Background/Objectives: Clostridioides difficile is a “One Health” pathogen and a cause of antibiotics-associated diarrhea and pseudomembranous colitis. Mobile genetic elements (MGEs) have been documented in the genomes of clinical C. difficile strains; however, the presence of MGEs in environmental strains remains poorly characterized. Thus, the present study was conducted with the objective of identifying the prevalence of MGEs, including mobilizable transposons (MTns), conjugative transposons (CTns), plasmids, and insertion sequences, in whole genome sequences (WGSs) of environmental C. difficile isolates. Methods: The analysis of MGEs was conducted using 166 WGSs obtained from C. difficile strains isolated from various environmental sources contaminated with feces. The MGEs were identified using bioinformatic tools. Results: A total of 48.2% (80/166) of the studied genomes were identified to harbor nine transposons, including Tn916, Tn6194-like, Tn5397, Tn6215, Tn4001, Tn6073, Tn6110, Tn6107, or Tn5801-like. The majority of MTns and CTns could be found within C. difficile sequence types ST11, ST3, and ST35. The results demonstrated close genetic relatedness among the studied genomes, the array of antimicrobial resistance (AMR) genes, such as tetM, ermB, and aac(6′)-aph(2″), and the presence of CTns. Furthermore, the analysis revealed that 24.7% (41/166) of the genome sequences of isolates were associated with various predominant plasmid groups, including pCD6, pCD-ECE4-6, pCD-WTSI1-4, pCDBI1, and pCd1_3, which belonged to 16 different sequence types. Furthermore, several plasmids were identified as harboring the prophage phiCDHM19. Conclusions: The results of the current study suggest that the identified plasmids are abundant and may encode functions that are relevant to C. difficile physiology. The genomes of C. difficile strains examined contain closely related CTns, suggesting that horizontal transfer of AMR is important in this species or other bacterial species. Further research is required to ascertain the effect of these genetic elements and their transferability on the biology of C. difficile.

SUMMARY This research documents the initial finding of Chloris pycnothrix Trin., frequently referred to as spiderweb chloris, in Libya, particularly from the Ariggiba region close to Sabha city. This grass, originating from tropical and subtropical regions, is versatile and may become invasive. It was recognized via morphological examination during field surveys in 2023–2024, with samples stored in a herbarium. The species flourishes in alluvial plains as well as in disturbed habitats like road shoulders. It can be mistaken for Chloris virgata, although they differ in some important morphological aspects. It was previously believed that only one species of Chloris existed in Libya. This study provides a revision and an identification key for them in the context of Libyan flora. The tolerant nature of C. pycnothrix to drought, combined with its ability to colonize poorly maintained land, makes its potential introduction—most likely through anthropogenic means—alarming ecologically and emphasizes the need for management strategies to contain its spread and monitor its impact. 

ABSTRACT

Argemone ochroleuca Sweet, Brit. Fl. Gard. Ser.1 3(2): pl. 242 (1828) of the family Papaveraceae is reported here for the first time from Libya. This species is native to Mexico. Flowering specimens of A. ochroleuca were collected from the region Ariggiba (26°35’11.8”N 13°29’24.8”E), 110 km southwest of the city of Sabha, which lies about 700 km south of Tripoli. It is presented with updated nomenclature, taxonomic description, geographical distribution, key to genera, location in Libya, and colour photographs taken in the field. The voucher specimens are deposited in the Herbarium of Botany Department (ULT), University of Tripoli, Libya.

A new record for Leptadenia arborea (Forssk.) Schweinf. is reported for the first time in the flora of Libya. This species was collected from Ariggiba region (110 km southwest Sabha city). A full description and habitat information on the plant are provided. A brief discussion about the most important traits of this species is presented.

Abstract

For the first time in Libyan flora, a new record for Boerhavia repens L is recorded. This widespread wild weed was collected from the Ariggiba region (110 km west-south Sabha city). A full description, habitat information, distribution map, and distribution data are provided to facilitate further identification and future detection. A brief discussion about the most important threats this species poses is presented.

Clostridioides difficile is widely recognised as one of the most common causes of healthcare-associated C. difficile infections due to the ability of spores to survive for prolonged periods in the hospital environment. This study aimed to evaluate the efficacy of UV-C 254 nm light in the inactivation of the spores of different C. difficile ribotypes on brain heart infusion (BHI) agar plates or in phosphate-buffered saline (PBS) with varying spore densities. Furthermore, the effectiveness of a sporicidal surface disinfectant against C. difficile spores was determined on different surfaces. Spore suspensions of different C. difficile strains in the range of 10⁵–10⁷ colony-forming units (CFUs) mL⁻¹ were inoculated on BHI agar plates or in PBS and exposed to UV-C light for up to 30 min. Additionally, a spore suspension of 10³–10⁵ CFUs was spread over a 1 cm² test area on different surfaces, and sporicidal surface wipes were used according to the manufacturer’s instructions. The findings demonstrated that spores of C. difficile ribotypes exhibited a complete reduction in log₁₀ CFU on BHI agar plates and PBS following 20 min of exposure to a UV-C dose of 2208 mJ cm⁻². The surface wipes with sporicidal properties demonstrated efficacy in reducing the number of C. difficile spores on the Formica, stainless steel, and plastic surfaces by 2.03–3.53 log₁₀. The present study demonstrates that moist surfaces or liquids can enhance the efficacy of UV-C treatment in reducing C. difficile spores. This approach may be applicable to the surfaces of healthcare facilities and to water disinfection systems.

