Parasitoid Wasp Research Articles

Effect of wheat cultivars, with varying resistance levels to Rhyzopertha dominica, on life history parameters of the ectoparasitoid wasp Anisopteramalus calandrae

The ectoparasitoid wasp Anisopteromalus calandrae (Howard) (Hymenoptera: Pteromalidae), as a widely distributed ectoparasitoid wasp, can parasitize several Coleopteran insect pests such as the lesser grain borer Rhyzopertha dominica (F.) (Coleoptera, Bostrichidae). In the current study, the plant-herbivore-parasitoid interactions with A. calandrae and R. dominica on six cultivars of wheat Triticum aestivum L. (Poals: Poaceae), including Aftab, Araz, Ghabos, Kalateh, Meraj, and Tirgan, were assessed. The largest egg and larval-pupal periods of R. dominica were observed on cultivars Tirgan and Aftab. Rhyzopertha dominica reared on cultivar Araz had the largest total developmental period. The highest oviposition period and fecundity of R. dominica were observed on cultivar Meraj. The pest had the brief life and the low fecundity duration on cultivar Tirgan. The longest developmental time of A. calandrae was observed on the cultivar Tirgan. The parasitic wasp lived longer on cultivar Ghabos. The developmental time of A. calandrae was positively affected by phenolic content of wheat cultivars. The fecundity and intrinsic rate of increase (r) of the parasitic wasp were significantly decreased by increasing the phenolic content of the cultivars. In general, the cultivar Meraj was sensitive, the cultivars Aftab and Kalateh were semi-resistant, and the cultivars Araz, Ghabos, and Tirgan were resistant to R. dominica. The cultivar Tirgan was found as an unsuitable host compared to others for performance of A. calandrae, while Aftab, Kalateh, Araz, and Meraj were suitable ones. Consequently, the use of cultivar Araz as a resistant host to R. dominica and the suitable cultivar to A. calandrae can be considered as an effective integrated management choice against the pest.

Density-dependent fitness and mass-rearing of Habrobracon hebetor (Say) (Hymenoptera: Braconidae) incorporating life table variability

The ectoparasitoid Habrobracon hebetor (Say) (Hymenoptera: Braconidae) is an important natural enemy insect deployed for controlling lepidopteran larvae in warehouses and fields. However, there is limited information on mass-rearing methods of H. hebetor. This study investigated the effects of different Ephestia elutella (Hübner) (Lepidoptera: Pyralidae) and parasitic egg densities on the fitness and mass-rearing efficiency of H. hebetor, aiming to determine the optimal conditions for developing an economically viable and sustainable mass-rearing system for this parasitoid. Our results showed that the average number of eggs laid per host dropped below 7.54 eggs when the number of hosts exceeded five. H. hebetor exhibited significantly increased preadult survival rate (67.41%-81.00%), female and male body size (2224.55-2379.83 μm, 2213.39-2324.65 μm), fecundity (443.98-492.43 eggs), net reproductive rate (R0=154.57-216.67 offspring), intrinsic rate of increase (r=0.2776-0.3223 d-1), net paralyzed rate (C0=72.37-93.17 larvae), and net parasitism rate (Z0=43.55-55.04 larvae) at a density of 5-15 eggs per host than at 20 eggs per host. Meanwhile, considering rearing costs and production rate, a density of 5-15 eggs per host proved to be the most economical for mass-rearing H. hebetor, achieving a daily harvest rate of 10,000 newly emerged female adults. Therefore, for an efficient mass-rearing system for H. hebetor, maintaining a density of 5-15 eggs per host is recommended considering variability in life table and discarding aged individuals (after 36 days old). In summary, providing 5-10 5th instar larvae of E. elutella per day per pair of male and female parasitoid wasps is advised for effective mass-rearing of H. hebetor.

Unraveling the venom constituents of the endoparasitoid Aphidius gifuensis with an emphasis on the discovery of a novel insecticidal peptide.

