Biological attributes of the species, such as its polyphagic regime, multivoltine pattern of reproduction, and seasonal changes in population size, are discussed in an attempt to explain the low variability.     

Assortative mating among Anastrepha fraterculus (Diptera: Tephritidae) hybrids as a possible route to radiation of the fraterculus cryptic species complex

Reproductive isolation among populations of Anastrepha fraterculus has been found acting at the pre‐ and post‐zygotic levels. Differences in timing of sexual activity and male sexual pheromone composition among populations could partially account for prezygotic isolation. Hybrid males were found to produce a novel pheromone, which is a mix of parental pheromones. In the present study, we found that the hybrid females showed a significant preference to mate with hybrid males than with parental males. Male location during pheromone emission is associated with its reproductive success and, thus, differences in the location of males during courtship could also play a role in isolation. We found evidence that reproductive isolation is also related to the location of males during courtship. Hybrid male behaviour regarding location during pheromone release was found to be influenced by the maternal lineage. If these populations hybridized in the field, the hybrid females would tend to mate with hybrid males probably leading to the formation of a new entity within the A. fraterculus complex. This simple and fast process could be one reason explaining the high number of taxonomic entities within this complex. Further studies on other members of the fraterculus species group may reveal whether this can be considered as an example of homoploid hybrid speciation. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102 , 346–354.     

The Identity of Specimens of the Anastrepha fraterculus Complex (Diptera, Tephritidae) with Atypical Aculeus Tip

Several specimens collected in Paraguay along with Anastrepha fraterculus (sensu lato) have an aculeus tip similar to species from the fraterculus complex, but the teeth of the aculeus of these specimens are poorly defined. As Anastrepha species identification is based mostly on subtle differences in the aculeus tip, we studied these specimens with atypical aculeus tips (with poorly defined teeth) that slightly differs from the aculeus tip of species of the fraterculus complex (with well-developed blunt teeth), to determine if this is due to intraspecific variation or if it can characterize a full species. The Paraguayan specimens were separated in six groups under stereomicroscope according to variation in their aculeus tip. Specimens within each group were studied by means of morphometrics (traditional and geometric) and gene sequence analysis (COI and ITS1). Morphometric analyses were significant, but no clear groups were formed by the discriminant analyses of the aculeus and wing, and the COI and ITS1 sequence analysis clustered specimens with all six aculeus variations. Therefore, the subtle morphological differences observed in the aculeus tip of Paraguayan specimens are intraspecific variations and the Paraguayan specimens were more genetically closely related to Anastrepha sp. 3 from the fraterculus complex.     

Reproductive behaviour of the South American fruit fly Anastrepha fraterculus (Diptera: Tephritidae): laboratory and field studies

Abstract. The mate choice, courtship and oviposition behaviour of laboratory-reared and field-collected Anastrepha fraterculus (Wied.) were compared. In laboratory cultures in Southampton the duration of male calling activity in small leks increased gradually from 1-2h at 5 days old to up to 7 h at 10 days. This finding correlates with previous reports on the time at which male salivary glands, which are believed to produce sex pheromone, are fully developed. Wild flies which emerged from infested fruits in Brazil began to oviposit on the day they mated, whereas in laboratory flies oviposition began 1 day following the first mating. Both types of fly usually defended their position on a particular fruit throughout the day, and re-mated with either virgin or mated males. There was no significant difference in mating duration. Females did not copulate before the mean age (±SE) of 16.8±0.9 days. For both types of flies mating initiation occurred in the first 2h of photophase, with virgin females choosing mainly mated males. The average number of matings in the laboratory was three for females and four for males, and the interval between matings in females was significantly increased after the second mating. It is suggested that the tendency of virgin females to mate with mated males will lead to increased fitness, as males are on average 48 days old at their second mating. The potential life span of around 200 days for both sexes would allow adults to bridge the gap between seasonally available fruits in warm-temperate and sub-tropical South America.     

Wolbachia strains in cryptic species of the Anastrepha fraterculus complex (Diptera,Tephritidae) along the Neotropical Region

Infection by Wolbachia was described previously in eleven species of Anastrepha fruit flies some of which are important pests of fruticulture. One such species is the nominal Anastrepha fraterculus, the South American fruit fly, which actually comprises a complex of cryptic species. The suggestions of using Wolbachia for the control of these pest species, make imperative a more precise characterization of the existing strains of the bacteria. In this study, population samples of the A. fraterculus complex from Brazil, Argentina, Peru, Ecuador, Colombia, Guatemala and Mexico were analyzed for Wolbachia infection. The bacteria were genotyped by the MLST and WSP Typing methodologies. All samples were infected with Wolbachia of supergroup “A”. For each of the five MLST genes, unique as well as already known alleles were detected. Nineteen sequence types for the concatenated sequences of the five MLST genes, and twenty wsp alleles were found in the samples. Host-specific haplotypes, shared strains among distinct hosts, and more than one strain of Wolbachia were found in some population samples. Recombination among the MLST genes and intragenic recombination between wsp haplotypes was rare. Phylogenetic analysis showed a great similarity among the Wolbachia strains in the A. fraterculus complex. However, some strains of Wolbachia are found throughout the Neotropical Region and there are specific strains in determined geographical areas.     

