Arshad Ismail

Ability To Develop Broadly Neutralizing HIV-1 Antibodies Is Not Restricted by the Germline Ig Gene Repertoire

The human Ig repertoire is vast, producing billions of unique Abs from a limited number of germline Ig genes. The IgH V region (IGHV) is central to Ag binding and consists of 48 functional genes. In this study, we analyzed whether HIV-1–infected individuals who develop broadly neutralizing Abs show a distinctive germline IGHV profile. Using both 454 and Illumina technologies, we sequenced the IGHV repertoire of 28 HIV-infected South African women from the Centre for the AIDS Programme of Research in South Africa (CAPRISA) 002 and 004 cohorts, 13 of whom developed broadly neutralizing Abs. Of the 259 IGHV alleles identified in this study, approximately half were not found in the International Immunogenetics Database (IMGT). This included 85 entirely novel alleles and 38 alleles that matched rearranged sequences in non-IMGT databases. Analysis of the rearranged H chain V region genes of mAbs isolated from seven of these women, as well as previously isolated broadly neutralizing Abs from other donors, provided evidence that at least eight novel or non-IMGT alleles contributed to functional Abs. Importantly, we found that, despite a wide range in the number of IGHV alleles in each individual, including alleles used by known broadly neutralizing Abs, there were no significant differences in germline IGHV repertoires between individuals who do and do not develop broadly neutralizing Abs. This study reports novel IGHV repertoires and highlights the importance of a fully comprehensive Ig database for germline gene usage prediction. Furthermore, these data suggest a lack of genetic bias in broadly neutralizing Ab development in HIV-1 infection, with positive implications for HIV vaccine design.

Genome Sequencing of Extended-Spectrum β-Lactamase (ESBL)-Producing Klebsiella pneumoniae Isolated from Pigs and Abattoir Workers in Cameroon

Background and objectives: Extended-spectrum β-lactamase (ESBL)-producing Klebsiella pneumoniae is a serious public health issue globally. In this study, the antibiotic resistance genes, virulence factors, mobile genetic elements, and genetic lineages of circulating ESBL-producing K. pneumoniae strains isolated from pigs and humans in Cameroonian abattoirs were investigated using whole genome sequencing (WGS), in order to ascertain zoonotic transmission (viz. from animals to humans and/or vice-versa) in the food chain. Methods: During March–October 2016, 288 nasal and rectal pooled samples from 432 pigs as well as nasal and hand swabs from 82 humans were collected from Cameroon and South Africa. Seven ESBL-producing K. pneumoniae circulating in Cameroonian pig abattoirs were selected and their genomic DNA sequenced using an Illumina MiSeq platform. Generated reads were de novo assembled using the Qiagen CLC Genomics Workbench and SPAdes. The assembled contigs were annotated using RAST and antibiotic resistance genes, virulence factors, plasmids, and bacteriophages were identified with ResFinder, Virulence Finder, PlasmidFinder, and PHAST, respectively. Results: ESBL-producing K. pneumoniae were detected in pigs (34/158; 21.52%) and exposed workers (8/71; 11.26%) in Cameroon only. The circulating K. pneumoniae strains were dominated principally by the sequence type (ST) 14 and 39. In addition, the “high-risk” ST307 clone and two novel STs assigned ST2958 and ST2959 were detected. Genomic analysis identified various antibiotic resistance genes associated with resistance to β-lactams, aminoglycosides, fluoroquinolones, macrolide, lincosamide and streptogramins, rifampicin, sulfonamides, trimethoprim, phenicols and tetracycline. None of the ESBL-producing K. pneumoniae harbored virulence genes. Intermingled K. pneumoniae populations were observed between pig- and human-source within and across abattoirs in the country. Conclusion: Our study shows that ESBL-producing K. pneumoniae is actively disseminating in pigs and occupationally exposed workers in Cameroonian pig abattoirs and is probably underestimated in the absence of molecular epidemiological studies. It suggests pigs, abattoir workers and food products as potential reservoirs and sources of zoonotic transmission in Cameroon. Our findings underline the existence of a potential unheeded food safety and public health threat associated with these resistant strains and reinforce the crucial importance of implementing appropriate food safety measures and promoting rational antibiotic use.

