Safety, regulatory and environmental issues related to breeding and international trade of edible insects in Africa.

Insect breeding or farming for food and feed is an emerging enterprise that can address the ever-growing demand for protein and curb high unemployment rates in Africa and beyond. However, for the sector to prosper, its value chain needs to be regulated to ensure sustainability and safety for consumers and the environment. Although a few African countries, such as Kenya, Uganda and Rwanda, have promulgated standards on the use of insects as food and feed, greater efforts are needed in other countries, and relevant policies governing the sector need to be formulated. All over the globe, attention to the regulation of the edible insect sector is increasing, and more investment in the industry is foreseen. Safety issues such as identifying which species should be reared, substrate quality and traceability imposed by importing countries will be critical for expansion of the sector. This paper analyses safety, regulatory and environmental issues related to breeding and international trade of edible insects in Africa and provides case studies and recommendations for sustainable use of insects for food and feed.

Les élevages et les fermes d'insectes destinés à l'alimentation humaine et animale sont de nouvelles entreprises qui pourraient répondre à la hausse continue de la demande en protéines tout en réduisant les taux élevés du chômage en Afrique et ailleurs. Néanmoins, pour que ce secteur puisse prospérer, sa chaîne de création de valeur doit être réglementée afin de garantir sa durabilité et son innocuité pour les consommateurs et l'environnement. Si un petit nombre de pays africains dont le Kenya, l'Ouganda et le Rwanda ont élaboré des normes applicables à l'utilisation des insectes pour l'alimentation humaine et animale, dans d'autres pays les efforts doivent se poursuivre et des politiques appropriées doivent être mises en place pour régir ce secteur. Partout dans le monde, la réglementation du secteur des insectes comestibles fait désormais l'objet d'une attention considérable et des investissements accrus dans la filière sont attendus. Les questions de sécurité telles que l'identification des insectes à élever, la qualité du substrat et la traçabilité exigée par les pays importateurs seront cruciales pour le développement du secteur. Les auteurs analysent les questions de sécurité et les enjeux réglementaires et environnementaux liés à l'élevage et au commerce international d'insectes comestibles en Afrique et présentent des études de cas et des recommandations pour une utilisation durable des insectes destinés à l'alimentation humaine et animale.

La cría o producción de insectos con fines de alimentación humana o animal es una actividad incipiente que puede ayudar a responder a la siempre creciente demanda de proteínas y a contener las elevadas tasas de desempleo de África y otras regiones. Para que el sector prospere, no obstante, es preciso reglamentar su cadena de valor a fin de asegurar su sostenibilidad y su inocuidad para el consumidor y el medio ambiente. Aunque unos pocos países africanos, como Kenia, Uganda o Ruanda, tienen promulgadas normas sobre el uso de insectos para la alimentación humana o la producción de piensos, aún hay que redoblar esfuerzos en otros países y formular políticas que ordenen el sector. En todo el planeta se presta hoy una atención sin precedentes a la regulación del sector de los insectos comestibles, un sector que previsiblemente va a recibir cada vez más inversiones. Las cuestiones ligadas a la inocuidad, como la identidad de los insectos que se van a producir, la calidad del substrato o la rastreabilidad impuesta por los países importadores, serán fundamentales para el desarrollo del sector. Los autores analizan las cuestiones reglamentarias, ambientales y de inocuidad que se plantean en relación con la cría y el comercio internacional de insectos comestibles en África, presentan ejemplos concretos y formulan recomendaciones para un uso sostenible de los insectos con fines de alimentación humana o animal.

Distribution, abundance and natural enemies of the invasive tomato leafminer, Tuta absoluta (Meyrick) in Kenya.

