Dynamics of genetic variability in <Emphasis Type="Italic">Anastrepha fraterculus</Emphasis>(Diptera: Tephritidae) during adaptation to laboratory rearing conditions

Background

Anastrepha fraterculus is one of the most important fruit fly plagues in the American continent and only chemical control is applied in the field to diminish its population densities. A better understanding of the genetic variability during the introduction and adaptation of wild A. fraterculus populations to laboratory conditions is required for the development of stable and vigorous experimental colonies and mass-reared strains in support of successful Sterile Insect Technique (SIT) efforts.

Methods

The present study aims to analyze the dynamics of changes in genetic variability during the first six generations under artificial rearing conditions in two populations: a) a wild population recently introduced to laboratory culture, named TW and, b) a long-established control line, named CL.

Results

Results showed a declining tendency of genetic variability in TW. In CL, the relatively high values of genetic variability appear to be maintained across generations and could denote an intrinsic capacity to avoid the loss of genetic diversity in time.

Discussion

The impact of evolutionary forces on this species during the adaptation process as well as the best approach to choose strategies to introduce experimental and mass-reared A. fraterculus strains for SIT programs are discussed.

     

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.     

Courtship in Two Morphotypes of the <Emphasis Type="Italic">Anastrepha fraterculus</Emphasis> (Diptera: Tephritidae) Cryptic Species Complex and their Implications for Understanding Mate Recognition

Ritualized courtship behaviors are used to recognize potential mates and behavioral patterns are inevitably different among populations that demonstrate reproductive incompatibility. We characterized and compared the courtship behaviors of two morphotypes of the cryptic species complex Anastrepha fraterculus: Brazil-1 morphotype and Brazil-3 morphotype. Courtship behaviors were filmed to analyze the behavioral sequences of these two morphotypes during homotypic crossings. The behavioral units Alignment (AL) and Abdominal movements (AB and AB-call) were newly recognized in the courtship ethogram of Anastrepha fraterculus males. The two morphotypes show distinct behavioral sequences leading up to copulation. Some behaviors were repeated frequently during the courtship process, while others were more restricted to the final moments of courtship. The three behavioral units that contributed most to copulation success were Contact, Alignment, and Arrowhead 1 in the Brazil-1 morphotype and Alignment, Arrowhead 1, and Fanning in the Brazil-3 morphotype. Some behavioral routines differed across the two morphotypes. Significant differences were also noted between the frequencies of the behavioral units displayed during courtship in the two morphotypes. The relationships between the pre-zygotic incompatibilities of the Brazil-1 and Brazil-3 morphotypes and the differences between the courtship behaviors of their males are discussed. Our results indicate that behavioral isolation is involved in the process of pre-zygotic reproductive isolation of Brazil-1 and Brazil-3 morphotypes.     

Development of a genetic sexing strain in <Emphasis Type="Italic">Bactrocera carambolae</Emphasis> (Diptera: Tephritidae) by introgression of sex sorting components from <Emphasis Type="Italic">B. dorsalis</Emphasis>, Salaya1 strain

Background

The carambola fruit fly, Bactrocera carambolae Drew & Hancock is a high profile key pest that is widely distributed in the southwestern ASEAN region. In addition, it has trans-continentally invaded Suriname, where it has been expanding east and southward since 1975. This fruit fly belongs to Bactrocera dorsalis species complex. The development and application of a genetic sexing strain (Salaya1) of B. dorsalis sensu stricto (s.s.) (Hendel) for the sterile insect technique (SIT) has improved the fruit fly control. However, matings between B. dorsalis s.s. and B. carambolae are incompatible, which hinder the application of the Salaya1 strain to control the carambola fruit fly. To solve this problem, we introduced genetic sexing components from the Salaya1 strain into the B. carambolae genome by interspecific hybridization.

