Integrated pest management relies on the combined use of different practices in time and/or space. The main objectives are to better control pests, not to induce too much selective pressure on resistance mechanisms present in pest populations and to minimize non-targeted effects on the ecosystem [1]. The efficiency of such a strategy requires at least additional or synergistic effects of chosen tools against targeted pest population in a specific environment. Any antagonistic effect on targeted or non-targeted organisms might reduce control effort to nil even worst.

Van Oudenhove et al [2] raised the question of the interaction between botanical pesticides (BPs) and egg parasitoids. Each of these two strategies used for pest management present advantages and are described as eco-friendly. First, the use of parasitoids is a great example of biological control and is massively used in a broad range of crop production in different ecological settings. Second, BPs, especially essential oils (EOs) used for a wide range of activities on pests (repellent, antifeedant, antiovipositant, ovicidal, larvicidal and simply pesticidal) present low-toxicity to non-target vertebrates and do not last too long in the environment. Combining these two strategies might be considered as a great opportunity to better pest control with minimized impact on environment. However, EOs used to target a wide range of pest might directly or indirectly affect parasitoids.

Van Oudenhove et al [2] focused their study on non-target effects of 10 essentials oils with pesticide potential on larval development and egg-seeking behaviour of five strains of the biocontrol agent Trichogramma evanescens. Within two laboratory experiments mimicing EOs fumigation (i.e. contactless EOs exposure), the authors evaluated (1) the toxicity of EOs on parasitoid development and (2) the repellent effect of these EOs on adult wasps. They confirmed that contactless exposure of EOs can (1) induce mortality during pre-imaginal development (more acute at the pupal stage) and (2) induce behavioural avoidance of EOs odour plume. These experiments ran onto five strains of T. evanescens also highlighted the variation of the effects of EOs among parasitoid strains.

The complex and dynamic interaction between pest, plant, parasitoid (a natural enemy) and their environment is disturbed by EOs. EOs plumes are also dynamic and variable upon the environmental conditions. The results of van Oudenhove et al. experimentally illustrate such a complexity by describing opposite effects (repellent and attractive) of the same EO on the behaviour of two T. evanescens strains. These contrasting results led us to question more broadly the non-target effects of pest management programs based on EOs fumigation on natural enemies.

Finally, the limits of this experimental study as discussed in the paper draw research avenues taking into account biotic variables such as plant chemical cues, odour plume dynamics, individual behavioural experiences and abiotic variables such as temperature, light and gravity [3] in laboratory, semi-field and field experiments. Facing such a complexity, modelling studies at fine scale in time and space have the operational objective to help farmers to choose the best IPM strategy regarding their environment (as illustrated for aphid population management in the recent review by Stell et al. [4]). But before such research effort to be undertaken, Van Oudenhove et al study [2] sounds like an alert for a cautious use of EOs in pest control programs that integrate biological control with parasitoids.

References

[1] Fauvergue, X. Biocontrôle Elements Pour Une Protection Agroecologique des Cultures; Éditions Quae: Versailles, France, 2020.

[2] van Oudenhove L, Cazier A, Fillaud M, Lavoir AV, Fatnassi H, Pérez G, Calcagno V. Non-target effects of ten essential oils on the egg parasitoid Trichogramma evanescens. bioRxiv 2022.01.14.476310, ver. 4 peer-reviewed and recommended by PCI Zoology. https://doi.org/10.1101/2022.01.14.476310

[3] Victor Burte, Guy Perez, Faten Ayed, Géraldine Groussier, Ludovic Mailleret, Louise van Oudenhove and Vincent Calcagno (2022) Up and to the light: intra- and interspecific variability of photo- and geo-tactic oviposition preferences in genus Trichogramma, Peer Community Journal, 2: e3. https://doi.org/10.24072/pcjournal.78

[4] Stell E, Meiss H, Lasserre-Joulin F, Therond O. Towards Predictions of Interaction Dynamics between Cereal Aphids and Their Natural Enemies: A Review. Insects 2022, 13, 479. https://doi.org/10.3390/insects13050479

Molecular markers have drastically changed and improved our understanding of biological processes. In combination with PCR, markers revolutionized the study of all organisms, even tiny insects, and eukaryotic pathogens amongst others. Microsatellite markers were the most prominent and successful ones. Their success started in the early 1990s. They were used for population genetic studies, mapping of genes and genomes, and paternity testing and inference of relatedness. Their popularity is based on some of their characteristics as codominance, the high polymorphism information content, and their ease of isolation (Schlötterer 2004). Still, microsatellites are the marker of choice for a range of non-model organisms as next-generation sequencing technologies produce a huge amount of single nucleotide polymorphisms (SNPs), but often at expense of sample size and higher costs.
 
