Why Riaz et al. (2023) should be retracted
A few weeks ago, I stumbled across Riaz et al. (2023), “Symbiotic association between ants and fungus” in the Annals of the Entomological Society of America. A potentially interesting paper as I study interactions (even though the title is a little off) but I wasn’t particularly blown away. On the contrary, I was rather surprised that this “review” made it through peer review and got published in a journal with impact factor. Anyway, it didn’t bother me, except for the fact that it inacurrately cited some of my work.
A few days ago, the first author of a recent paper in Scientific Reports that I was invited to contribute to, András Tartally, emailed me and the other co-authors about Riaz et al. (2023). I had not expected to have to read their paper again but when I did, I realized that things were much worse than I had initially realized. There were several cases of plagiarism, both using illustrations while not citing the original sources and copy-pasting text from other papers.
I made a tweet about two plagiarism issues specifically, but was pointed to more issues by other tweeps (thank you!) and so below I am giving an overview of all cases of plagiarism, issues with citations, and anything else that seems problematic. With all this evidence, I do not understand at all how the reviewers did not point to these errors and how the handling editor accepted this monstrosity of a paper. Being an editor myself, doing this above all other responsibilities of an academic, unpaid, I understand the time pressure issues—but still. I only have to read the first sentence of the abstract to feel something is off with this paper.
Let’s start with the abstract, very first sentence:
Ants are eusocial insects belong to family Formicidae and live in symbiotic association with different species of the fungus.
Riaz S, Farooq F, Manzoor F. 2023. Symbiotic association between ants and fungus. Annals of the Entomological Society of America 116(1): 2-9. https://doi.org/10.1093/aesa/saac019
Wait, what? There is something seriously wrong … with the reviewers not pointing this out and the editor not taking action to see this improved. Personally, I am of the school of researchers that thinks that poor English in first instance should be disregarded. I am interested in the science. But reviewers and especially the handling editor are welcomed to improve the English. Sometimes, authors are requested to get help from someone with “full professional proficiency in English.” (Adriana Romero-Olivares wrote a very nice Working Life piece about reviewing with care in Science and a solution for breaking the language barrier was recently published in Trends in Ecology & Evolution.) Anyway, none of that seems to have happened here. (Throughout the paper, the English remains poor; I have only shown this first sentence as an example of the quality of writing.)
Next, Figure 3.
Figure 2 from Báthori et al. (2017) in Journal of Hymenoptera Research. Original caption: Photograph of a slide-mounted Rickia wasmannii thallus (deposition number: G00562301) from Myrmica hellenica host, recorded on the AntWeb (specimen: CASENT0907653). 
Figure 1 from Tartally et al. (2021) in Scientific Reports. Original caption: Illustrations of study organisms. (a) Phengaris alcon female laying eggs on Gentiana pneumonanthe flower buds. (b) Overwintered P. alcon caterpillars in a Myrmica scabrinodis nest. (c) Drawing of an adult Rickia wasmannii thallus. (d) Myrmica scabrinodis worker uninfected with R. wasmannii. (e) Myrmica scabrinodis worker with numerous R. wasmannii thalli on its cuticle; see also Supplementary Video S1. Photo: David R. Nash (a), Ádám Bakos (b, d, e); the drawing (c) is in the public domain, from the original illustrations by Roland Thaxter, courtesy of the Farlow Reference Library of Cryptogamic Botany, Harvard University.
First, the authors talk about Phengaris butterflies but the pictures in Fig. 3a, b are Papilio machaon. Huge fail for what claims to be a review paper on this topic. Next, Fig. 3c is indeed a thallus of Rickia wasmannii. It’s not a drawing but I am happy to let that pass. Worse is that the image was taken from a paper by collaborators of mine: Báthori et al. (2017) in Journal of Hymenoptera Research. This paper is not cited by Riaz et al. Note that the authors edited the original picture, making it weirdly broader and altering its color. One is only left to wonder why. Third, the pictures in Fig. 3d, e were taken from Tartally et al. (2021) in Scientific Reports. Again, not cited by Riaz et al.
What is funny (I don’t know if “funny” is the right word here but I am so glabbergasted at all this that I cannot think of any better word) is the following quote:
Among others, the ectoparasitic Rickia wasmannii fungus and the parasitic caterpillars of myrmecophilous Phengaris butterflies often infect the same Myrmica colonies (Shapiro-Ilan et al. 2012, Haelewaters et al. 2020).
Riaz S, Farooq F, Manzoor F. 2023. Symbiotic association between ants and fungus. Annals of the Entomological Society of America 116(1): 2-9. https://doi.org/10.1093/aesa/saac019
What is really odd here, are the citations. Shapiro-Ilan et al. (2012) do not mention Myrmica ants, Rickia wasmannii or other microfungi in the same order, or Phengaris (Maculinea) butterflies. They also do not talk about the exploitation of hosts by multiple natural enemies simultaneously. Haelewaters et al. (2020) is, uh, my paper about ladybird-associated Laboulbeniales. Again, we did not mention Myrmica ants, Rickia microfungi, or Phengaris butterflies. If anything, Riaz et al. should have cited Tartally et al. (2021), from which they took the pictures for their Fig. 3d, e, as pointed out above. I find it very likely that the authors used this paper to write their quote on Rickia wasmannii and Phengaris caterpillars co-infecting Myrmica colonies, but this cannot be inferred with certainty from the paper.
