Ceratitis Species Research Articles

Loop-mediated isothermal amplification of economically important Ceratitis species (Diptera: Tephritidae).

Ceratitis is an economically important genus of fruit flies that originated in Africa, has a wide host range, and causes serious economic losses due to its invasive damage. As a result, it is critical to identify them accurately and quickly in the world. Loop-mediated isothermal amplification (LAMP), as one of the representatives of isothermal amplification technology, has been widely used in the rapid nucleic acid detection of human pathogens and has shown its advantages in the identification of insect agricultural pests. In this study, using the mitochondrial cox1 and cob genes as target genes, the rapid molecular identification of the Ceratitis FARQ complex, C. cosyra, and C. capitata was realized based on LAMP. The experimental conditions optimization results showed that F3/B3:FIP/BIP = 1:8 was the optimal primer concentration ratio and 63 °C was the optimal reaction temperature. The sensitivity of the primers obtained in this study can reach up to 0.01 ng/μl DNA. A loop-mediated isothermal amplification identification technology system was established based on rapid, rough DNA extraction and visual detection of Ceratitis economically important fruit flies. The positive reaction system changed from pink to khaki by visual detection. The identification flow can be completed within 1 hour, including sample processing, DNA extraction, and LAMP visual detection.

Response of some mango-infesting fruit flies to aqueous solutions of the basil plant Ocimum tenuiflorum L

IntroductionHorticulture is an important sub-sector for many African countries. Tephritid fruit flies continue to cause serious direct and indirect damage to the sub-sector. The male annihilation technique (MAT) using methyl eugenol (ME) is among the most revered techniques in fruit fly management. However, ME-based lures are only available to some farmers in Africa. Several basils of the genus Ocimum produce ME.MethodsIn laboratory experiments, we used a dual choice olfactometer to assess the attraction of adult oriental fruit fly Bactrocera dorsalis to intact plants, 100 µL commercial methyl eugenol (ME) (positive control), 10 mg/mL of O. tenuiflorum aqueous extract (Otae), and a negative control (blank). Large arena experiments were conducted to compare the responses of four fruit fly species of economic importance in Africa to Otae-baited traps.ResultsIn dual choice olfactometer assay, more B. dorsalis males (78.6%) than females (23.8%) were attracted to Otae. Most B. dorsalis males were attracted to the positive control (90.5%) and 10 mg/mL Otae (88.5%) compared to the whole plant (84.0%). In large arena experiments, Otae attracted the most B. dorsalis males (47.7%), followed by Zeugodacus cucurbitae males (25.1%), while males of Ceratitis species were the least attracted. The analysis of headspace volatiles from aerial plants, Otae and essential oil using gas chromatography-mass spectrometry detected 97 compounds. The intact plant without flowers, intact plant with flowers, Otae, and the O. tenuiflorum essential oil contributed 76, 62, 33 and 28 compounds, respectively. ME comprised the top three most abundant compounds, especially in the essential oil.DiscussionOur study confirms the potential of O. tenuiflorum as a source of ME for tephritid fruit fly integrated management in Africa. Otae, a readily available alternative to commercial ME, can be used to develop fruit fly-baited traps, particularly for smallholder farmers.

Tephritid Fruit Fly Species Composition, Seasonality, and Fruit Infestations in Two Central African Agro-Ecological Zones.

Bactrocera dorsalis and several Africa-native Ceratitis species are serious constraints to fruit production in sub-Saharan Africa. A long-term trapping and fruit collection study was conducted (2011-2016) in two contrasting agro-ecological zones (AEZs) of Cameroon to determine fruit fly species composition, seasonality, attraction to various lures and baits, and fruit infestation levels. Ten tephritid species from genera Bactrocera, Ceratitis, Dacus, and Perilampsis were captured in traps. Bactrocera dorsalis was the most dominant of the trapped species and persisted throughout the year, with peak populations in May-June. Ceratitis spp. were less abundant than B. dorsalis, with Ceratitis anonae dominating in the western highland zone and Ceratitis cosyra in the humid forest zone. Methyl eugenol and terpinyl acetate captured more B. dorsalis and Ceratitis spp., respectively than Torula yeast. The latter was the most effective food bait on all tephritid species compared with BioLure and Mazoferm. Bactrocera dorsalis was the dominant species emerging from incubated fruits, particularly mango, guava, and wild mango. Four plant species-I. wombolu, Dacryodes edulis, Voacanga Africana and Trichoscypha abut-were new host records for B. dorsalis. This study is the first long-duration and comprehensive assessment of frugivorous tephritid species composition, fruit infestations, and seasonality in Central Africa.

