Bacteroides pyogenes isolated from appendiceal abscess in a patient without animal contact.

Bacteroides pyogenes is a member of the oral flora of cats and dogs. We report a case of B. pyogenes isolated from an appendiceal abscess in a patient without a history of animal contact. This species was identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and sequencing of 16S rRNA, rpoB, gyrB and hsp60 genes.

Extensive genetic diversity with novel mutations in spike glycoprotein of severe acute respiratory syndrome coronavirus 2, Bangladesh in late 2020.

In Bangladesh, coronavirus disease 2019 (COVID-19) has been highly prevalent during late 2020, with nearly 500 000 confirmed cases. In the present study, the spike (S) protein of severe acute respiratory coronavirus 2 (SARS-CoV-2) circulating in Bangladesh was genetically investigated to elucidate the diversity of mutations and their prevalence. The nucleotide sequence of the S protein gene was determined for 15 SARS-CoV-2 samples collected from eight divisions in Bangladesh, and analysed for mutations compared with the reference strain (hCoV-19/Wuhan/WIV04/2019). All the SARS-CoV-2 S genes were assigned to B.1 lineage in G clade, and individual S proteins had 1-25 mutations causing amino acid substitution/deletion. A total of 133 mutations were detected in 15 samples, with D614G being present in all the samples; 53 were novel mutations as of January 2021. On the receptor-binding domain, 21 substitutions including ten novel mutations were identified. Other novel mutations were located on the N-terminal domain (S1 subunit) and dispersed sites in the S2 subunit, including two substitutions that remove potential N-glycosylation sites. A P681R substitution adjacent to the furin cleavage site was detected in one sample. All the mutations detected were located on positions that are functionally linked to host transition, antigenic drift, host surface receptor binding or antibody recognition sites, and viral oligomerization interfaces, which presumably related to viral transmission and pathogenic capacity.

Clonal diversity of Acinetobacter baumannii clinical isolates in Myanmar: identification of novel ST1407 harbouring blaNDM-1.

Recent Acinetobacter baumannii clinical isolates in a teaching hospital in Myanmar comprised three major sequence types (ST2, ST16 and ST23) and two sporadic STs, showing a high resistance rate to carbapenem associated with blaOXA-23 . The NDM-1 encoding gene was identified in only one isolate exhibiting novel ST1407 (a triple-locus variant of ST16).

Predominance of Leptospira wolffii in north-central Bangladesh, 2019.

Leptospira was detected in 48.9% of blood samples from 182 febrile patients in north-central Bangladesh in 2019. Most Leptospira were classified as L. wolffii (93%) on the basis of phylogenetic analysis of 16S ribosomal RNA genes, while others were assigned to L. borgpetersenii and L. meyeri.

Pacemaker-associated infection caused by ST81/SCCmec IV methicillin-resistant, vancomycin-intermediate Staphylococcus aureus in Japan.

A 76-year-old Japanese man was admitted to hospital for treatment of fever and skin lesion at the implantation site of his pacemaker. During his hospitalization, vancomycin-intermediate Staphylococcus aureus (MIC 4 µg/mL) with reduced susceptibility to daptomycin was isolated from venous blood. This isolate was identified as methicillin-resistant S. aureus with SCCmec IV and was genotyped as sequence type 81, coa VIIa and spa type t7044, harbouring blaZ, aac(6')-aph(2″) and enterotoxin(-like) genes sea, seb, sek, sel, selx and selw. The patient was successfully treated with daptomycin, minocycline and sulfamethoxazole/trimethoprim. We describe the identification of sequence type 81/SCCmec IV vancomycin-intermediate S. aureus from pacemaker-associated septicaemia.

Resurgence and predominance of G3P[8] human rotaviruses in north-central Bangladesh, 2018-2019.

Predominance of genotype G3P[8] rotavirus was revealed for children and adults with diarrhoea in north-central Bangladesh for a 1-year period from September 2018. The G3P[8] rotaviruses were phylogenetically close to recent Indian strains, having antigenic variation in VP7 and VP4 compared with old Bangladeshi strains.

Co-circulation of dengue virus type 3-genotype I and type 2-Cosmopolitan genotype in 2018 outbreak in Dhaka, Bangladesh.