Venom serves as a pivotal parasitic factor employed by parasitoid wasps to manipulate their hosts, creating a favorable environment for the successful growth of their progeny, and ultimately kill the host. The bioactive molecules within parasitoid venoms exhibit insecticidal activities with promising prospects for agricultural applications. However, knowledge regarding the venom components of parasitoids and the discovery of functional biomolecules from them remains limited. In this study, 30 venom proteins were identified from the endoparasitoid Aphidius gifuensis through the application of a transcriptomic approach. These proteins were categorized into five groups: hydrolase, molecular chaperone, transferase, other functional protein, and hypothetical protein with unknown function. Particularly noteworthy is the abundant expression of the peptide Vn1 in the venom apparatus of A. gifuensis, indicating its pivotal role in venom activity. Consequently, Vn1 was chosen for further functional analysis, exhibiting insecticidal activity against Tenebrio molitor pupae. Further assessment for revealing its mode of action disclosed that Vn1 impacts genes related to immune response, environmental information processing, metabolism, and response to external stimuli in T. molitor, suggesting its involvement in the intricate parasitoid wasp-host interaction. The findings of this study significantly contribute to our knowledge of the composition and functionality of A. gifuensis venom, establishing a foundation for further investigation into the biological roles of the identified venom constituents. The insecticidal Vn1 isolated from the venom of this parasitoid represents a valuable resource for the development of innovative biocontrol agents with potential applications in agriculture. © 2024 Society of Chemical Industry.

Selection of a δ-latroinsectotoxin fragment from Latrodectus tredecimguttatus venom for efficient bacterial expression

Toxins from the venom of predatory and parasitoid arthropods, such as scorpions, spiders, and parasitoid wasps, are used as promising tools in plant protection against insect pests. In many cases, polyclonal antibodies can be created to study a single molecule in the laboratory, which allows for specific methods of detection and purification. In the present study, heterologous expression of δ-latroinsectotoxin from the venom of the black widow spider Latrodectus tredecimguttatus, as well as its N- and C- terminal fragments, was carried out in Escherichia coli cells at various temperatures and aeration conditions. Regardless of the expression conditions, the accumulation of the recombinant protein in large quantities in the bacterial cells as inclusion bodies was observed only for the C-terminal fragment of the toxin. The high efficiency of expression of this fragment allowed the use of the insoluble protein fraction of the bacterial homogenate for the immunization of mice and the production of antibodies to δ-latroinsectotoxin. The antigen-binding activity of the immunoglobulins was confirmed by immunoblotting of the synthesized toxin fragment in bacteria using a vector with removed leader sequences. The antibodies obtained can be used to detect the black widow spider toxin during its expression in various systems, such as in the creation of genetically modified entomopathogenic fungi or viruses.

The first record of hyperparasitism in mermithid nematodes

Summary The Mermithidae are primarily simple parasitoids of insects. In this study, we report the first observation of hyperparasitism by mermithid nematodes. Two Mermithidae individuals emerged from a specimen of the parasitoid wasp Coleocentrus incertus collected in Hokkaido, Japan. Although the nematodes were in the juvenile stage and many morphological characteristics were underdeveloped, we were able to observe cross fibre layers on the cuticles. A molecular sequence analysis of the 18S ribosomal DNA suggested that the nematode belonged to the genus Hexamermis, which was supported by the presence of the cross fibre layers. The phylogenetic analysis also suggested a close relationship between our specimens and ‘Mermithidae sp. NM1’, a taxon found in bumblebees, as differences between the sequences of the two taxa were minimal; however, further research is required to confirm the species’ identity. Hyperparasitism by nematodes is rare and is typically unsuccessful in non-host species, yet the nematodes in this study successfully parasitised a parasitoid wasp. Our study is the first to demonstrate that mermithid nematodes can exhibit hyperparasitism in addition to their known parasitic behaviour. The parasitic strategies of Mermithidae nematodes appear to be more flexible than previously assumed, indicating a high degree of adaptability as parasites.

Immature stages and natural history of Eruga unilabiana (Hymenoptera: Ichneumonidae), an ectoparasitoid of two linyphiid spiders

AbstractWasps from the Polysphincta genus group (Hymenoptera: Ichneumonidae: Pimplinae) are currently the only parasitoid wasps that are ectoparasitoids of spiders. The species of the group exhibit several morphological and behavioral adaptations, which allow exploiting their host spiders. Here, we describe the external morphology, behavior and natural history of the immature stages of a Darwin wasp Eruga unilabiana Pádua & Sobczak, 2018, a parasitoid of sheet‐weaving spiders, Sphecozone sp. and Eurymorion sp. (Linyphiidae). The egg of E. unilabiana is a white mass that is deposited in the anterior region of the spider's abdomen. First stage larvae partially emerge from the egg's chorion. Second stage larvae have a body divided into 13 segments and induce spiders to build a modified web. The third stage larvae have eight pairs of retractable dorsal tubercles with tiny little hooks that help the larvae hang and move on the web threads after the spider dies. These results indicate that the immature stages of E. unilabiana present the same basic morphological and behavioral characteristics already known for the species of the group. However, the last stage larva's strategy of moving first to hang on the web threads before killing the spider is the first record in the group and more studies are needed to assess whether this behavior is adaptive for the wasp.

