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The Mediterranean fruit fly (medfly), Ceratitis capitata, is a pest of worldwide substantial economic importance, as well as a Tephritidae model for sterile insect technique (SIT) applications. The latter is partially due to the development and utilization of genetic sexing strains (GSS) for this species, such as the Vienna 8 strain, which is currently used in mass rearing facilities worldwide. Improving the performance of such a strain both in mass rearing facilities and in the field could significantly enhance the efficacy of SIT and reduce operational costs. Recent studies have suggested that the manipulation of gut symbionts can have a significant positive effect on the overall fitness of insect strains. We used culture-based approaches to isolate and characterize gut-associated bacterial species of the Vienna 8 strain under mass rearing conditions. We also exploited one of the isolated bacterial species, Enterobacter sp., as dietary supplement (probiotic) to the larval diet, and we assessed its effects on fitness parameters under the standard operating procedures used in SIT operational programs. Probiotic application of Enterobacter sp. resulted in improvement of both pupal and adult productivity, as well as reduced rearing duration, particularly for males, without affecting pupal weight, sex ratio, male mating competitiveness, flight ability and longevity under starvation.

In insects, rapidly evolving primary sex-determining signals are transduced by a conserved regulatory module controlling sexual differentiation. In the agricultural pest Ceratitis capitata (Mediterranean fruit fly, or Medfly), we identified a Y-linked gene, Maleness-on-the-Y (MoY), encoding a small protein that is necessary and sufficient for male development. Silencing or disruption of MoY in XY embryos causes feminization, whereas overexpression of MoY in XX embryos induces masculinization. Crosses between transformed XY females and XX males give rise to males and females, indicating that a Y chromosome can be transmitted by XY females. MoY is Y-linked and functionally conserved in other species of the Tephritidae family, highlighting its potential to serve as a tool for developing more effective control strategies against these major agricultural insect pests.

The Mediterranean fruitfly Ceratitis capitata (medfly) is an invasive agricultural pest of high economic impact and has become an emerging model for developing new genetic control strategies as an alternative to insecticides. Here, we report the successful adaptation of CRISPR-Cas9-based gene disruption in the medfly by injecting in vitro pre-assembled, solubilized Cas9 ribonucleoprotein complexes (RNPs) loaded with gene-specific single guide RNAs (sgRNA) into early embryos. When targeting the eye pigmentation gene white eye ( we ), a high rate of somatic mosaicism in surviving G0 adults was observed. Germline transmission rate of mutated we alleles by G0 animals was on average above 52%, with individual cases achieving nearly 100%. We further recovered large deletions in the we gene when two sites were simultaneously targeted by two sgRNAs. CRISPR-Cas9 targeting of the Ceratitis ortholog of the Drosophila segmentation paired gene ( Ccprd ) caused segmental malformations in late embryos and in hatched larvae. Mutant phenotypes correlate with repair by non-homologous end-joining (NHEJ) lesions in the two targeted genes. This simple and highly effective Cas9 RNP-based gene editing to introduce mutations in C. capitata will significantly advance the design and development of new effective strategies for pest control management.

Tephritid fruit fly pests pose an increasing threat to the agricultural industry due to their global dispersion and a highly invasive nature. Here we showcase the feasibility of an early-detection SEPARATOR sex sorting approach through using the non-model Tephritid pest, Ceratitis capitata. This system relies on female-only fluorescent marker expression, accomplished through the use of a sex-specific intron of the highly-conserved transformer gene from C. capitata and Anastrepha ludens. The herein characterized strains have 100% desired phenotype outcomes, allowing accurate male–female separation during early development. Overall, we describe an antibiotic and temperature-independent sex-sorting system in C. capitata , which, moving forward, may be implemented in other non-model Tephritid pest species. This strategy can facilitate the establishment of genetic sexing systems with endogenous elements exclusively, which, on a wider scale, can improve pest population control strategies like sterile insect technique.