Dengue virus (DENV) that caused an outbreak in Dhaka, Bangladesh during 2018 was analysed phylogenetically. DENV samples were classified into type 2-Cosmopolitan genotype (54%) and type 3-genotype I (46%), indicating co-circulation of two DENV types and resurgence of type 3 associated with genotype replacement.

Molecular characterization of Orientia tsutsugamushi causing scrub typhus among febrile patients in north-central Bangladesh.

Scrub typhus is a mite-borne rickettsial disease caused by Orientia tsutsugamushi, which is endemic in Asia Pacific region. In this study, infection rate and molecular epidemiologic traits of O. tsutsugamushi was investigated in Mymensingh, located in north-central Bangladesh. Among the blood samples from 453 febrile patients who visited Mymensingh medical college hospital in 2018, the 47 kDa protein gene of O. tsutsugamushi was detected in 78 samples (17.2%) by nested PCR. Phylogenetic analysis of the O. tsutsugamushi 56 kDa protein gene (18 samples) revealed a predominance of Karp-related genotype (89%), while the remaining belonged to Gilliam genotype. Samples of the Karp-related genotype mostly clustered with those of China, Taiwan, Thailand and India, etc., in emergent subgroups clades 2 and 4, which were distinct from clade 1, including prototype Karp strains. Among the 18 samples, three variable domains (VD) of 56 kDa type-specific antigen had different types of sequence diversity; VDI contained two or three repeats of eight amino acid units, while VDII and VDIII had amino acid substitution, deletion or insertion. The present study documented a potentially high prevalence of genetically diverse O. tsutsugamushi in north-central Bangladesh.

First molecular identification of two Leptospira species (Leptospira interrogans and Leptospira wolffii) in Bangladesh.

Leptospiral 16S rRNA genes were detected in 13 blood samples from 74 febrile patients in north-central Bangladesh, and their sequences phylogenetically clustered with those of Leptospira interrogans or Leptospira wolffii. Genetic diversity in O-antigen polymerase (wzy) was found in an L. interrogans sample.

First detection of the mcr-1 colistin resistance gene in Escherichia coli from a patient with urinary tract infection in Myanmar.

Colistin-resistance gene mcr-1 was detected in an Escherichia coli sample among 442 clinical isolates collected in a tertiary-care hospital in Yangon, Myanmar, in 2018. This isolate was classified into phylogroup A-ST23 complex and harboured bla CTX-M-15 and bla TEM-1, associated with multiple mutations in quinolone-resistance-determining regions in gyrA and parC.

Molecular characterization of chikungunya virus causing the 2017 outbreak in Dhaka, Bangladesh.

Chikungunya viruses from the 2017 outbreak in Dhaka, Bangladesh, were analysed phylogenetically. E1 sequences from 21 strains belonged to the Indian Ocean clade of the East/Central/South African (ECSA) genotype, forming a novel cluster with latest South Asian strains. They lacked the A226V substitution.

Genetic characteristics of VanA-type vancomycin-resistant Enterococcus faecalis and Enterococcus faecium in Cuba.

VanA-type vancomycin-resistant enterococci isolates from bloodstream infections in Cuba were genetically characterized. Enterococcus faecalis isolates were assigned to sequence type (ST) 28, closely related to Eastern Europe, while Enterococcus faecium belonged to ST262, ST656 and ST1349, and showed different genetic profiles.

Nationwide prevalence of Rickettsia felis infections in patients with febrile illness in Bangladesh.

From July 2015 to December 2016, the presence of rickettsial pathogens was investigated for 414 patients with unknown fever in eight places in all the divisions of Bangladesh. Rickettsia felis was identified in blood samples from all the regions (overall detection rate, 19.6%), suggesting nationwide prevalence of R. felis infections.

Septic arthritis caused by an emerging ST121 methicillin-susceptible, PVL-negative Staphylococcus aureus harbouring a variant of bone sialoprotein-binding protein gene.

ST121/agr-IV methicillin-susceptible Staphylococcus aureus was isolated from a patient of septic arthritis (synovial fluid, blood, skin and nasal cavity). Although the Panton-Valentine leukocidin (PVL) gene was negative, this isolate harboured a gene encoding a variant of bone sialoprotein-binding protein with a shortened SD-repeat region.