Patterns of Remating Behaviour in Ceratitis (Diptera: Tephritidae) Species of Varying Lifespan.

Trade-offs between life-history traits offset the energetic costs of maintaining fitness in complex environments. Ceratitis species have been recorded to have long lifespans, which may have evolved in response to seasonal resource fluctuation. It is thus likely that reproductive patterns have evolved concomitantly as part of the trade-off between lifespan and reproduction. In this study, we investigated how reproductive patterns differ between Ceratitis cosyra (Walker) and Ceratitis capitata (Wiedemann; Diptera: Tephritidae), two species with different average and maximum lifespans. Females of both species were mated and patterns of female survival, fecundity, remating and sperm storage were tested. Ceratitis cosyra had a higher rate of survival and a lower fecundity when compared with the shorter-lived C. capitata, suggesting that both species exhibit a trade-off between lifespan and reproduction. Both species showed a similar and consistent willingness to remate, despite declines in sperm storage, suggesting that sperm alone does not fully inhibit remating. As expected, C. cosyra transferred high numbers of sperm during the first mating. However, sperm stores declined unexpectedly by 14 days. This indicates that males might transfer large ejaculates as a nuptial gift, that females then later degrade as a source of nutrients. Large declines in sperm storage may also indicate that females discard excess sperm stores due to the toxicity involved with storing sperm. These results do not suggest that patterns of sperm storage and remating align with lifespan and resource seasonality in these species, but a wider range of species needs to be assessed to better understand variation in Ceratitis mating systems.

A loop-mediated isothermal amplification (LAMP) assay for rapid identification of Ceratitis capitata and related species.

True fruit flies (Tephritidae) are among the most destructive agricultural pests in the world, attacking a wide range of fruits and vegetables. The Mediterranean fruit fly Ceratitis capitata is a highly polyphagous species but, being widely established in the Mediterranean region, is not considered as a EU quarantine pest. Hence, it is important to discriminate Ceratitis capitata from non-EU tephritid species, present in imported fruit and vegetables, as non-EU species have a quarantine status. However, morphological identification of tephritid larvae, the most frequently intercepted stage in non-EU produce, is difficult and an easy-to-use molecular diagnostic tool would be helpful for rapid species identification. Therefore, a loop-mediated isothermal amplification (LAMP) method was developed for C. capitata and non-EU tephritids Ceratitis cosyra group1 and Ceratitis species from the FARQ complex, C. fasciventris, C. anonae, C. rosa and C. quilicii. LAMP assays were run with DNA from ILVO collected specimens and DNA samples collected during previous research surveys. LAMP primers were species-specific, with LAMP amplification occurring within 45 minutes for the targeted species. In addition, LAMP assays were successful for all C. capitata life stages or a limited amount of tissue. To conclude, the LAMP assays developed in this study were able to distinguish C. capitata from non-EU Tephritidae species and could be a useful tool for the rapid identification of C. capitata.

Are Yeast Autolysate Attractants for Ceratitis Species (Diptera: Tephritidae) in South Africa More Attractive and Palatable Than a Currently Used Protein Attractant?

Due to increasing usage of copper fungicides in citrus orchards shortly before the application of protein baits for fruit fly control and the risk of this combination causing fruit phytotoxicity, an investigation of attractants containing higher amounts of yeast autolysate rather than protein hydrolysate was conducted. A commercial yeast autolysate from Australia was less attractive than a South African product HymLure (with a mixture of autolysate and hydrolysate) to three Ceratitis species: Ceratitis capitata (Wiedemann), Ceratitis rosa s.s. Karsch, and Ceratitis cosyra (Walker). Other custom-formulated autolysates were also inferior to HymLure for these species. Consumption of autolysates during the first hour of feeding was similar to that of HymLure. The inferior attraction of these three species of fruit flies to autolysates, relative to one containing hydrolysate, did not warrant further investigation of autolysates for use in baits to control these quarantine pests.