The high level of polymorphism of microsatellite markers, which consist of one to six base-pair nucleotide motifs replicated up to 10 or 20 times, results from slippage events during DNA replication. Short hairpin loops might shorten the template strand or extend the new strand. However, such slippage events might occur during PCR amplification resulting in additional bands or peaks. Such stutter alleles often appear to differ by one repeat unit and might be hard to interpret but definitively reduce automated scoring of microsatellite results.
 
A standalone software package available to handle stuttering is Microchecker (van Oosterhout et al., 2004, which nowadays faces incompatibilities with updated versions of different operating systems. Thus, de Meeûs and Noûs (2022), in their manuscript, tackled the stuttering issue by developing an OS-independent analysis pipeline based on standard spreadsheet software such as Microsoft Office (Excel) or Apache Open Office (Calc). The authors use simulated populations differing in the mating system (pangamic, selfing (30%), clonal) and a different number of subpopulations and individuals per subpopulation to test for differences among the null model (no stuttering), a test population with 2 out of 20 loci (10%) with stuttering, and the latter with stuttering cured. Further to this, the authors also re-analyse data from previous studies utilising organisms differing in the mating system to understand whether control of stuttering changes major parameter estimates and conclusions of those studies.
 
Stuttering of microsatellite loci might result in increased heterozygote deficits. The authors utilise the FIS (inbreeding coefficient) as a tool to compare the different treatments of the simulated populations. Their method detected stuttering in pangamic and selfing populations, while the detection of stuttering in clonal organisms is more difficult. The cure for stuttering resulted in FIS values similar to those populations lacking stuttering. The re-analysis of four previously published studies indicated that the new method presented here is more accurate than Microchecker (van Oosterhout et al., 2004) in a direct comparison. For the Lyme disease-transmitting tick Ixodes scapularis (De Meeûs et al., 2021), three loci showed stuttering and curing these resulted in data that are in good agreement with pangamic reproduction. In the tsetse fly Glossina palpalis palpalis (Berté et al., 2019), two out of seven loci were detected as stuttering. Curing them resulted in decreased FIS for one locus, while the other showed an increased FIS, an indication of other problems such as the occurrence of null alleles. Overall, in dioecious pangamic populations, the method works well, and the cure of stuttering improves population genetic parameter estimates, although FST and FIS might be slightly overestimated. In monoecious selfers, the detection and cure work well, if other factors such as null alleles do not interfere. In clonal organisms, only loci with extremely high FIS might need a cure to improve parameter estimates.
 
This spreadsheet-based method helps to automate microsatellite analysis at very low costs and thus improves the accuracy of parameter estimates. This might certainly be very useful for a range of non-model organisms, parasites, and their vectors, for which microsatellites are still the marker of choice. 
 
References

Berté D, De Meeus T, Kaba D, Séré M, Djohan V, Courtin F, N'Djetchi KM, Koffi M, Jamonneau V, Ta BTD, Solano P, N’Goran EK, Ravel S (2019) Population genetics of Glossina palpalis palpalis in sleeping sickness foci of Côte d'Ivoire before and after vector control. Infection Genetics and Evolution 75, 103963. https://doi.org/0.1016/j.meegid.2019.103963

de Meeûs T, Chan CT, Ludwig JM, Tsao JI, Patel J, Bhagatwala J, Beati L (2021) Deceptive combined effects of short allele dominance and stuttering: an example with Ixodes scapularis, the main vector of Lyme disease in the U.S.A. Peer Community Journal 1, e40. https://doi.org/10.24072/pcjournal.34

de Meeûs T, Noûs C (2022) A simple procedure to detect, test for the presence of stuttering, and cure stuttered data with spreadsheet programs. Zenodo, v5, peer-reviewed and recommended by PCI Zoology. https://doi.org/10.5281/zenodo.7029324

Schlötterer C (2004) The evolution of molecular markers - just a matter of fashion? Nature Reviews Genetics 5, 63-69. https://doi.org/10.1038/nrg1249

van Oosterhout C, Hutchinson WF, Wills DPM, Shipley P (2004) MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Molecular Ecology Notes 4, 535-538. https://doi.org/10.1111/j.1471-8286.2004.00684.x

Genetic (bio)diversity is one of three recognised levels of biodiversity, besides species and ecosystem diversity. Its importance for species survival and adaptation is increasingly highlighted and its monitoring recommended (e.g. O’Brien et al 2022). Especially the management of ex-situ populations has a long history of taking into account genetic aspects (through pedigree analysis but increasingly also by applying molecular tools). As in-situ and ex-situ efforts are nowadays often aligned (in a One-Plan-Approach), genetic management is becoming more the standard (supported by quickly developing genomic techniques). However, rarely quantitative genetic aspects are raised in this issue, while its relevance cannot be underestimated. Hence, the current manuscript by Sauve et al (2022) is a welcome contribution, in order to improve conservation efforts. The authors give a clear overview on how quantitative genetic analysis can aid the measurement, monitoring, prediction and management of adaptive genetic variation. The main tools are pedigrees (mainly of ex-situ populations) and the Animal Model. The main goal is to prevent adaption to captivity and altered genetics in general (in reintroduction projects). The confounding factors to take into account (like inbreeding, population structure, differences between facilities, sample size and parental/social effects) are well described by the authors. As such, I fully recommend this manuscript for publication, hoping increased interest in quantitative analysis will benefit the quality of species conservation management.