First report of Panton-Valentine leukocidin-positive methicillin-susceptible Staphylococcus aureus ST88 harbouring ΦSa2usa isolated from refractory breast abscesses in Japan.

A methicillin-susceptible Staphylococcus aureus with Panton-Valentine leukocidin (PVL) genes was isolated from refractory breast abscesses of 12-year-old girl in Japan, and classified into ST88, spa-t1245 and coa-IIIa. This strain harboured PVL phage ΦSa2usa, which is usually found in ST8 community-acquired methicillin-resistant S. aureus clone USA300.

Drug resistance and genetic characteristics of clinical isolates of staphylococci in Myanmar: high prevalence of PVL among methicillin-susceptible Staphylococcus aureus belonging to various sequence types.

Prevalence, drug resistance and genetic characteristics were analysed for a total of 128 clinical isolates of staphylococci obtained from a tertiary hospital in Myanmar. The dominant species were S. aureus (39%) and S. haemolyticus (35%), followed by S. epidermidis (6%) and S. saprophyticus (5%). The majority of S. haemolyticus isolates (71.1%) harboured mecA, showing high resistance rates to ampicillin, cephalosporins, erythromycin and levofloxacin, while methicillin-resistant S. aureus (MRSA) was only 8% (four isolates) among S. aureus with type IV SCCmec. Panton-Valentine leukocidin (PVL) genes were detected in 20 isolates of S. aureus (40%), among which only one isolate was MRSA belonging to sequence type (ST) 88/agr-III/coa-IIIa, and the other 19 methicillin-susceptible S. aureus (MSSA) isolates were classified into six STs (ST88, ST121, ST1153, ST1155, ST1930, ST3206). An ST1153 MSSA isolate with PVL was revealed to belong to a novel coa type, XIIIa. ST121 S. aureus was the most common in the PVL-positive MSSA (47%, 9/19), harbouring genes of bone sialoprotein and variant of elastin binding protein as a distinctive feature. Although PVL-positive MSSA was susceptible to most of the antimicrobial agents examined, ST1930 isolates were resistant to erythromycin and levofloxacin. ST59 PVL-negative MRSA and MSSA had more resistance genes than other MRSA and PVL-positive MSSA, showing resistance to more antimicrobial agents. This study indicated higher prevalence of mecA associated with multiple drug resistance in S. haemolyticus than in S. aureus, and dissemination of PVL genes to multiple clones of MSSA, with ST121 being dominant, among hospital isolates in Myanmar.

Detection of ST772 Panton-Valentine leukocidin-positive methicillin-resistant Staphylococcus aureus (Bengal Bay clone) and ST22 S. aureus isolates with a genetic variant of elastin binding protein in Nepal.

Genetic characteristics were analysed for recent clinical isolates of methicillin-resistant and -susceptible Staphylococcus aureus (MRSA and MSSA respectively) in Kathmandu, Nepal. MRSA isolates harbouring Panton-Valentine leukocidin (PVL) genes were classified into ST1, ST22 and ST88 with SCCmec-IV and ST772 with SCCmec-V (Bengal Bay clone), while PVL-positive MSSA into ST22, ST30 and ST772. ST22 isolates (PVL-positive MRSA and MSSA, PVL-negative MRSA) possessed a variant of elastin binding protein gene (ebpS) with an internal deletion of 180 bp, which was similar to that reported for ST121 S. aureus previously outside Nepal. Phylogenetic analysis indicated that the ebpS variant in ST22 might have occurred independently of ST121 strains. This is the first report of ST772 PVL-positive MRSA in Nepal and detection of the deletion variant of ebpS in ST22 S. aureus.

Emerging non-PCV13 serotypes of noninvasive Streptococcus pneumoniae with macrolide resistance genes in northern Japan.