Clostridioides difficile is an important human pathogen causing antibiotic-associated diarrhoea worldwide. Besides using antibiotics for treatment, the interest in bacteriophages as an alternative therapeutic option has increased. Prophage abundance and genetic diversity are well-documented in clinical strains, but the carriage of prophages in environmental strains of C. difficile has not yet been explored. Thus, the prevalence and genetic diversity of integrated prophages in the genomes of 166 environmental C. difficile isolates were identified. In addition, the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems were determined in the genomes of prophage regions. Predicted prophages and CRISPR-Cas systems were identified by using the PHASTER web server and CRISPRCasFinder, respectively. Phylogenetic relationships among predicated prophages were also constructed based on phage-related genes, terminase large (TerL) subunits and LysM. Among 372 intact prophages, the predominant prophages were phiCDHM1, phiCDHM19, phiMMP01, phiCD506, phiCD27, phiCD211, phiMMP03, and phiC2, followed by phiMMP02, phiCDKM9, phiCD6356, phiCDKM15, and phiCD505. Two newly discovered siphoviruses, phiSM101- and phivB_CpeS-CP51-like Clostridium phages, were identified in two C. difficile genomes. Most prophages were found in sequence types (STs) ST11, ST3, ST8, ST109, and ST2, followed by ST6, ST17, ST4, ST5, ST44, and ST58. An obvious correlation was found between prophage types and STs/ribotypes. Most predicated prophages carry CRISPR arrays. Some prophages carry several gene products, such as accessory gene regulator (Agr), putative spore protease, and abortive infection (Abi) systems. This study shows that prophage carriage, along with genetic diversity and their CRISPR arrays, may play a role in the biology, lifestyle, and fitness of their host strains.

Clostridioides difficile is the most important pathogen causing antimicrobial-associated diarrhea and has recently been recognized as a cause of community-associated C. difficile infection (CA-CDI). This study aimed to characterize virulence factors, antimicrobial resistance (AMR), ribotype (RT) distribution and genetic relationship of C. difficile isolates from diverse fecally contaminated environmental sources. C. difficile isolates were recovered from different environmental samples in Northern Germany. Antimicrobial susceptibility testing was determined by E-test or disk diffusion method. Toxin genes (tcdA and tcdB), genes coding for binary toxins (cdtAB) and ribotyping were determined by PCR. Furthermore, 166 isolates were subjected to whole genome sequencing (WGS) for core genome multi-locus sequence typing (cgMLST) and extraction of AMR and virulence-encoding genes. Eighty-nine percent (148/166) of isolates were toxigenic, and 51% (76/148) were positive for cdtAB. Eighteen isolates (11%) were non-toxigenic. Thirty distinct RTs were identified. The most common RTs were RT127, RT126, RT001, RT078, and RT014. MLST identified 32 different sequence types (ST). The dominant STs were ST11, followed by ST2, ST3, and ST109. All isolates were susceptible to vancomycin and metronidazole and displayed a variable rate of resistance to moxifloxacin (14%), clarithromycin (26%) and rifampicin (2%). AMR genes, such as gyrA/B, bla_CDD-1/2, aph(3′)-llla-sat-4-ant(6)-la cassette, ermB, tet(M), tet(40), and tetA/B(P), conferring resistance toward fluoroquinolone, beta-lactam, aminoglycoside, macrolide and tetracycline antimicrobials, were found in 166, 137, 29, 32, 21, 72, 17, and 9 isolates, respectively. Eleven “hypervirulent” RT078 strains were detected, and several isolates belonged to RTs (i.e., RT127, RT126, RT023, RT017, RT001, RT014, RT020, and RT106) associated with CA-CDI, indicating possible transmission between humans and environmental sources pointing out to a zoonotic potential.

The goal of this study was to investigate the vegetation composition and structure of the Al-Orban area in Gharyan district, Libya over two continues growing seasons from the first of January 2020 to the first of Jaunary 2022. During this period, a total of 309 plant species were identified and collected in the field. These species belonged to 43 families, with 39 families of dicotyledons and 4 families of monocotyledons. The most dominant family was Asteraceae with 60 species, followed by Fabaceae with 32 species, Poaceae with 33 species, and Brassicaceae with 35 species. The most dominant genera were Plantago and Euphorbia, each represented by 6 species, followed by Erodium and Chenopodiun with 5 species each, and Astragalus, Centaurea, Rumex, Concolvulus, and Stipa each represented by 4 species. Life-form spectrum analysis indicated that Therophytes were the most predominant with 189 species, followed by Hemicryptophytes with 47 species, and geophytes with 40 species. Furthermore, chorotype spectrum analysis showed the dominance of Mediterranean species with 128 species, followed by Mediterranean/Iranian-Turanian with 66 species. These findings provide valuable insight into the plant diversity and distribution in the Al-Orban area of Gharyan district, Libya.