Results

Morphological characteristics, mating competitiveness, male pheromone profiles, and genetic relationships revealed consistencies that helped to distinguish Salaya1 and B. carambolae strains. A Y-autosome translocation linking the dominant wild-type allele of white pupae gene and a free autosome carrying a recessive white pupae homologue from the Salaya1 strain were introgressed into the gene pool of B. carambolae. A panel of Y-pseudo-linked microsatellite loci of the Salaya1 strain served as markers for the introgression experiments. This resulted in a newly derived genetic sexing strain called Salaya5, with morphological characteristics corresponding to B. carambolae. The rectal gland pheromone profile of Salaya5 males also contained a distinctive component of B. carambolae. Microsatellite DNA analyses confirmed the close genetic relationships between the Salaya5 strain and wild B. carambolae populations. Further experiments showed that the sterile males of Salaya5 can compete with wild males for mating with wild females in field cage conditions.

Conclusions

Introgression of sex sorting components from the Salaya1 strain to a closely related B. carambolae strain generated a new genetic sexing strain, Salaya5. Morphology-based taxonomic characteristics, distinctive pheromone components, microsatellite DNA markers, genetic relationships, and mating competitiveness provided parental baseline data and validation tools for the new strain. The Salaya5 strain shows a close similarity with those features in the wild B. carambolae strain. In addition, mating competitiveness tests suggested that Salaya5 has a potential to be used in B. carambolae SIT programs based on male-only releases.

     

The plant growth promoting bacterium <Emphasis Type="Italic">Azospirillum brasilense</Emphasis> is vertically transmitted in <Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> (common bean)

Bactrocera dorsalis (Diptera: Tephritidae) is a serious menace to agricultural production worldwide. In order to prevent further damage, it is of paramount important that cost-effective strategies should be developed for their management. Gut bacteria has established diverse relationships with their insect hosts, which could be exploited in pest management programs to improve on control efficiency. In this study, gut bacteria isolates identified by culture dependent technique were incorporated into larval diets in an attempt to understand the roles they play in the development and survival of oriental fruit fly. From our results, the isolated bacteria belonged to four different phyla including the Firmicutes, Proteobacteria, Bacteroidetes and Actinobacteria. The response of the fly to different gut isolates varied greatly. Diets enriched with Enterococcus phoeniculicola had lower larval developmental duration, higher pupal weight, and an increased percentage survival. On the other hand, diets supplemented with Lactobacillus lactis had negative effects on B. dorsalis development. This study provides clues on how symbiotic bacteria could be exploited in mass rearing for an efficient implementation of the Sterile Insect Technique (SIT) in pest management programs.     

Combining paratransgenesis with SIT: impact of ionizing radiation on the DNA copy number of <Emphasis Type="Italic">Sodalis glossinidius in tsetse flies</Emphasis>

Background

Tsetse flies (Diptera: Glossinidae) are the cyclical vectors of the causative agents of African Trypanosomosis, which has been identified as a neglected tropical disease in both humans and animals in many regions of sub-Saharan Africa. The sterile insect technique (SIT) has shown to be a powerful method to manage tsetse fly populations when used in the frame of an area-wide integrated pest management (AW-IPM) program. To date, the release of sterile males to manage tsetse fly populations has only been implemented in areas to reduce transmission of animal African Trypanosomosis (AAT). The implementation of the SIT in areas with Human African Trypanosomosis (HAT) would require additional measures to eliminate the potential risk associated with the release of sterile males that require blood meals to survive and hence, might contribute to disease transmission. Paratransgenesis offers the potential to develop tsetse flies that are refractory to trypanosome infection by modifying their associated bacteria (Sodalis glossinidius) here after referred to as Sodalis. Here we assessed the feasibility of combining the paratransgenesis approach with SIT by analyzing the impact of ionizing radiation on the copy number of Sodalis and the vectorial capacity of sterilized tsetse males.