Niche shift of tephritid species after the Oriental fruit fly (Bactrocera dorsalis) invasion in La Réunion

AbstractAimIn a context of successive fruit fly invasions (Tephritidae), this study investigated how the top invader, Bactrocera dorsalis, displaced established fruit fly populations. We focused, particularly, on how this invasion impacted the host range and climatic niche of each resident species.LocationLa Réunion, France, Indian Ocean.MethodsWe collected fruit from more than 100 plant species across the island, in cultivated and non‐cultivated areas at different altitudes in order to monitor the emergence of fruit flies. Fruit collection was conducted over two field campaigns: from 2001 to 2009, before the B. dorsalis invasion; and from 2018 to 2019, after the B. dorsalis invasion. We compared the distribution and host range of fruit fly species for the two periods.ResultsOur results confirmed the generalist character of B. dorsalis, with the infestation of 52 out of 112 of the fruit species collected in the field. After the B. dorsalis invasion, we observed a shift in the host range and spatial distribution of established tephritids. The host range of specialist species that only share a few host species with B. dorsalis did not change significantly. On the contrary, we observed a significant shift in diversity or proportion of host range and climatic niches for the generalist species, such as Bactrocera zonata, Ceratitis quilicii and Ceratitis capitata.Main conclusionsWe provide evidence of the competitive displacement induced by B. dorsalis on other established species. The coexistence between B. dorsalis and generalist Ceratitis species seems possible because they have different responses to climatic conditions or the capacity to exploit other host fruit species. In contrast, the coexistence of B. zonata with B. dorsalis seems to be compromised because both species have similar ecological requirements. This research provides useful information for managing invasions, particularly since understanding competitive displacements is essential for the identification of empty niches and for modelling potential species distribution.

Plant Chemicals and the Sexual Behavior of Male Tephritid Fruit Flies

Plant compounds affect insects in many different ways. In addition to being a food source, plants also contain secondary metabolites that may have positive and negative impacts on insects. The influence of these compounds on sexual behavior, in particular, has been the focus of many recent studies. Here, we review the existing literature on the effects of plant compounds on the sexual behavior of tephritid fruit fly males. We put special focus on polyphagous species whose males congregate in leks, where females exert strong mate selection. We first summarize the main findings related to plant compounds that increase male signaling behavior and attraction of females and consequently increase mating frequency, a phenomenon that has been recorded mainly for species of Anastrepha and Ceratitis. In other tephritid species, males are attracted to phenylpropanoids produced by plants (such as methyl eugenol or raspberry ketone) that, upon encounter, are consumed and sequestered by males. These compounds, or metabolic derivatives, which normally have negligible nutritional value, are included in the pheromone and also confer advantages in a sexual context: enhanced female attraction and improved male mating success. These phenomena have been reported for several Bactrocera species as well as for Zeugodacus cucurbitae. Because many tephritid species are serious pests, the effect of plant compounds on male behavior has been explored for potential incorporation into control strategies such as the sterile insect technique (SIT). We conclude noting several factors, such as age and nutrition during larval and adult stage, that modulate the effect of plant compounds on male mating behavior as well as some prominent gaps that preclude a thorough understanding of the plant-mediated enhancement of male sexual performance and hence limit our ability to effectively utilize phytochemicals in pest control strategies.

Diversity and seasonal abundance of tephritid fruit flies in three agro‐ecosystems in Benin, West Africa

AbstractThe horticulture industry in Benin is under threat due to fruit fly infestation on important crops such as cucurbits and mango. Given the economic losses due to fruit fly infestations, population dynamics of fruit flies on these crops were monitored over 6 years (2004–2010) using three parapheromones—cuelure, methyl eugenol and terpinyl acetate. A total of 2,714,186 specimens belonging to 14 species in four genera (Bactrocera, Ceratitis, Dacus and Zeugodacus) were collected. Bactrocera dorsalis was the dominant species that accounted for 89%, 89.6% and 77.2% of captures in the Forest Savannah Mosaic (FSM), Southern Guinea Savannah (SGS) and Northern Guinea Savannah (NGS) agro‐ecosystems, respectively. Shannon index values of species diversity differed significantly among agro‐ecosystems, with highest diversity in FSM followed by SGS and NGS. Temperature, rainfall and relative humidity (RH) positively influenced the abundance of Dacus species and Zeugodacus cucurbitae across the three agro‐ecosystems. Monthly mean temperatures were in a range suitable for development and reproduction of fruit flies, although minimum temperatures could be below such range, which may explain the lower abundance of fruit fly populations during this period of the year. The abundance of B. dorsalis was negatively correlated with maximum temperature and positively correlated with RH and rainfall. Ceratitis species assemblages were significantly affected by minimum temperatures, rainfall and agro‐ecosystem. Populations of B. dorsalis and Ceratitis cosyra dropped drastically in August, while those of Z. cucurbitae and Dacus punctatifrons increased sharply from August. Results of this study provide a platform for developing a comprehensive and sustainable management strategy for B. dorsalis and other fruit flies in the three agro‐ecosystems to protect the horticulture industry in Benin.