Tuta absoluta (Meyrick) has become a serious menace to sustainable production of tomato in Kenya. A survey was conducted between April 2015 and June 2016 to determine its distribution, abundance, infestation, and damage levels on tomato, and associated natural enemies. Trap counts of T. absoluta moths were recorded in all surveyed 29 counties, which indicated its nationwide distribution irrespective of altitude. Tuta absoluta was present in both open fields and greenhouses. The highest moth/trap/day was 115.38 ± 15.90. Highest leaf infestation was 92.22% and the highest number of mines and larvae per leaf were 3.71 ± 0.28 and 2.16 ± 0.45, respectively. Trap captures in terms of moth/trap/day were linearly and positively related to leaf infestations in open fields (R2 = 0.81) and greenhouses (R2 = 0.61). Highest fruits' infestation and damage were 60.00 and 59.61%, respectively, while the highest number of mines per fruit was 7.50 ± 0.50. Nesidiocoris tenuis (Reuter) and Macrolophus pygmaeus (Rambur) were identified as predators of T. absoluta larvae. Nine species of larval parasitoids were recovered from infested foliage, with a combined parasitism of 7.26 ± 0.65%. Hockeria species was the most dominant (31.25%) and accounted for 12.88 ± 1.47% parasitism. Two species of larval parasitoids, Hockeria and Necremnus were obtained from sentinel plants with an average parasitism of 1.13 ± 0.25. The overall abundance and parasitism rates of recovered natural enemies were low to effectively control the field populations of T. absoluta. These findings form the basis of researching and developing effective and sustainable management strategies for the pest.

Temperature-dependent development, survival and reproduction of Apanteles hemara (Nixon) (Hymenoptera: Braconidae) on Spoladea recurvalis (F.) (Lepidoptera: Crambidae).

The temperature-dependent development of Apanteles hemara (Nixon), a larval endoparasitoid of the devastating amaranth pest Spoladea recurvalis (F.) was studied in the laboratory at six constant temperatures (10, 15, 20, 25, 30 and 35 °C), a photoperiod of 12L:12D and a relative humidity of 60-70%. Developmental time decreased significantly with increasing temperature within the range of 15-30 °C. The parasitoid's pupal mortality, successful parasitism rate, adult emergence rate and longevity, sex ratio and fecundity were affected by temperature. The population of A. hemara failed to develop at 10 and 35 °C. The development threshold (Tmin) and the thermal constant (K) were calculated by the linear model while the lethal temperature (Tmax) was determined by the Lactin-1 model. The estimated values of Tmin, Tmax and K by the two models were 10.3 °C, 35.0 °C and 185.18 DD respectively for the total immature development. The estimated value of the optimum temperature using the Taylor model was 30.8 °C. This is the first study to report on the effect of temperature on the developmental parameters of A. hemara giving an insight into its biology. The implications of these findings for the use of A. hemara in biological control are discussed.

Effects of Host Age and Density on the Performance of Apanteles hemara (Hymenoptera: Braconidae), a Larval Endoparasitoid of Spoladea recurvalis (Lepidoptera: Crambidae).

The amaranth leaf-webber, Spoladea recurvalis (Fabricius; Lepidoptera: Crambidae), is a serious pest of Amaranthus sp. in Africa and Asia. Apanteles hemara (Nixon; Hymenoptera: Braconidae) is by far the most important larval endoparasitoid of the amaranth leaf-webber. We examined the effects of host density and age on the biological characteristics of A. hemara. The regression model of the number of hosts supplied to A. hemara against the number of larvae parasitized resulted in a curve corresponding to type II functional response, with a significant increase in the number of hosts parasitized up to the density of 30 hosts before being constant up to 40 hosts. In contrast, the parasitism rate decreased linearly with increasing host densities. Development time, sex ratio, and adult longevity were not significantly affected by host density. The immature parasitoid mortality was significantly higher at higher host densities. Apanteles hemara did not parasitize 7-d-old larvae and beyond, while parasitism was significantly higher among 1- to 2-d-old compared with 3- to 4-d-old larvae. Immature parasitoid mortality was 2.6 times higher in 1- to 2-d-old larvae compared with 5- to 6-d-old larvae. The developmental period of the parasitoid from egg to adult was longest among 1- to 2-d-old larvae and least among 5- to 6-d-old larvae. Nonreproductive mortality was markedly higher among 1- to 2-d-old larvae compared with the older larvae. Adult female A. hemara were significantly larger on 3- to 4-d-old larvae compared with either 1- to 2-d-old or 5- to 6-d-old larvae. We discuss the implications of our results for the interpretation of functional response in parasitoids, mass rearing, conservation, and augmentative biological control of S. recurvalis.

African Citrus Greening Disease in East Africa: Incidence, Severity, and Distribution Patterns.