Results

Adult Glossina morsitans morsitans that emerged from puparia irradiated on day 22 post larviposition did not show a significant decline in Sodalis copy number as compared with non-irradiated flies. Conversely, the Sodalis copy number was significantly reduced in adults that emerged from puparia irradiated on day 29 post larviposition and in adults irradiated on day 7 post emergence. Moreover, irradiating 22-day old puparia reduced the copy number of Wolbachia and Wigglesworthia in emerged adults as compared with non-irradiated controls, but the radiation treatment had no significant impact on the vectorial competence of the flies.

Conclusion

Although the radiation treatment significantly reduced the copy number of some tsetse fly symbionts, the copy number of Sodalis recovered with time in flies irradiated as 22-day old puparia. This recovery offers the opportunity to combine a paratransgenesis approach – using modified Sodalis to produce males refractory to trypanosome infection – with the release of sterile males to minimize the risk of disease transmission, especially in HAT endemic areas. Moreover, irradiation did not increase the vector competence of the flies for trypanosomes.

     

Male-specific Y-linked transgene markers to enhance biologically-based control of the Mexican fruit fly, <Emphasis Type="Italic">Anastrepha ludens</Emphasis>(Diptera: Tephritidae)

Background

Reliable marking systems are critical to the prospective field release of transgenic insect strains. This is to unambiguously distinguish released insects from wild insects in the field that are collected in field traps, and tissue-specific markers, such as those that are sperm-specific, have particular uses such as identifying wild females that have mated with released males. For tephritid fruit flies such as the Mexican fruit fly, Anastrepha ludens, polyubiquitin-regulated fluorescent protein body markers allow transgenic fly identification, and fluorescent protein genes regulated by the spermatocyte-specific β2-tubulin promoter effectively mark sperm. For sterile male release programs, both marking systems can be made male-specific by linkage to the Y chromosome.

Results

An A. ludens wild type strain was genetically transformed with a piggyBac vector, pBXL{PUbnlsEGFP, Asβ2tub-DsRed.T3}, having the polyubiquitin-regulated EGFP body marker, and the β2-tubulin-regulated DsRed.T3 sperm-specific marker. Autosomal insertion lines effectively expressed both markers, but a single Y-linked insertion (Y^EGFP strain) expressed only PUbnlsEGFP. This insertion was remobilized by transposase helper injection, which resulted in three new autosomal insertion lines that expressed both markers. This indicated that the original Y-linked Asβ2tub-DsRed.T3 marker was functional, but specifically suppressed on the Y chromosome. The PUbnlsEGFP marker remained effective however, and the Y^EGFP strain was used to create a sexing strain by translocating the wild type allele of the black pupae (bp⁺) gene onto the Y, which was then introduced into the bp^- mutant strain. This allows the mechanical separation of mutant female black pupae from male brown pupae, that can be identified as adults by EGFP fluorescence.

Conclusions

A Y-linked insertion of the pBXL{PUbnlsEGFP, Asβ2tub-DsRed.T3} transformation vector in A. ludens resulted in male-specific expression of the EGFP fluorescent protein marker, and was integrated into a black pupae translocation sexing strain (T(Y^EGFP/bp⁺), allowing the identification of male adults when used in sterile male release programs for population control. A unique observation was that expression of the Asβ2tub-DsRed.T3 sperm-specific marker, which was functional in autosomal insertions, was specifically suppressed in the Y-linked insertion. This may relate to the Y chromosomal regulation of male-specific germ-line genes in Drosophila.

     

Biology and Thermal Requirements of <Emphasis Type="Italic">Fopius arisanus</Emphasis> (Sonan, 1932) (Hymenoptera: Braconidae) Reared on <Emphasis Type="Italic">Ceratitis capitata</Emphasis> Eggs (Wiedemann) (Diptera: Tephritidae)