Sensitivity of an enriched ginger oil based trapping system for Ceratitis fruit fly pests (Diptera: Tephritidae)

Enriched ginger oil (EGO) is a new male attractant for Ceratitis (Diptera: Tephritidae) species. The relative sensitivity of three Ceratitis pest species, C. capitata, C. rosa and C. cosyra, and their distance-dependent responses to EGO were determined using mark-release-recapture trials in three commercial fruit orchards in Mpumalanga, South Africa. Mature males of the three species (9–14 days after adult emergence) marked with fluorescent pigments were released at four distances: 25 m, 50 m, 100 m and 200 m from a centrally located white Delta trap baited with EGO. Different pigment colours were used for the different distances. Two releases were conducted in each orchard with an interval of one month between the releases. Traps were checked the following day, one week, two weeks and a month after release. Specimens captured were examined under a UV light to determine pigment colour on males of C. capitata, C. rosa and C. cosyra. There were no significant differences in recapture rates of the three Ceratitis species in the EGO baited trap. Most of the recaptures of all species occurred within a 50 m distance from the EGO baited trap. Most of the recaptures also occurred on the day following release. Based on the recapture rates obtained in this study, the probability of a trapping grid of 5 EGO baited traps per 2.59 km2 capturing one or more flies of C. capitata, C. rosa and C. cosyra for a population consisting of 1000 males for each species was estimated at over 95%. The EGO based trapping system would be an effective detection method for C. capitata, C. rosa and C. cosyra in pest free areas and in areas of low pest prevalence.

Host species and vegetable fruit suitability and preference by the parasitoid wasp Fopius arisanus

AbstractParasitoids that oviposit in a concealed host inside a plant part need to be able to find both the plant and the host. Egg parasitoids of fruit‐infesting Tephritidae need to assess the oviposition site based both on the host egg and the infested fruit. Infestation by Tephritidae fruit flies threatens fruit and vegetable production. Management methods have been implemented including biological control, using Fopius arisanus Sonan (Hymenoptera: Braconidae). The parasitism by F. arisanus in three Tephritidae flies in vegetable fruits was investigated. Laboratory assays were conducted to assess the parasitoid's preference and survival. Zucchini, sweet pepper, and tomato were artificially infested with eggs of Bactrocera dorsalis Hendel, Ceratitis capitata Wiedemann, and Ceratitis cosyra Walker (all Diptera: Tephritidae), then exposed to mated naïve F. arisanus females in a 20:1 egg:parasitoid ratio. Parasitoid behavioral activities (resting, antennating, probing, ovipositing) were observed on the infested fruits. Parasitism rate was determined by dissection of fruit fly eggs under a stereomicroscope. Behavioral activities of F. arisanus differed between all the fruits when infested with B. dorsalis or C. cosyra eggs but differed only between some of the fruits when infested with C. capitata. Fopius arisanus preferred B. dorsalis over C. capitata and C. cosyra, with a parasitism rate 2× higher on B. dorsalis compared to the Ceratitis species. Preference for fruits was dependent on the infesting fruit fly. The emergence of F. arisanus was higher with B. dorsalis than with Ceratitis spp. Although B. dorsalis completed its development earlier than Ceratitis spp., host fly species did not affect the developmental time of F. arisanus. We discuss the significance of F. arisanus preference in relation to naturally occurring Tephritidae infestations. We also discuss whether some fruits might constitute a refuge for Tephritidae flies and whether this will affect the current biological control efforts against B. dorsalis.