African citrus greening disease (ACGD) is considered as one of the major diseases of citrus threatening citrus production in East Africa. Our study aimed for the first time to assess the incidence, severity, and distribution patterns of ACGD in Kenya and Tanzania. In total, 105 citrus orchards were assessed in 13 regions representing low, mid, and high altitude areas. In each backyard and orchard, trees were randomly selected and rated for visual ACGD symptoms; then leaves and insect samples collected for analysis of 'Candidatus Liberibacter africanus' (CLaf), the presumptive causal agent of ACGD. Endpoint PCR, sequencing, and molecular phylogenetic tools were employed to confirm the identity of potential circulating pathogens. Incidence and severity of ACGD varied significantly among the different regions. Both Trioza erytreae (Del Guerico) (Hemiptera: Triozidae) and the invasive Asian citrus psyllid vector Diaphorina citri (Kuwayama) (Hemiptera: Liviidae) were found to co-occur in upper and lower midland regions. Molecular characterization identified 'Candidatus Liberibacter africanus spp. Clausenae' (CLafCl) as the main causal agent of ACGD in most of the citrus plants and insect samples. No instances of Candidatus Liberibacter asiaticus infection were found. These findings provide valuable insights into understanding and management of ACGD by employing stringent and early disease detection tools to curb the spread of the disease.

Postharvest processes of edible insects in Africa: A review of processing methods, and the implications for nutrition, safety and new products development.

In many African cultures, insects are part of the diet of humans and domesticated animals. Compared to conventional food and feed sources, insects have been associated with a low ecological foot print because fewer natural resources are required for their production. To this end, the Food and Agriculture Organization of the United Nations recognized the role that edible insects can play in improving global food and nutrition security; processing technologies, as well as packaging and storage techniques that improve shelf-life were identified as being crucial. However, knowledge of these aspects in light of nutritional value, safety, and functionality is fragmentary and needs to be consolidated. This review attempts to contribute to this effort by evaluating the available evidence on postharvest processes for edible insects in Africa, with the aim of identifying areas that need research impetus. It further draws attention to potential postharvest technology options for overcoming hurdles associated with utilization of insects for food and feed. A greater research thrust is needed in processing and this can build on traditional knowledge. The focus should be to establish optimal techniques that improve presentation, quality and safety of products, and open possibilities to diversify use of edible insects for other benefits.

Infestation Levels and Molecular Identification Based on Mitochondrial COI Barcode Region of Five Invasive Gelechiidae Pest Species in Kenya.

Invasive Gelechiidae pest species, namely Tuta absoluta, Phthorimaea operculella, Aproaerema simplixella, Sitotroga cerealella, and Pectinophora gossypiella are among the major constraints hampering agricultural economy in Kenya. Infestation levels were determined on respective host crops sampled from different localities and P. operculella recorded the highest infestation of 68.00 ± 4.92% on stored potato. Aproaerema simplixella and T. absoluta accounted for 61.33 ± 5.35% and 51.56 ± 5.22% maximal infestation on groundnuts and tomato leaves, respectively. Stored maize was significantly infested by S. cerealella (54.33 ± 5.31%) while no infestation was observed on the freshly harvested grains. Infestation on open bolls by P. gossypiella was relatively low (6.11 ± 3.46%) compared to Anatrachyntis simplex (45.67 ± 7.84%) that emerged as the key pest of cotton. The species were discriminated based on sequence similarities, evolutionary divergences, and phylogenetic analyses. A 658-bp fragment of mitochondrial cytochrome c oxidase subunit I (COI) gene was obtained from 302 specimens. Generally, genetic variations were low within and between Gelechiid populations, with an average of 0.02% and all intraspecific divergences were less than 2% except for S. cerealella. The Gelechiids data set generated eight Barcode Index Numbers (BINs), five of which were concordant and three belonging to S. cerealella were singleton. All species were separated into distinct clusters on a maximum likelihood tree. Data on infestation levels will be useful in defining the pest status of these Gelechiids in Kenya. DNA barcoding is also presented as a valuable tool to complement traditional taxonomy for rapid and accurate identification of these species of agronomic interest.

Insects for Income Generation Through Animal Feed: Effect of Dietary Replacement of Soybean and Fish Meal With Black Soldier Fly Meal on Broiler Growth and Economic Performance.