A naturally occurring mitochondrial genome variant confers broad protection from infection in Drosophila.

The role of mitochondria in immunity is increasingly recognized, but it is unclear how variation in mitochondrial DNA (mtDNA) contributes to variable infection outcomes. To quantify the effect of mtDNA variation on humoral and cell-mediated innate immune responses, we utilized a panel of fruit fly Drosophila melanogaster cytoplasmic hybrids (cybrids), where unique mtDNAs (mitotypes) were introgressed into a controlled isogenic nuclear background. We observed substantial heterogeneity in infection outcomes within the cybrid panel upon bacterial, viral and parasitoid infections, driven by the mitotype. One of the mitotypes, mtKSA2 protected against bacterial, parasitoid, and to a lesser extent, viral infections. Enhanced survival was not a result of improved bacterial clearance, suggesting mtKSA2 confers increased disease tolerance. Transcriptome sequencing showed that the mtKSA2 mitotype had an upregulation of genes related to mitochondrial respiration and phagocytosis in uninfected flies. Upon infection, mtKSA2 flies exhibited infection type and duration specific transcriptomic changes. Furthermore, uninfected mtKSA2 larvae showed immune activation of hemocytes (immune cells), increased hemocyte numbers and ROS production, and enhanced encapsulation response against parasitoid wasp eggs and larvae. Our results show that mtDNA variation acts as an immunomodulatory factor in both humoral and cell-mediated innate immunity and that specific mitotypes can provide broad protection against infections.

Inhibition of the Notch signal transducer CSL by Pkc53E-mediated phosphorylation to fend off parasitic immune challenge in Drosophila

Notch signalling activity regulates hematopoiesis in Drosophila and vertebrates alike. Parasitoid wasp infestation of Drosophila larvae, however, requires a timely downregulation of Notch activity to allow the formation of encapsulation-active blood cells. Here, we show that the Drosophila CSL transcription factor Suppressor of Hairless [Su(H)] is phosphorylated at Serine 269 in response to parasitoid wasp infestation. As this phosphorylation interferes with the DNA binding of Su(H), it reversibly precludes its activity. Accordingly, phospho-deficient Su(H)S269A mutants are immune-compromised. A screen for kinases involved in Su(H) phosphorylation identified Pkc53E, required for normal hematopoiesis as well as for parasitoid immune response. Genetic and molecular interactions support the specificity of the Su(H)-Pkc53E relationship. Moreover, phorbol ester treatment inhibits Su(H) activity in vivo and in human cell culture. We conclude that Pkc53E targets Su(H) during parasitic wasp infestation, thereby remodelling the blood cell population required for wasp egg encapsulation.

Inhibition of the Notch signal transducer CSL by Pkc53E-mediated phosphorylation to fend off parasitic immune challenge in Drosophila.

Notch signalling activity regulates hematopoiesis in Drosophila and vertebrates alike. Parasitoid wasp infestation of Drosophila larvae, however, requires a timely downregulation of Notch activity to allow the formation of encapsulation-active blood cells. Here, we show that the Drosophila CSL transcription factor Suppressor of Hairless [Su(H)] is phosphorylated at Serine 269 in response to parasitoid wasp infestation. As this phosphorylation interferes with the DNA binding of Su(H), it reversibly precludes its activity. Accordingly, phospho-deficient Su(H)S269A mutants are immune-compromised. A screen for kinases involved in Su(H) phosphorylation identified Pkc53E, required for normal hematopoiesis as well as for parasitoid immune response. Genetic and molecular interactions support the specificity of the Su(H)-Pkc53E relationship. Moreover, phorbol ester treatment inhibits Su(H) activity in vivo and in human cell culture. We conclude that Pkc53E targets Su(H) during parasitic wasp infestation, thereby remodelling the blood cell population required for wasp egg encapsulation.