Fopius arisanus (Sonan) is a solitary parasitoid of eggs and the first instar larvae of Tephritidae. Due to the occurrence of Ceratitis capitata (Wiedemann) in various regions and under several climatic conditions, this study aimed to evaluate the effect of different temperatures on the embryonic development (egg–adult) and determine thermal requirements and the number of annual generations F. arisanus on eggs of C. capitata. In the laboratory, eggs of C. capitata (24 h) were submitted to parasitism of F. arisanus during 6 h. Later, the eggs were placed in plastic containers (50 mL) (50 eggs/container) on a layer of artificial diet and packed in chambers at temperatures 15, 18, 20, 22, 25, 28, 30, and 32 ± 1°C, RH 70 ± 10%, and a photophase of 12 h. The largest number of offspring, emergence rate, and weight of adults of F. arisanus were observed at 25°C. The highest sex ratios (sr > 0.75) were recorded at 15 and 18°C, being statistically higher than the temperatures 20°C (0.65), 22°C (0.64), 25°C (0.65), 28°C (0.49), and 30°C (0.47). At 32°C, there was no embryonic development of F. arisanus. The egg–adult period was inversely proportional to temperature. Based on the development of the biological cycle (egg–adult), the temperature threshold (T _t) was 10.3°C and thermal constant (K) of 488.34 degree-days, being the number of generations/year directly proportional to the temperature increase. The data show the ability of F. arisanus to adapt to different thermal conditions, which is important for biological control programs of C. capitata.     

<Emphasis Type="Italic">Metarhizium bibionidarum</Emphasis> and <Emphasis Type="Italic">M. purpureogenum</Emphasis>: new species from Japan

Two new species of Metarhizium, M. bibionidarum and M. purpureogenum are described from Japan. Metarhizium bibionidarum is the phylogenetic sister species of M. pemphigi and a member of the M. flavoviride species complex. It is distinguished morphologically from M. pemphigi by its larger conidia. The species is based on a collection of an infected March fly larva (Diptera: Bibionidae) but is also known to occur on fruit beetle (Coleoptera: Scarabaeidae) encountered in France. Metarhizium purpureogenum was isolated from soil by plating and insect baiting methods and represents a unique phylogenetic lineage placed outside the M. anisopliae and M. flavoviride species complexes. Three isolates of M. purpureogenum excreted a distinctive red-purple pigment into agar medium when co-cultured with M. robertsii or Aspergillus oryzae.     

Subtractive and differential hybridization molecular analyses of <Emphasis Type="Italic">Ceratitis capitata</Emphasis> XX/XY versus XX embryos to search for male-specific early transcribed genes

The agricultural pest Ceratitis capitata, also known as the Mediterranean fruit fly or Medfly, is a fruit crop pest of very high economic relevance in different continents. The strategy to separate Ceratitis males from females (sexing) in mass rearing facilities is a useful step before the sterilization and release of male-only flies in Sterile Insect Technique control programs (SIT). The identification of genes having early embryonic male-specific expression, including Y-linked genes, such as the Maleness factor, could help to design novel and improved methods of sexing in combination with transgenesis, aiming to confer conditional female-specific lethality or female-to-male sexual reversal.We used a combination of Suppression Subtractive Hybrydization (SSH), Mirror Orientation Selection (MOS) and differential screening hybridization (DSH) techniques to approach the problem of isolating corresponding mRNAs expressed in XX/XY embryos versus XX-only embryos during a narrow developmental window (8-10 hours after egg laying, AEL ). Here we describe a novel strategy we have conceived to obtain relatively large amounts of XX-only embryos staged at 8-10 h AEL and so to extract few micrograms of polyA+ required to apply the complex technical procedure. The combination of these 3 techniques led to the identification of a Y-linked putative gene, CcGm2, sharing high sequence identity to a paralogous gene, CcGm1, localized either on an autosome or on the X chromosome.We propose that CcGm2 is a first interesting putative Y-linked gene which could play a role in sex determination. The function exterted by this gene should be investigated by novel genetic tools, such as CRISPR-CAS9, which will permit to target only the Y-linked paralogue, avoiding to interfere with the autosomal or X-linked paralogue function.     

