Abstract

The family Agromyzidae or leaf-miner flies (Diptera, Acalyptratae) is widely distributed in the Palaearctic Region, in which it comprises more than 1,303 species. They are considered as the third largest of Acalyptratae families which possess entirely phytophagous larvae that develop in a wide range of hosts in the plant kingdom.

Our knowledge of Moroccan agromyzid fauna is still poor and many gaps remain for this diverse family of Diptera where many regions in the country are still unexplored such as the Rif region in northern Morocco. This study aims to present new faunistic records as an attempt to fill gaps in our knowledge of Moroccan leafminers.

The specimens used in the present study were captured by sweep net and by Malaise trap at different localities throughout Morocco from 2012 to 2023. Of the 140 sites surveyed, 38 were home to the agromyzid species mentioned in this study. Specimens were kept in 70% ethanol until identification. Each specimen was examined in detail before and after its genitalia were mounted in glycerin on glass-slides. All species have been identified based on the male genitalia.

We provide new records of the family Agromyzidae (leaf-miner flies) from Morocco. Thirty agromyzid species are newly recorded, seven of these in Agromyzinae (Agromyza anthracina, A. conjuncta, A. mobilis, A. myosotidis, A. nigrescens, Hexomyza simplex and Ophiomyia pinguis). The other 23 new records are in Phytomyzinae (Amauromyza (Cephalomyza) karli, Calycomyza flavomaculata, C. solidaginis, Cerodontha (Butomomyza) eucaricis, C. (Dizygomyza) luctuosa, C. (Icteromyza) geniculata, Chromatomyia centaurii, Liriomyza amoena, L. brassicae, L. strigata, Phytoliriomyza arctica, P. perpusilla, P. pteridii, Phytomyza anemones, Ph. clematidis, Ph. crassiseta, Ph. notata, Ph. plantaginis, Ph. rufipes, Ph. vitalbae, Pseudonapomyza confusa, Ps. palavae and Ps. vota). Calycomyza Hendel is newly reported from the country. The present study has increased the total number of agromyzid species in Morocco to 92.

Tomato (Solanum lycopersicum L.) is a vital horticultural crop with economic and nutritional benefits. However, it is vulnerable to pests like the two-spotted spider mite (Tetranychus urticae Koch), which causes significant yield loss and resists many acaricides. Biological control using the phytoseiid mite Neoseiulus longispinosus (Evans) is an eco-friendly alternative. However, the fungicides and pesticides used in tomato cultivation may impact its effectiveness.

The present study was conducted to assess the toxicity of four fungicides viz., copper oxychloride 50% WP (3000 ppm), thiophanate methyl 70% WP (1000 ppm), mancozeb 75% WP (2500 ppm), and metalaxyl 4% + mancozeb 64% WP (2500 ppm) to N. longispinosus and its progeny's fitness against T. urticae. Ten gravid females of N. longispinosus were released onto tomato leaf discs (3cm × 3cm) treated with fungicides in eight replicates by the leaf dip method under laboratory conditions. After 48 hours, cumulative mortality data of N. longispinosus females showed that the fungicide combination metalaxyl + mancozeb caused the highest mortality (60%), followed by mancozeb (52.50%), copper oxychloride (35%), and thiophanate methyl (17.50%). From each treatment and control, 10 surviving predator females were selected and a cohort of 100 predator eggs was collected to further investigate the life table parameters of N. longispinosus on T. urticae. Metalaxyl + mancozeb and mancozeb treatments significantly impacted the life table parameters of N. longispinosus compared to the control, while the effects of copper oxychloride and thiophanate methyl were moderate to negligible.

Thus, N. longispinosus exhibited the ability to withstand the effects of copper oxychloride and thiophanate methyl, while metalaxyl + mancozeb and mancozeb affected its fitness. However, controlled laboratory experiments may not fully account for the influence of biotic and abiotic factors that affect invertebrate fitness under field conditions. Therefore, further field investigations will ascertain the response of the predatory mite to fungicide exposure.

Fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a highly damaging pest of agricultural crops, particularly of maize. As screening of the available crop germplasm is an integral part of integrated pest management programs, the main objective of this research was the screening of available maize varieties, including both hybrid and non-hybrid varieties, against S. frugiperda larval infestation. The seeds of available maize varieties were sown in the field under RCBD, and one-month old plants were artificially infested with laboratory-reared 3^rd instar larvae of S. frugiperda. Data regarding larval infestation and plant damage were assessed using a literature-based damage scale. Moreover, the comparative feeding preference and performance of S. frugiperda larvae were determined in the laboratory under controlled conditions. The overall highest and lowest larval weight gain was observed for Dekalb-6321(27.12 mg) and YH-5427 (15.55 mg), respectively. Maximum and minimum leaf consumption was recorded for Dekalb-6321 (378 mg) and Gohar-19 (297 mg), respectively. The maximum and minimum relative consumption rate was recorded for the varieties YH-5427 (66.79 mg/mg/day) and Mercury (16.27 mg/mg/day), respectively. Regarding relative growth rate (RGR), the larvae of S. frugiperda showed maximum growth when offered leaves of maize varieties Super Green (0.71 mg/mg/day) and Dekalb-6321 (0.48 mg/mg/day). Similarly, maximum ingested food conversion efficiency (ECI) was observed in Super Green (7.47%), followed by Pioneer-1429 (7.43%) and G-7 (7.33%). According to William’s leaf damage scale, the highest levels of in situ leaf damage by lab-reared larvae were recorded on the Mercury (7.33) and Afgoi (6.48) varieties, while the minimum levels of damage were observed on Pioneer (5.20) and YH-5427 (5.41). The findings of this laboratory and field experiment showed the differential feeding preferences of S. frugiperda for the evaluated maize varieties.

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a destructive invasive pest that has emerged as a major threat to agricultural production and food security in India. Understanding its biological parameters and feeding behavior on different host plants is essential for developing sustainable pest management strategies. This study assessed the development, survival, and food consumption of S. frugiperda on five crops: maize (Zea mays L.), popcorn (Zea mays everta Sturt), sweet corn (Zea mays saccharata Sturt), sorghum (Sorghum bicolor L.), and soybean (Glycine max L. Merr.).

Life table analysis and consumption rates were modeled using computer simulations. The results indicated that larvae completed development most rapidly on popcorn (28.02 days), maize (28.04 days), and sweet corn (28.31 days), whereas soybean led to the slowest growth (34.83 days). The highest reproductive and population growth rates were recorded on maize, which also supported the greatest food consumption. The sixth-instar larvae consumed a maximum of 19,470.47 mm² of maize leaf area, whereas the lowest consumption was observed on soybean (9,033.67 mm²). Over a simulated 90-day period, population growth was fastest on maize, popcorn, and sweet corn, while soybean restricted development.

Significant larval and pupal survival on non-maize hosts such as sorghum and soybean suggest that S. frugiperda could pose a risk to their cultivation and may impact maize–soybean intercropping systems. Thus, this study highlights the adaptability of S. frugiperda to the tested crops and lays the groundwork for evaluating its potential threat to other intercropping systems.

The insecticidal action of the entomopathogenic fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) has been documented for a wide range of pests, and in recent years, its formulated application has been tested in several crops. In this work, two different application methods of B. Bassiana PPRI 5339 Velifer^® were studied over two years. In one case, B. bassiana was applied as a coating to the cotton seed (treatment A) Gossypium hirsutum (Malvales: Malvaceae), and in the other case, it was applied via syringe inoculation to the kiwi trunk (treatment B) Actinidia deliciosa “Hayward” (Ericales: Actinidiaceae). For each case, plants without the application of B. bassiana were used as a control (treatment C). The sucking insects Aphis gossypii (Hemiptera: Aphididae) on cotton plants and Halyomorpha halys (Hemiptera: Pentatomidae) on kiwi trees were counted. In addition, the total chlorophyll contents and the leaf area were measured during the experiment. The presence of H. halys was significantly reduced in kiwi trees, and the total chlorophyll content in the leaves was increased. In the cotton plants, the population of A. gossypii was reduced, and the leaf area appeared larger. The presence of B. bassiana on treatments A and B, on the one hand, reduced the sucking pests, while on the other hand, it enhanced the total chlorophyll content and the leaf area. The results of both application methods are encouraging reasons to investigate B. bassiana and similar endophytes in other crops, given the biotic stresses and yield losses that can be caused by aphids or new exotic pests in Greece, such as H. halys.

