A thorough assessment of the phylogenetic diversity and community structure of halophilic archaea from three halite-crystal salts, processed from two separated saline systems of Southern Tunisia has been performed using culture dependent and independent methods targeting different regions of 16S rRNA gene sequences including DGGE, 16S rRNA clone libraries and Illumina Miseq sequencing. Two samples, CDR (red halite-crystal salts) and CDW (white halite-crystal salts), were collected from Chott-Eljerid and one sample CDZ (white halite-crystal salts) from Chott Douz. Fourteen isolates were identified as Halorubrum, Haloferax, Haloarcula, and Halogeometricum genera members. Culture-independent approach revealed a high diversity of archaeal members present in all samples, represented by the Euryarchaeal phylum and the dominance of the Halobacteria class. Nanohaloarchaea were also identified only in white halite samples based on metagenomic analysis. In fact, a total of 61 genera were identified with members of the Halorhabdus, Halonotius, Halorubrum, Haloarcula, and unclassified. Halobacteriaceae were shared among all samples. Unexpected diversity profiles between samples was observed where the red halite crust sample was considered as the most diverse one. The highest diversity was observed with Miseq approach, nevertheless, some genera were detected only with 16S rRNA clone libraries and cultured approaches.

The aim of this study was to characterize the prevalence of fecal carriage of extended-spectrum beta-lactamases and carbapenemase-producing Gram-negative bacteria among healthy humans in Tunisia. Fifty-one rectal swabs of healthy volunteers were plated on MacConkey agar plates supplemented with cefotaxime or imipenem. The occurrences of resistance genes, integrons, and phylogroup typing were investigated using PCR and sequencing. The genetic relatedness of isolates was determined by pulsed-field-gel-electrophoresis (PFGE) and multilocus-sequence-typing (MLST). Whole-genome-sequencing (WGS) was performed for the carbapenem-resistant isolate. Sixteen ESBL-producing Escherichia coli isolates and one carbapenem-resistant Enterobacter bugandensis were detected out of the fifty-one fecal samples. The ESBL-producing E. coli strains contained genes encoding CTX-M-15 (n = 9), CTX-M-1 (n = 3), CTX-M-27 (n = 3), and CTX-M-55 (n = 1). Three CTX-M-1-producers were of lineages ST131, ST7366, and ST1158; two CTX-M-15-producers belonged to lineage ST925 and ST5100; one CTX-M-27-producer belonged to ST2887, and one CTX-M-15-producer belonged to ST744. Six isolates contained class 1 integrons with the following four gene cassette arrangements: dfrA5 (two isolates), dfrA12-orf-aadA2 (two isolates), dfrA17-aadA5 (one isolate), and aadA1 (one isolate). E. bugandensis belonged to ST1095, produced IMI-2 carbapenemase, and contained qnrE1 and fosA genes. A genome-sequence analysis of the E. bugandensis strain revealed new mutations in the bla_ACT and qnr genes. Our results reveal an alarming rate of ESBL-E. coli in healthy humans in Tunisia and the first description of IMI-2 in E. bugandensis.

Here, we report the osmoadaptation strategies adopted by the halotolerant species Halomonas titanicae BH1(T) inferred from genome sequence analysis. BH strain was isolated in 2010 from a rusticated sample collected in 1991 from the wreck of the Titanic, genome deposited in the database under the accession number (CP059082.1). It showed a high salt tolerance ranging from 0.5 to 25% NaCl (w/v) (optimal growth at 10% NaCl) with no growth in the absence of NaCl. The phylogenomic analysis showed that the BH1 strain is more closely related to the Halomonas sedementi QX-2, a strain isolated from deep-sea sediments. The RAST (Rapid Annotation using Subsystem Technology) annotation revealed divergent mechanisms involved in the primary and secondary response to osmotic stress citing protein implicated in potassium transport, periplasmic glucan synthesis, choline and betaine upake system, biosynthesis of glycine-betaine, ectoine, and proline. These findings provide an overview of the osmoadaptive mechanisms of H. titanicae BH1, and could offer helpful information to future biotechnological applications like osmolyte synthesis and related applications.

Open access peer-reviewed chapter

Bemisia tabaci (Aleyrodidae family) is one of the most damaging pests of numerous crops worldwide. Insecticides, namely pyrethroids and organophosphates, have long been the primary control tools against this pest, resulting in several resistance cases. In Tunisia, the two most damaging biotypes of B. tabaci, MEAM1-B and MED-Q, are sympatric, and more concerns about developing resistance keep rising due to the extensive use of insecticides. Here, we aimed to elucidate the molecular mechanism of resistance to pyrethroids and organophosphorus insecticides in two Tunisian populations of B. tabaci, collected respectively on Capsicum annuum and Lantana camara, and then determine the bacterial community associated with insecticide resistance and susceptible biotypes based on 16S rRNA Illumina sequencing. The results showed that the population collected on Capsicum annuum belonged to the MEAM1-B biotype with an insecticide resistance profile. In contrast, the population collected on the Lantana camara belonged to the MED-Q biotype with a sensitive profile. The bacterial communities of the two biotypes were predominantly structured by the Proteobacteria phylum and three genera, including Candidatus Portiera, the secondary facultative symbiont, and Hamiltonella, which were unevenly distributed between the two biotopes. Our results provide the first evidence for insecticide resistance alleles in Tunisian MEAM1-B populations and suggest an association between bacterial community composition within susceptible biotypes and insecticide resistance.