In Japan, the 7-valent pneumococcal conjugate vaccine (PCV7) was introduced to the nation's routine immunization program in April 2013 and was replaced by the 13-valent pneumococcal conjugate vaccine (PCV13) in November 2013. Distribution of serotypes and macrolide resistance genotypes was investigated for a total of 1097 (975 children, 122 adults) and 960 (873 children, 87 adults) clinical isolates of Streptococcus pneumoniae from noninvasive infections in Hokkaido (northern main island of Japan) in the routine immunization periods for PCV7 and PCV13 (April-October 2013 and November 2013-November 2014, respectively). Serotype was determined by sequential multiplex PCR and additional genetic analyses. Macrolide resistance genes erm(B) and mef(A/E) were detected by multiplex PCR. Although the most prevalent serotypes in children were 23A and 6C in the PCV7 period, after replacement with PCV13, 19A became the most common, followed by 6C, 15A and 23A. Among adults, serotype 3 was consistently the most frequent throughout the study periods. Compared with values from the pre-PCV7 routine immunization period, PCV7 serotypes decreased from 48.3 to 3.3% in the PCV13 period among children, while the rates of non-PCV13 serotypes (particularly 15A, 23A, 11A, 10A and 35B) increased from 39.7 to 75.1% (p < 0.001). In the PCV13 period, erm(B), mef(A/E) and both of these genes were detected in 75.8, 31.6 and 11.3% of all isolates, respectively. Serotype 19A accounted for 76.9% of the isolates with both the macrolide resistance genes, and emerging non-PCV13 serotypes 15A, 15C and 23A mostly harboured erm(B).

High prevalence of bla OXA-23 in Acinetobacter spp. and detection of bla NDM-1 in A. soli in Cuba: report from National Surveillance Program (2010-2012).

As a first national surveillance of Acinetobacter in Cuba, a total of 500 Acinetobacter spp. isolates recovered from 30 hospitals between 2010 and 2012 were studied. Acinetobacter baumannii-calcoaceticus complex accounted for 96.4% of all the Acinetobacter isolates, while other species were detected at low frequency (A. junii 1.6%, A. lwoffii 1%, A. haemolyticus 0.8%, A. soli 0.2%). Resistance rates of isolates were 34-61% to third-generation cephalosporins, 49-50% to ß-lactams/inhibitor combinations, 42-47% to aminoglycosides, 42-44% to carbapenems and 55% to ciprofloxacin. However, resistance rates to colistin, doxycycline, tetracycline and rifampin were less than 5%. Among carbapenem-resistant isolates, 75% harboured different bla OXA genes (OXA-23, 73%; OXA-24, 18%; OXA-58, 3%). The bla NDM-1 gene was identified in an A. soli strain, of which the species was confirmed by sequence analysis of 16S rRNA gene, rpoB, rpoB-rpoC and rpoL-rpoB intergenic spacer regions and gyrB. The sequences of bla NDM-1 and its surrounding genes were identical to those reported for plasmids of A. baumannii and A. lwoffi strains. This is the first report of bla NDM-1 in A. soli, together with a high prevalence of OXA-23 carbapenemase for carbapenem resistance in Acinetobacter spp. in Cuba.

Automated anatomical demarcation using an active shape model for videofluoroscopic analysis in swallowing.

The current gold standard method in the clinical assessment of swallowing is the visual inspection of videofluoroscopic frames. Specific clinical measurements are estimated based on various anatomical and bolus positional information with respect to time (or frame number). However, due to the subjective nature of visual inspection clinicians face intra- and inter-observer repeatability issues and bias when making these estimations. The correct demarcations of reference lines highlighting the positions of important anatomical landmarks would serve as a visual aid and could also be used in conjunction with bolus detection methods to objectively determine these desirable measurements. In this paper, we introduce and test the reliability of applying a 16-point Active Shape Model as a deformable template to demarcate the boundaries of salient anatomical boundaries with minimal user input. A robust end and corner point detection algorithm is also used to provide image information for the suggested movement of the template during the fitting stage. Results show the model deformation constraints calculated from a training set of images are clinically coherent. The Euclidean distances between the fitted model points against their corresponding target points were measured. Test images were taken from two different data sets from frames acquired using two different videofluoroscopy units. Overall, fitting was found to be more reliable on the vertebrae and inferior points of the larynx compared to the superior laryngeal points and hyoid bone, with the model always fitting the C7 vertebra with discrepancies no higher than a distance of 23 pixels (3.2% of the image width, approximately 7.6mm).