The ever-increasing animal feed costs are driving many vulnerable communities involved in animal husbandry out of business. The high cost is mainly driven by the protein source, which represents the most expensive component in animal feed. In conventional feed, protein is obtained mainly from soybean and fish meal (SFM). The present study explored potential of partially replacing this SFM with black soldier fly prepupae meal (BSFPM) in Cobb 500 broiler chicken diets. A SFM-based diet was compared to three experimental diets formulated by partially substituting SFM with BSFPM at 13.8, 27.4, and 42.0% of the crude protein (CP) in the starter feed and 11.0, 37.2, and 55.5% of the CP in the finisher feed of diets D1, D2, and D3, respectively. Dietary effects on average daily feed intake, average daily body weight gain, feed conversion ratio, carcass characteristics, breast meat sensory attributes, and the economic implication of their use in broiler production were evaluated. Replacement of SFM with BSFPM did not affect daily feed intake, daily body weight gain, feed conversion ratio, aroma or taste of cooked breast meat. A 16.0% higher Cost Benefit Ratio and 25.0% better Return on Investment was recorded when the birds were reared on the highest concentration of black soldier fly (D3) compared to the conventional diet which was 19.0% more expensive. The implication of these findings for the promotion of insect mass production enterprises for animal feed protein, and their potential for income generation and job creation particularly in developing countries is discussed.

Moisture adsorption properties and shelf-life estimation of dried and pulverised edible house cricket Acheta domesticus (L.) and black soldier fly larvae Hermetia illucens (L.).

Edible insects are part of the diets of a significant proportion of rural populations in the tropics especially Africa and Asia, and their use as source of key nutrients for better nutrition is re-emerging. Indigenously, elemental methods are used to process the insects before they are consumed or sold in retail outlets. In recent years, better knowledge of processing, packaging and storage has become necessary because of commercialisation needs. A common processing approach involves drying after a brief heat-treatment step, and then milling into a powdered product which is sold to manufacturers or consumers as ingredient for processing final products. The hydration properties of dried powders of edible house cricket and black soldier fly larvae (BSFL) were studied with the aim of predicting shelf-life stability under typical packaging and storage temperatures experienced in the tropics. Moisture adsorption isotherms were determined gravimetrically at 25, 30 and 35 °C, over 0.11-0.97 water activity (aW) range, and the data fitted to various models. Sorption isotherms were of type II according to Brunauer classification indicating monolayer-multilayer sorption behaviour. Cricket powder exhibited higher hydration capacity, and aW of this product was less sensitive to temperature variation as compared to BSFL powder. In the two products, water exhibited transitions from bound- to free- state at ~5 g/100 g moisture content. Based on Heiss-Eichner model, a shelf-life of 7 months at 25 °C can be achieved if the cricket and BSFL powders are dried to ca. 5 g/100 g moisture content and packaged in 80 µm thick polyethylene films. At 35 °C the shelf-life of the cricket product is shortened three- to four-fold whereas the BSFL powder is unable to store.

Performance of Apanteles hemara (Hymenoptera: Braconidae) on two Amaranth Leaf-webbers: Spoladea recurvalis and Udea ferrugalis (Lepidoptera: Crambidae).

Amaranth is an African indigenous vegetable that is gaining popularity due to its nutritional, medicinal, and economic values. In East Africa, frequent outbreaks of Lepidopteran leaf-webbers, Spoladea recurvalis F. (Lepidoptera: Crambidae) and Udea ferrugalis Hübner (1976) are reported on this crop, causing up to 100% foliage loss. The larval endoparasitoid Apanteles hemara Nixon is also frequently found associated with these pests during the outbreaks, however, its performance on both pests has never been documented. Laboratory studies were therefore carried out to assess the acceptability and suitability of S. recurvalis and U. ferrugalis to A. hemara. Both leaf-webber species were accepted by and suitable for the parasitoid. The mean host searching time and oviposition attempts were neither affected by rearing host nor test host. The total developmental time of A. hemara ranged between 10.6 ± 0.16 and 12.8 ± 0.30 days on both hosts. The sex ratio of the parasitoid was female biased when reared on S. recurvalis but male biased on U. ferrugalis. When offered 50 larvae of leaf-webbers for 24 h, a single female A. hemara achieved parasitism rates from 42.63 ± 5.80 to 44.55 ± 5.95, while a cohort of five females resulted in parasitism rates between 87.25 ± 2.70 and 94.67 ± 1.98 %. There was no significant difference between hosts in regard to progeny fitness at each parasitoid density. The parasitoid also caused significant nonreproductive larval mortalities in the hosts. The implications of these findings for mass rearing of the parasitoid as well as for conservation and augmentative biological control of amaranth lepidopteran leaf-webbers in East Africa are discussed.