Development,genetic and cytogenetic analyses of genetic sexing strains of the Mexican fruit fly, <Emphasis Type="Italic">Anastrepha ludens</Emphasis> Loew (Diptera: Tephritidae)

Background

Anastrepha ludens is among the pests that have a major impact on México's economy because it attacks fruits as citrus and mangoes. The Mexican Federal government uses integrated pest management to control A. ludens through the Programa Nacional Moscas de la Fruta [National Fruit Fly Program, SAGARPA-SENASICA]. One of the main components of this program is the sterile insect technique (SIT), which is used to control field populations of the pest by releasing sterile flies.

Results

To increase the efficiency of this technique, we have developed a genetic sexing strain (GSS) in which the sexing mechanism is based on a pupal colour dimorphism (brown-black) and is the result of a reciprocal translocation between the Y chromosome and the autosome bearing the black pupae (bp) locus. Ten strains producing wild-type (brown pupae) males and mutant (black pupae) females were isolated. Subsequent evaluations for several generations were performed in most of these strains. The translocation strain named Tapachula-7 showed minimal effect on survival and the best genetic stability of all ten strains. Genetic and cytogenetic analyses were performed using mitotic and polytene chromosomes and we succeeded to characterize the chromosomal structure of this reciprocal translocation and map the autosome breakpoint, despite the fact that the Y chromosome is not visible in polytene nuclei following standard staining.

Conclusions

We show that mitotic and polytene chromosomes can be used in cytogenetic analyses towards the development of genetic control methods in this pest species. The present work is the first report of the construction of GSS of Anastrepha ludens, with potential use in a future Moscafrut operational program.

     

Conditional embryonic lethality to improve the sterile insect technique in Ceratitis capitata (Diptera: Tephritidae)

Background  

The sterile insect technique (SIT) is an environment-friendly method used in area-wide pest management of the Mediterranean fruit fly Ceratitis capitata (Wiedemann; Diptera: Tephritidae). Ionizing radiation used to generate reproductive sterility in the mass-reared populations before release leads to reduction of competitiveness.     

Metabolomics reveals immunomodulation as a possible mechanism for the antibiotic effect of <Emphasis Type="Italic">Persicaria capitata</Emphasis> (Buch.-Ham. ex D. Don) H.Gross

Introduction

In spite of advances in antibiotics, urinary tract infection (UTI) is still among the most common reasons for antibiotic medication worldwide. Persicaria capitata (Buch.-Ham. ex D. Don) H.Gross (P. capitata) is a herbal medicine used by the Miao people in China to treat UTI. However studies of its mechanism are challenging, owing to the complexity of P. capitata with multiple constituents acting on multiple metabolic pathways.

Objective

The objective of this study was to explore the working mechanism of P. capitata on urinary tract infection.

Methods

Relinqing® granule, which is solely made from aqueous extracts of the whole P. capitata plant, was used in this study. Urine metabolomics based on gas chromatography-mass spectroscopy was employed to assess the metabolic changes caused by administration of Relinqing® granule in a UTI mouse model. Female specific-pathogen-free Kunming mice were divided into control group (mock infection, saline treatment), model group (E.coli infection, saline treatment), Relinqing® group (E.coli infection, Relinqing® granule treatment), ciprofloxacin group (E.coli infection, ciprofloxacin treatment), and sham-Relinqing® group (no surgery, Relinqing® granule treatment).

Results

The results showed that after the treatments, urine levels of itaconic acid in Relinqing® group increased by 4.9 fold and 11.3 fold compared with model and ciprofloxacin groups respectively. Itaconic acid is an endogenous antibacterial metabolite produced by macrophages, which also functions as a checkpoint for metabolic reprogramming of macrophage.

Conclusion

Our findings suggest that this herbal medicine can cure urinary tract infection through modulation of immune system.