A novel adenovirus isolated from the Egyptian fruit bat in South Africa is closely related to recent isolates from China

Recently a number of novel adenoviruses have been isolated from diverse bat species and from diverse geographical locations. We describe the isolation of a novel adenovirus (Family Adenoviridae, genus Mastadenovirus) from a pool of liver and spleen tissue of an apparently healthy wild-caught Egyptian fruit bat (Rousettus aegyptiacus) in South Africa. Genetically the virus is most closely related to four mastadenoviruses recently isolated in China, from Miniopterus schreibersi and Rousettus leschenaultii bats, which are highly divergent from previously identified bat adenoviruses. The length of the Rousettus aegyptiacus adenovirus-3085 (RaegAdV-3085) genome, at 29,342 bp is similar to its closest relatives, and contains 27 open reading frames. The RaegAdV-3085 genome has a low G + C content (36.4%) relative to other viruses in the genus (between 43.6 and 63.9%) but similar to its closest relatives. The inverted terminal repeat (ITR) of RaegAdV-3085 is only 40 bp compared to between 61 and 178 bp of its closest relatives. The discovery of RaegAdV-3085 expands the diversity of known adenoviruses in bats and might represent a member of a new mastadenovirus species in bats.

Biological crusts of serpentine and non-serpentine soils from the Barberton Greenstone Belt of South Africa

Climate and geography can influence biological soil crust (BSC) community composition, but local heterogeneity in variables such as soil characteristics or microclimate gradients can also impact cryptogamic diversity. Heavy metals and nutrient imbalances in serpentine soils are known to influence the distributions of higher plants, but cryptogamic species appear to be more tolerant of substrate. The aim of this study was to compare the cryptogamic composition of serpentine and non-serpentine soils by using integrative taxonomy, which combines morphological and DNA barcoding data, to determine how soil characteristics in combination with rainfall can influence BSC community composition. Samples from serpentine and non-serpentine soils were enumerated and total genomic DNA was isolated from the soil samples. Analyses of the 16S rRNA gene and ITS sequences were done using the quantitative insights into microbial ecology (QIIME) workflow to determine which eukaryotic microorganisms were present in the samples. Sixty genera from the Cyanophyceae (38), Chlorophyceae (10), Bacillariophyceae (6), Eustigmatophyceae (4), Trebouxiophyceae (1) and Xanthophyceae (1) classes were detected with this approach. Results confirm that algae and cyanobacteria are tolerant of most substrates and can even colonize environments with high levels of heavy metal and nutrient imbalances, if moisture is present. Genera such as Acaryochloris, Annamia, Brasilonema, Chrocosphaera, Halomicronema, Planktothricoides, Rubidibacter, and Toxopsis are reported for the first time for South African soil.

Draft genome sequences of extended-spectrum β-lactamase-producing Enterobacter aerogenes isolated from swine and human

OBJECTIVES The draft genome sequences of two Enterobacter aerogenes strains (HN503E2II and PN108E5IIB) isolated from two Cameroonian abattoirs are reported. METHODS Bacterial genomic DNA of the two isolates was sequenced using an Illumina MiSeq platform. Generated reads were de novo assembled using CLC Genomics Workbench and SPAdes. The assembled contigs were annotated, and antibiotic resistance genes, virulence factors and plasmids were identified. RESULTS Whole-genome sequencing revealed that both strains were similar, with genomes of 4878638bp and 4794257bp, encoding several resistance genes associated with resistance to β-lactams, fluoroquinolones, aminoglycosides, fosfomycin, phenicols, sulphonamides, trimethoprim, macrolides and tetracycline. In silico analysis also revealed chromosomal integration of one plasmid in the genome of PN108E5IIB. CONCLUSION The genome sequences reported here will provide useful information for a better understanding of the genetic structure of E. aerogenes in Africa.