DNA Barcode Reference Library for the African Citrus Triozid, Trioza erytreae (Hemiptera: Triozidae): Vector of African Citrus Greening.

Citrus (Citrus spp.) production continues to decline in East Africa, particularly in Kenya and Tanzania, the two major producers in the region. This decline is attributed to pests and diseases including infestation by the African citrus triozid, Trioza erytreae (Del Guercio) (Hemiptera: Triozidae). Besides direct feeding damage by adults and immature stages, T. erytreae is the main vector of 'Candidatus Liberibacter africanus', the causative agent of Greening disease in Africa, closely related to Huanglongbing. This study aimed to generate a novel barcode reference library for T. erytreae in order to use DNA barcoding as a rapid tool for accurate identification of the pest to aid phytosanitary measures. Triozid samples were collected from citrus orchards in Kenya, Tanzania, and South Africa and from alternative host plants. Sequences generated from populations in the study showed very low variability within acceptable ranges of species. All samples analyzed were linked to T. erytreae of GenBank accession number KU517195. Phylogeny of samples in this study and other Trioza reference species was inferred using the Maximum Likelihood method. The phylogenetic tree was paraphyletic with two distinct branches. The first branch had two clusters: 1) cluster of all populations analyzed with GenBank accession of T. erytreae and 2) cluster of all the other GenBank accession of Trioza species analyzed except T. incrustata Percy, 2016 (KT588307.1), T. eugeniae Froggatt (KY294637.1), and T. grallata Percy, 2016 (KT588308.1) that occupied the second branch as outgroups forming sister clade relationships. These results were further substantiated with genetic distance values and principal component analyses.

Identification of aphid (Hemiptera: Aphididae) species of economic importance in Kenya using DNA barcodes and PCR-RFLP-based approach.

Aphids are among pests of economic importance throughout the world. Together with transmitting plant viruses, aphids are capable of inflicting severe crop production losses. They also excrete honeydew that favours the growth of sooty mold which reduces the quality of vegetables and fruits and hence their market values. Rapid and accurate identification of aphids to the species level is a critical component in effective pest management and plant quarantine systems. Even though morphological taxonomy has made a tremendous impact on species-level identifications, polymorphism, morphological plasticity and immature stages are among the many challenges to accurate identification. In addition, their small size, presence of cryptic species and damaged specimens dictate the need for a strategy that will ensure timely and accurate identification. In this study, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)-based on mitochondrial cytochrome c oxidase subunit I gene and DNA barcoding were applied to identify different aphid species collected from different agro-ecological zones of Kenya. Three restriction enzymes RsaI, AluI and Hinf1 produced patterns that allowed unambiguous identification of the species except Aphis craccivora and Aphis fabae. Analyses of the barcode region indicated intraspecific and interspecific sequence divergences of 0.08 and 6.63%, respectively. DNA barcoding identified all species, including the morphologically indistinguishable A. craccivora and A. fabae and separated two subspecies of A. fabae. Based on these results, both PCR-RFLPs and DNA barcoding could provide quick and accurate tools for identification of aphid species within Aphididae subsequently aiding in effective pest management programmes and enhance plant quarantine systems.

A major host plant volatile, 1-octen-3-ol, contributes to mating in the legume pod borer, Maruca vitrata (Fabricius) (Lepidoptera: Crambidae).

Previous studies on the legume pod borer, Maruca vitrata Fabricius (Lepidoptera: Crambidae), a serious pest of cowpea, Vigna unguiculata (L.) Walp. (Fabales: Fabaceae), in sub-Saharan Africa have focused on sex pheromones, but the role of the host plant on sexual behavior has not been explored. We investigated this interaction in the laboratory using behavioral assays and chemical analyses. We found that the presence of cowpea seedlings and a dichloromethane extract of the leaf increased coupling in the legume pod borer by 33 and 61 %, respectively, compared to the control, suggesting the involvement of both contact and olfactory cues. We used coupled gas chromatography-electroantennographic detection (GC/EAD) and GC-mass spectrometry (GC/MS) to identify compounds from the cowpea leaf extract, detected by M. vitrata antenna. We found that the antennae of the insect consistently detected four components, with 1-octen-3-ol identified as a common and dominant component in both the volatiles released by the intact cowpea plant and leaf extract. We therefore investigated its role in the coupling of M. vitrata. In dose-response assays, 1-octen-3-ol increased coupling in M. vitrata with increasing dose of the compound compared to the control. Our results suggest that the cowpea volatile 1-octen-3-ol contributes to M. vitrata sexual behavior.