Regulation of melanization in aphids by parasitoid wasp venom proteins enhances mummification.

Interactions between parasitic insects and their hosts demonstrate the complexity of evolutionary processes. Specifically, the parasitoid wasp Aphidius ervi manipulates its host, the pea aphid Acyrthosiphon pisum, through strategic venom injection to enhance mummification. This study explores how this venom affects the aphid's immune system, particularly targeting the activity of the phenoloxidase (PO) enzyme. Following the injection of venom from A. ervi, significant changes were observed in the expression of immune-related genes in A. pisum, especially notable expression changes of ApPPOs and a reduction of PO activity. Multi-omics sequencing identified 74 potential venom proteins in the venom gland of A. ervi, including serine protease homolog 1 (AeSPH1) and serine protease inhibitor (AeSPN1), hypothesized to regulate PO activity. The injection of recombinant protein AeSPH1 and AeSPN1 into the A. pisum hemocoel selectively reduced the expression of ApPPO1, without affecting ApPPO2, and effectively suppressed melanization. Moreover, RNAi targeting AeSPH1 significantly reduced the mummification rate in A. pisum population parasitized by A. ervi. Our findings clarify the complex biochemical mechanisms underlying host-wasp interactions and highlight potential avenues for developing targeted biological control strategies. © 2024 Society of Chemical Industry.

New species of braconid parasitoid wasps from the subfamilies Doryctinae and Brachistinae from late Eocene Baltic amber

Three new fossil species of braconid wasps are described and illustrated from late Eocene Baltic amber: Palaeorhoptrocentrus tenuicornis sp. nov. (Doryctinae), Taphaeus obscurus sp. nov. (Brachistinae: Diospilini), and Eubazus electrus sp. nov. (Brachistinae: Brachistini). Keys to the known species of Palaeorhoptrocentrus Belokobylskij, 2023 and to species of Eubazus Nees, 1812 described from Baltic amber are provided. An illustrated redescription of Blacus (Electroblacus) facialis Brues, 1933 (Brachistinae: Blacini) is also provided, along with digital photographs for the first time of the female of B. (Electroblacus) facialis, whose type material was lost during the World War II. We also provide morphological notes and digital photographs of a male belonging to an undescribed species of Meteorus (Euphorinae).

Sodalis praecaptivus subsp. spalangiae subsp. nov., a nascent bacterial endosymbiont isolated from the parasitoid wasp, Spalangia cameroni.

An endosymbiotic bacterium of the genus Sodalis, designated as strain HZT, was cultured from the parasitoid wasp Spalangia cameroni, which develops on the pupae of various host flies. The bacterium was detected in S. cameroni developed on houseflies, Musca domestica, in a poultry facility in Hazon, northern Israel. After culturing, this bacterium displayed no surface motility on Luria-Bertani agar and was rod-shaped and irregular in size, ~10-30 nm in diameter and 5-20 µm in length. Phylogenetic analyses revealed that strain HZT is closely related to Sodalis praecaptivus strain HST, a free-living species of the genus Sodalis that includes many insect endosymbionts. Although these bacteria maintain >98% sequence identity in shared genes, genomic characterization revealed that strain HZT has undergone substantial reductive evolution, such that it lacks many gene functions that are maintained in S. praecaptivus strain HST. Based on the results of phylogenetic, genomic and chemotaxonomic analyses, we propose that this endosymbiont should be classified in a new subspecies as S. praecaptivus subsp. spalangiae subsp. nov. The type strain for this new subspecies is HZT (=ATCC TSD-398T=NCIMB 15482T). The subspecies Sodalis praecaptivus subsp. praecaptivus strain HST is created automatically with the type strain ATCC BAA-2554T (=DSMZ 27494T).

Strength of enemy release from parasitoids is context-dependent in the invasive African Fig Fly, Zaprionus indianus.

