Antimicrobial susceptibility and pulsed – Field Gel Electrophoretic analysis of Salmonella in a tertiary hospital in northern Malaysia

SummaryBackground and aimsSalmonella infections remain a major public health problem in developing countries. The occurrence of infections caused by antimicrobial-resistant Salmonella has been on the rise complicating the available therapeutic options. The study aimed to determine the antibiograms and genotypes of prevalent Salmonella serotypes.MethodsA retrospective study involving 80 stool and extra-intestinal Salmonella strains collected over a 18-month period (January 2005–June 2006) from a tertiary hospital in Penang, Malaysia was conducted. Isolates were examined for resistance to 14 antimicrobial drugs and the clonality of the strains was determined by PFGE.ResultsTwenty-one serotypes were identified, the most common being S. enteritidis (42.5%) followed by S. corvallis (11.25%) and S. braenderup (11.25%). S. enteritidis was significantly more common amongst the extra-intestinal isolates compared to stool isolates (74.2% versus 22.4%, p<0.0001). Overall, the highest resistance was observed for tetracycline (66.3%), sulphonamides (56.3%), streptomycin (32.5%), trimethoprim (28.8%) and nalidixic acid (27.5%). Amongst the 31 invasive extra-intestinal isolates, resistance towards therapeutically relevant antibiotics was as follows: co-trimoxazole (38.7%), ampicillin (29%) and ceftriaxone (3.2%). Although there was no detectable resistance towards chloramphenicol and ciprofloxacin, 29% strains showed nalidixic acid resistance. About 41% of the 80 isolates were multidrug-resistant. PFGE subtyped the 78 Salmonella isolates to 33 distinct XbaI-pulsotypes. Isolates within the serotypes S. enteritidis, S. corvallis, S. branderup and S. fasta were more homogeneous while S. typhi and S. weltervden were genetically more diverse.ConclusionsThe high percentage of multidrug-resistant Salmonella strains is worrying and is of public health concern. PFGE was a useful and discriminative method for assessing the genetic diversity of Salmonellae

Whole-genome sequences of two carbapenem-resistant Klebsiella quasipneumoniae strains isolated from a tertiary hospital in Johor, Malaysia

We report the whole-genome sequences of two carbapenem-resistant clinical isolates of Klebsiella quasipneumoniae subsp. similipneumoniae obtained from two different patients. Both strains contained three different extended-spectrum &beta;-lactamase genes and showed strikingly high pairwise average nucleotide identity of 99.99% despite being isolated 3 years apart from the same hospital

MicroRNA profiling of mouse liver in response to DENV-1 infection by deep sequencing

Background Dengue caused by dengue virus (DENV) serotypes −1 to −4 is the most important mosquito-borne viral disease in the tropical and sub-tropical countries worldwide. Yet many of the pathophysiological mechanisms of host responses during DENV infection remain largely unknown and incompletely understood. Methods Using a mouse model, the miRNA expressions in liver during DENV-1 infection was investigated using high throughput miRNA sequencing. The differential expressions of miRNAs were then validated by qPCR, followed by target genes prediction. The identified miRNA targets were subjected to gene ontology (GO) annotation and pathway enrichment analysis to elucidate the potential biological pathways and molecular mechanisms associated with DENV-1 infection. Results A total of 224 and 372 miRNAs out of 433 known mouse miRNAs were detected in the livers of DENV-1-infected and uninfected mice, respectively; of these, 207 miRNAs were present in both libraries. The miR-148a-3p and miR-122-5p were the two most abundant miRNAs in both groups. Thirty-one miRNAs were found to have at least 2-fold change in upregulation or downregulation, in which seven miRNAs were upregulated and 24 miRNAs were downregulated in the DENV-1-infected mouse livers. The miR-1a-3p was found to be the most downregulated miRNA in the DENV-1-infected mouse livers, with a significant fold change of 0.10. To validate the miRNA sequencing result, the expression pattern of 12 miRNAs, which were highly differentially expressed or most abundant, were assessed by qPCR and nine of them correlated positively with the one observed in deep sequencing. In silico functional analysis revealed that the adaptive immune responses involving TGF-beta, MAPK, PI3K-Akt, Rap1, Wnt and Ras signalling pathways were modulated collectively by 23 highly differentially expressed miRNAs during DENV-1 infection. Conclusion This study provides the first insight into the global miRNA expressions of mouse livers in response to DENV-1 infection in vivo and the possible roles of miRNAs in modulating the adaptive immune responses during DENV-1 infection

Major surgery in an osteosarcoma patient refusing blood transfusion: case report

We describe an unusual case of osteosarcoma in a Jehovah's Witness patient who underwent chemotherapy and major surgery without the need for blood transfusion. This 16-year-old girl presented with osteosarcoma of the right proximal tibia requiring proximal tibia resection, followed by endoprosthesis replacement. She was successfully treated with neoadjuvant chemotherapy and surgery with the support of haematinics, granulocyte colony-stimulating factor, recombinant erythropoietin and intraoperative normovolaemic haemodilution. This case illustrates the importance of maintaining effective, open communication and exploring acceptable therapeutic alternative in the management of these patients, whilst still respecting their beliefs