Indigenous hosts of economically important fruit fly species (Diptera: Tephritidae) in South Africa

AbstractFruit flies are among the most economically important fruit pests worldwide. The larvae are phytophagous and constitute a major production constraint in the horticultural industry. Various indigenous fruits were sampled from 2010 to 2015 in Limpopo and Mpumalanga Provinces of South Africa to determine the presence of fruit fly species. The Oriental fruity fly, Bactrocera dorsalis (Hendel), Mediterranean fruit fly, Ceratitis capitata (Wiedemann), the Marula fruit fly, Ceratitis cosyra (Walker) and the Natal fruit fly, Ceratitis rosa sensu lato Karsch were reared from the indigenous fruits. B. dorsalis was reared from one marula, Sclerocarya birrea (A.Rich.) Hochst. (Anacardiaceae) sample. All three Ceratitis species were reared from the Mobola plum, Parinari curatellifolia Planch. ex Benth. (Chrysobalanaceae) and Kei‐apple, Dovyalis caffra (Hook.f. & Harv.) Sim (Salicaceae). Wild plum, Harpephyllum caffrum Bernh. (Anacardiaceae), African mangosteen, Garcinia livingstonei T.Anderson (Clusiaceae), Cape ash, Ekebergia capensis Sparrm. (Meliaceae), waterberry, Syzygium cordatum Hochst. ex Krauss (Myrtaceae) and stemfruit, Englerophytum magalismontanum (Sond.) T.D.Penn. (Sapotaceae) were hosts to both C. capitata and C. rosa s.l. C. capitata was also reared from num‐num, Carissa bispinosa (L.) Desf. ex Brenan (Apocynaceae), simple‐spined num‐num, Carissa spinarum L. (Apocynaceae), brown ivory, Berchemia discolor (Klotzsch) Hemsl. (Rhamnaceae), wild coffee, Coffea racemosa Lour. (Rubiaceae) and common red milkwood, Mimusops zeyheri Sond. (Sapotaceae). C. cosyra was also found in the fruit of S. birrea and the wild mango, Cordyla africana Lour. (Leguminosae). C. rosa s.l. was also reared from Natal plum, Carissa macrocarpa (Eckl.) A.DC. (Apocynaceae), waterpear, Syzygium guineense (Willd.) DC. (Myrtaceae), Natal dovyalis, Dovyalis longispina (Harv.) Warb. (Salicaceae) and wild apricot, Dovyalis zeyheri (Sond.) Warb. (Salicaceae). No fruit flies were reared from Ficus spp. (Moraceae).Various indigenous hosts could serve as a reservoir for infesting cultivated crops.

The complex case of Ceratitis cosyra (Diptera: Tephritidae) and relatives. A DNA barcoding perspective

AbstractThe mango fruit fly, Ceratitis (Ceratalaspis) cosyra, is a major agricultural pest affecting mango production in sub‐Saharan Africa. Morphological differences between C. cosyra and five closely related Ceratitis (Ceratalaspis) species (C. discussa, C. pallidula, C. quinaria, C. silvestrii, C. striatella) are subtle, so that their identification often requires specialized expertise. A previous study based on nuclear molecular markers (microsatellites) showed cryptic genetic variation within C. cosyra with separate microsatellite genotypic clusters also occurring in sympatry. This study aimed at verifying whether DNA barcoding can (a) separate C. cosyra from morphologically similar Ceratitis (Ceratalaspis) species and (b) resolve the cryptic genetic variation previously observed within C. cosyra. A subset of 62 C. cosyra specimens previously genotyped at 16 microsatellite loci was subjected to DNA barcoding. This data set was integrated with 130 DNA barcodes of C. cosyra, C. discussa, C. quinaria, C. silvestrii and C. striatella. Neighbor Joining, maximum likelihood and Bayesian tree reconstructions confirmed the presence of two main C. cosyra genotypic groups and resolved well‐supported groups corresponding to C. discussa, C. pallidula, C. striatella and to C. quinaria/C. silvestrii. The analysis of morphological characters did not show consistent morphological differences between the two groups of C. cosyra and called into question the morphological characters currently implemented in the identification of C. striatella. These results further support the hypothesis of cryptic speciation within the mango fruit fly and suggest that DNA barcoding represents a suitable complementary tool for the problematic diagnosis of C. cosyra and C. striatella.