Resolution of three cryptic agricultural pests (Ceratitis fasciventris, C. anonae, C. rosa, Diptera: Tephritidae) using cuticular hydrocarbon profiling.

Discrimination of particular species within the species complexes of tephritid fruit flies is a very challenging task. In this fruit-fly family, several complexes of cryptic species have been reported, including the African cryptic species complex (FAR complex). Cuticular hydrocarbons (CHCs) appear to be an excellent tool for chemotaxonomical discrimination of these cryptic species. In the present study, CHC profiles have been used to discriminate among three important agricultural pests from the FAR complex, Ceratitis fasciventris, Ceratitis anonae and Ceratitis rosa. Hexane body surface extracts of mature males and females were analyzed by two-dimensional gas chromatography with mass spectrometric detection and differences in CHC profiles between species and sexes tested through multivariate statistics and compared with species identification by means of microsatellite markers. Quantitative as well as qualitative CHC profile differences between sexes and species are reported. The CHC profiles consisted of a mixture of linear, internally methyl-branched and mono-, di- and tri-unsaturated alkanes. Twelve compounds were pinpointed as potential chemotaxonomical markers. The present study shows that presence or absence of particular CHCs might be used in the chemical diagnosis of the FAR complex. Moreover, our results represent an important first step in the development of a useful chemotaxonomic tool for cryptic species identification of these important agricultural pests.

Selection of promising fungal biological control agent of the western flower thrips Frankliniella occidentalis (Pergande).

AIMS: Larval stages of Frankliniella occidentalis are known to be refractory to fungal infection compared with the adult stage. The objective of this study was to identify promising fungal isolate(s) for the control of larval stages of F. occidentalis. METHODS AND RESULTS: Ten isolates of Metarhizium anisopliae and eight of Beauveria bassiana were screened for virulence against second-instar larvae of F. occidentalis. Conidial production and genetic polymorphism were also investigated. Metarhizium anisopliae isolates ICIPE 7, ICIPE 20, ICIPE 69 and ICIPE 665 had the shortest LT(50) values of 8.0-8.9 days. ICIPE 69, ICIPE 7 and ICIPE 20 had the lowest LC(50) values of 1.1 × 10(7), 2.0 × 10(7) and 3.0 × 10(7) conidia ml(-1), respectively. Metarhizium anisopliae isolate ICIPE 69 produced significantly more conidia than M. anisopliae isolates ICIPE 7 and ICIPE 20. Internally transcribed spacers sequences alignment showed differences in nucleotides composition, which can partly explain differences in virulence. CONCLUSION: These results coupled with the previous ones on virulence and field efficacy against other species of thrips make M. anisopliae isolate ICIPE 69 a good candidate. SIGNIFICANCE AND IMPACT OF THE STUDY: Metarhizium anisopliae isolate ICIPE 69 can be suggested for development as fungus-based biopesticide for thrips management.

Cold susceptibility and disinfestation of Bactrocera invadens (Diptera: Tephritidae) in oranges.

To develop a cold disinfestation treatment for the fruit fly Bactrocera invadens Drew, Tsuruta & White (Diptera: Tephritidae) that is rapidly spreading across Africa, research was conducted in Nairobi, Kenya, using flies from a laboratory culture and 'Valencia' orange (Citrus sinensis L. Osbeck) as the host. The developmental rate of B. invadens in Valencia oranges was determined at 28 degrees C, and the third instar was found to be the least susceptible of the egg and larval life stages to cold treatment at 1.1 degrees C in oranges. When 22,449 B. invadens third instars were exposed in oranges to a cold treatment with an approximate midpoint of 1.1 +/- 0.5 degrees C, the results suggested that a period of 16 d would be worthwhile verifying on a larger scale in oranges. Results from the first replicate of 16,617 larvae showed no survivors, but the second replicate of 23,536 larvae had three survivors. Because a longer cold treatment based on a mean temperature of 1.1 degrees C would create logistical difficulties for some export markets, further replicates were conducted at an approximate midpoint of 0.5 degrees C and at mean hourly maximum of 0.9 +/- 0.5 degrees C, for 16 d. After three replicates, in which 65,752 B. invadens third instars in total were treated with no survivors, the Japanese requirement of 99.99% mortality at the 95% confidence level was surpassed. The following treatment protocol for B. invadens larvae in oranges can therefore be recommended: fruit pulp to be maintained at temperatures of 0.9 degrees C or lower for 16 consecutive days.