Native and introduced host plants of Anastrepha fraterculus and Ceratitis capitata (Diptera: Tephritidae) in northwestern Argentina

Wild or commercially grown, native and exotic fruit were collected in 30 localities in the Tucumán province (NW Argentina) from January 1990 to December 1995 to determine their status as hosts of Anastrepha fraterculus (Wiedemann) and/or Ceratitis capitata (Wiedemann), the only two fruit fly species of economic and quarantine importance in Argentina. A total of 84,094 fruit (3,466.1 kg) representing 33 species (7 native and 26 exotic) in 15 plant families were sampled. We determined the following 17 host plant associations: Annona cherimola Miller (Annonaceae), Citrus paradisi Macfadyn (Rutaceae), Diospyros kaki L. (Ebenaceae), Eugenia uniflora L., Psidium guajava L., Myrcianthes pungens (Berg) Legrand (Myrtaceae), Ficus carica L. (Moraceae), Juglans australis Grisebach (Juglandaceae), Mangifera indica L. (Anacardiaceae), Eriobotrya japonica (Thunb.) Lindl., Prunus armeniaca L., P. domestica L., and P. persica (L.) Batsch (Rosaceae) were infested by both A. fraterculus and C. capitata. Citrus aurantium L., Citrus reticulata Blanco, Citrus sinensis (L.) Osbeck (Rutaceae), and Passiflora caerulea L. (Passifloraceae) were only infested by Ceratitis capitata. Out of a total of 99,627 adults that emerged from pupae, 69,180 (approximately 69.5%) were Anastrepha fraterculus, 30,138 (approximately 30.2%) were C. capitata, and 309 (approximately 0.3%) were an unidentified Anastrepha species. Anastrepha fraterculus predominated in native plant species while C. capitata did so in introduced species. Infestation rates (number of larvae/kg of fruit) varied sharply from year to year and between host plant species (overall there was a significant negative correlation between fruit size and infestation level). We provide information on fruiting phenology of all the reported hosts and discuss our findings in light of their practical (e.g., management of A. fraterculus and C. capitata in citrus groves) implications.     

An integrative multidisciplinary approach to understanding cryptic divergence in Brazilian species of the Anastrepha fraterculus complex (Diptera: Tephritidae)

An integrative multidisciplinary approach was used to delimit boundaries among cryptic species within the Anastrepha fraterculus complex in Brazil. Sexual compatibility, courtship and sexual acoustic behaviour, female morphometric variability, variation for the mitochondrial gene COI, and the presence of Wolbachia were compared among A. fraterculus populations from the Southern (Vacaria, Pelotas, Bento Gonçalves, São Joaquim) and Southeastern (Piracicaba) regions of Brazil. Our results suggest full mating compatibility among A. fraterculus populations from the Southern region and partial pre‐zygotic reproductive isolation of these populations when compared with the population from the Southeastern region. A. fraterculus populations from both regions differed in the frequency of courtship displays and aspects of the calling phase and mounting acoustic signal. Morphometric analysis showed differences between Southern region and Southeastern region samples. All populations analyzed were infected with Wolbachia. The trees generated from the COI sequencing data are broadly congruent with the behavioural and morphometric data with the exception of one Southern population. The likely mechanisms by which A. fraterculus populations might have diverged are discussed in detail based on behavioural, morphometric, molecular genetics, and biogeographical studies.     

Karyotypic and molecular identification of laboratory stocks of the South American fruit fly Anastrepha fraterculus (Wied) (Diptera: Tephritidae)

The taxonomic status of the tephritid pest Anastrepha fraterculus (Wied.) is a controversial subject, mainly because of misinterpretation of the observed genetic variation. In this work, the different karyotypes and DNA polymorphism of a geographically defined population from Northeastern Argentina were studied, using derived stocks maintained in the laboratory during 25 generations. The karyotypes were analyzed using C-banding and N-banding techniques, while DNA analysis was performed through the DNA polymerase chain reaction (PCR). The variants isolated from both the wild Montecarlo population and the derived laboratory stocks were fully compatible and are present in other wild populations from South Brazil (lat 31 degrees 30' S) to Mid-Argentina (lat 34 degrees 30' S). Single-pair crosses among stocks carrying different chromosomal variants demonstrated the absence of isolation barriers. The polymorphic fragments isolated by RAPDs/PCR showed polymorphisms among stocks whereas the analysis of rDNA ITS1 exhibit a unique ITS1 length. Our results seem to indicate that all the examined variants belong to a single species with extended polymorphism and therefore do not support the hypothesis that the extended chromosomal polymorphism in A. fraterculus implies the existence of a complex of cryptic species.     