Efficacy of trapping systems for monitoring of Afrotropical fruit flies

AbstractThe efficacy of different attractant trap combinations for monitoring of Afrotropical fruit flies was evaluated in commercial fruit production areas and natural areas in the northern parts of South Africa. Five food‐based attractants: three‐component Biolure, a combination of ammonium acetate and trimethylamine, a combination of ammonium acetate and putrescine, torula yeast and Questlure, were evaluated in McPhail‐type traps. Two male lures targeting Ceratitis species: EGO Pherolure and trimedlure, were evaluated in Sensus bucket traps. Three male lures targeting Dacine fruit flies: methyl eugenol, cue lure and zingerone, were evaluated in Moroccan bucket traps. The three‐component Biolure was among the most effective food‐based attractant for females of Ceratitis capitata, C. rosa (R1 and R2), C. cosyra and Bactrocera dorsalis. EGO Pherolure attracted a wider range of Ceratitis species than trimedlure but was equally as effective as trimedlure for males of C. capitata and C. rosa (R1 and R2). Dacine flies were specific in their responses to male lures. Bactrocera dorsalis males responded mainly to methyl eugenol. Males of Dacus species responded to either cue lure or zingerone. Zingerone attracted two cucurbit‐infesting Dacus species: D. frontalis and D. durbanensis. For B. dorsalis, C. capitata and C. cosyra in commercial fruit production areas, patterns of female catches in traps containing food‐based attractants were different to patterns of male catches in male lure‐based traps. Implications of these trapping results are discussed.

Diversity of Frugivore Fruit Fly Species (Diptera: Tephritidae) in Niger State, Southern Guinea Savannah Zone of Nigeria

The diversity of frugivore fruit fly (Diptera: Tephritidae) species was studied from May-October, 2014 in orchards of mango, cashew, guava and water melon situated in Paikoro and Lapai Local Government Areas of Niger State, Nigeria, The flies were trapped using Ball and Steiner traps baited with torula yeast and a para-pheromone, methyl eugenol, respectively. Traps were cleared at 7-day intervals and serviced monthly. Trapped flies were collected in zip-packs, transferred into 75% alcohol and identified in the laboratory. Twenty-three fly species within the genera Bactrocera, Ceratitis, Dacus, Perilampsis and Trirhithrum were collected, with Ceratitis species being the richest. The trap with para-pheromone trapped higher number of species than that with torula yeast bait. Bactrocera invadens was the most abundant species at all sites. Alpha diversity index showed that the mango orchard was richer in species than other sites while Beta diversity index revealed high similarities among sampling sites.

Do thermal tolerances and rapid thermal responses contribute to the invasion potential of Bactrocera dorsalis (Diptera: Tephritidae)?

Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) has shown remarkable range expansion over the past 10years and invaded several new continents including Africa. Here we report results of a detailed assessment of acute high and low temperature survival ability and the plasticity thereof, to test the hypothesis that traits of the thermal niche have contributed to the species’ invasion ability. We also assess life-stage-related variation of thermal tolerances to determine potential stage-related environmental sensitivity. The temperatures at which c. 20% of the population survived of B. dorsalis were determined to be −6.5°C and 42.7°C, respectively, when using 2h exposures. Further, four life stages of B. dorsalis (egg, 3rd instar larvae, pupae and adults) were exposed to high and low discriminating temperatures to compare their thermal survival rates. The egg stage was found to be the most resistant life stage to both high and low temperatures, since 44±2.3% survived the low and 60±4.2% survived the high discriminating temperature treatments respectively. Finally, the potential for adult hardening responses to mediate tolerance of extremes was also considered using a diverse range of acute conditions (using 2h exposures to 15°C, 10°C and 5°C and 30°C, 35°C, 37°C and 39°C as hardening temperatures, and some treatments with and without recovery periods between hardening and discriminating temperature treatment). These showed that although some significant hardening responses could be detected in certain treatments (e.g. after exposure to 37°C and 39°C), the magnitude of this plasticity was generally low compared to two other wide-spread and more geographically-range-restricted con-familial species, Ceratitis capitata and C. rosa. In other words, Bactrocera dorsalis adults were unable to rapidly heat- or cold-harden to the same extent as the other Ceratitis species examined to date. These results suggest a narrower thermal niche in B. dorsalis compared to these Ceratitis species – in both basal and plastic terms – and suggests that its geographic distribution might be more restricted in consequence.