A community-based prospective cohort study of dengue viral infection in Malaysia: the study protocol

Daniel Reidpath - ORCID: 0000-0002-8796-0420 https://orcid.org/0000-0002-8796-0420Background Globally, dengue infections constitute a significant public health burden. In recent decades, Malaysia has become a dengue hyper-endemic country with the co-circulation of the four dengue virus serotypes. The cyclical dominance of sub-types contributes to a pattern of major outbreaks. The consequences can be observed in the rising incidence of reported dengue cases and dengue related deaths. Understanding the complex interaction of the dengue virus, its human hosts and the mosquito vectors at the community level may help develop strategies for addressing the problem. Methods A prospective cohort study will be conducted in Segamat district of Johor State in Peninsular Malaysia. Researchers received approval from the Malaysian Medical Research Ethics Committee and Monash University Human Research Ethics Committee. The study will be conducted at a Malaysian based health and demographic surveillance site over a 1 year period in three different settings (urban, semi-urban and rural). The study will recruit healthy adults (male and female) aged 18 years and over, from three ethnic groups (Malay, Chinese and Indian). The sample size calculated using the Fleiss method with continuity correction is 333. Sero-surveillance of participants will be undertaken to identify asymptomatic, otherwise healthy cases; cases with dengue fever who are managed as out-patients; and cases with dengue fever admitted to a hospital. A genetic analysis of the participants will be undertaken to determine whether there is a relationship between genetic predisposition and disease severity. A detailed medical history, past history of dengue infection, vaccination history against other flaviviruses such as Japanese encephalitis and Yellow fever, and the family history of dengue infection will also be collected. In addition, a mosquito surveillance will be carried out simultaneously in recruitment areas to determine the molecular taxonomy of circulating vectors. Discussion The research findings will estimate the burden of asymptomatic and symptomatic dengue at the community level. It will also examine the relationship between virus serotypes and host genotypes, and the association of the clinical manifestation of the early phase with the entire course of illness.https://doi.org/10.1186/s40249-016-0172-35pubpub

Seroprevalence of dengue among healthy adults in a rural community in Southern Malaysia: a pilot study

Daniel Reidpath - ORCID: 0000-0002-8796-0420 https://orcid.org/0000-0002-8796-0420Background The frequency and magnitude of dengue epidemics continue to increase exponentially in Malaysia, with a shift in the age range predominance toward adults and an expansion to rural areas. Despite this, information pertaining to the extent of transmission of dengue virus (DENV) in the rural community is lacking. This community-based pilot study was conducted to establish DENV seroprevalence amongst healthy adults in a rural district in Southern Malaysia, and to identify influencing factors. Methods In this study undertaken between April and May 2015, a total of 277 adult participants were recruited from households across three localities in the Sungai Segamat subdistrict in Segamat district. Sera were tested for immunoglobulin G (IgG) (Panbio® Dengue Indirect IgG ELISA/high-titer capture) and immunoglobulin M (IgM) (Panbio®) antibodies. The plaque reduction neutralization test (PRNT) was conducted on random samples of IgG-positive sera for further confirmation. Medical history and a recall of previous history of dengue were collected through interviews, whereas sociodemographic information was obtained from an existing database. Results The overall seroprevalence for DENV infection was 86.6% (240/277) (95% CI: 83–91%). Serological evidence of recent infection (IgM/high-titer capture IgG) was noted in 11.2% (31/277) of participants, whereas there was evidence of past infection in 75.5% (209/277) of participants (indirect IgG minus recent infections). The PRNT assay showed that the detected antibodies were indeed specific to DENV. The multivariate analysis showed that the older age group was significantly associated with past DENV infections. Seropositivity increased with age; 48.5% in the age group of 45 years (P < 0.001). No associations with occupation, study site, housing type, comorbidity, educational level, and marital status were observed, although the latter two were statistically significant in the univariate analysis. None of the studied factors were significantly associated with recent DENV infections in the multivariate analysis, although there was a pattern suggestive of recent outbreak in two study sites populated predominately by Chinese people. The majority of infections did not give rise to recognizable disease (either asymptomatic or nonspecific symptoms) as only 12.9% of participants (31/240) recalled having dengue in the past. Conclusions The predominantly rural community under study had a very high previous exposure to dengue. The finding of a high proportion of unreported cases possibly due to subclinical infections underscores the need for enhanced surveillance and control methods. This finding also has implications for measuring disease burden, understanding transmission dynamics, and hypothesizing effects on DENV vaccine efficacy and uptake.https://doi.org/10.1186/s40249-017-0384-17pubpub

Can Salmonella enteritidis cause fatal necrotising fasciitis? A case report

This is a presentation of a case of mono microbial necrotising fasciitis due to the unusual organism Salmonella enteritidis. The patient presented with swelling and blistering of the right calf. There are only five other such cases reported in the literature. This was the only case that had positive blood cultures for the organism. Prompt and appropriate treatment was intuited but the patient died because of multi-organ failure