The fruit fly Ceratitis capitata is a successful generalist due to the trade-off between different variables in its life history traits. The present study aimed to assess some life history traits of C. capitata recovered from peach, plum and fig, three key host species, under the climatic conditions of an irrigation oasis in the arid province of San Juan, Argentina. Pupal abundance, sex ratio and morphometric parameters such as pupal and adult weight, pupal volume, wingspan and head-caudal length were influenced by the host in which the larvae were reared. Sexual maturity, measured as the peak of calling (pheromone emission), was homogeneous on the sixth and seventh days after emergence, but males recovered from fig showed early calling activity on the third day after emergence. The plasticity of C. capitata to adapt its life history traits to specific nutrient variations within the larval environment allows it to colonise and establish populations where key hosts, as those studied here, are available. The knowledge of the interactions between host fruit trees and C. capitata populations provides helpful information for planning the management of local orchards and urban fruit plantations within irrigation oases, taking into consideration the concept of key hosts as peach and fig.

Ceratitis capitata (medfly) is one of the most devastating crop pests worldwide. The Sterile Insect Technique (SIT) is a control method that is based on the mass rearing of males, their sterilization, and release in the field. However, the effectiveness of the technique depends on the quality of the released males and their fitness. We previously isolated and selected a probiotic bacteria ( Enterobacter sp.), from wild-caught medflies, according to criteria that improved biological quality traits of reared medfly males.We firstly evaluated the impact of the irradiation on the expression of different immune and stress genes in the medfly sterile males. Expression was measured at differents time points ranging from 0 to 168 h after irradiation to capture the response of genes with distinct temporal expression patterns. Then, we supplemented the larval diet with previously isolated Enterobacter sp.strain, live and autoclaved at various concentrations to see whether the probiotic treatments affect, through their protective role, the gene expression level, and quality traits. The irradiation had significant effect on the genes attacin , cecropin , PGPR-LC , hsp23 , and hsp70 level expression. The expression of attacin and PGPR-LC was up-regulated while that of cecropin was down-regulated. Hsp genes showed decreased levels between 0 and 18 h to peak at 72 h. However, the supplementation of the probiotic strain, either live or autoclaved, was statistically significant only for attacin gene. However, significant interaction time x probiotic was noticed for attacin , cecropin , hsp23 and hsp70 . The probiotic treatments also improved the quality control parameters like pupal weight. From this work we can conclude that a consortium of parabiotics (autoclaved probiotics) treatment will be recommended in insectaries considering both the beneficial effects on mass reared insects and its general safety for insectary workers and for environment.

The Mediterranean fruit fly ( Ceratitis capitata Wied.) is an agricultural pest of significant economic importance. This species has been globally managed using the Sterile Insect Technique (SIT). Insects, including tephritid flies, harbor a diverse gut microbiota that plays critical roles in their physiology, behavior, and overall fitness, suggesting that microbial communities may profoundly influence the biology of this pest. The aim of this study was to characterize the fungal and bacterial gut microbial communities of C. capitata from Tucumán, Argentina, and to assess their response to antimicrobial treatment using amplicon-based 16S rRNA gene and ITS region sequencing. Both control and treated flies were dominated by Proteobacteria (bacteria) and Zygosaccharomyces (fungi). Antimicrobial treatment induced significant shifts in bacterial and fungal composition, reducing diversity and altering gut community structure. Untreated flies exhibited a diverse and structured bacterial gut community dominated by the family Enterobacteriaceae, while antibiotic-treated communities were dominated by Rhizobiaceae. Despite these shifts, fungal communities in both treated and untreated groups were consistently dominated by the genus Zygosaccharomyces . Functional predictions revealed notable changes in metabolic pathways following antibiotic treatment, including increased gene abundance for ABC transporters and the phosphotransferase system, and decreased representation of genes involved in antibiotic biosynthesis and two-component systems. These results indicate significant alterations in bacterial metabolism and stress response mechanisms induced by the treatment. Such changes may help explain the underperformance of irradiated, mass-reared males within the context of SIT. This study provides new insights into the structural and functional dynamics of the C. capitata gut microbiome under disturbance. These findings have implications for understanding the ecological roles of microbial communities in this pest and their potential impact on fly health and fitness. Identification of dominant gut bacterial and fungal groups may support the development of probiotic diets, enhancing the efficiency of SIT application.