Understanding the mechanisms underlying the success of biological invasions is essential to employ effective prediction and management strategies. Escape from natural enemies in invaded regions (enemy release hypothesis, ERH) and increased competitive ability are hallmarks of invasive species; however, these two processes are rarely studied within the same context. Here, we examined the effect of enemy release on the competition outcomes of a successful invasive insect pest in North America, the African fig fly (Zaprionus indianus). Parasitoid wasps such as Leptopilina heterotoma that parasitize drosophilid larvae may seek out established species with known host suitability over a novel species, so we hypothesized Z. indianus may have low susceptibility to parasitoids, giving them a competitive advantage over co-occurring drosophilids. We tested this hypothesis by comparing the adult emergence rates from Z. indianus larvae reared alone or in competition with Drosophila hydei or D. simulans larvae in the presence and absence of parasitoid wasps under low and high larval densities. At low larval densities, Z. indianus emerged at equal rates to D. hydei but outcompeted D. simulans, and these outcomes were not affected by parasitoids. However, at high densities, the addition of parasitoids shifted competition outcomes in favor of Z. indianus, suggesting enemy release provides a competitive advantage under some circumstances. These results indicate that the strength of enemy release in Z. indianus is widely dependent on contextual factors such as density and competitor species. This study emphasizes how a community approach to testing the ERH is vital as the overall interpretation of the presence and strength of enemy release differed between intraspecific and interspecific experiments. Further investigation of how these results apply to field environments could offer insight into how Z. indianus alters ecosystems and how productive biological control may limit the spread of Z. indianus.

Host Plants for the Lanternfly Saiva formosana Kato, 1929 (Hemiptera, Fulgoridae) Endemic to Taiwan, and Parasitism of Its Eggs by Wasps.

Since Saiva formosana Kato, 1929 was first reported as a new species in Taiwan; there have been few published reports on its ecology, and fundamental knowledge of this species is still lacking. The objectives of this study are to (1) determine the preferred plants of adults, egg-laying, and nymphs, (2) analyze the change in host plants with time and development, and (3) explore the relationship between the survival of eggs and parasitism by their wasps. We detected the adults of this species between May and September 2023, and again in April 2024, mainly on Elaeocarpus decipiens F. B. Forbes & Hemsl. During our investigation, we frequently observed parasitic wasps ovipositing on the egg masses. We established that most egg masses produced only Saiva nymphs or Anastatus adults. This lanternfly might better survive parasitic pressure by laying lower numbers of eggs per mass across a broader range of plant species. The first batch of hatching nymphs was found on Magnolia compresssa Maxim. on 20 June 2023; however, most nymphs in the second and third instars were detected on Ficus fistulosa Reinw. ex Blume, between August and October 2023. After the autumn, the occurrences of nymphs increased on Heptapleurum heptaphyllum (L.) Y. F. Deng which probably served as a shelter for overwintering.

Preliminary observations on the physiology and behavior of Sytretomorpha szaboi Papp

Sytretomorpha szaboi Papp (Hymenoptera: Braconidae) is a parasitic wasp that specializes in parasitizing the Apis cerana and is widely distributed in the Eastern Ocean region, particularly in southern China and northern India. The parasitization rate of this parasitic wasp can reach as high as 20%, which poses a significant threat to the health of bee colonies. This study reports the results of observations of S. szaboi in Chongqing, China. Current research on the physiology and behavior of S. szaboi is limited; thus, this report provides a valuable contribution to the existing body of knowledge on this type of parasitic wasp. Consequently, during the summers of 2023 and 2024, we observed the physiological and behavioral characteristics of S. szaboi and their hosts in the field at the Bee Breeding Center of Chongqing, along with additional incubated observations conducted in the laboratory.

Hymenopterous parasitoids of the fig leaf roller moth, Phycodes radiata (Ochsenheimer, 1808) (Lepidoptera: Brachodidae) in Iran

ABSTRACT The fig leaf roller (FLR), Phycodes radiata (Ochsenheimer, 1808) (Lepidoptera: Brachodidae), is an Oriental defoliator pest of Ficus spp. In the Oriental region, only two parasitic wasps, belonging to the Chalcididae and Eulophidae (Hymenoptera) families have been identified as natural enemies of FLR. Therefore, attempts are being made to discover parasitoids for biological control in Iran. In the laboratory condition (25 ± 1°C, 50%RH), two parasitoid species were successfully reared from FLR pupae. A chalcid species, Psilochalcis ceratoniae Delvare, 2011 (Hymenoptera: Chalcididae) was obtained for the first time on FLR, previously known to parasitise the carob moth, Ectomyelois ceratoniae (Lepidoptera: Pyralidae). Additionally, an ichneumonid species, Goryphus nursei (Cameron, 1907) (Hymenoptera: Ichneumonidae), was also recorded parasitising FLR for the first time. These newly discovered associations bring the total count of parasitoids of FLR to four species from three different families.