Let’s continue. Riaz et al. claim to be a review. When I was reading through their text more rigourously, I came across an interesting section. Read with me:
I don’t know why but this was an immediate red flag to me. “We show that …” sounds like this is not an original piece of writing. I copied both sentences and pasted them into Google Scholar: no result. Strange. Next I tried only the second sentence, and I found a direct hit with Bruner-Montero et al. (2021). And it became clear why my first attempt was unsuccessful: the word “aposymbiotic” is in between double quotation marks in Bruner-Montero et al. while Riaz et al. placed it in between single quotation marks. Did they make this change on purpose to make it more difficult for the plagiarism to be detected? One is left wondering.
Update: I just realized that Riaz et al. copy-pasted much more than I indicated above. No less than six sentences from the abstract of Bruner-Montero et al. were copy-pasted word for word in Riaz et al. If that is not a red flag to halt the publication of a manuscript (or in this case: to support the retraction of this paper), I don’t know what is.
ABSTRACT
Many fungus-growing ants engage in a defensive symbiosis with antibiotic-producing ectosymbiotic bacteria in the genus Pseudonocardia, which help protect the ants’ fungal mutualist from a specialized mycoparasite, Escovopsis. Here, using germfree ant rearing and experimental pathogen infection treatments, we evaluate if Acromyrmex ants derive higher immunity to the entomopathogenic fungus Metarhizium anisopliae from their Pseudonocardia symbionts. We further examine the ecological dynamics and defensive capacities of Pseudonocardia against M. anisopliae across seven different Acromyrmex species by controlling Pseudonocardia acquisition using ant-nonnative Pseudonocardia switches, in vitro challenges, and in situ mass spectrometry imaging (MSI). We show that Pseudonocardia protects the ants against M. anisopliae across different Acromyrmex species and appears to afford higher protection than metapleural gland (MG) secretions. Although Acromyrmex echinatior ants with nonnative Pseudonocardia symbionts receive protection from M. anisopliae regardless of the strain acquired compared with Pseudonocardia-free conditions, we find significant variation in the degree of protection conferred by different Pseudonocardia strains. Additionally, when ants were reared in Pseudonocardia-free conditions, some species exhibit more susceptibility to M. anisopliae than others, indicating that some ant species depend more on defensive symbionts than others. In vitro challenge experiments indicate that Pseudonocardia reduces Metarhizium conidiospore germination area. Our chemometric analysis using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) reveals that Pseudonocardia-carrying ants produce more chemical signals than Pseudonocardia-free treatments, indicating that Pseudonocardia produces bioactive metabolites on the Acromyrmex cuticle. Our results indicate that Pseudonocardia can serve as a dual-purpose defensive symbiont, conferring increased immunity for both the obligate fungal mutualist and the ants themselves.
IMPORTANCE In some plants and animals, beneficial microbes mediate host immune response against pathogens, including by serving as defensive symbionts that produce antimicrobial compounds. Defensive symbionts are known in several insects, including some leaf-cutter ants where antifungal-producing Actinobacteria help protect the fungal mutualist of the ants from specialized mycoparasites. In many defensive symbioses, the extent and specificity of defensive benefits received by the host are poorly understood. Here, using “aposymbiotic” rearing, symbiont switching experiments, and imaging mass spectrometry, we explore the ecological and chemical dynamics of the model defensive symbiosis between Acromyrmex ants and their defensive symbiotic bacterium Pseudonocardia. We show that the defensive symbiont not only protects the fungal crop of Acromyrmex but also provides protection from fungal pathogens that infect the ant workers themselves. Furthermore, we reveal that the increased immunity to pathogen infection differs among strains of defensive symbionts and that the degree of reliance on a defensive symbiont for protection varies across congeneric ant species. Taken together, our results suggest that Acromyrmex-associated Pseudonocardia have evolved broad antimicrobial defenses that promote strong immunity to diverse fungal pathogens within the ancient fungus-growing ant-microbe symbiosis.
Bruner-Montero G, Wood M, Horn HA, Gemperline E, Li L, Currie CR. 2021. Symbiont-mediated protection of Acromyrmex leaf-cutter ants from the entomopathogenic fungus Metarhizium anisopliae. Mbio 12(6): e01885-21. https://doi.org/10.1128/mBio.01885-21
Sadly, there is more. The authors made a graphical abstract but haven’t done themselves (or the readers, for that matter) any favors. I agree that I do not quite understand why they went through the ordeal of designing one because (1) it does not fullfill the function of a graphical abstract, at all, and (2) it contains parts that were shameslessly copy-pasted from other, uncited papers. It was actually Mariane Ronque (@MarianeRonque) who made me aware of her 2019 paper in Insectes Sociaux of which Fig. 1 was used in part by Riaz et al., without being cited.