The Mediterranean fruit fly ( Ceratitis capitata ) is a globally invasive species and an economically significant pest of fruit crops. Understanding the evolutionary history and local climatic adaptation of this species is crucial for developing effective pest management strategies. We conducted a comprehensive investigation using whole genome sequencing to explore (i) the invasion history of C. capitata with an emphasis on historical admixture and (ii) local climatic adaptation across African, European, Central, and South American populations of C. capitata . Our results suggest a stepwise colonization of C. capitata in Europe and Latin America in which Mediterranean and Central American populations share an ancestral lineage. Conversely, South American invasion history is more complex, and our results partly suggest an old secondary invasion into South America from Europe or a colonization of South America directly from Africa, followed by admixture with an European lineage. Throughout its invasive range, C. capitata is challenged with diverse climatic regimes. A genome wide association study identified a relationship between allele frequency changes and specific bioclimatic variables. Notably, we observed a significant allele frequency shift related to adaptation to cold stress (BIO6), highlighting the species’ ability to rapidly adapt to seasonal variations in colder climates.

Biotic and abiotic factors influence how insects respond to stimuli. This can make it challenging to interpret captures in traps used to monitor pest abundance in management programmes. To address this, the lure response of three pest fruit flies (Diptera: Tephritidae) was evaluated in a semi-field setting with respect to several physiological and environmental factors. Using standardised methods with known fly numbers in field cages, the response to Biolure (food-based lure) was evaluated for Ceratitis capitata, Ceratitis cosyra and Bactrocera dorsalis. Response to the male lures was tested: E.G.O PheroLure for C. capitata and C. cosyra, Trimedlure for C. capitata, and methyl eugenol for B. dorsalis. The physiological variables evaluated were fly age, sex, weight, and total body nutritional composition. The environmental effects of temperature, relative humidity and light intensity were also assessed. Protein-deprived adults responded more strongly to Biolure. The response to Biolure was not sex-specific. Fly age influenced the response of all species to all tested lures. However, this effect was species and lure specific. Temperature was the most influential environmental factor, with response generally increasing with temperature. Lower thresholds for lure response, despite the proximity of responsive flies, range from 12.21 to 22.95 °C depending on the species and lure tested. These results indicate that trapping systems and management activity thresholds must take physiological and environmental variation into account to increase their accuracy.

The sterile insect technique (SIT) is a method of biological control whereby millions of factory reared sterile male insects are released into the field. This technique is commonly used to combat the Mediterranean fruit fly ( Ceratitis capitata , Diptera: Tephritidae). Sterile medfly males are less competent in attracting and mating with wild females, a property commonly linked to the irradiation process responsible for the sterilization. As bacteria are important partners in the fly's life cycle, we used molecular analytical methods to study the community structure of the gut microbiota in irradiated male medflies. We find that the sterilizing irradiation procedure affects the gut bacterial community structure of the Mediterranean fruit fly. Although the Enterobacteriaceae family remains the dominant bacterial group present in the gut, the levels of Klebsiella species decreases significantly in the days after sterilization. In addition, we detected substantial differences in some bacterial species between the mass rearing strain Vienna 8 and the wild strain. Most notable among these are the increased levels of the potentially pathogenic species Pseudomonas in the industrial strain. Testing the hypothesis that regenerating the original microbiota community could result in enhanced competitiveness of the sterile flies, we found that the addition of the bacterial species Klebsiella oxytoca to the postirradiation diet enables colonization of these bacteria in the gut while resulting in decreased levels of the Pseudomonas sp. Feeding on diets containing bacteria significantly improved sterile male performance in copulatory tests. Further studies will determine the feasibility of bacterial amelioration in SIT operations.