The global antibiotic resistance crisis has prompted the search for new therapeutic alternatives. In this context, the aquaculture industry has been focused on screening natural alternative sources for developing innovative therapeutics with antibacterial activity that can act as alternatives to conventional antibiotics without promoting resistance. Recently, researchers aimed to design novel strategies for infection management that reduce negative consequences for fish, humans, and aquatic ecosystems. Antimicrobial peptides (AMPs) have gained attention as potential alternatives to conventional antimicrobial agents, primarily due to their effectiveness in suppressing bacterial pathogens and, secondly, for the advantage that microorganisms are unable to develop resistance against these substances. Silkworms B. mori, as representative lepidopteran economic insects, are a promising natural source of AMPs defined as amphipathic small protein molecules, generally containing fewer than fifty amino acid residues. Most of these peptides are recognised for their immunostimulatory effects and broad-spectrum activities against various infectious and non-infectious microorganisms including bacteria, fungi, viruses, and parasites. Therefore, the synthesis and release of AMPs occur as a fundamental aspect of the innate immune defence mechanism against pathogens. In summary, this review synthesizes current knowledge on the various families of AMPs identified in silkworms, their mechanisms of action, antimicrobial properties, and the potential health benefits of AMPs as natural antimicrobial sources in aquaculture.

Culicoides kingi and Culicoides oxystoma belong to the Schultzei group of biting midges, which are significant vectors of livestock diseases in India. The morphological identification of these species remains challenging due to overlapping characteristics, making species-specific identification crucial for resolving taxonomic ambiguities and understanding their roles in disease transmission. This study aims to develop accurate tools for differentiating C. oxystoma from C. kingi using traditional morphometry and DNA-based identification assays which can be effectively utilized. Specimens were collected from various regions of India. A total of 143 C. oxystoma/C. kingi individuals were identified using a combination of traditional morphometric analyses and PCR targeting the cytochrome c oxidase subunit I gene. Thirteen morphometric measurements were taken from the head, wings, and abdomen of slide-mounted specimens for species discrimination. Five morphometric characteristics such as the antennae, wings, spermathecae, maxillary palp with size and shape of the sensory pit, and the hind tibial comb proved to be effective for species differentiation. Molecular phylogenetic analysis further validated the accuracy of the DNA-based identification approach, demonstrating its efficiency as a simple and rapid molecular identification tool. This study provides the first comprehensive differentiation of both species in India using a combination of morphological and molecular techniques. The application of identification from female genitalia and the DNA-based identification method offers a reliable approach for species identification, which can be instrumental in future epidemiological studies at national and international levels.

Introduction
Varroa destructor is currently the most serious parasitic threat to honey bee health. The intensive use of synthetic acaricides has led to the emergence of resistant mite populations, creating an urgent need for alternative control strategies. This study evaluates the acaricidal activity of essential oils (EOs) from Origanum vulgare subsp. viridulum, Thymus capitatus, Thymus longicaulis, and Salvia rosmarinus against V. destructor through in vitro testing.

Methods
Each EO was dissolved in HPLC-grade acetone (2 mg/ml) and applied at a volume of 50 µl inside Eppendorf tubes. The open tubes were placed in an incubator at 35 °C and rotated multiple times to facilitate the EO impregnation of the tube walls while allowing for acetone evaporation. Subsequently, five Varroa mites were introduced into each tube, which was then resealed and returned to the incubator. Control groups included a negative control (acetone only) and a positive control (Amitraz diluted in acetone). Mortality was assessed manually after 1 hr in the incubator by probing individual mites with an entomological pin. Mites showing no movement were classified as dead, while those unable to walk but still capable of moving at least one leg were classified as inactive.