Draft Genome Sequence of Methicillin Resistant Staphylococcus epidermidis Isolate from Swine

OBJECTIVES Here we report the draft genome sequence of a methicillin-resistant Staphylococcus epidermidis strain (sequence type 59) isolated from a pooled rectal sample from pigs collected in an abattoir in South Africa. METHODS Genomic DNA of S. epidermidis PR246B0 was sequenced using an Illumina MiSeq platform. Generated reads were de novo assembled using CLC Genomics Workbench (QIAGEN). The assembled contigs were annotated and antimicrobial resistance genes, plasmids and the sequence type were identified. RESULTS The genome comprised a circular chromosome of 2537769bp, with a G-C content of 32.32% and various antimicrobial resistance genes associated with resistance to β-lactams, fluoroquinolones, aminoglycosides, fosfomycin, macrolides, lincosamides and tetracycline. Genome analysis also revealed the presence of seven plasmid replicon types. CONCLUSION The genome sequence reported herein will provide useful information for a better understanding of the genetic structure of the S. epidermidis genome in Africa.

Draft genome sequence of an extended-spectrum β-lactamase (CTX-M-15)-producing Escherichia coli ST10 isolated from a nasal sample of an abattoir worker in Cameroon

OBJECTIVES Here we report the draft genome sequence of Escherichia coli strain HN503E1II isolated from a nasal sample of an abattoir worker in Cameroon. METHODS Bacterial genomic DNA of E. coli HN503E1II was sequenced using an Illumina MiSeq platform. Generated reads were de novo assembled using the Qiagen CLC Genomics Workbench. The assembled contigs were annotated and antibiotic resistance genes, virulence factors, plasmids and the sequence type (ST) were identified. RESULTS The genome comprised a circular chromosome of 4 674201bp, with a 50.78% G+C content and several resistance genes associated with resistance to β-lactams, fluoroquinolones, aminoglycosides, sulphonamides, trimethoprim, macrolides and tetracycline. The isolate was assigned to ST10 with 100% identity among the seven housekeeping genes. In silico analysis also revealed the presence six plasmid types and one virulence factor. CONCLUSION The genome sequence reported here will provide valuable information for a better understanding of the genetic structure of the E. coli genome in Africa.

Whole-genome sequence of a Mycobacterium goodii isolate from a pediatric patient in South Africa

ABSTRACT We describe here the draft genome sequence of a Mycobacterium goodii isolate from a pediatric patient in Western Cape, South Africa. To our knowledge, this is the second reported genome of this rapidly growing nontuberculous mycobacterial species.

Draft genome sequence of Bacillus oleronius DSM 9356 isolated from the termite Reticulitermes santonensis

Bacillus oleronius strain DSM 9356 isolated from the termite Reticulitermes santonensis was sequenced to gain insights in relation to its closest phylogenetic neighbor Bacillus sporothermodurans. The draft genome of strain DSM 9356 contains 5,083,966 bp with an estimated G + C content of 35%, 4899 protein-coding genes, 116 tRNAs and 18 rRNAs. The RAST annotation assigned these genes into 462 subsystems, with the maximum number of genes associated with amino acids and derivatives metabolism (14.84%), followed by carbohydrates (13.89%) and protein metabolism subsystems (9.10%). The draft genome sequence and annotation has been deposited at NCBI under the accession number MTLA00000000.

Draft Genome Sequence of a Multidrug-Resistant Serratia marcescens Strain, Isolated from a Patient with Peritoneal Cancer in South Africa

ABSTRACT We report here the draft genome sequence of Serratia marcescens ML2637, isolated from a South African pediatric patient in the intensive care unit with peritoneal cancer. The genome comprised 5,718,350 bp, with a 59.1% G+C content. There were 5,594 predicted genes, including 5,301 protein-coding genes, 199 pseudogenes, and 94 RNA genes.