Description of new Ceratitis species (Diptera: Tephritidae) from Africa, or how morphological and DNA data are complementary in discovering unknown species and matching sexes

This paper describes five new Ceratitis species from the eastern and southern parts of the Afrotropical Region: C. (Pterandrus) quilicii De Meyer, Mwatawala & Virgilio sp. nov.; C. (Ceratalaspis) pallidula De Meyer, Mwatawala & Virgilio sp. nov.; C. (Ceratalaspis) taitaensis De Meyer & Copeland sp. nov.; C. (Ceratalaspis) sawahilensis De Meyer & Virgilio sp. nov.; and C. (Ceratalaspis) flavipennata De Meyer & Virgilio sp. nov. Their relationships with closely allied species within their respective subgenera are discussed where appropriate, and diagnostic characters are given. DNA barcodes are provided for all new species. In addition, the hitherto unknown male of C. (Pardalaspis) serrata De Meyer, 1996 is described, based on material collected in the Democratic Republic of Congo. Recognition of these new species and sexes is the result of an integrative approach using morphological characters and DNA data.

Description of third instar larvae of Ceratitis fasciventris, C. anonae, C. rosa (FAR complex) and C. capitata (Diptera, Tephritidae).

Third instar larvae of members of the Ceratitis FAR complex, including Ceratitis fasciventris (Bezzi), Ceratitis anonae Graham, and Ceratitis rosa Karsch are described and compared with those of Ceratitis capitata (Wiedemann). Diagnostic characters, such as presence vs. absence of a secondary tooth on the mandibles, previously used to separate Ceratitis capitata from Ceratitis rosa, are shown to vary in each species. Significant variation in diagnostic morphological characters among populations of Ceratitis rosa from east and south Africa is documented; however, the differences are not simply congruent with the R1 and R2 designations based on other studies. Quantitative measures of numerous morphological characters are consistently smaller in the larvae of Ceratitis fasciventris and distinguish them from other species of the FAR complex. Larvae of Ceratitis capitata can be distinguished from those of the FAR complex by characters such as absence of accessory plates of the oral ridges, the shape of the anterior spiracle, and the pattern of dorsal spinules. Previous studies indicated that absence of accessory lobes separate the genus Ceratitis from Bactrocera, but this is shown to be incorrect, as accessory lobes are in fact present in several species of Ceratitis.

Molecular Identification of Tephritid Fruit Flies (Diptera: Tephritidae) Infesting Sweet Oranges in Nsukka Agro-Ecological Zone, Nigeria, Based on PCR-RFLP of COI Gene and DNA Barcoding

The exotic tephritid Bactrocera invadens (syn. Bactrocera dorsalis) and the native Ceratitis species constitute serious pests and major impediment to Citrus production in Nigeria. Lack of accurate and reliable diagnostic methods independent of the life stages intercepted for both genera hampers implementation of appropriate early management programmes to avoid economic losses and delays in quarantine decision regarding these pests. We used a PCR amplification technique to successfully establish RFLP patterns of the COI coding gene to differentiate the exotic B. invadens from the native Ceratitis species infesting sweet oranges in Nigeria. The universal barcoding primer pair LCO1490/HCO2198 was used to amplify 658 bp-long fragment of the mitochondrial COI coding gene. The amplified fragment was analysed by automated direct sequencing and RFLP. Intraspecific variation in nucleotide sequences of B. invadens was very low with not more than 2 bp substitutions in four out of 31 individuals sequenced. Among the ...

Molecular Identification ofCeratitis capitata(Diptera: Tephritidae) using DNA Sequences of the COI Barcode Region

AbstractThe utility of the cytochrome oxidase I gene barcode region for diagnosis of the Mediterranean fruit fly, Ceratitis capitata (Weidemann), is evaluated using African fruit fly collections. The method fails to discern C. capitata from its close relative Ceratitis caetrata Munro, based on genetic distances, parsimony networks, or nucleotide diagnostic characters observed in the DNA barcode sequences. When treated as a single taxon, it is possible to discern the C. capitata + C. caetrata lineage from other Ceratitis species. Levels of intraspecific diversity vary within the genus Ceratitis and multiple copies of the mitochondrial gene are reported for Ceratitis cosyra (Walker). The DNA barcoding method based on genetic distance is compared with a molecular identification method using restriction fragment length polymorphism. The DNA barcode and restriction fragment-length polymorphism methods provide similar identification results, but the DNA sequence information is more suitable for quantitative analysis of the information.