References

O'Brien D, Laikre L, Hoban S, Bruford MW et al. (2022) Bringing together approaches to reporting on within species genetic diversity. Journal of Applied Ecology, 00, 1–7. https://doi/10.1111/1365-2664.14225

Sauve D., Spero J., Steiner J., Wheeler H., Lynch C., Chabot A.A. (2022) Improving species conservation plans under IUCN’s One Plan Approach using quantitative genetic methods. EcoEvoRxiv, ver. 9 peer-reviewed and recommended by Peer Community in Zoology. https://doi.org/10.32942/osf.io/n3zxp

Because of the inability of eggs to move, the fitness of oviparous organisms is particularly dependent on the oviposition site. The choice of oviposition site by mothers is therefore the result of trade-offs between exposure to risk factors or favorable conditions such as the presence/absence of predators, the threat of extreme temperatures, the risk of desiccation, the presence and quality of nutritional resources... In addition to these trade-offs between different biotic and abiotic factors that determine oviposition site selection, the ability of mothers to move their eggs after oviposition is a game-changer in insect strategies to optimize egg development and survival [1]. Oviposition site selection combined with egg transport has been explored in insects in relation to the risk of exposure to egg parasitoids [2] or needs for oxygenation [3] but surprisingly has not been investigated in regards to temperatures. Considering egg transport in the ability of insects to adapt their behavior to environmental conditions and in particular to potential extreme temperatures is yet inherent in providing a complete picture of the diversity of behaviors that shape adaptation to temperature and potential tolerance to climate change. In this sense, the study presented by Tourneur et al. [4], explores whether insects capable of egg-care might use egg transport as an adaptive behavior to protect them from suboptimal or extreme temperatures. The study was conducted in the European earwig, Forficula auricularia Linnaeus, 1758, which is known to practice egg-care in a variety of ways, that presumably includes egg-transportation, for several weeks or months during winter until hatching. The authors characterized different life-history traits related to egg-laying, egg-transport, and egg-development in two device systems with three experimental temperature regimes in two populations of European earwigs from Canada. The inclusion of two populations, which turned out to belong to two clades, allowed the identification of a diversity of behaviors although this did not allow to attribute the differences between the two populations to specific population differences, genetic differences, or to their geographical origins. Interestingly, the study showed that oviposition site selection in the European earwig is driven by temperature and that in winter temperatures, female earwigs may move their eggs to warmer temperatures that are adequate for hatching. These results are original in the sense that they highlight new adaptive strategies in female insects used during the post-oviposition stage to protect their eggs from temperature changes.

In the current context of climate change and potential changes in selective pressures, the study contributes to the understanding of the wide range of strategies deployed by insects to adapt to the temperature. This appears essential to predict and anticipate the consequences of global instability, it also describes from an academic point of view a new and fascinating adaptive strategy in an overlooked biological system. 

References

[1] Machado G, Trumbo ST (2018) Parental care. In: Insect Behavior, pp. 203–218. Oxford University Press, Oxford. https://doi.org/10.1093/oso/9780198797500.003.0014

[2] Carrasco D, Kaitala A (2009) Egg-laying tactic in Phyllomorpha laciniata in the presence of parasitoids. Entomologia Experimentalis et Applicata, 131, 300–307. https://doi.org/10.1111/j.1570-7458.2009.00857.x

[3] Smith RL (1997) Evolution of paternal care in the giant water bugs (Heteroptera: Belostomatidae). In: The Evolution of Social Behaviour in Insects and Arachnids (eds Crespi BJ, Choe JC), pp. 116–149. Cambridge University Press, Cambridge. https://doi.org/10.1017/CBO9780511721953.007

[4] Tourneur J-C, Cole C, Vickruck J, Dupont S, Meunier J (2022) Pre- and post-oviposition behavioural strategies to protect eggs against extreme winter cold in an insect with maternal care. bioRxiv, 2021.11.23.469705, ver. 3 peer-reviewed and recommended by Peer Community in Zoology. https://doi.org/10.1101/2021.11.23.469705