Afrotropical Centistidea Rohwer, 1914 (Hymenoptera, Braconidae) with description of four new species.

The braconid parasitoid wasp genus Centistidea Rohwer, 1914 is revised for the Afrotropical region, with four new species described; Centistideaareolaris Liu & Polaszek, sp. nov., C.linearis Liu & Polaszek, sp. nov., C.longipedes Liu & Polaszek, sp. nov., and C.turneri Liu & Polaszek, sp. nov. are described based on specimens from the Natural History Museum, United Kingdom, and the Royal Museum for Central Africa, Belgium. An illustrated key to species in the Afrotropical region is provided.

Infection with pathogenic fungi is a major threat to the mass production of the parasitoid wasp Habrobracon hebetor: 20 generations under surveillance

Pathogen infection is one of the most important challenges threatening the success of mass rearing of natural enemies either in insectaries or in augmentative biological control programs. In this investigation, we studied the biological and population growth parameters of a naturally infected colony of the parasitoid wasp Habrobracon hebetor (Say) (Hymenoptera: Braconidae) over 20 generations (G1inf − G20inf) of rearing on Ephestia kuehniella (Zeller) (Pyralidae: Lepidoptera) under laboratory conditions. Based on morphological characteristics and ITS sequences, two types of fungal colonies were isolated from the diseased females and identified as Thermothielavioides terrestris and Aspergillus flavus. The results showed that although no significant differences were found in the female longevity and fecundity of H. hebetor on E. kuehniella up to G15inf, the duration of the immature period was extended as the number of generations increased. In addition, the intrinsic rate of increase (r) and finite rate of increase (λ) had no significant differences up to G8inf. The net reproductive rate (R0) of G15inf had no significant differences with G2inf, G8inf, and G10inf. On the other hand, in G15inf, R0 was significantly higher than G4inf, G6inf, and G20inf. The findings underscore the negative impact of fungal infection on population growth parameters compared with uninfected population. This study showed that naturally fungal infection of the parasitoid affected the population growth and life table parameters through mass rearing, compared and discussed with the healthy parasitoid.

Host adaptability of Microplitis manilae (Hymenoptera: Braconidae) in Noctuidae.

Biological control is an effective and sustainable method of integrated pest management. Microplitis manilae (Hymenoptera: Braconidae) is a common internal parasitoid wasp of the order Lepidoptera. To determine a suitable host species for the reproduction of parasitoids, it is essential to assess host suitability and parasitic potential of the parasitized pests. Herein, we investigated host selection and exploitation of M. manilae. We evaluated the parasitic efficacy of M. manilae in 4 pest species in the Spodoptera and Mythimna genera of the Noctuidae family. The results indicated that the parasitism rate of M. manilae on 3 species in the Spodoptera genus is higher than on Mythimna separata in the Mythimna genus, with M. manilae exhibiting a higher parasitism rate and shorter development duration on Spodoptera litura compared to other species. The parasitism rate for 1st instars hosts was 86.67 ± 0.04%, while the development duration was 14.1 ± 0.03 days. However, when parasitizing the 3rd instar of Spodoptera frugiperda, parasitoids showed a higher sex ratio, of 0.71 ± 0.05. Additionally, M. manilae had parasitic effect on M. separata, providing a new choice for its parasitism. The results identify the optimal host, which could enhance and reproduction rate and survival rate of M. manilae, thus facilitating their large-scale propagation. Understanding the parasitic effects of M. manilae on pests can further its field application, also plays a major role in promoting the development of biological control technologies and sustainable agricultural production.

Three new species of parasitoid wasps of the genus Mnioes Townes, 1946 (Ichneumonidae: Banchinae) in a humid forest in Peru

Mnioes is a predominantly Neotropical genus of Banchinae (Ichneumonidae), currently including 26 species. Three new species of Mnioes are described, found in a humid montane forest in southern Peru, M. chunka sp. nov, M. chusaq sp. nov. and M. isqun sp. nov, and the male of M. huk Alvarado 2020 is also described. DNA barcoding for all the species treated here is included and provides information to match females with males, especially for M. chunka sp. nov., M. chusaq sp. nov., and M. soqta Alvarado 2020 where females are similar to each other and occur in sympatry. Additionally, an updated key to the Peruvian species is provided.