Results and conclusions

The control groups confirmed assay reliability, with 100% survival in the negative control and 100% mortality in the Amitraz-treated group. Among the tested EOs, O. vulgare subsp. viridulum and T. longicaulis exhibited the highest acaricidal activity, each killing 94% of mites. T. capitatus showed slightly lower but still significant efficacy (92%). In contrast, S. rosmarinus demonstrated a more moderate effect, killing 32% of mites. These findings suggest that oregano and thyme essential oils have strong acaricidal properties and could be promising candidates for Varroa control in beekeeping.

The cryptic bumblebee species of Bombus lucorum-complex pose challenges for conservation management. They are morphologically indistinguishable in much of their range,leading to misidentification in most cases. However, total DNA barcoding for such a widespread species is impossible due to financial and labor costs. Among the regions of Northern Europe, reliable data on their distribution exist for Britain, Ireland and Finland. Regarding Finland, only one paper (Pamilo et al. 1997) has been published which provides some data on the distribution of the cryptic species B. lucorum-complex. Concerning the Russian part of Fennoscandia (Murmansk Region and Karelia), no information is available.

In this research, we studied the distribution of the cryptic species B. lucorum-complex in Eastern Fennoscandia (within the administrative boundaries of two regions of the Russian Federation, i.e., Murmansk Region and Karelia). Morphological characters were used for initial species identification. After that, we used the PRC-RFLP method. This approach allows for accurate, quick and relatively cheap identification of the species of Bombus lucorum-complex.

We found that two species, B. cryptarum and B. lucorum, are present in Eastern Fennoscandia. Two other species of B. lucorum-complex, i.e., B. terrestris and B. magnus, have not yet been found in this region. B. cryptarum is recorded in the whole territory of the studied region. Only this species is recorded in the northernmost localities near Norway and the northern part of Finland. In Karelia, the higher proportion of B. lucorum in the samples is in the southern part of this region. B. cryptarum and B. lucorum are associated with a wide range of habitats. The typical sites of concentration of foraging bumblebees are different types of meadows. However, the range of habitats of B. cryptarum is broader. Only this species is found in tundra in the north of Eastern Fennoscandia.

Anthropogenic causes like habitat destruction, pollution, species invasion, and excessive urbanization are major reasons for biodiversity loss. Butterflies are good bioindicators for biodiversity assessments, as habitat destruction can affect butterfly communities. This study was conducted across four types of habitats, rural areas near agricultural fields, suburban areas near grassland, urban parks, and reserve forests, seasonally for a year. A maximum of five, 500 m permanent transect lines were considered in each study area. Butterflies were surveyed following the ‘Pollard Walk’ method. Photographs of the observed species were taken for further identification. Temperature and humidity were recorded for each study site on each sampling day. Butterfly diversity was estimated using the Shannon and Simpson’s indexes. Species richness was calculated using Margalef’s index. A comparative analysis of the species richness, habitat preferences, and frequency of individual species at different sites across all seasons was conducted based on the diversity indices. According to the analysis, there was a statistically significant difference in species numbers and individuals between rural areas (species number: 60; individuals: 3656) and reserve forests (species number: 38; individuals: 985), as well as between rural areas and suburban grassland (species number: 50; individuals: 1832). This study indicated that rural areas exhibited high butterfly diversity and richness, likely due to the presence of various nectaring host plants and lower anthropogenic pressure. This study also signified a statistically significant difference in the species number between the post-monsoon (species number: 58; individuals: 3145) and winter(species number: 45; individuals: 1150) seasons. Correlation and regression analyses showed a positive correlation between individual butterfly numbers and humidity, while species evenness and humidity were correlated negatively. No significant relationship was found between temperature and species diversity. These insights can assist in protecting the butterfly population by considering areas with high butterfly diversity as butterfly conservation zones in the future.