Review of Proxima DeLong & Freytag (Hemiptera: Cicadellidae: Gyponini) with description of two new species from Brazil and identification key to species

The previously monotypic genus Proxima DeLong & Freytag, 1975 is reviewed and new diagnostic characters are added. The type-species Proxima ocellata DeLong & Freytag, 1975 described from the State of Espírito Santo, Brazil, is redescribed and newly recorded from the State of Minas Gerais. Also, two new species are described and illustrated: Proxima nigromaculata sp. nov. from the states of Espírito Santo, Maranhão, Minas Gerais, Rio de Janeiro and Rio Grande do Sul and Proxima meloi sp. nov. from the states of Bahia and Minas Gerais. An identification key to the species of the genus is provided.

Asiacantha gen. nov., a new genus of the orb-weaver spiders from China (Araneae, Araneidae, Gasteracanthinae)

Asiacantha gen. nov., a new genus of the subfamily Gasteracanthinae is established with the type species A. pengi sp. nov. (♀♂) from Guangxi and two other new species: A. jianfeng sp. nov. (♂) from Hainan and A. yinae sp. nov. (♀♂) from Yunnan and Guizhou. Diagnostic drawn and detailed description, as well as a distributional map, are provided.

Catalog of the horse-flies (Diptera: Tabanidae) of Chile

A catalog of the Tabanidae from Chile is provided. All valid names and synonyms for the 116 species and 25 genera known for the country including information about name, author, year of publication, page number, type species, type locality and references are listed. The species Dasybasis albohirta (Walker) is cited for the first time for Chile. The type locality of Dasybasis nigrifrons (Philippi) is corrected to Chile (Todos Los Santos Lake). The dates of the description of Tabanus nigrifrons Philippi (now Dasybasis), and Pangonia australis Philippi (now Pseudoscione) are corrected from their original description by Philippi; the original description dates for Tabanus andicola (now Dasybasis), and Tabanus magellanicus (now synonym junior of Dasybasis trita (Walker)) are corrected from 1865 to 1862. Three species previously recorded from Chile are deemed to have been misidentified or misrecorded from Chile: Esenbeckia (E.) cisandeana Wilkerson & Fairchild and Esenbeckia (E.) enderleini Kröber, and Esenbeckia (E.) testaceiventris (Macquart, 1848). The genus Dasybasis Macquart is the richest genus in Chile, with 33 species. Seven genera (Mycteromyia Philippi, Promycteromyia Coscarón & Philip, Archeomyotes Philip & Coscarón, Austromyans Philip & Coscarón, Chaetopalpus Philippi, Pseudomelpia Enderlein, and Sixtomyia Krolow, Henriques & González), three subgenera (Esenbeckia (Astomyia) Burger, E. (Palassomyia) Fairchild, and Protodasyapha (Protodasyapha) Enderlein), and 64 species are known only from Chile. There are also one subgenus (Protodasyapha (Curumyia) Coscarón), and 36 specie with distribution shared only between Chile and Argentina.

Kerevata Belokobylskij (Hymenoptera: Braconidae: Rogadinae) is no longer a Papua New Guinean endemic with descriptions of three new species from the Indomalayan Region

The rogadine genus Kerevata Belokobylskij is newly reported from the Indomalayan region. We describe and illustrate three new species, two from India (K. kethai sp. nov. and K. orientalia sp. nov.) and one from Vietnam (K. longi sp. nov.) and provide an illustrated key to the extant species of the genus along with the photographic illustration of the type species of K. pacifica Belokobylskij. Range extension and morphological characters of Kerevata are discussed.

Delimitation of the segmented trapdoor spider genus Luthela gen. nov., with comments on the genus Sinothela from northern China (Araneae, Mesothelae, Liphistiidae)

A new genus of the primitively segmented trapdoor spiders, which is endemic to the north of China, is described, Luthela gen. nov., and the status of Sinothela Haupt, 2003 and Sinothela sinensis (Bishop & Crosby, 1932) is discussed and both are treated as nomina dubia. The new genus Luthela gen. nov. is erected based on morphology and molecular data of the type species Luthela yiyuan sp. nov. A taxonomic revision of the new genus is given. Three Sinothela species are transferred to the new genus, L. luotianensis comb. nov. and L. schensiensis comb. nov. are redescribed using our newly collected specimens, include L. heyangensis comb. nov. as a junior synonym of L. schensiensis comb. nov., and describe six new species based on both male and female morphological characters: L. badong sp. nov., L. dengfeng sp. nov., L. handan sp. nov., L. taian sp. nov., L. yiyuan sp. nov., and L. yuncheng sp. nov.