I recommend the Van Steenberge et al. study. With over 2000 endemic species, the East African cichlids are a well-established model system in speciation research (Salzburger 2018) and several models have been proposed and tested to explain how these radiations formed (Kocher 2004). Hybridization was shown to be a main driver of the rapid speciation and adaptive radiations of the East African Cichlid fishes (Seehausen 2004). However, it is obvious that unrestrained hybridization also has the potential to reduce taxonomic diversity by erasing species barriers. In the classical model of cichlid evolution, special emphasis was placed on mate preference (Kocher 2004). However, no attention was placed on species recognition, which was implicitly assumed. There is, however, more research needed on what species recognition means, especially in radiating lineages such as cichlids. In a previous study, Nevado et al. 2011 found traces of asymmetrical hybridization between members of the Lake Tanganyika radiation: the genus Ophthalmotilapia. This recommended study by Van Steenberge et al. is based on Nevado et al. (2011), which detected that in one genus of Ophthalmotilapia mitochondrial DNA ‘typical’ for one of the four species (O. nasuta) was also found in three other species (O. ventralis, O. heterodonta, and O. boops). The authors suggested that this could be explained by the fact that females of the three other species accepted O. nasuta males, but that O. nasuta females were more selective and accepted only conspecifc males. This could hence be due to asymmetric mate preferences, or by asymmetric abilities for species recognition. 

This is exactly what the current study by Van Steenberge et al. did. They tested the latter hypothesis by presenting females of two different Ophthalmotilapia species with con- and heterospecific males. This was tested through experiments, making use of wild specimens of two species: O. nasuta and O. ventralis. The authors assumed that if they performed classical “choice-experiments”, they would not notice the recognition effects, given that females would just select preferred, most likely conspecific, males. Instead, specimens were only briefly presented to other fishes since the authors wanted to compare differences in the ability for ‘species recognition’. In this, the authors followed Mendelson and Shaw (2012) who used “a measurable difference in behavioural response towards conspecifics as compared to heterospecifics’’ as a definition for recognition. Instead of the focus on selection/preference, they investigated if females of different species behaved differently, and hence detected the difference between conspecific and heterospecific males. This was tested by a short (15 minutes) exposure to another fish in an isolated part of the aquarium. Recognition was defined as the ‘difference in a particular behaviour between the two conditions’. What was monitored was the swimming behaviour and trajectory (1 image per second) together with known social behaviours of this genus. The selection of these behaviours was further facilitated based on experimental set-ups of reproductive behaviour or the same species previously described by the same research team (Kéver et al. 2018).

The result was that O. nasuta females, for which it was expected that they would not hybridize, showed a different behaviour towards a con- or a heterospecific male. They interacted less with males of the other species. What was unexpected is that there was no difference in behaviour of the females whether they recognized a male or (control) female of their own species. This suggests that they did not detect differences in reproductive behaviour, but rather in the interactions between conspecifics. For females of O. ventralis, for which there are indications for hybridization in the wild, they did not find a difference in behaviour. Females of this species behaved identically with respect to the right and wrong males as well as towards the control females. Interestingly is thus that a complex pattern between species in the wild could be (partially) explained by the behaviour/interaction at first impression of the individuals of these species. 

References

Kéver L, Parmentier E, Derycke S, Verheyen E, Snoeks J, Van Steenberge M, Poncin P (2018) Limited possibilities for prezygotic barriers in the reproductive behaviour of sympatric Ophthalmotilapia species (Teleostei, Cichlidae). Zoology, 126, 71–81. https://doi.org/10.1016/j.zool.2017.12.001

Kocher TD (2004) Adaptive evolution and explosive speciation: the cichlid fish model. Nature Reviews Genetics, 5, 288–298. https://doi.org/10.1038/nrg1316

Mendelson TC, Shaw KL (2012) The (mis)concept of species recognition. Trends in Ecology & Evolution, 27, 421–427. https://doi.org/10.1016/j.tree.2012.04.001

Nevado B, Fazalova V, Backeljau T, Hanssens M, Verheyen E (2011) Repeated Unidirectional Introgression of Nuclear and Mitochondrial DNA Between Four Congeneric Tanganyikan Cichlids. Molecular Biology and Evolution, 28, 2253–2267. https://doi.org/10.1093/molbev/msr043

Salzburger W (2018) Understanding explosive diversification through cichlid fish genomics. Nature Reviews Genetics, 19, 705–717. https://doi.org/10.1038/s41576-018-0043-9

Seehausen O (2004) Hybridization and adaptive radiation. Trends in Ecology & Evolution, 19, 198–207. https://doi.org/10.1016/j.tree.2004.01.003

Steenberge MV, Jublier N, Kéver L, Gresham S, Derycke S, Snoeks J, Parmentier E, Poncin P, Verheyen E (2022) The initial response of females towards congeneric males matches the propensity to hybridise in Ophthalmotilapia. bioRxiv, 2021.08.07.455508, ver. 3 peer-reviewed and recommended by Peer Community in Zoology. https://doi.org/10.1101/2021.08.07.455508