Wolbachia in Anastrepha Fruit Flies (Diptera: Tephritidae)

Endosymbiotic bacteria of the genus Wolbachia are widespread among arthropods and cause a variety of reproductive abnormalities, such as cytoplasmic incompatibility, thelytokous parthenogenesis, male-killing, and host feminization. In this study, we used three sets of Wolbachia-specific primers (16S rDNA, ftsZ, and wsp) in conjunction with the polymerase chain reaction (PCR), cloning and sequencing to study the infection of fruit flies (Anastrepha spp. and Ceratitis capitata) by Wolbachia. The flies were collected at several localities in Brazil and at Guayaquil, Ecuador. All of the fruit flies studied were infected with Wolbachia supergroup A, in agreement with the high prevalence of this group in South America. Phylogenetic analysis showed that the wsp gene was the most sensitive gene for studying the relationships among Wolbachia strains. The Wolbachia sequences detected in these fruit flies were similar to those such as wMel reported for other fruit flies. These results show that the infection of Anastrepha fruit flies by Wolbachia is much more widespread than previously thought.     

X-ray doses to safely release the parasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae) reared on Anastrepha fraterculus larvae (Diptera: Tephritidae)

Diachasmimorpha longicaudata is a koinobiont larval parasitoid that is currently used to control fruit flies of the genera Anastrepha, Ceratitis and Bactrocera. In the rearing process, a fraction of the host larvae that are exposed to parasitoids escape from parasitism and develop into viable and fertile flies. This creates the need to eliminate emerging flies before the parasitoids are shipped for release, increasing costs due to additional handling steps. Exposure of fly eggs or larvae to gamma-irradiation before they are parasitised has been used to reproductively sterilise hosts, or even inhibit their emergence. Our aim was to determine whether X-ray radiation applied to Anastrepha fraterculus third instar larvae before they are exposed to parasitoids, inhibits fly emergence in non-parasitised larvae without affecting the performance of the parasitoids that emerge from parasitised larvae. Three X-ray doses: 6250.2 R, 8333.6 R and 10417 R (equivalent to 60, 80 and 100 Gy, respectively) and one γ-ray dose (100 Gy) were tested. Fly emergence decreased with increasing doses of radiation, showing null values for the higher X-ray dose and the dose of 100 Gy. Irradiation showed either no impact or a positive effect on parasitism rate and fecundity. Sex rate was biased towards females in almost every dose. We conclude that the two types of radiation evaluated here were equally effective in suppressing fly emergence with no detrimental effects on the biological quality of the produced parasitoids. X-rays offer an alternative method of irradiation than the conventional radiation source, i.e. γ-rays. These results represent a significant improvement in the development of a biological control programme against A. fraterculus.     

Consumption of sugars by Anastrepha suspensa (Diptera: Tephritidae)

We were interested in determining the feeding response of the Caribbean fruit fly, Anastrepha suspensa Loew (Diptera: Tephritidae), to various sugar concentrations to develop an improved bait for adults. We compared the consumption of 0.01-1.00 M concentrations of glucose, fructose, raffinose, and sucrose in no-choice tests for 24-h- and 6-d-old male and female flies. Sucrose was the most consumed sugar or within the most consumed group of sugars at 0.02-0.20 M concentrations. There were no differences in consumption among sugars at 0.01, 0.40, and 1.00 M. Consumption generally increased with increasing sugar concentration except that sucrose consumption peaked at 0.20 M. Twenty-four-hour females consumed less fructose than other sugars; 24-h males consumed more sucrose than fructose or raffinose, with an intermediate response to glucose. Females in the 6-d group consumed more sucrose than the other three sugars, whereas 6-d males exhibited no difference in consumption among sugars. In choice tests, flies consumed more sugar solution than water, but the difference between 0.20 M fructose and water was not significant for 24-h males or 24-h females. In choice tests between 0.20 M fructose and 0.20 M sucrose, both 24-h and 6-d females showed a preference for fructose. Males of both age classes showed no preference. These results indicate that the responses of flies to different sugars can vary by sugar, gender, and age.     

New mariner elements in Anastrepha species (Diptera: Tephritidae)

Mariner-like elements (MLE) are members from class II of transposable elements also known as DNA transposons. These elements have a wide distribution among different groups of organisms, including insects, which can be explained by horizontal and vertical gene-transfer. MLE families have been described in tephritid flies and other genera. During screening for Wolbachia bacteria in fruit flies of the genus Anastrepha, we discovered two sequences related to mariner-like elements. Based on these sequences, we designed primers that allowed us to isolate and characterize two new mariner-like elements (Anmar1 and Anmar2) in Anastrepha flies. These elements, which belong to the mellifera and rosa subfamilies have a low nucleotide diversity, and are probably inactive and acquired by vertical transfer. This is the first report of mariner-like transposons in flies found in South America.