Genomic evolution of the globally disseminated multidrug-resistant Klebsiella pneumoniae clonal group 147

The rapid emergence of multidrug-resistant Klebsiella pneumoniae is being driven largely by the spread of specific clonal groups (CGs). Of these, CG147 includes 7-gene multilocus sequence typing (MLST) sequence types (STs) ST147, ST273 and ST392. CG147 has caused nosocomial outbreaks across the world, but its global population dynamics remain unknown. Here, we report a pandrug-resistant ST147 clinical isolate from India (strain DJ) and define the evolution and global emergence of CG147. Antimicrobial-susceptibility testing following European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines and genome sequencing (Illumina and Oxford Nanopore Technologies, Unicycler assembly) were performed on strain DJ. Additionally, we collated 217 publicly available CG147 genomes [National Center for Biotechnology Information (NCBI), May 2019]. CG147 evolution was inferred within a temporal phylogenetic framework (beast) based on a recombination-free sequence alignment (Roary/Gubbins). Comparative genomic analyses focused on resistance and virulence genes and other genetic elements (BIGSdb, Kleborate, PlasmidFinder, phaster, ICEfinder and CRISPRCasFinder). Strain DJ had a pandrug-resistance phenotype. Its genome comprised the chromosome, seven plasmids and one linear phage-plasmid. Four carbapenemase genes were detected: bla NDM-5 and two copies of bla OXA-181 in the chromosome, and a second copy of bla NDM-5 on an 84 kb IncFII plasmid. CG147 genomes carried a mean of 13 acquired resistance genes or mutations; 63 % carried a carbapenemase gene and 83 % harboured bla CTX-M. All CG147 genomes presented GyrA and ParC mutations and a common subtype I-E CRISPR-Cas system. ST392 and ST273 emerged in 2005 and 1995, respectively. ST147, the most represented phylogenetic branch, was itself divided into two main clades with distinct capsular loci: KL64 (74 %, DJ included, emerged in 1994 and disseminated worldwide, with carbapenemases varying among world regions) and KL10 (20 %, emerged in 2002, predominantly found in Asian countries, associated with carbapenemases NDM and OXA-48-like). Furthermore, subclades within ST147-KL64 differed at the yersiniabactin locus, OmpK35/K36 mutations, plasmid replicons and prophages. The absence of IncF plasmids in some subclades was associated with a possible activity of a CRISPR-Cas system. K. pneumoniae CG147 comprises pandrug-resistant or extensively resistant isolates, and carries multiple and diverse resistance genes and mobile genetic elements, including chromosomal bla NDM-5. Its emergence is being driven by the spread of several phylogenetic clades marked by their own genomic features and specific temporo–spatial dynamics. These findings highlight the need for precision surveillance strategies to limit the spread of particularly concerning CG147 subsets.

An evaluation of risk factors for Staphylococcus aureus colonization in a pre-surgical population

Staphylococcus aureus (SA) colonization has significant implications in healthcare-associated infections. Here we describe a prospective study conducted in pre-surgical outpatients, done with the aim of identifying demographic and clinical risk factors for SA colonization. We found younger age to be a potential predictor of SA colonization.

Environment in the lung of cystic fibrosis patients stimulates the expression of biofilm phenotype in Mycobacterium abscessus

Introduction. Pulmonary infections caused by organisms of the Mycobacterium abscessus complex are increasingly prevalent in populations at risk, such as patients with cystic fibrosis, bronchiectasis and emphysema. Hypothesis. M. abscessus infection of the lung is not observed in immunocompetent individuals, which raises the possibility that the compromised lung environment is a suitable niche for the pathogen to thrive in due to the overproduction of mucus and high amounts of host cell lysis. Aim. Evaluate the ability of M. abscessus to form biofilm and grow utilizing in vitro conditions as seen in immunocompromised lungs of patients. Methodology. We compared biofilm formation and protein composition in the presence and absence of synthetic cystic fibrosis medium (SCFM) and evaluated the bacterial growth when exposed to human DNA. Results. M. abscessus is capable of forming biofilm in SCFM. By eliminating single components found in the medium, it became clear that magnesium works as a signal for the biofilm formation, and chelation of the divalent cations resulted in the suppression of biofilm formation. Investigation of the specific proteins expressed in the presence of SCFM and in the presence of SCFM lacking magnesium revealed many different proteins between the conditions. M. abscessus also exhibited growth in SCFM and in the presence of host cell DNA, although the mechanism of DNA utilization remains unclear. Conclusions. In vitro conditions mimicking the airways of patients with cystic fibrosis appear to facilitate M. abscessus establishment of infection, and elimination of magnesium from the environment may affect the ability of the pathogen to establish infection.

