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Analysis of the Bacterial Microbiota in Wild Populations of Prickly Pear Cochineal, Dactylopius opuntiae in Morocco
by
, , , , , , , and
Imane Remmal
1,2,3
,
Youssef El Yamlahi
1,2,3
,
Naima Bel Mokhtar
3,
Ioannis Galiatsatos
3
,
Dimitrios Loukovitis
4,
Eva Dionyssopoulou
3,
Mohammed Reda Britel
1,
Panagiota Stathopoulou
3,5
,
Amal Maurady
1,2 and
George Tsiamis
3,5,*
1
Laboratory of Innovative Technologies, National School of Applied Sciences of Tangier, Abdelmalek Essaadi University, Tetouan 93000, Morocco
2
Faculty of Sciences and Technology of Tangier, Abdelmalek Essaadi University, Tetouan 93000, Morocco
3
Laboratory of Systems Microbiology and Applied Genomics, Department of Sustainable Agriculture, University of Patras, 2 Seferi St, 30100 Agrinio, Greece
4
Department of Fisheries and Aquaculture, School of Agricultural Sciences, University of Patras, New Buildings, 30200 Mesolongi, Greece
5
BioDetect P.C., Stadiou St., Platani, 26504 Rio, Greece
*
Author to whom correspondence should be addressed.
Insects 2025, 16(12), 1184; https://doi.org/10.3390/insects16121184
Submission received: 7 October 2025
/
Revised: 11 November 2025
/
Accepted: 18 November 2025
/
Published: 21 November 2025
Simple Summary
The cochineal Dactylopius opuntiae (Cockerell) (Hemiptera: Dactylopiidae) is a significant pest affecting prickly pears in Morocco and globally. This study focused on the bacterial community associated with Dactylopius opuntiae. Through targeted PCR screening and high-throughput sequencing of the full-length 16S rRNA gene, we successfully identified low-abundance endosymbionts, specifically Wolbachia and Spiroplasma, and identified Candidatus Dactylopiibacterium as the predominant bacterium. Following the exclusion of this taxon from the dataset during bioinformatic analysis, a more abundant and varied microbiota was observed, highlighting specific patterns related to developmental stage and geographic location. These findings deepen our understanding of the D. opuntiae microbiome and provide valuable insights into possible biological control strategies.
Abstract
Dactylopius opuntiae (Cockerell) (Hemiptera: Dactylopiidae), the wild cochineal scale, is a major pest of prickly pear crops worldwide. This study characterized the bacterial community structure of D. opuntiae from four Moroccan regions using targeted PCR and full-length 16S rRNA MinION sequencing. We report the first detection of Wolbachia (16.6% prevalence) in D. opuntiae, with infection rates varying geographically from 0% (Rabat) to 53.3% (Ouazzane). Spiroplasma was detected at a lower prevalence (3.3%) and exclusively in males. Phylogenetic analysis showed that Wolbachia sequences likely belong to supergroup B, based on their similarity to reference sequences, while Spiroplasma sequences were placed within the poulsonii–citri complex. MinION sequencing revealed Candidatus Dactylopiibacterium as the dominant taxon (97.7%), consistent with its role as an obligate symbiont. After removing this dominant species, we uncovered a diverse bacterial community, including Flavisolibacter, Pseudomonas, Phyllobacterium, Acinetobacter, and Brevibacillus. Beta diversity analysis showed significant geographic variation (PERMANOVA p < 0.008), with distinct communities across regions. Females harbored a more specialized microbiome dominated by Flavisolibacter (except in Agadir), whereas males and nymphs showed Pseudomonas dominance. Core microbiome analysis revealed no universal genera across all groups, with females displaying a more restricted core than males and nymphs. The detection of reproductive symbionts, combined with geographic and sex-specific microbiome patterns, provides valuable insights into the potential roles of these bacteria in host adaptation and their implications for microbiome-based pest management strategies. The complementary use of targeted and untargeted sequencing methods is essential for comprehensive microbiome characterization in this economically important pest.