     

Feasibility assessment for the classical biological control of <Emphasis Type="Italic">Tamarix</Emphasis> in Argentina

Saltcedars are woody plants in the genus Tamarix L. (Caryophyllales: Tamaricaceae) and are native to Eurasia and Africa. Several species have become invasive in the Americas, Australia and South Africa. In Argentina there are four species of Tamarix distributed in arid, semi-arid and coastal areas of most provinces. The taxonomic isolation of Tamarix spp. in Argentina, their widespread distribution, negative impact to natural areas and lack of impact from existing natural enemies all indicate that Tamarix is an ideal candidate for classical biological control in Argentina. Biological control of Tamarix spp. has been rapid and highly successful in the USA after the introduction of four Diorhabda spp. (Coleoptera: Chrysomelidae). Biological control of Tamarix spp. in Argentina could be implemented easily, rapidly, and at a low cost by utilizing the information developed in the USA.     

Mexican Fruit Fly Populations in the Semi-Arid Highlands of the Sierra Madre Oriental in Northeastern Mexico

The Mexican fruit fly, Anastrepha ludens Loew (Diptera: Tephritidae), is one of the most important pests of citrus in Mexico. We report the results of an analysis of A. ludens populations that inhabit the semi-arid highlands of the Sierra Madre Oriental in northeastern Mexico. This study aimed to provide information on population fluctuation of A. ludens and how it relates to climate variables, as well as insights into habitat and native parasitoids. Population peaked in the period July–November when ripe fruits of the wild host, Casimiroa pubescens Ramírez, were available. No adults were captured the rest of the year, suggesting that high populations depend on the availability of wild host fruit. No significant relationships between population fluctuation and climatic variables were observed, except for minimum temperature. Fruit samples of citron (Citrus medica L.), pomegranate (Punica granatum L.), and C. pubescens were collected to determine degree of infestation. Infestation levels (pupae/g) ranged between 0.0006 for citron, 0.0047 for pomegranate, and 0.0240 for C. pubescens. A native parasitoid of Tephritidae, Doryctobracon crawfordii (Viereck) (Braconidae), was identified. Parasitism percentage was calculated at 12.5% on C. pubescens fruits. No parasitoids were observed on citron or pomegranate fruit samples. These results contribute to knowledge on behavior of A. ludens native to temperate environments where no commercial hosts are available. Further research on host expansion of this pest in light of scenarios of global climate change is suggested.     

Development and validation of KASP markers for the greenbug resistance gene <Emphasis Type="Italic">Gb7</Emphasis> and the Hessian fly resistance gene <Emphasis Type="Italic">H32</Emphasis> in wheat

Key message

Greenbug and Hessian fly are important pests that decrease wheat production worldwide. We developed and validated breeder-friendly KASP markers for marker-assisted breeding to increase selection efficiency.

Abstract

Greenbug (Schizaphis graminum Rondani) and Hessian fly [Mayetiola destructor (Say)] are two major destructive insect pests of wheat (Triticum aestivum L.) throughout wheat production regions in the USA and worldwide. Greenbug and Hessian fly infestation can significantly reduce grain yield and quality. Breeding for resistance to these two pests using marker-assisted selection (MAS) is the most economical strategy to minimize losses. In this study, doubled haploid lines from the Synthetic W7984 × Opata M85 wheat reference population were used to construct linkage maps for the greenbug resistance gene Gb7 and the Hessian fly resistance gene H32 with genotyping-by-sequencing (GBS) and 90K array-based single nucleotide polymorphism (SNP) marker data. Flanking markers were closely linked to Gb7 and H32 and were located on chromosome 7DL and 3DL, respectively. Gb7-linked markers (synopGBS773 and synopGBS1141) and H32-linked markers (synopGBS901 and IWB65911) were converted into Kompetitive Allele Specific PCR (KASP) assays for MAS in wheat breeding. In addition, comparative mapping identified syntenic regions in Brachypodium distachyon, rice (Oryza sativa), and sorghum (Sorghum bicolor) for Gb7 and H32 that can be used for fine mapping and map-based cloning of the genes. The KASP markers developed in this study are the first set of SNPs tightly linked to Gb7 and H32 and will be very useful for MAS in wheat breeding programs and future genetic studies of greenbug and Hessian fly resistance.

     

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.