As I was writing this post, I could no longer believe that the authors made the illustrations in the graphical abstract about amylase (the enzyme responsible for the biodegradation of organic material into glucose) themselves. I decided to see if I could find the image elsewhere. I googled for images using “amylase” and “glucose” as search terms: no luck. Next I tried “amylase”, “glucose”, and “ants”: again, no luck. Finally, I searched for “amylase”, “glucose”, and “biodegradation”, and there it was: a figure from Araújo et al. (2022) in Insects. I knew it! Note how the authors edited the figure: they made it narrower overall not respecting the original aspect ratio (the egg-shaped enzyme depictions were round in the original figure) and they made an effort of adding “Amylase” twice but neglected to remove the original “Amylase” annotations in the background. I’d call this very lazy editing.
Graphical abstract from Riaz et al. (2023). 
Fig. 1 from Ronque et al. (2019) in Insectes Sociaux. Original caption: External and internal appearance of nests of the five species of fungus-farming ants studied in an area of Atlantic rainforest at the Parque Estadual Serra do Mar, SE Brazil. Yellow arrows indicate the nest entrance. Note the fungus garden suspended from the ceiling in the M. smithii chamber (yellow arrow). 
Fig. 1 from Araújo et al. (2022) in Insects. Original caption: Symbiotic relationship between leaf-cutting ants and Leucoagaricus gongylophorus symbiotic fungus: (a) Gongylidia produced in the fungus garden are the main source of food for the ant larvae; (b) fungus enzymes such as amylase are responsible for the biodegradation of organic material into glucose, the main food source for the worker ants; (c) leaf-cutting ants produce antibiotic compounds that protect the fungus garden against harmful agents; (d) pathogenic microorganisms such Escovopsis fungal genera.
I do not know if there is more. I haven’t actually read the entire paper, given I already amassed quite some evidence that should be enough for the editorial team at Annals of the Entomological Society of America to go sit around the table and discuss what has gone wrong with this paper by Riaz et al. Based on what we know, its publication in the Annals is a disgrace. It is my hope that the paper is retracted, that the authors are prohibited from publishing in ESA journals, and perhaps that a statement is put out by the Editor-in-chief or entire editorial board about the need for authors to adhere to ethical guidelines (and what those guidelines are).
References
Araújo S, Seibert J, Ruani A, Alcántara-de la Cruz R, Cruz A, Pereira A, Zandonai D, Forim M, Silva MF, Bueno O, Fernandes J. 2022. The symbiotic fungus Leucoagaricus gongylophorus (Möller) Singer (Agaricales, Agaricaceae) as a target organism to control leaf-cutting ants. Insects 13(4): 359. https://doi.org/10.3390/insects13040359
Báthori F, Pfliegler WP, Zimmerman CU, Tartally A. 2017. Online image databases as multi-purpose resources: discovery of a new host ant of Rickia wasmannii Cavara (Ascomycota, Laboulbeniales) by screening AntWeb.org. Journal of Hymenoptera Research 61: 85-94. https://doi.org/10.3897/jhr.61.20255
Bruner-Montero G, Wood M, Horn HA, Gemperline E, Li L, Currie CR. 2021. Symbiont-mediated protection of Acromyrmex leaf-cutter ants from the entomopathogenic fungus Metarhizium anisopliae. Mbio 12(6): e01885-21. https://doi.org/10.1128/mBio.01885-21
Haelewaters D, Hiller T, Kemp EA, van Wielink PS, Shapiro-Ilan DI, Aime MC, Nedvěd O, Pfister DH, Cottrell TE. 2020. Mortality of native and invasive ladybirds co-infected by ectoparasitic and entomopathogenic fungi. PeerJ 8: e10110. https://doi.org/10.7717/peerj.10110
Khelifa R, Amano T, Nuñez MA. 2022. A solution for breaking the language barrier. Trends in Ecology & Evolution 37(2): 109-112. https://doi.org/10.1016/j.tree.2021.11.003
Romero-Olivares AL. 2019. Review with care. Science 366(6461): 146-146. https://doi.org/10.1126/science.366.6461.146
Ronque MU, Feitosa RM, Oliveira PS. 2019. Natural history and ecology of fungus-farming ants: a field study in Atlantic rainforest. Insectes Sociaux 66(3): 375-387. https://doi.org/10.1007/s00040-019-00695-y
Shapiro-Ilan DI, Bruck DJ, Lacey LA. 2012. Principles of epizootiology and microbial control. pp. 29-72. In: Vega FE, Kaya HK (eds.) Insect Pathology. 2nd edition. Elsevier, San Diego, California. https://doi.org/10.1016/B978-0-12-384984-7.00003-8
Tartally A, Szabó N, Somogyi AÁ, Báthori F, Haelewaters D, Mucsi A, Fürjes-Mikó Á, Nash D. 2021. Ectoparasitic fungi of Myrmica ants alter the success of parasitic butterflies. Scientific Reports 11: 24031. https://doi.org/10.1038/s41598-021-02800-3
Thanks Danny for pointing out the different issues related to this paper! It is sad to see such scientific misconduct !
I hope this will be considered as a wake-up call for the Journals to pay more attention on the revision process!