Ecological niche and potential geographic distribution of the invasive fruit fly Bactrocera invadens (Diptera, Tephritidae).

Two correlative approaches to the challenge of ecological niche modeling (genetic algorithm, maximum entropy) were used to estimate the potential global distribution of the invasive fruit fly, Bactrocera invadens, based on associations between known occurrence records and a set of environmental predictor variables. The two models yielded similar estimates, largely corresponding to Equatorial climate classes with high levels of precipitation. The maximum entropy approach was somewhat more conservative in its evaluation of suitability, depending on thresholds for presence/absence that are selected, largely excluding areas with distinct dry seasons; the genetic algorithm models, in contrast, indicate that climate class as partly suitable. Predictive tests based on independent distributional data indicate that model predictions are quite robust. Field observations in Benin and Tanzania confirm relationships between seasonal occurrences of this species and humidity and temperature.

Uncovering the tracks of a recent and rapid invasion: the case of the fruit fly pest Bactrocera invadens (Diptera: Tephritidae) in Africa.

Phytophagous insects of the genus Bactrocera are among the most economically important invasive fruit fly pests. In 2003, an unknown Bactrocera species was found in Kenya. First identified as an 'aberrant form' of the Asian B. dorsalis complex, it was later recognized as a new species, Bactrocera invadens. Within 2 years of its discovery, the species was recorded in several African countries, becoming an important quarantine pest. As this invasive fly was discovered only recently, no data are available on its invasion pattern in Africa. This pilot study attempts to infer from genetic data the dynamic aspects of the African invasion of this pest. Using microsatellite markers, we evaluated the level of genetic diversity and the extent of common ancestry among several African populations collected across the invaded areas. A sample from the Asian Sri Lankan population was analysed to confirm the Asian origin of this pest. Genetic data cast no doubt that Sri Lanka belongs to the native range, but only a small percentage of its genotypes can be found in Africa. African populations display relatively high levels of genetic diversity associated with limited geographical structure and no genetic footprints of bottlenecks. These features are indicative of processes of rapid population growth and expansion with possible multiple introductions. In the span of relatively few years, the African invasion registered the presence of at least two uncorrelated outbreaks, both starting from the East. The results of the analyses support that invasion started in East Africa, where B. invadens was initially isolated.

Isolation and characterization of microsatellite markers in the newly discovered invasive fruit fly pest in Africa, Bactrocera invadens (Diptera: Tephritidae).

We describe the isolation and characterization of 11 polymorphic microsatellite loci from the recently discovered fruit fly pest, Bactrocera invadens. The polymorphism of these loci was tested in individual flies from two natural populations (Sri Lanka and Democratic Republic of Congo). Allele number per locus ranged from three to 15 and eight loci displayed a polymorphic information content greater than 0.5. These microsatellite loci provide useful markers for studies of population dynamics and invasion history of this pest species.

Field infestation, life history and demographic parameters of the fruit fly Bactrocera invadens (Diptera: Tephritidae) in Africa.

Field infestation rates of an invasive fruit fly species, Bactrocera invadens Drew Tsuruta & White on mango was determined at different localities in Kenya. At most of the locations and especially at low elevations, B. invadens frequently shared the same fruit with the indigenous fruit fly species Ceratitis cosyra (Walker) but often occurred at higher numbers than C. cosyra. The level of infestation varied with location ranging from 3.0 to 97.2 flies per kg of fruit. There was a significant inverse relationship between numbers of flies per kg of fruit and elevation at which fruit was collected, suggesting that B. invadens is a predominantly lowland pest. On an artificial diet, development of B. invadens immatures lasted 25 days; egg incubation required 1.2 days, larval development 11.1 days and puparia-adult development 12.4 days. About 55% of eggs developed to the adult stage. Life expectancy at pupal eclosion was 75.1 days in females and 86.4 days in males. Average net fecundity and net fertility were 794.6 and 608.1 eggs, respectively, while average daily oviposition was 18.2 eggs. Daily population increase was 11% and mean generation time was 31 days. Results are discussed in relation to the biology and ecology of the insect and in the development of mass rearing and control measures for B. invadens.