Four New Members of The Family Cytophagaceae: Cryseosolum Histidinii gen. nov., sp. nov., Cryseosolum Indiensis gen. nov., sp. nov., Reichenbachia Cretensis, gen. nov., sp. nov., and Reichenbachia Soli, gen. nov., sp. nov. Isolated From Diverse Habitat

Four strains isolated, PWU4T, PWU20T, PWU5T and PWU37T were from both of soil in Germany, India and a faces sheep collected in Crete Island, respectively. Cells were Gram-negative, strictly aerobic, rod shaped, grew optimally between 28oC and 34oC, pH between 7.0 and 8.0 without the addition of NaCl. Catalase and oxidase-negative and grew on most mono- and disaccharides, a few polysaccharides and organic acid. The predominant menaquinone was MK-7. Major fatty acid was c16:1 ω7c (PWU4T and PWU20T) and c16:1 ω5c (PWU5T and PWU37T). The DNA G+C content of them were 50.2 mol %; 51.6 mol %; 39.8 mol % and 53.8 mol %, respectively. The 16S rRNA gene sequence analysis revealed that the closest relatives of them are less than 93.8% compared to Ohtaekwangia koreensis 3B-2T and Ohtaekwangia kribbensis 10AOT. It classified in two groups, where PWU4T, PWU20T shared 93.0% and PWU5T, PWU37T shared 97.5% sequence similarity. However, in both groups represent different species on the low average nucleotide identity (ANI) of their genomes, 69.7% and 83.8%, respectively. We proposed that the four strains represent four novel species of two new genera in the family Cytophagaceae. The type species of the novel genus Cryseosolum are Cryseosolum histdinii gen. nov., sp. nov. strain PWU4T (=DSM 111594T=NCCB 100798T), Cryseosolum indiensis sp. nov. strain PWU20T (=DSM 111597T=NCCB 100800T). The type species of the novel genus Reichenbachia are Reichenbachia cretensis gen. nov., sp. nov. strain PWU5T (=DSM 111596T=NCCB 100799T), Reichenbachia soli sp. nov. strain PWU37T (=DSM 111595T=NCCB 100801T).

Hackflex: low-cost, high-throughput, Illumina Nextera Flex library construction

We developed a low-cost method for the production of Illumina-compatible sequencing libraries that allows up to 14 times more libraries for high-throughput Illumina sequencing to be generated for the same cost. We call this new method Hackflex. The quality of library preparation was tested by constructing libraries from Escherichia coli MG1655 genomic DNA using either Hackflex, standard Nextera Flex (recently renamed as Illumina DNA Prep) or a variation of standard Nextera Flex in which the bead-linked transposase is diluted prior to use. In order to test the library quality for genomes with a higher and a lower G+C content, library construction methods were also tested on Pseudomonas aeruginosa PAO1 and Staphylococcus aureus ATCC 25923, respectively. We demonstrated that Hackflex can produce high-quality libraries and yields a highly uniform coverage, equivalent to the standard Nextera Flex kit. We show that strongly size-selected libraries produce sufficient yield and complexity to support de novo microbial genome assembly, and that assemblies of the large-insert libraries can be much more contiguous than standard libraries without strong size selection. We introduce a new set of sample barcodes that are distinct from standard Illumina barcodes, enabling Hackflex samples to be multiplexed with samples barcoded using standard Illumina kits. Using Hackflex, we were able to achieve a per-sample reagent cost for library prep of A$7.22 (Australian dollars) (US $5.60; UK £3.87, £1=A$1.87), which is 9.87 times lower than the standard Nextera Flex protocol at advertised retail price. An additional simple modification and further simplification of the protocol by omitting the wash step enables a further price reduction to reach an overall 14-fold cost saving. This method will allow researchers to construct more libraries within a given budget, thereby yielding more data and facilitating research programmes where sequencing large numbers of libraries is beneficial.